Analysis of corneal surface evolution after moderate alkaline burns by using impression cytology.

• Lopez-Garcia JS,
• Rivas L,
• Garcia-Lozano I,
• Murube J.

Ophthalmology Services, Hospital Cruz Roja, Madrid, Spain. docsantilopez@hotmail.com

PURPOSE: To compare corneal surface evolution after moderate alkaline burns by impression cytology in patients treated with medical therapy or with amniotic membrane transplantation (AMT). METHODS: A prospective study of 24 eyes from 18 patients (13 men and 5 women) with moderate alkaline burns was performed. All patients were divided according to the clinical ocular severity and the therapy used. Twelve eyes were treated surgically with AMT and the other 12 eyes received only medical therapy. Corneal cytology was obtained immediately after the burns, and 1, 2, 5, and 9 months later. We differentiated between samples obtained from affected areas and areas not affected by the burns. Cellular size, nuclear size, and nuclear-cytoplasmic (N:C) ratio were examined in corneal epithelial cells, as was the presence of goblet cells in corneal epithelium. RESULTS: Nuclear size, cellular size, and N:C ratio in non-burn-affected corneal areas had no significant alterations in comparison with normal eyes. In contrast, in burn-affected corneal areas, these parameters were significantly worse, and the presence of goblet cells in corneal epithelium was frequent 1 month after severe burns. Cellular size, nuclear size, N:C ratio, and corneal conjunctivalization improved during the study in all patients, but corneal reepithelialization occurred earlier in patients treated with AMT than in patients with only medical therapy. CONCLUSION: Morphologic and morphometric analysis of corneal cells by impression cytology after ocular burns permits the establishment of cellular reepithelialization patterns in relation with limbal deficiency level and with clinical ocular severity. AMT improves corneal reepithelialization earlier than medical therapy in moderate alkaline burns.

PMID: 17102665 [PubMed - indexed for MEDLINE]

Antimicrobial activity of antibiotic-treated amniotic membrane: An in vitro study.

• Mencucci R,
• Menchini U,
• Dei R.

Department of Oto-Neuro-Opthalmological Surgical Sciences, University of Florence, Florence, Italy. rita.mencucci@unifi.it

PURPOSE: To investigate if amniotic membrane incubated with antibiotics could inhibit bacterial growth in vitro. METHODS: Amniotic membrane fragments were incubated with the antibiotics (netilmicin) solution; the washed and drained fragments were either tested after treatment or further incubated in antibiotic-free medium. The antibacterial activity of both amniotic membrane and elution media was carried out by the Agar diffusion method, with Staphylococcus epidermidis as indicator, measuring the inhibition zone after overnight incubation. RESULTS: The amniotic membrane fragments soaked in antibiotics inhibited bacterial growth. Antibiotic uptake was dose-dependent and occurred rapidly. The drug was released from the membrane, and the antibacterial effect was present in the elution media at least 3 days after treatment. CONCLUSIONS: Our preliminary in vitro data show that amniotic membrane can absorb the antibiotic netilmicin and in the future may be used to deliver antibiotics, as reported for collagen shields and other medical prosthetic devices.

PMID: 16670480 [PubMed - indexed for MEDLINE]

Induced endoplasmic reticulum (ER) stress and binding of over-expressed ER specific chaperone GRP78/BiP with dimerized epidermal growth factor receptor in mammalian cells exposed to low concentration of N-methyl-N'-nitro-N-nitrosoguanidine.

• Liu G,
• Shang Y,
• Yu Y.

Department of Pathology and Pathophysiology, Zhejiang University School of Medicine, Hangzhou 310031, China.

Previously we have shown that alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) can induce the clustering of epidermal growth factor receptor (EGFR) in human amnion FL cells. However, the biological consequence of MNNG-induced clustering is different from that of epidermal growth factor (EGF)-induced clustering. In addition, MNNG strongly blocks the autophosphorylation of EGFR in response to its ligand, we speculate it might be due to the altered conformation of EGFR by MNNG alkylation, or the binding of some unknown suppressive molecules to EGFR, which could lead to the down-regulation of EGFR pathway. In this study, we further demonstrated that EGFR could not be phosphorylated by EGF in lysates prepared from MNNG-pretreated cell. In addition, it was found that the clustering of EGFR induced by low concentration (<or=1 microM) of MNNG on cell surface was indeed the dimerization of EGFR; however, unlike EGF treatment, the dimerization initiated by MNNG was irreversible upon mild-acid washing. Besides, in accordance with our previous results, the recruitment of adaptor proteins Grb-2/Sos1, which play key roles in activating ensuing RAS-MAPK pathway, was also suppressed. Interestingly, we found that endoplasmic reticulum (ER) stress participates in MNNG-induced down-regulation of EGFR signaling. It was demonstrated that the ER specific chaperone, glucose-regulated protein 78 (GRP78/BiP) formed a stable complex with EGFR in MNNG-treated cell. However, in the presence of 1mM ATP, EGF induced phosphorylation of tyrosine residues of EGFR can be revitalized in lysates prepared from MNNG pretreated cells. We also found that MNNG can induce ER stress or unfolded protein response (UPR) which is characterized by induced expression of ER-stress response proteins, such as GRP78/BiP, GADD153/CHOP, and activation of ER-localized caspase-12. Therefore, it is concluded MNNG is also an ER stress inducer. In MNNG-exposed cells, ER stress plays an important role in the blockage of EGFR-signaling pathway by forming a stable complex of EGFR/BiP.

PMID: 16488447 [PubMed - indexed for MEDLINE]

[Staphylococcal endophthalmitis following cataract surgery in a patient with ocular rosacea]

• Gicquel JJ,
• Quinton J,
• Salama B,
• Pommeraud D,
• Dighiero P.

Service d'Ophtalmologie, CHU de Poitiers, Poitiers, France. gicquelophtha@aol.com

PURPOSE: To report staphylococcal endophthalmitis following cataract surgery in a patient with ocular rosacea. OBSERVATION: A 69-year-old man presented with decreased visual acuity in the right eye, a corneal abscess on the incision and hypopyon 2 weeks after phacoemulsification with intraocular lens placement. The patient was hospitalized. A diagnostic vitreous tap was performed. Antibiotic therapy was initiated both locally (two intravitreal shots + topical administration) and intravenously. RESULTS: Vitreal tap cultures indicated Staphylococcus aureus. The patient's clinical status improved 48 hours after the second intravitreal shot. The same bacterium was cultured from the patient's eyelids and incision abscess. Multilayer amniotic membrane transplantation enabled satisfactory healing of the corneal abscess, which had become perforative. CONCLUSIONS: The source of the S. aureus was the skin lesions on the patient's eyelids. Rosacea is a skin disease that frequently affects the face, nose and eyelids. Although good surgical techniques were performed, with the incision requiring suture, the risk of endophthalmitis after intraocular surgery was increased in the absence of proper preoperative treatment of the dermatological condition.

PMID: 16395213 [PubMed - in process]

Amniotic membrane transplantation in ocular surface disorders.

• Chandra A,
• Maurya OP,
• Reddy B,
• Kumar G,
• Pandey K,
• Bhaduri G.

Department of Ophthalmology, Institute of Medical Sciences, Banaras Hindu University, Varanasi.

In this prospective study, 81 eyes of 70 patients diagnosed with various ocular surface disorders were enrolled to document the use of amniotic membrane transplantation in various ocular surface disorders. Detailed history and ocular examination was done. Ocular photographs and consent from all patients were taken. Fluorescein staining and impression cytology was done preoperatively and postoperatively in selected cases. Amniotic membrane was prepared from the placenta of a donor (consent taken and negative for infectious disorders), after separating amnion from chorion. It was washed with antibiotic solutions, transferred over nitrocellulose paper and stored in Dulbecco's modified Eagle's minimum essential medium at -80 degrees C. Recipient bed was prepared by removing the fibrovascular pannus and necrosed conjunctiva. Amniotic membrane was transplanted with the epithelial side up and sutured. Sixty-four eyes had good result by clinical evaluation or impression cytology findings, 5 eyes later required limbal stem cell culture and transplantation. All the 3 eyes had failure of the fornix reconstruction and 5 eyes had recurrence of the pterygium. Amniotic membrane provides lower recurrence rate in cases of recurrent pterygium. Alkali injuries are more dangerous but showed good response to amniotic membrane transplantation combined with limbal autografting or ex-vivo expansion and later transfer. Initial proper assessment of limbal involvement, conjunctival necrosis and corneal involvement is the key to the management of acute cases. Contracted sockets showed no improvement. Shield ulcers and persistent epithelial defect and ocular surface defects secondary to tumour excision showed excellent results.

PMID: 16366188 [PubMed - indexed for MEDLINE]

Evaluation of amniotic membrane transplantation as an adjunct to medical therapy as compared with medical therapy alone in acute ocular burns.

• Tamhane A,
• Vajpayee RB,
• Biswas NR,
• Pandey RM,
• Sharma N,
• Titiyal JS,
• Tandon R.

Rajendra Prasad Centre for Ophthalmic Sciences, All India Institute of Medical Sciences, New Delhi, India.

PURPOSE: To evaluate the role of amniotic membrane transplantation (AMT) in acute ocular burns. DESIGN: Prospective, randomized, controlled clinical trial. PARTICIPANTS AND CONTROLS: Patients with grade II to IV ocular burns within 3 weeks of injury were recruited. Thirty-seven patients, 7 of whom had bilateral involvement (total, 44 eyes), participated in the trial. Twenty eyes were included in group A (AMT) and 24 eyes were included in group B (controls). INTERVENTION: The eyes in the AMT group underwent AMT in addition to conventional medical therapy. In the control group, only conventional medical therapy was instituted. MAIN OUTCOME MEASURES: The patients' subjective assessments of relief of ocular discomfort, healing of the corneal epithelial defect, visual acuity, extent of corneal vascularization, formation of symblepharon, and tear function tests. RESULTS: At day 1, subjective ocular discomfort scores were reduced significantly in eyes with moderate burns in the AMT group compared with controls (P = 0.05), but there was no difference between the 2 groups in eyes with severe burns. The log mean percentage reduction in size of epithelial defect by day 7 was 7.43+/-0.89 after AMT and 6.23+/-1.10 with medical treatment alone in moderate grade burns at day 7 (P = 0.01), but there was no difference between the 2 groups in eyes with severe burns. There was no overall difference in the final visual acuity, symblepharon formation, corneal vascularization, and tear function tests between the 2 groups over the next 3 months and further follow-up. There was a high dropout rate for long-term follow-up. CONCLUSIONS: Amniotic membrane transplantation in eyes with acute ocular burns has advantages in terms of reduction of pain and promotion of early epithelialization in patients with moderate grade burns, but not so in severe burns. There seems to be no definite advantage to AMT over medical therapy alone in terms of improvement in visual acuity, appearance of symblepharon, corneal vascularization, and results of tear function tests on short-term follow-up.

PMID: 16198422 [PubMed - indexed for MEDLINE]

[Amniotic membrane transplantation after surgical resection in conjunctival Kaposi's sarcoma]

• Rodriguez-Perez C,
• Gris-Castellon O,
• Soler-Lluis N,
• Del Campo-Carrasco Z,
• Wolley-Dod C.

Servicio de Oftalmologia, Hospital de la Santa Creu i Sant Pau, Universidad Autonoma de Barcelona, Spain. carmen_rp_deruibal@msn.com

OBJECTIVES/METHODS: To describe a case of isolated conjunctival Kaposi's sarcoma treated with surgical resection and amniotic membrane graft. After surgical resection of the lesion an amniotic membrane graft was used to reconstruct the conjunctival surface. Histological studies were performed. RESULTS/CONCLUSIONS: Rapid regeneration of the conjunctiva, with minimal scarring, occurred after the surgical procedure and no recurrence of the lesion was observed at 18 months of follow-up. Surgical resection with amniotic membrane grafting appears to be a useful therapeutic option in isolated conjunctival Kaposi's sarcoma.

PMID: 16193435 [PubMed - indexed for MEDLINE]

Application of biological dressings from radiosterilized amnios with cobalt 60 and serologic studies on the handling of burns in pediatric patients.

• Ley-Chavez E,
• Martinez-Pardo ME,
• Roman R,
• Oliveros-Lozano Fde J,
• Canchola-Martinez E.

Hospital Materno Infantil, ISSEMyM, Toluca, Mexico. eley_chavez@yahoo.com.mx

We present the use of biological dressing (amnios) as an alternative for skin replacement due to burn accidents. With the use of serologic tests, it is guaranteed to be free of the possibility of transmitting infectious diseases. The amnios is sterilized with 60Co gamma radiation. In this way, amnios is free of bacteria and fungi. In addition, with the use of serologic tests at the moment of the childbirth and 6 months later, we can be sure that it won't transmit the syphilis, AIDS, hepatitis b and c viruses. This treatment was applied to 12 children with burns of 1 degree and 2 degrees degree (7 girls and 5 boys) that required hospitalization. The application is very simple. It does not require of anesthesic procedure, or the use of surgical room. The pain decreased notoriously, and the procedure can be repeated as frequently as necessary. Once the dressing is applied, it does not require any type of surgical cleaning. The scaring process begins at the borders and under the dressing, where there is a new tissue. It decreases: 1) the possibility of infection, and therefore reduces the consumption of antibiotic; 2) the use of analgesics; 3) the time of scaring and 4) the number of days of hospitalization.

PMID: 15171007 [PubMed - indexed for MEDLINE]

Lactosylceramide-induced apoptosis in primary amnion cells and amnion-derived WISH cells.

• Moore RM,
• Lundgren DW,
• Silver RJ,
• Moore JJ.

Department ofPediatrics, Cleveland, Ohio 44109, USA. jmoore@metroheath.org

Amnion apoptosis is part of a programmed process of fetal membrane remodeling leading to weakening and rupture. The apoptotic agent lactosylceramide is elevated in amniotic fluid of premature infants with rupture of membranes. We have shown that apoptosis in WISH cells, induced by staurosporine, cycloheximide, or actinomycin D, can be blocked by cyclooxygenase inhibitors, suggesting a relationship between prostaglandin production and apoptosis. Cyclic adenosine monophosphate (cAMP) is known to inhibit prostaglandin release in amnion and WISH cells. This study was undertaken to determine the apoptotic potential of lactosylceramide and the effect of cyclooxygenase inhibitors and cAMP activators on lactosylceramide-induced apoptosis in primary amnion and WISH cells.Primary amnion cells and WISH cells were incubated with lactosylceramide to determine apoptosis and prostaglandin E(2) (PGE(2)) release. Apoptosis was confirmed by agarose gel electrophoretic DNA fragmentation analysis, nuclear matrix protein (NMP), and nucleosome enzyme-linked immunosorbent assay. In some studies, cells were preincubated with cyclooxygenase inhibitors or cAMP activators.Lactosylceramide induced a 20-fold increase in NMP (measure of cell death) in both cell types. Apoptosis was confirmed by the studies listed in methods. Lactosylceramide increased PGE(2) release in parallel with apoptosis. Cyclooxygenase inhibitors as well as cAMP activators inhibited both PGE(2) release and apoptosis.Lactosylceramide-induced apoptosis in both amnion and WISH cells. Parallel PGE(2) release was demonstrated with apoptosis. Cyclooxygenase inhibitors and cAMP activators blocked both processes.

PMID: 12383912 [PubMed - indexed for MEDLINE]

Antibiotic regimens for management of intraamniotic infection.

• Hopkins L,
• Smaill F.

Department of Pathology and Molecular Medicine, Faculty of Health Sciences, McMaster University, Room 2N29, 1200 Main Street West, Hamilton, Ontario, Canada, L8N 3Z5.

BACKGROUND: Intraamniotic infection is associated with maternal morbidity and neonatal sepsis, pneumonia and death. Although antibiotic treatment is accepted as the standard of care, few studies have been conducted to examine the effectiveness of different antibiotic regimens for this infection and whether to administer antibiotics intrapartum or postpartum. OBJECTIVES: To study the effects of different maternal antibiotic regimens for intraamniotic infection on maternal and perinatal morbidity and mortality. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register (May 2002) and the Cochrane Controlled Trials Register (The Cochrane Library, Issue 2, 2002). SELECTION CRITERIA: Trials where there was a randomized comparison of different antibiotic regimens to treat women with a diagnosis of intraamniotic infection were included. The primary outcome was perinatal morbidity. DATA COLLECTION AND ANALYSIS: Data were extracted from each publication independently by the authors. MAIN RESULTS: Two eligible trials (181 women) were included in this review. No trials were identified that compared antibiotic treatment with no treatment. Intrapartum treatment with antibiotics for intraamniotic infection was associated with a reduction in neonatal sepsis (relative risk (RR) 0.08; 95% confidence interval (CI) 0.00, 1.44) and pneumonia (RR 0.15; CI 0.01, 2.92) compared with treatment given immediately postpartum, but these results did not reach statistical significance (number of women studied = 45). There was no difference in the incidence of maternal bacteremia (RR 2.19; CI 0.25, 19.48). There was no difference in the outcomes of neonatal sepsis (RR 2.16; CI 0.20, 23.21) or neonatal death (RR 0.72; CI 0.12, 4.16) between a regimen with and without anaerobic activity (number of women studied = 133). There was a trend towards a decrease in the incidence of post-partum endometritis in women who received treatment with ampicillin, gentamicin and clindamycin compared with ampicillin and gentamicin alone, but this did not reach statistical significance (RR 0.54; CI 0.19, 1.49). REVIEWER'S CONCLUSIONS: The conclusions that can be drawn from this meta-analysis are limited due to the small number of studies. For none of the outcomes was a statistically significant difference seen between the different interventions. Current consensus is for the intrapartum administration of antibiotics when the diagnosis of intraamniotic infection is made; however, the results of this review neither support nor refute this although there was a trend towards improved neonatal outcomes when antibiotics were administered intrapartum. No recommendations can be made on the most appropriate antimicrobial regimen to choose to treat intraamniotic infection.

PMID: 12137684 [PubMed - indexed for MEDLINE]

Amniotic membrane, tear film, corneal and aqueous levels of ofloxacin in rabbit eyes after amniotic membrane transplantation.

• Panda A,
• Nainiwal SK,
• Sundan R,
• Pangtey MS.

PMID: 12131050 [PubMed - indexed for MEDLINE]

Toxic shock syndrome after laminaria insertion.

• Sutkin G,
• Capelle SD,
• Schlievert PM,
• Creinin MD.

Department of Obstetrics, Gynecology, and Reproductive Sciences, University of Pittsburgh School of Medicine and Magee-Womens Hospital, Pittsburgh, Pennsylvania, USA. obggs@ttuhsc.edu

BACKGROUND: Laminaria tents used to facilitate surgical abortion are rarely associated with significant infectious morbidity. CASE: A parous woman in midpregnancy had laminaria placed in her cervix followed by a second set after 24 hours. Eight hours later, she presented with dyspnea, hives, fever, tachycardia, and hypotension. Antibiotic treatment was initiated and a dilation and evacuation procedure was performed. Amniotic membrane cultures showed a heavy growth of Staphylococcus aureus with staphylococcal enterotoxin C expression, compatible with toxic shock syndrome. CONCLUSION: Laminaria cervical dilation might be associated with toxic shock syndrome.

PMID: 11704221 [PubMed - indexed for MEDLINE]

Early onset mixed Morganella and Klebsiella sepsis in a neonate.

• Maheshwari A,
• Dutta S,
• Kumar P,
• Narang A.

Neonatology Unit, Department of Pediatrics, Post Graduate Institute of Medical Education and Research (PGIMER), Chandigarh-160012, India.

A premature baby girl was delivered vaginally to a mother who had an otherwise normal pregnancy, and spontaneous premature onset of labour. She had early onset neonatal sepsis with pneumonia. The baby's blood culture as well as the amniotic membrane culture grew Morganella and Klebsiella. She recovered on appropriate antibiotics. This is only the second reported case of early onset neonatal sepsis due to Morganella. The literature is reviewed.

PMID: 11519292 [PubMed - indexed for MEDLINE]

Amniotic membrane, tear film, corneal, and aqueous levels of ofloxacin in rabbit eyes after amniotic membrane transplantation.

• Kim HS,
• Sah WJ,
• Kim YJ,
• Kim JC,
• Hahn TW.

Department of Ophthalmology, College of Medicine, The Catholic University of Korea, Seoul, Korea.

PURPOSE: We evaluated ocular penetration and drug levels in tears after topical ofloxacin instillation in rabbit eyes with amniotic membrane transplantation (AMT). METHODS: Forty-eight New Zealand White rabbits were used. In the first set of experiments, 24 rabbits (24 eyes) were divided into four groups according to the epithelial removal or AMT. Topical ofloxacin was instilled four times every 15 minutes. One hour after the last eyedrop, the concentration of ofloxacin in the amniotic membrane, cornea, and aqueous humor was evaluated. In the second set of experiments, 24 rabbits were divided into six groups according to AMT (transplantation of lyophilized or fresh amniotic membrane) or duration of application. Ofloxacin ointment or two drops of ofloxacin were applied to the right eye, and then tear samples were collected after 0.5, 1, 2, 4, and 6 hours for the analysis of ofloxacin concentration. RESULTS: Mean ofloxacin concentrations in the cornea and aqueous humor were statistically higher in deepithelialized cornea regardless of AMT (p < 0.05). The mean tear levels of ofloxacin in the AMT groups were statistically higher than those in non-AMT groups (p < 0.05). There was no statistical significance in the tear level of ofloxacin between lyophilized amniotic membrane groups and fresh amniotic membrane groups nor between 1-hour amniotic membrane-attached groups and 6-hour amniotic membrane-attached groups. CONCLUSION: Amniotic membrane transplantation seems to interfere with the ocular penetration of topical ofloxacin in normal rabbit corneas but enhances ofloxacin penetration in corneas with epithelial defects. The ofloxacin level in tears was higher in eyes with AMT up to 1 hour after topical ofloxacin use. Therefore, it seems that amniotic membrane has some potential to act as an effective drug delivery system.

PMID: 11473165 [PubMed - indexed for MEDLINE]

Epiphora caused by conjunctivochalasis: treatment with transplantation of preserved human amniotic membrane.

• Georgiadis NS,
• Terzidou CD.

Department of Ophthalmology, Aristotle University of Thessaloniki, Thessaloniki, Greece. gnick@med.auth.gr

PURPOSE: To present our experience with the use of preserved human amniotic membrane on patients with epiphora caused by conjunctivochalasis. METHODS: Twelve patients, seven women and five men, ages ranging from 56 to 72 years (mean, 61 years) were referred to our Cornea Service with chronic epiphora. In all patients, no punctal ectropion was present, the lacrimal pathway was patent, and the dye disappearance test was abnormal. All patients had already undergone various therapies including multiple irrigations of the lacrimal system, antibiotic drops, steroid drops, and artificial tear drops. In all patients, conjunctivochalasis, which was not previously diagnosed, was evident on slit-lamp examination. After surgical removal of the excess conjunctiva, preserved human amniotic membrane was placed over and sutured with 10-0 nylon continuous suture to the free conjunctival edges. During the postoperative period, artificial tear drops and steroid/antibiotic drops were applied. RESULTS: Improvement of the epiphora was evident from the first postoperative day. After removal of the suture 10 to 15 days (mean, 12 days) after surgery, no patient complained of epiphora. The dye disappearance test was normal. During the follow-up period, which ranged from 6 to 11 months (mean, 8 months), no patient complained of epiphora and no conjunctivochalasis was detected in the area in which human amniotic membrane was transplanted. CONCLUSION: In our experience, transplantation of preserved human amniotic membrane greatly improved symptoms of epiphora caused by conjunctivochalasis. Continued education of the general ophthalmologists concerning this condition is required.

PMID: 11473163 [PubMed - indexed for MEDLINE]

Multilayer amniotic membrane transplantation in severe ocular graft versus host disease.

• Peris-Martinez C,
• Menezo JL,
• Diaz-Llopis M,
• Avino-Martinez JA,
• Navea-Tejerina A,
• Risueno-Reguillo P.

Department of Ophthalmology, University Hospital La Fe, Valencia, Spain. cperis@ctv.es

PURPOSE: To clarify the usefulness of multilayer amniotic membrane transplantation in an unusual case of calcareous corneal degeneration in a patient with graft-versus-host disease. METHODS: A 20-year-old Caucasian woman had bilateral calcareous corneal degeneration of one year of evolution, secondary to graft-versus-host disease. Treatment for both eyes with topical steroids and antibiotic ointment was not successful. Right eye had a spontaneous corneal perforation, and a three-layer circle amniotic membrane graft was applied to the whole cornea. RESULTS: During a follow-up period of 20 months we observed stability of the corneal epithelium and stroma. The amniotic membrane-covered area showed rapid epithelization, reduced inflammation and suppressed fibrosis formation. CONCLUSIONS: Multilayer amniotic membrane transplantation may be considered an alterantive for reconstructing the ocular surface in a patient with severe dry eyes and calcareous corneal degeneration, even with little perforation.

PMID: 11456023 [PubMed - indexed for MEDLINE]

[Antibiotics for preterm and premature rupture of amniotic membranes]

• Aguilera C,
• Sagala J.

Fundacio Institut Catala de Farmacologia. Servicios de Farmacologia Clinica. Hospital Vall d'Hebron. Barcelona.

PMID: 11412700 [PubMed - indexed for MEDLINE]

Systematic review of the use of honey as a wound dressing.

• Moore OA,
• Smith LA,
• Campbell F,
• Seers K,
• McQuay HJ,
• Moore RA.

Pain Research Unit and Nuffield Department of Anaesthetics, University of Oxford, Oxford, Radcliffe Hospital, The Churchill, Headington, Oxford OX3 7LJ, UK. omoore@doctors.org.uk

OBJECTIVE: To investigate topical honey in superficial burns and wounds though a systematic review of randomised controlled trials. DATA SOURCES: Cochrane Library, MEDLINE, EMBASE, PubMed, reference lists and databases were used to seek randomised controlled trials. Seven randomised trials involved superficial burns, partial thickness burns, moderate to severe burns that included full thickness injury, and infected postoperative wounds. REVIEW METHODS: Studies were randomised trials using honey, published papers, with a comparator. Main outcomes were relative benefit and number-needed-to-treat to prevent an outcome relating to wound healing time or infection rate. RESULTS: One study in infected postoperative wounds compared honey with antiseptics plus systemic antibiotics. The number needed to treat with honey for good wound healing compared with antiseptic was 2.9 (95% confidence interval 1.7 to 9.7). Five studies in patients with partial thickness or superficial burns involved less than 40% of the body surface. Comparators were polyurethane film, amniotic membrane, potato peel and silver sulphadiazine. The number needed to treat for seven days with honey to produce one patient with a healed burn was 2.6 (2.1 to 3.4) compared with any other treatment and 2.7 (2.0 to 4.1) compared with potato and amniotic membrane. For some or all outcomes honey was superior to all these treatments. Time for healing was significantly shorter for honey than all these treatments. The quality of studies was low. CONCLUSION: Confidence in a conclusion that honey is a useful treatment for superficial wounds or burns is low. There is biological plausibility.

PMID: 11405898 [PubMed - indexed for MEDLINE]

Multilayered amniotic membrane transplantation for severe ulceration of the cornea and sclera.

• Hanada K,
• Shimazaki J,
• Shimmura S,
• Tsubota K.

Department of Ophthalmology, Tokyo Dental College, Chiba, Japan. hanada@asahikawa-med.ac.jp

PURPOSE: To examine the efficacy of amniotic membrane transplantation in the treatment of deep corneal and scleral ulcers. PATIENTS: A total of 11 patients were recruited for this study: four patients (four eyes) with corneal perforation, five patients (five eyes) with a deep corneal ulcer and descemetocele, and two patients (two eyes) with a scleral ulcer. METHODS: Ulcers were treated by amniotic membrane transplantation. Separate amniotic membranes were transplanted as material to fill the stromal layer (amniotic membrane filling), as a basement membrane (amniotic membrane graft), and as a wound cover (amniotic membrane patch). After surgery, all cases were treated with artificial tears, autologous serum drops, antibiotic eyedrops, topical corticosteroids, and sodium hyaluronate eyedrops. RESULTS: Eight eyes (72.7%) healed with epithelialization in 16.5 +/- 8.0 days (range, 7 to 29 days), with five and three eyes showing corneal epithelialization and conjunctival epithelialization, respectively. A persistent epithelial defect was noted in one eye with corneal ulcer after limbal allograft transplantation for a chemical burn and in two eyes with corneal ulcers as a complication of rheumatoid arthritis. CONCLUSION: Multilayered amniotic membrane transplantation may be effective for the treatment of deep ulceration of the cornea and sclera. In some eyes with total corneal limbal dysfunction or autoimmune disorders, amniotic membrane transplantation alone is not effective.

PMID: 11239864 [PubMed - indexed for MEDLINE]

Antibacterial properties of human amnion and chorion in vitro.

• Kjaergaard N,
• Hein M,
• Hyttel L,
• Helmig RB,
• Schonheyder HC,
• Uldbjerg N,
• Madsen H.

Department of Obstetrics and Gynecology, Aalborg Hospital, DK-9000 Aalborg, Denmark. n.kjaergaard@dadlnet.dk

OBJECTIVE: The purpose of the present study was to explore the direct effects of amnion and chorion on bacterial growth in vitro including the antibacterial spectrum. Chorioamniotic membranes were obtained under sterile conditions from 13 healthy women undergoing elective cesarean section at term. Likewise, chorioamniotic membranes were obtained from 10 healthy women with spontaneous vaginal delivery at term. Five strains of Hemolytic streptococci group B (GBS) were tested and one clinical isolate of the following species or bacterial groups: Hemolytic streptococcus group A, Staphylococcus aureus, Staphylococcus saprophyticus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter calcoaceticus and Lactobacillus species. Bacteriological media included (1) blood-agar medium; (2) a transparent agar medium for submerged cultures; and (3) a nutrient broth medium. RESULTS: An inhibitory effect of fetal membranes against a range of bacteria was found. Consistent results were obtained in experiments with cultures on agar and cultures suspended in agar (membranes from eight women in both studies). In experiments with liquid cultures (seven women) only chorion showed a marginal inhibitory effect. All strains were inhibited, but the most pronounced inhibition was obtained for streptococcus group A, S. aureus and S. saprophyticus by both chorion and amnion. CONCLUSION: This study demonstrated an inhibitory effect of the fetal membranes on a diverse panel of bacteria

PMID: 11165729 [PubMed - indexed for MEDLINE]

Treatment of persistent corneal epithelial defect with extended wear of a fluid-ventilated gas-permeable scleral contact lens.

• Rosenthal P,
• Cotter JM,
• Baum J.

The Boston Foundation for Sight (Drs Rosenthal and Cotter), Boston, Massachusetts 02467, USA. perry-rosenthal@polymer.com

PURPOSE: To report treatment of persistent corneal epithelial defects unresponsive to other therapies by extended wear of a fluid-ventilated gas-permeable scleral contact lens.METHODS: In this retrospective study, 14 eyes of 13 consecutive patients referred for the treatment of persistent corneal epithelial defects that failed to heal with conventional therapies or developed epithelial defects after penetrating keratoplasty for persistent corneal epithelial defects were fitted with an extended-wear gas-permeable scleral lens. These included seven eyes of six patients with Stevens-Johnson syndrome and seven eyes of seven patients who did not have Stevens-Johnson syndrome. Twelve eyes had undergone recent penetrating keratoplasty. All 14 eyes were fitted with a gas-permeable scleral contact lens designed to avoid the intrusion of air bubbles under its optic. An antibiotic and corticosteroid were added to the lens fluid reservoir or instilled before each lens insertion in 12 of 14 eyes. The lenses were worn continuously except for brief periods of removal for purposes of cleaning, replacement of the lens fluid reservoir, and examination and photography of the cornea.RESULTS: Five of the seven persistent corneal epithelial defects associated with Stevens-Johnson syndrome healed. The persistent corneal epithelial defects of four of these eyes re-epithelialized within 7 days, and a fifth healed in 27 days of gas-permeable scleral lens extended wear. A sixth persistent corneal epithelial defect that failed to heal initially re-epithelialized after a subsequent penetrating keratoplasty and gas-permeable scleral lens extended wear. The seventh eye healed after 3 days of gas-permeable scleral lens extended wear, but the persistent corneal epithelial defect subsequently recurred. Three of seven non-Stevens-Johnson syndrome persistent corneal epithelial defects re-epithelialized within 36 hours, 6 days, and 36 days, respectively. Of the six (six of 14) persistent corneal epithelial defects that failed to heal with a gas-permeable scleral lens extended wear, one subsequently healed after multiple amniotic membrane grafts. Microbial keratitis occurred in four eyes (four of 14) and graft failure in one eye, all of which required repeat penetrating keratoplasty.CONCLUSION: Extended wear of an appropriately designed gas-permeable scleral contact lens was effective in promoting the healing of persistent corneal epithelial defects in some eyes that failed to heal after other therapeutic measures. Re-epithelialization appears to be aided by a combination of oxygenation, moisture, and protection of the fragile epithelium afforded by the scleral lens. However, microbial keratitis represents a significant risk.

PMID: 11004257 [PubMed - indexed for MEDLINE]

Comment in:

Ophthalmology. 2001 Oct;108(10):1714-5.

Hypopyon after repeated transplantation of human amniotic membrane onto the corneal surface.

• Gabler B,
• Lohmann CP.

University Eye Clinic, Regensburg, Germany. bernhard.gabler@klinik.uni-regensburg.de

OBJECTIVE: The authors describe a localized probable immunoreaction after repeated transplantation of amniotic membrane (AM) onto the corneal surface. DESIGN: Interventional case report. INTERVENTION: Amniotic membrane was transplanted onto the corneal surface of a 78-year-old female with a deep trophic corneal ulcer resulting in temporary epithelial closure. A second and finally third amniotic membrane transplantation (AMT) was performed because of recurrent ulcerations. All three AMs were obtained from the same donor. RESULTS: The first transplantation of the AM was without complication. However, a hypopyon developed 2 days after the second and 2 days after the third AMT, but the patient responded immediately to topical and systemic corticosteroids. CONCLUSIONS: Immunologic, toxic, and hypersensitivity effects could have contributed to the hypopyon iritis that appeared after the second and third AMT, but not after the initial transplantation. In case of a repeated AMT, the use of AM from different donors may help to minimize the risk of an immediate postoperative intraocular inflammation.

PMID: 10889109 [PubMed - indexed for MEDLINE]

[Apoptotic cell death induced by actinomycin D arisen at different levels and cell cycle stages according to human cultured cell species]

• Ohyama K,
• Sakai N,
• Maruhashi Y,
• Enn P,
• Uchide N,
• Yamakawa T.

Department of Biochemistry, School of Pharmacy, Tokyo University of Pharmacy & Life Science, Japan.

Nine kinds of human cultured cells, including fetus cells (smooth chorion trophoblast cells, amnion epithelial cells and HE-21), adult non-carcinoma cells (HCF), and carcinoma cells (KATO-III, COLO 201, Lu-134-AH, SK-OV-3 and SKG-3a) were stimulated with Actinomycin (Act.) D for 24 h. Apoptosis induction was investigated by agarose gel electrophoresis for DNA fragmentation analysis and by flow cytometric analysis of stained cells using in situ terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick-end labeling TUNEL) staining techniques for the quantification of apoptosis, and simultaneously using propidium iodide for the gain of some information about cell cycle. By agarose gel electrophoresis, DNA fragmentation of these cells except amnion epithelial and SKG-3a cells was detected, depending on concentration of Act. D. Using flow cytometric analysis, these cells were separated into four groups according to the information about cell cycle. Group 1 included amnion epithelial and SKG-3a cells, which were TUNEL negative. In group 2, all cell populations at G0/G1 and G2/M phases of HCC, KATO-III and SK-OV-3 were TUNEL staining positive. A portion of each G0/G1 or G2/M phase cell of Lu-134-AH and COLO 201 in group 3 was TUNEL stain positive. In group 4, G2/M phase cells of smooth chorion trophoblast cells and HE-21 were mostly stained and a small population of G0/G1 phase cells were also TUNEL stain positive. These results show that the stages of the cell cycle at which apoptosis was arisen by Act. D stimulation were significantly different depending on the cells types.

PMID: 10825812 [PubMed - indexed for MEDLINE]

Cyclooxygenase inhibitors decrease apoptosis initiated by actinomycin D, cycloheximide, and staurosporine in amnion-derived WISH cells.

• Moore RM,
• Lundgren DW,
• Moore JJ.

Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, Ohio, USA.

Apoptosis is a process by which external or developmental factors induce a specific series of events leading to cell death. Recently, apoptotic cells have been described in rat amnion membrane at late gestation, suggesting apoptosis may be involved in membrane rupture. Mechanisms controlling amnion cell apoptosis are unknown. The objective of this study was to investigate whether cyclooxygenase and prostaglandins are integral to apoptosis in amnion, as reported in intestinal epithelial cells and renal mesangial cells. Amnion-derived WISH cells underwent apoptosis in a dose- and time-dependent manner after incubation with actinomycin D, cycloheximide, or staurosporine, as determined by cell viability, DNA fragmentation analysis, and fluorescent in situ fragmentation analysis. Cells cultured with increasing doses of these agents also demonstrated concomitant increases in prostaglandin E2 output. WISH cell coincubation with these agents and the cyclooxygenase inhibitors indomethacin or piroxicam resulted in dose-dependent decreases in both prostaglandin E2 and apoptosis. Cultures incubated with 0.5 microgram/mL actinomycin D showed 80.7% cell apoptosis after 12 hours compared with 1.1% in untreated cultures. After 24 hours incubation with actinomycin D, 0.8% of the original cell number remained attached to the plate. In cultures coincubated with 0.5 microgram/mL actinomycin D and 100 mumol/L indomethacin, only 19.2%, 24.7%, and 39.3% of the cells were found to be apoptotic after 12, 24, and 48 hours in culture, respectively. Similar trends were observed after the use of cycloheximide or staurosporine in combination with indomethacin or prioxicam. These data suggest that cyclooxygenase and/or prostaglandins play a role in programmed cell death of amnion-derived WISH cells in culture.

PMID: 10554762 [PubMed - indexed for MEDLINE]

Tyrosine kinase inhibitors block the glucocorticoid stimulation of prostaglandin endoperoxide H synthase expression in amnion cells.

• Zakar T,
• Mijovic JE,
• Bhardwaj D,
• Olson DM.

University of Alberta Perinatal Research Centre, Department of Obstetrics and Gynaecology, Edmonton, Canada.

Human amnion cells in primary culture respond to glucocorticoids in a characteristic fashion by the increased expression of the inducible prostaglandin endoperoxide H synthase isoenzyme, PGHS-2. Since PGHS-2 induction by agonists generally involves tyrosine kinases, we examined the possibility that the glucocorticoid stimulation of PGHS-2 in the amnion cells is tyrosine kinase dependent. PGHS-2 expression was stimulated in confluent, serum-starved amnion cells with dexamethasone, and the effect of the tyrosine kinase inhibitors herbimycin A and tyrphostins AG126, AG1288, and A1 on enzyme activity induction was determined. All four inhibitors blocked the increase of PGHS activity in a concentration-dependent manner with IC50 values of 0.077 +/- 0.05, 15.38 +/- 5.14, 20.91 +/- 3.1, and 29.77 +/- 8.21 microM, respectively (mean +/- SE, n = 4). Dexamethasone increased (approximately twofold) the tyrosine phosphorylation of 120-, 110-, and 77-kDa proteins in cell extracts, and herbimycin A selectively blocked the phosphorylation of the 110-kDa phosphoprotein. The stimulation of the steady-state level of PGHS-2 mRNA by dexamethasone was also inhibited by herbimycin A. These results suggest that glucocorticoids induce PGHS-2 expression in amnion cells with the involvement of tyrosine kinase(s). The role of tyrosine kinase dependent mechanisms in the control of amnion cell responsiveness to corticosteroids remains to be established.

PMID: 10535705 [PubMed - indexed for MEDLINE]

Effect of different classes of antibiotics on amniotic prostaglandin E release.

• Vesce F,
• Pavan B,
• Buzzi M,
• Pareschi MC,
• Bianciotto A,
• Iorizzo G,
• Biondi C.

Department of Biomedical Sciences, University of Ferrara, Italy.

Our purpose was to investigate the effects of different classes of antibiotics, namely beta-lactamines, aminoglicosides, tetracyclines, macrolides, on amniotic prostaglandin E release to clarify their role in the treatment of premature labor. The effects of these antibiotics were tested also in combination with ampicillin, whose antiprostaglandinergic action had been demonstrated previously. Ceftriaxone and gentamicin significantly and reversibly inhibit both basal and arachidonic acid- or oxytocin-stimulated prostaglandin E release from amnion, although to a different extent. On the contrary, tetracycline and erythromycin do not influence prostaglandin E output. The inhibitory effect of ampicillin is potentiated, in an additive manner, by ceftriaxone, reduced by gentamycin, and eliminated by tetracycline and erythromycin. The finding that diverse classes of antibiotics and their combinations affect amniotic prostaglandin E release should be taken into account in the management of premature labor.

PMID: 10402215 [PubMed - indexed for MEDLINE]

Amnioinfusion in the evaluation of fetal obstructive uropathy: the effect of antibiotic prophylaxis on complication rates.

• Feldman B,
• Hassan S,
• Kramer RL,
• Kasperski SB,
• Evans MI,
• Johnson MP.

Division of Reproductive Genetics, Department of Obstetrics and Gynecology, Center for Fetal Diagnosis and Therapy, Hutzel Hospital, Detroit, Mich., USA.

OBJECTIVE: Amnioinfusion plays an important role in the intrauterine evaluation and treatment of fetal obstructive uropathy. However, it may significantly increase the risk for chorioamnionitis, premature rupture of membranes and premature labor. We evaluated the impact of prophylactic antibiotics on postamnioinfusion complications. METHODS: Thirty pregnancies complicated by fetal obstructive uropathy, treated by amnioinfusion and with documentation of pregnancy outcome were identified from our database. Pregnancy outcomes were compared between patients who were treated with prophylactic antibiotics and those with no prophylaxis. RESULTS: Chorioamnionitis was diagnosed in 3 out of 15 (20%) patients who did not receive prophylactic antibiotics as compared to 2 (13%) in the treated group. The overall rate of serious obstetrical complication was significantly higher in the untreated group (66 vs. 20%; p = 0. 021). Patients receiving prophylactic antibiotics delivered at a significantly greater gestational age than those who did not receive antibiotics (34.0 +/- 3.7 vs. 31.3 +/- 1.9 weeks, respectively; p = 0.018). CONCLUSIONS: Our study supports the use of oral prophylactic antibiotics as being effective in reducing the previously observed significant risks associated with amnioinfusion in fetal obstructive uropathy.

PMID: 10364669 [PubMed - indexed for MEDLINE]

Prophylactic amnioinfusion in preganancies complicated by chorioamnionitis: a prospective randomized trial.

• Parilla BV,
• McDermott TM.

Department of Obstetrics and Gynecology, Northwestern University Medical School and Northwestern Memorial Hospital, Chicago, Illinois, USA.

The objective of this article is to prospectively investigate the efficacy of amnioinfusion as a means to reduce febrile morbidity in pregnancies complicated by chorioamnionitis. All laboring patients with a temperature > or =100.1 degrees F were approached for study participation. Exclusion criteria included amnionitis diagnosed at greater than 8 cm dilation, multiple gestation, placental abruption, or a nonreassuring fetal heart rate tracing. Consenting patients were randomized to receive antibiotics (ampicillin or penicillin with gentamicin) and acetaminophen with or without amnioinfusion. All patients received intrauterine pressure catheter placement. For study patients, normal saline at room temperature was infused at 10 mL/min for 60 min, then 3 mL/min until delivery. Postpartum endometritis was defined as a temperature = 100.4 degrees F accompanied by uterine tenderness more than 12 hr after delivery. Statistical analysis was performed using the Student's t-test for continuous data and Chi-square for discrete variables. Thirty-six patients were enrolled, and complete data were available for 34 patients (17 in each group). There were no differences between groups with respect to maternal age, gravidity, race, or gestational age. There were also no differences between groups in duration of rupture of membranes, temperature at randomization, interval from randomization to delivery, cesarean section rate, or umbilical cord arterial pH. The mean temperature at the time of delivery was 99.8+/-0.9 degrees F for the amnioinfusion group versus 100.5+/-1.0 degrees F for the control group (p=0.046). Three of 17 amnioinfusion patients and 3 of 17 control patients had postpartum endometritis. There was 1 neonatal infection in the treatment group and no neonatal infections among the control patients. Prophylactic amnioinfusion was associated with a decline in temperature at the time of delivery. No untoward effects from the amnioinfusion were identified.

PMID: 10333390 [PubMed - indexed for MEDLINE]

Pathophysiology, diagnosis, and management of intraamniotic infection.

• Riggs JW,
• Blanco JD.

Department of Obstetrics and Gynecology, University of Texas Medical School-Houston, Lyndon B. Johnson General Hospital 77026, USA.

Intraamniotic infection (IAI) is a term used to describe a clinically diagnosed infection of the contents of the uterus. It is found most often after rupture of the membranes. The most useful diagnostic tests are physical examination, amniotic fluid glucose determination, and amniotic fluid Gram's stain. There is no clearly established means for the prevention of IAI, but cervical examinations and cervical manipulation can increase the risk, so caution with their use is still warranted. Treatment for this infection should be initiated when the diagnosis is made to provide the lowest risk of neonatal and maternal complications. Ampicillin or penicillin plus gentamicin are the most extensively tested antibiotics for treatment before delivery. Clindamycin or metronidazole should be added if a cesarean section is performed. As a general rule, antibiotics should be continued postpartum until the patient has been afebrile and asymptomatic for a minimum of 24 hours. Neonatal complications of IAI may be substantial especially for the premature fetus. Women with this infection have a greater risk for dysfunctional labor and cesarean section.

PMID: 9738989 [PubMed - indexed for MEDLINE]

Induction of oxytocin receptor gene expression in rabbit amnion cells.

• Jeng YJ,
• Lolait SJ,
• Soloff MS.

Department of Obstetrics and Gynecology, University of Texas Medical Branch, Galveston 77555-1062, USA.

Oxytocin (OT)-stimulated PGE2 release by rabbit amnion is enhanced by the up-regulation of oxytocin receptors (OTR), which increase about 200-fold at the end of pregnancy. As recent studies have shown that PGs are essential for parturition, the rise in amnion OTR and associated PGE2 synthesis are probably essential for labor initiation. The present work was directed toward understanding the mechanisms of OTR up-regulation. Levels of agents that stimulate adenylyl cyclase activity and cortisol are increased in amniotic fluid at the end of pregnancy. Addition of either forskolin or cortisol to cultured amnion cells caused an increase in OTR ligand-binding sites and steady state OTR messenger RNA (mRNA) levels. Forskolin treatment elevated OTR mRNA levels rapidly, but transiently, whereas cortisol's effects were slower and sustained. Actinomycin or cycloheximide, added 3 h after forskolin, led to a sustained elevation in OTR mRNA levels, suggesting that forskolin increases the activities of OTR mRNA-destabilizing factors along with increasing OTR mRNA concentration. Cortisol did not appear to affect OTR mRNA stability. Measurement of OTR mRNA transcription rates showed that forskolin's effects were maximal within 1 h of treatment. In contrast, cortisol-induced transcription was not apparent until 8 h. The effects of forskolin and cortisol on OTR gene transcription were synergistic. Thus, the increase in OTR mRNA levels occurring after either forskolin or cortisol treatments is the result of induction of OTR gene expression, but the effects of the two agents appear to occur at separate sites.

PMID: 9681495 [PubMed - indexed for MEDLINE]

Inhibition of amniotic prostaglandin E release by ampicillin.

• Vesce F,
• Buzzi M,
• Ferretti ME,
• Pavan B,
• Bianciotto A,
• Jorizzo G,
• Biondi C.

Department of Biomedical Sciences and A.T., University of Ferrara, Italy.

OBJECTIVE: The effect of antibiotics in the prevention of preterm labor needs to be further investigated. The aim of this study was to determine the effect of ampicillin on prostaglandin E release from amnion as a possible explanation for its ability to retard preterm labor. STUDY DESIGN: The effect of the beta-lactam antibiotic ampicillin on prostaglandin E release from human amnion was tested under basal and stimulated conditions. RESULTS: Ampicillin dose dependently inhibits basal prostaglandin E release from amnion in both static and dynamic conditions. In our experiments, 10(-7) mol/L ampicillin (a concentration able to significantly inhibit prostaglandin E output) leaves the microbiologic features of the medium substantially unmodified up to 5 hours of incubation. Moreover, the drug reversibly counteracts the prostaglandin E elevation induced by arachidonic acid or oxytocin. CONCLUSION: This finding (i.e., that ampicillin inhibits prostaglandin E release from amnion) may offer an explanation for a beneficial response to ampicillin therapy in the case of preterm labor even in the absence of bacterial infection.

PMID: 9579440 [PubMed - indexed for MEDLINE]

Regulation of prostaglandin H2 synthase-2 expression in primary human amnion cells by tyrosine kinase dependent mechanisms.

• Zakar T,
• Mijovic JE,
• Eyster KM,
• Bhardwaj D,
• Olson DM.

Perinatal Research Centre, Department of Obstetrics and Gynaecology, University of Alberta, Edmonton, Alberta, Canada. tzakar@gpu.srv.ualberta.ca

Prostaglandin H2 synthase (PGHS)-1 and PGHS-2 expression was examined in primary cultures of human amnion cells, an in vitro model of amnion tissue. Epidermal growth factor (EGF), the protein kinase C (PKC) activating phorbol ester TPA, and the protein phosphatase inhibitor, okadaic acid (OA), stimulated PGHS activity and the level of PGHS-2 mRNA, but did not affect the level of PGHS-1 mRNA. In situ hybridization suggested that the same population of cells responded to EGF, TPA and OA. Okadaic acid promoted PGHS activity independently of PKC. EGF stimulated the activity of extracellular signal-regulated protein kinase (Erk) and N-terminal c-Jun kinase (Jnk). OA increased Jnk activity but had no effect on Erk activity, while TPA had no influence on either Erk or Jnk activity. PD098059, a selective inhibitor of the Erk-activating kinase MEK, blocked the stimulation of PGHS expression by EGF, but did not decrease stimulation in response to OA. Herbimycin A, a tyrosine kinase inhibitor, suppressed the stimulation of PGHS activity and PGHS-2 mRNA abundance by all three stimulants, and blocked signalling via the Erk and Jnk mitogen-activated protein kinase pathways. Thus, growth factor stimulation, PKC activation and protein phosphatase inhibition induced the expression of PGHS-2 in primary amnion cells by distinct regulatory mechanisms involving tyrosine kinase(s). Tyrosine kinase inhibitors may constitute a new category of PGHS-2 inhibitors that act by blocking the expression of the enzyme. Copyright 1998 Elsevier Science B.V.

PMID: 9518544 [PubMed - indexed for MEDLINE]

Hypotonic stress increases cyclooxygenase-2 expression and prostaglandin release from amnion-derived WISH cells.

• Lundgren DW,
• Moore RM,
• Collins PL,
• Moore JJ.

Labor Focus Research Group, Case Western Reserve University School of Medicine, MetroHealth Medical Center, Cleveland, Ohio 44109, USA.

This report examines the effect of cell volume expansion on cyclooxygenase-2 (COX-2) mRNA expression, COX-2 protein expression, and prostaglandin E2 release from human amnion-derived WISH cells. Earle's balanced salts solution (EBSS) with limited NaCl concentration was utilized as the induction medium. COX-2 mRNA was elevated 6-fold in cells incubated for 1 h in hypotonic EBSS. COX-2 mRNA expression was not increased when raffinose or sucrose were used to reconstitute low NaCl. Actinomycin D blocked COX-2 mRNA increase by hypotonic stress, while cycloheximide enhanced COX-2 mRNA expression. COX-2 mRNA and protein concentrations increased as a function of decreasing media osmolarity and incubation time in hypotonic EBSS. Hypotonic EBSS induced a 3-fold increase in prostaglandin E2 release. WISH cells transiently transfected with a luciferase expression vector driven by the human COX-2 promoter for the COX-2 gene show a 3-fold increase in luciferase activity when incubated in hypotonic EBSS. COX-2 mRNA levels in primary human amnion cells were also increased by hypotonic stress. This study suggests that amnion cell COX-2 gene expression is regulated by cell volume expansion and/or increased plasma membrane tension.

PMID: 9242685 [PubMed - indexed for MEDLINE]

Interactions of the amino-terminal noncollagenous (NC1) domain of type VII collagen with extracellular matrix components. A potential role in epidermal-dermal adherence in human skin.

• Chen M,
• Marinkovich MP,
• Veis A,
• Cai X,
• Rao CN,
• O'Toole EA,
• Woodley DT.

Department of Dermatology, Northwestern University Medical School, Chicago, Illinois 60611, USA.

Type VII collagen, the major component of anchoring fibrils, consists of a central collagenous triple-helical domain flanked by two noncollagenous domains, NC1 and NC2. The NC1 domain contains multiple submodules with homology to known adhesive molecules including fibronectin type III-like repeats and the A domain of von Willebrand factor. In this study, we produced the entire NC1 domain of human type VII collagen in the stably transfected human kidney 293 cell clones and purified large quantities of the recombinant NC1 protein from serum-free culture media. The recombinant NC1 formed interchain disulfide-bonded dimers and trimers and was N-linked glycosylated. Tunicamycin inhibited the cellular secretion of NC1, suggesting that N-linked glycosylation may play a role in NC1 secretion. The recombinant NC1 was indistinguishable from the authentic NC1 obtained from human amnions or WISH cells with respect to N-linked sugar content, electrophoretic mobility, rotary shadow imaging, and binding affinity to type IV collagen. Purified recombinant NC1, like authentic NC1, also bound specifically to fibronectin, collagen type I, and a laminin 5/6 complex. Both monomeric and trimeric forms of NC1 exhibited equal affinity for these extracellular matrix components, suggesting that the individual arms of NC1 can function independently. The multiple interactions of NC1 with other extracellular matrix components may support epidermal-dermal adhesion.

PMID: 9169408 [PubMed - indexed for MEDLINE]

Prostaglandin endoperoxide H synthase-2 expression in human amnion cells: involvement of tyrosine kinases in the regulation.

• Zakar T,
• Mijovic JE,
• Olson DM.

Perinatal Research Centre, University of Alberta, Edmonton, Canada.

PMID: 9561184 [PubMed - indexed for MEDLINE]

[Cytotoxic properties of the complex of the antineoplastic antibiotic bleomycetin and Bacillus intermedius ribonuclease]

• Kalacheva NV,
• Bulgakova RSh,
• Kazantseva NIu,
• Kurinenko BM.

It was shown possible to change the cytotoxic properties of the antitumor antibiotic bleomycetin by its binding to Bacillus intermedius RNAse. The complexing lowered the antibiotic effect on DNA in the cells of the human amnion. At the same time the experiments with human red blood cells indicated that RNAse of B. intermedius in complex with bleomycetin-Fe(II) increased the antibiotic capacity for the cell membrane break down.

PMID: 9182501 [PubMed - indexed for MEDLINE]

Constitutively enhanced nbl expression is associated with the induction of internucleosomal DNA cleavage by actinomycin D.

• Naora H,
• Nishida T,
• Shindo Y,
• Adachi M,
• Naora H.

Research School of Biological Sciences, Australian National University, Canberra, Australia.

Previous studies have found nbl expression to transiently rise and fall during glucocorticoid-induced thymic apoptosis. This induction of apoptosis is blocked by the transcriptional inhibitor actinomycin D. However, actinomycin D can trigger apoptosis in other cell types, e.g., HL-60 cells. This study found that internucleosomal DNA cleavage typical of apoptosis is induced by actinomycin D in cell lines such as HL-60 which constitutively express high levels of nbl above a certain "threshold." In contrast, "DNA ladder" formation was not induced by actinomycin D in cell lines with low constitutive nbl expression. Enhanced nbl expression therefore appears to be associated with apoptosis which is either blocked or induced by actinomycin D.

PMID: 8694823 [PubMed - indexed for MEDLINE]

Renin stimulates decidual prostaglandin production via a novel mechanism that is independent of angiotensin II formation.

• Mitchell MD,
• Edwin SS,
• Pollard JK,
• Trautman MS.

Department of Pharmacology and Clinical Pharmacology, University of Auckland, New Zealand.

Renin is a proteolytic enzyme that has been considered to have only one function which is to cleave angiotensinogen between the 10th and 11th amino acids to form angiotensin-1. This is then converted to angiotensin-II, a potent vasoconstrictor, antinatriuretic and antidiuretic by angiotensin-converting enzyme. We have investigated the action of renin to stimulate prostaglandin production by decidual cells and in so doing have generated data that challenge the prevailing dogma. Renin stimulates decidual prostaglandin production in a concentration-related fashion that is unaffected by saralasin treatment. This stimulatory action of renin is enhanced rather than reduced by arachidonic acid treatment but abolished by treatment with cycloheximide or actinomycin D. Renin caused a more rapid recovery of decidual prostaglandin biosynthesis from acetylsalicylic acid treatment than did control media. Moreover, renin treatment of both decidual and amnion cells induced increased levels of PGHS-2 within 2 h. Collectively, these results indicate that renin can act directly, separately from the generation of angiotensin-I and II. In this case renin can induce PGHS expression.

PMID: 8829212 [PubMed - indexed for MEDLINE]

Clinical role of amnio-infusion.

• Ouzounian JG,
• Paul RH.

Department of Obstetrics & Gynaecology, University of Southern California School of Medicine, Los Angeles 90033, USA.

Amnio-infusion is a simple, yet beneficial, technique for improving pregnancy outcome. Antepartum amnio-infusion has been shown to be beneficial as an aid to enhancing ultrasonographic fetal imaging and may have a role in the administration of antibiotic therapy or the prevention of pulmonary hypoplasia. There are considerable data to support the intrapartum use of amnio-infusion in the presence of oligohydramnios, variable decelerations or meconium. Numerous prospective clinical trials have shown a significant benefit of amnio-infusion in reducing the rate of emergency caesarean section for fetal distress and complications related to meconium when used for these indications. Additional research is needed to clarify further its intrapartum role in patients with premature rupture of membranes or chorio-amnionitis.

PMID: 8836484 [PubMed - indexed for MEDLINE]

The activity of azithromycin on the infectivity of Chlamydia trachomatis in human amniotic cells.

• Patton DL,
• Wang SK,
• Kuo CC.

Department of Obstetrics and Gynaecology, University of Washington, Seattle 98195, USA.

The effects of azithromycin on the infectivity and growth of Chlamydia trachomatis were investigated in primary human amniotic epithelial cells. Infection was prevented when cultures were exposed to the drug 6 h after inoculation and growth was completely inhibited when the drug was added to cultures 24, 48, 72 h or 7 days after infection. The same inhibition was observed at 0.5, 1.0 and 5.0 mg/L. Ultrastructural observations depicted interruption in the growth cycle of the chlamydia and ghost-like envelopes were present in the near empty inclusions. Azithromycin is effective in inhibiting chlamydial growth no matter when treatment is initiated after infection.

PMID: 8821594 [PubMed - indexed for MEDLINE]

Differential expression patterns of beta-actin mRNA in cells undergoing apoptosis.

• Naora H,
• Naora H.

Research School of Biological Sciences, Australian National University, Canberra.

Microfilaments play an important role in the formation of apoptotic bodies. This study examined expression patterns of beta-actin, a key component of microfilament, during apoptosis. The data suggest that continuous beta-actin expression is not required for formation of apoptotic bodies. beta-actin mRNA levels rapidly declined in HL-60 cells following induction of apoptosis by actinomycin D, but were transiently elevated by two other apoptosis-inducing agents, cycloheximide and ionophore A23187. Furthermore, changes in beta-actin mRNA levels were less marked in U937 cells in which apoptosis was also induced by actinomycin D, as well as in WISH cells which demonstrated no typical apoptotic characteristics following actinomycin D treatment.

PMID: 7794260 [PubMed - indexed for MEDLINE]

Glucocorticoids stimulate the expression of prostaglandin endoperoxide H synthase-2 in amnion cells.

• Zakar T,
• Hirst JJ,
• Mijovic JE,
• Olson DM.

Department of Obstetrics and Gynaecology, University of Alberta, Edmonton, Canada.

Corticosteroids increase the production of prostaglandin E2 (PGE2) and the activity of prostaglandin endoperoxide H synthase (PGHS) in cultured amnion cells, although they inhibit prostanoid biosynthesis in numerous other cell types. This suggests that glucocorticoids control the level of PGHS in amnion cells by a hitherto unexplored, positive regulatory mechanism. We have tested the possibility that corticosteroids act by stimulating the expression of messenger RNAs (mRNAs) encoding one or both isoforms of PGHS. Ribonuclease protection assays were used to determine the levels of PGHS-1 and -2 mRNAs and, for reference, gamma-actin mRNA levels in confluent primary cultures of human amnion cells. In untreated cultures, PGHS-1 and -2 mRNA levels were low, often not reaching the level of detection. Dexamethasone (DEX) treatment for 4 h resulted in a measurable level of PGHS-2 mRNA, which increased further 10-fold and 20-fold after incubation with the glucocorticoid for 8 h and 16 h, respectively. The stimulation was dependent on DEX concentration, and was concomitant with an increase in the capacity of the cells to metabolize arachidonic acid to PGE2. PGHS-1 mRNA levels remained low in DEX-treated cells, while the gamma-actin message level showed no change. Estradiol and progesterone had no influence on PGHS-2 mRNA expression, but cortisol increased the PGHS-2 mRNA abundance. The glucocorticoid antagonist RU486 blocked the effect of DEX. Conditioned media of DEX-treated cells did not contain steroid-induced factor(s) stimulating PGE2 production. Inhibition of protein synthesis by cycloheximide potentiated the effect of DEX, and raised the abundance of PGHS-1, PGHS-2, and gamma-actin mRNAs in untreated cells. DEX did not affect the stability of the PGHS-2 mRNA. These results show that glucocorticoids promote PGE2 synthesis by amnion cells by stimulating the expression of PGHS-2 mRNA in a receptor-dependent, selective, and immediate fashion.

PMID: 7895671 [PubMed - indexed for MEDLINE]

The regulation of arachidonate lipoxygenase metabolite formation in cells derived from intrauterine tissues.

• Edwin SS,
• Thai D,
• LaMarche S,
• Branch DW,
• Mitchell MD.

Department of Obstetrics and Gynecology, University of Utah School of Medicine, Salt Lake City 84132, USA.

Products of arachidonic acid (AA) metabolism via the lipoxygenase pathways may have key roles in the maintenance of pregnancy and the onset of labor. We have determined whether calcium ionophores can modulate the rate of biosynthesis within the uterus of five important arachidonate lipoxygenase metabolites, i.e. leukotriene B4 (LTB4), LTC4, 5-hydroxyeicosatetraenoic acid (5-HETE), 12-HETE, and 15-HETE. Amnion, chorion, and decidual cells were isolated, grown to confluence and incubated with ionomycin. The production of LTB4, LTC4, 5-HETE, 12-HETE, and 15-HETE was determined using specific radioimmunoassays. Cell-specific, concentration-related stimulatory actions of ionomycin on 5-HETE, 12-HETE, 15-HETE, and LTC4 but not LTB4 production were found. A23187 had effects similar to ionomycin. Hence elevation of intracellular calcium levels can result in enhanced intrauterine production of arachidonate lipoxygenase metabolites that may affect pregnancy outcome.

PMID: 7784469 [PubMed - indexed for MEDLINE]

Amnioinfusion.

• Strong TH Jr.

Phoenix Perinatal Associates, Division of Maternal-Fetal Medicine, Good Samaritan Regional Medical Center, Arizona, USA.

Amnioinfusion is a relatively new technique with a variety of uses. Its most common application is the treatment of variable decelerations in the fetal heart rate during labor. By artificially increasing the amniotic fluid volume, the umbilical cord is better protected from compression. Amnioinfusion can represent the difference between operative intervention and spontaneous vaginal delivery. This simple, inexpensive technique appears to pose little risk and warrants consideration for the properly selected patient.

PMID: 7738919 [PubMed - indexed for MEDLINE]

A comparison of aztreonam and two regimens of gentamicin in a rabbit model of intra-amniotic infection and sepsis.

• McDuffie RS Jr,
• Heddleston LN,
• Blanton SJ,
• Gibbs RS.

Department of Obstetrics and Gynecology, Kaiser Permanente and Saint Joseph Hospital, Denver, Colorado, USA.

OBJECTIVE: To compare aztreonam in a standard dose with two gentamicin doses in the early treatment of experimental intra-amniotic infection in rabbits induced by intracervical inoculation with Escherichia coli. METHODS: Timed pregnant rabbits on day 21 (70% of gestation) were inoculated intracervically with 10(4)-10(5) colony-forming units of E coli. After inoculation, the animals were treated with one of three regimens: 1) aztreonam at 90 mg/kg/day ("standard" dose in humans), 2) gentamicin at 4.5 mg/kg/day ("standard" dose in humans), or 3) higher-dose gentamicin at 6.0 mg/kg/day, each given in three divided doses daily. Outcomes included fever, delivery, and presence of a live fetus. At necropsy, cultures were taken from endometrium, amniotic fluid, and blood. Data were analyzed by Fisher exact test because the expected cell size was fewer than five. RESULTS: Compared with rabbits treated with aztreonam, those treated with gentamicin 4.5 mg/kg/day delivered significantly more often (P = .002), had more positive cultures (P < .001), and had significantly fewer live fetuses (P < .001). Compared with rabbits treated with gentamicin 6.0 mg/kg/day, those treated with gentamicin 4.5 mg/kg/day delivered more often (P = .003), had fewer live fetuses (P = .02), and had more positive cultures (P = .02). There were no significant differences between the aztreonam and gentamicin 6.0 mg/kg/day groups. CONCLUSIONS: This study demonstrates in an animal model that aztreonam and gentamicin at 6.0 mg/kg/day are more effective than gentamicin at 4.5 mg/kg/day (a dose that is widely used empirically in humans) in the early treatment of experimental intra-amniotic infection in rabbits. Aztreonam was as effective as gentamicin at 6.0 mg/kg/day. In this rabbit model, in which intra-amniotic infection is accompanied by maternal sepsis, 4.5 mg/kg/day of gentamicin was not adequate for the treatment of severe maternal infection.

PMID: 9420844 [PubMed - indexed for MEDLINE]

Amnioinfusion. A review.

• Paszkowski T.

Department of Gynecology, University School of Medicine, Lublin, Poland.

The recent obstetric literature reports on various applications of amnioinfusion that have been used in an attempt to improve maternal and neonatal outcome. This paper reviews the indications, techniques, clinical benefits and hazards associated with the use of this procedure.

PMID: 7996522 [PubMed - indexed for MEDLINE]

Intraamniotic infection with Candida albicans successfully treated with transcervical amnioinfusion of amphotericin.

• Shalev E,
• Battino S,
• Romano S,
• Blondhaim O,
• Ben-Ami M.

Department of Obstetrics and Gynecology, Central Emek Hospital, Afula, Israel.

We present a case in which a pregnant woman was seen at 27 weeks' gestation with premature rupture of membranes and intraamniotic infection with Candida albicans, which was treated with transcervical amnioinfusion of amphotericin B. After 7 days of treatment spontaneous vaginal labor developed. A female newborn of 1030 gm was delivered. The infant was normal and did well.

PMID: 8178851 [PubMed - indexed for MEDLINE]

[Intra-amnion infusion in the management of premature rupture of the membranes before the 28th week]

• Nagy P,
• Szabo R.

Zala Megyei Korhaz Szuleszet-Nogyogyaszati Osztaly, Zalaegerszeg.

The authors applied intraamnial infusion in 5 cases altogether on account of the premature rupture of the membranes happened between the 25th-27th weeks. The pregnancies managed to be prolonged for 13-43 days. At present after 2 years, all the 4 survival children are alive and healthy. This method is advised to institutes with suitable conditions.

PMID: 8290239 [PubMed - indexed for MEDLINE]

Regulation of prostaglandin endoperoxide H synthase by glucocorticoids and activators of protein kinase C in the human amnion.

• Zakar T,
• Teixeira FJ,
• Hirst JJ,
• Guo F,
• MacLeod EA,
• Olson DM.

University of Alberta Perinatal Research Centre, Department of Obstetrics and Gynaecology, Edmonton, Canada.

Since glucocorticoids decrease and protein kinase C (PKC) activators increase amniotic PGE2 production, the possibility that they regulate the activity of prostaglandin endoperoxide H synthase (PGHS), the rate-limiting enzyme of prostaglandin synthesis from arachidonate, was investigated. Glucocorticoids inhibited the production of PGE2 from exogenous arachidonate specifically and in a concentration dependent fashion. Furthermore, cortisol decreased PGHS activity and the amount of PGHS protein in amnion microsomes, and reduced the rate of recovery of PGHS after acetylsalicylic acid (ASA) pretreatment. Actinomycin D blocked the inhibition of PGHS recovery by cortisol, but did not suppress the spontaneous recovery of the enzyme, indicating that the glucocorticoid induced a post-transcriptional inhibitor of PGHS synthesis. PKC-activating phorbol esters, such as 12-tetradecanoyl phorbol 13-acetate (TPA) increased the synthesis of PGE2 from exogenous arachidonate, also in a specific and concentration dependent manner. PGHS recovery after ASA treatment was enhanced by TPA. PGHS activity and protein concentrations were increased by phorbol ester treatment; however, this was apparent only in tissues in which the concentrations of PGHS were initially low. These results show that the synthesis of PGHS is positively and negatively regulated in the human amnion by PKC and glucocorticoids, respectively, and suggest that effectors using these pathways may regulate the enzyme in vivo.

PMID: 8182609 [PubMed - indexed for MEDLINE]

Amnioinfusion: a review.

• Lameier LN,
• Katz VL.

UNC School of Medicine, Department of Obstetrics and Gynecology, Chapel Hill 27599-7570.

Amnioinfusion is a commonly practiced technique used for intrapartum improvement of the fetal condition. Room temperature normal saline (0.9 per cent) infused through an intrauterine pressure catheter has been used to alleviate variable decelerations, dilute thick meconium, and improve the intrauterine environment. Randomized studies comparing amnioinfusion to no therapy have shown that amnioinfusion is associated with lower cesarean delivery rates, decreased numbers of operative deliveries, and improved umbilical artery and venous blood gas values. Amnioinfusion also has been suggested as means to instill antibiotics into an infected uterine cavity, or the uterine cavity of a woman with preterm premature rupture of the membranes. Transabdominal amnioinfusion may be used to improved prenatal ultrasound evaluation in pregnancies associated with oligohydramnios. Complications of amnioinfusion include umbilical cord prolapse, uterine overdistention, fetal bradycardia, and one report of possible amniotic fluid embolism. Overall, amnioinfusion seems to be a safe and effective technique to improve the intrauterine milieu.

PMID: 8309665 [PubMed - indexed for MEDLINE]

Tumor necrosis factor alpha stimulates amnion prostaglandin biosynthesis primarily via an action on fatty acid cyclooxygenase.

• Pollard JK,
• Mitchell MD.

Department of Obstetrics and Gynecology, Medical Center Hospital of Vermont, Burlington.

The purpose of this study was to determine how tumor necrosis factor alpha (TNF alpha) stimulates prostaglandin E2 production in human amnion. Amnion cells were isolated from term placentae and grown to confluence in culture. Incubations were conducted in quadruplicate wells for 16 hours with TNF alpha and protein synthesis inhibitors cycloheximide and actinomycin D, or arachidonic acid, acetylsalicylic acid (ASA), or staurosporine or H7 which inhibit protein kinase C activity. Prostaglandin E2 (PGE2) was measured by radioimmunoassay and cellular protein determined. The stimulatory action of TNF alpha on amnion PGE2 production was blocked by protein synthesis inhibitors, and the addition of arachidonic acid always enhanced the stimulatory properties of TNF alpha. TNF alpha consistently induced more rapid recovery from ASA treatment, and protein kinase C inhibition attenuated the stimulatory effects of TNF alpha. These results suggest that the stimulatory action of TNF alpha on amnion PGE2 production is likely at the level of induction of fatty acid cyclooxygenase activity and is partially dependent upon activation of protein kinase C.

PMID: 8295980 [PubMed - indexed for MEDLINE]

Epidermal growth factor and transforming growth factor-alpha enhance the interleukin-1- and tumor necrosis factor-stimulated prostaglandin E2 production and the interleukin-1 specific binding on amnion cells.

• Bry K.

Department of Pediatrics, University of California, Irvine 92717.

Interleukin-1 (IL-1), tumor necrosis factor (TNF), epidermal growth factor (EGF), and transforming growth factor-alpha (TGF-alpha) stimulate prostaglandin E2 (PGE2) production by amnion cells whereas TGF-beta inhibits the PGE2 production. During labor occurring in the setting of infection, several of these cytokines may be simultaneously present in amniotic fluid. The aim of the present study was to examine whether these cytokines modify each others' effects on amnion cell PGE2 production. Amnion cells in monolayer culture were treated with IL-1, TNF, EGF, TGF-alpha, TGF-beta 1, their combination, or vehicle. The PGE2 production and the specific binding of radiolabeled IL-1 beta on the cells were measured. IL-1 or TNF in combination with EGF or TGF-alpha stimulated synergistically the production of PGE2 by amnion cells. TGF-beta 1 did not modify the PGE2-stimulatory effect of EGF/TGF-alpha. Untreated amnion cells expressed 1030 +/- 100 IL-1 beta receptors per cell with a binding affinity of 1.40 +/- 0.26 nM. Treatment with TGF-alpha increased the number of receptors to 3940 +/- 260 per cell with no change in binding affinity. The potentiation of the PGE2-stimulatory effect of IL-1 by TGF-alpha may be related to its ability to induce IL-1 receptors on amnion cells. The synergistic effects of cytokines on amnion cell PGE2 production may promote labor.

PMID: 8140119 [PubMed - indexed for MEDLINE]

Mechanism of interleukin-1 beta stimulation of human amnion prostaglandin biosynthesis: mediation via a novel inducible cyclooxygenase.

• Mitchell MD,
• Edwin SS,
• Lundin-Schiller S,
• Silver RM,
• Smotkin D,
• Trautman MS.

Department of Obstetrics and Gynecology, University of Utah School of Medicine, Salt Lake City 84132.

We have evaluated the mechanism by which interleukin-1 beta (IL-1 beta) increases amnion cell PGE2 production in a concentration-dependent manner. IL-1 beta-stimulated amnion cell PGE2 biosynthesis was time-dependent, and significant stimulation occurred within 2 h of incubation. IL-1 beta stimulation occurred in the presence of added arachidonic acid but was abrogated by treatment with cycloheximide and actinomycin D. Amnion cells treated with IL-1 beta recovered rapidly from aspirin pretreatment suggesting an action on fatty acid cyclooxygenase (COX). Increased amounts of COX protein were demonstrated by Western blot analysis within 2 h of IL-1 beta treatment of amnion cells. Northern blot analysis using a probe specific for a novel form of COX (COX-II) showed an increase in mRNA for this COX within 30 min. This finding using a homologous detection system and human cells of fetal origin in primary culture provides strong support for a physiological role for COX-II in man.

PMID: 8153084 [PubMed - indexed for MEDLINE]

Interleukin-1 binding and prostaglandin E2 synthesis by amnion cells in culture: regulation by tumor necrosis factor-alpha, transforming growth factor-beta, and interleukin-1 receptor antagonist.

• Bry K,
• Lappalainen U,
• Hallman M.

Department of Pediatrics, University of California, Irvine 92717.

Proinflammatory cytokines may promote preterm labor in the setting of intrauterine infection. Tumor necrosis factor (TNF) and interleukin-1 (IL-1) synergistically stimulate the production of prostaglandin E2 (PGE2) by amnion cells. Transforming growth factor-beta (TGF-beta) inhibits the cytokine-stimulated PGE2 production. In the present study, we investigated the binding of IL-1 beta on human amnion cells in culture. Untreated amnion cells possessed 540 +/- 60 IL-1 receptors per cell, with a dissociation constant of 1.4 +/- 0.4 nM. Cells treated with TGF-beta 1 (10 ng/ml) had 570 +/- 110 receptors per cell. TNF-alpha (50 ng/ml) increased the number of IL-1 receptors to 2930 +/- 590. TGF-beta 1 inhibited the receptor upregulation by TNF-alpha. Cells treated with TGF-beta 1 and TNF-alpha expressed 1140 +/- 590 receptors per cell. The binding affinity was not changed by the cytokines. IL-1 receptor antagonist (IL-1ra) inhibited the stimulation of amnion cell PGE2 production by IL-1 beta, but not by TNF-alpha. Amnion cells secreted large amounts of IL-1ra (1.1 +/- 0.3 ng/10(5) cells). Treatment of the cells with TGF-beta 1 or TNF-alpha did not affect the release of IL-1ra. We conclude that IL-1 receptor expression is an important step in the regulation of the effects of cytokines on amnion cell PGE2 production.

PMID: 8457602 [PubMed - indexed for MEDLINE]

Inhibition of binding of bacteria to amniochorionic membranes by amniotic fluid.

• Daunter B,
• Forbes KL,
• Sanderson BM,
• Morrison J,
• Wright G.

University of Queensland, Department of Obstetrics and Gynaecology, Royal Brisbane Hospital, Australia.

The immunological composition of amniotic fluids is shown to be of such a lower order of activity that its role in fetal protection may be limited. Also, amniotic fluids were found not to have classical antibiotic activity. Amniotic fluids (25/31), however, were found to inhibit, by 27.5% to 88.2%, three target bacteria from binding to discs of amniochorionic membranes. This inhibition is also demonstrable with the monosaccharides alpha-D(+)-fucose, D(+)-galactose, alpha-D-glucose, alpha-D-lactose and bovine serum albumin-lactose conjugate, whereas other glycoconjugates enhanced bacterial binding. This demonstrates that the test bacteria bind to the amniochorionic membranes using bacterial lectins. In intraamniotic infection bacterial lectins may be complexed by amniotic fluid glycoconjugates which prevent the bacteria from binding to the amniochorionic membranes. This would explain asymptomatic infection and in the absence or reduced levels of the glycoconjugates the bacteria would bind to the amniochorionic membranes giving rise to symptomatic infection.

PMID: 1459334 [PubMed - indexed for MEDLINE]

Evidence of a role for phosphatidylinositol synthesis in human amnion cell proliferation.

• Ohno T,
• Imai A,
• Furui T,
• Matsunami K,
• Matsuda T,
• Tamaya T.

Department of Obstetrics and Gynecology, Gifu University School of Medicine, Japan.

Phosphatidylinositol (PtdIns) is the key precursor of phosphoinositide-derived intracellular mediators. The effects of changing the rate of PtdIns synthesis on mitogenic activity of human amnion-derived WISH cells were investigated. Incubation of the cells with [3H]inositol caused a time- and dose-dependent PtdIns labeling. Exogenous Ca2+ inhibited [3H]inositol incorporation in a dose-dependent fashion; half-maximal inhibition occurred with 0.3-1.0 mM Ca2+. In contrast, removal of cytosolic Ca2+ by ionophore A23187 and 1 mM EGTA induced enhancement of the PtdIns labeling as a function of A23187 concentration, perhaps through release of inhibitory effects of endogenous Ca2+. The A23187-stimulated PtdIns labeling with [3H]inositol was not abolished by additional unlabeled inositol, suggesting that [3H]inositol labeling of PtdIns occurred mainly through de novo synthesis catalyzed by PtdIns synthase (EC 2.7.8.11). In cells with PtdIns synthase activity decreased by exogenous Ca2+, [3H]thymidine incorporation was also inhibited, while A23187 caused dose-dependent enhancement of thymidine incorporation. The changes in PtdIns synthase activity occurred in parallel with changes in mitogenic activity caused by increasing the dose of exogenous Ca2+ or A23187. A similar lowering of mitogenic activity was observed upon suppression of PtdIns synthase by pemirolast potassium (9-methyl-3-1H-tetrazol-5yl-4H-pyrido[1,2-a]pyridin-4-one potassium) via a Ca(2+)-independent mechanism. These data demonstrate that changes in PtdIns synthase activity by some agents acting via different mechanisms are associated with parallel changes in thymidine incorporation, and suggest that PtdIns production is tightly coupled to cell proliferation in human amnion cells.

PMID: 1335765 [PubMed - indexed for MEDLINE]

Amnion cell biosynthesis of interleukin-8: regulation by inflammatory cytokines.

• Trautman MS,
• Dudley DJ,
• Edwin SS,
• Collmer D,
• Mitchell MD.

Department of Pediatrics, University of Utah, Salt Lake City 84132.

The cellular constituents of the placenta are important participants in the recruitment and trafficking of inflammatory cells within the placenta. In infection-induced labor, gestational tissues synthesize and release a variety of inflammatory cytokines whose effects include increased prostaglandin biosynthesis and the initiation of uterine contractions. Interleukin-8 (IL-8), a potent neutrophil chemoattractant, has been recently described as being elevated in the amniotic fluid of mothers with chorioamnionitis. We investigated the biosynthesis of IL-8 by human amnion cells and its regulation by other inflammatory cytokines. Cultured amnion cells obtained from normal term placentae were found to produce IL-8 in response to pathophysiologic concentrations of interleukin 1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF-alpha). Treatment of amnion cells stimulated by IL-1 beta with cycloheximide resulted in increased IL-8 production, while incubation of IL-1 beta treated amnion cells with actinomycin D resulted in a concentration-dependent decrease in detectable amounts of IL-8. Northern blot analysis of cultured amnion cells stimulated with IL-1 beta demonstrated a rapid increase in IL-8 mRNA which peaked at 2-4 hr. These in vitro results suggest inflammation of gestational tissues in vivo may result in locally produced IL-8 and, in association with other inflammatory mediators, may be important in the pathophysiology of infection-induced labor.

PMID: 1522134 [PubMed - indexed for MEDLINE]

Effects of ascorbic acid on Chlamydia trachomatis infection and on erythromycin treatment in primary cultures of human amniotic cells.

• Wang SK,
• Patton DL,
• Kuo CC.

Department of Obstetrics and Gynecology, University of Washington, Seattle 98195.

Ascorbic acid (vitamin C) is an essential nutrient for humans. It may also be needed by Chlamydia trachomatis, an intracellular bacterium. We investigated the effects of vitamin C on the growth of C. trachomatis E/UW-5/Cx in a primary culture of human amniotic epithelial cells. The results showed that vitamin C enhances C. trachomatis infection at concentrations of 0.2, 0.6, and 1.2 mg/dl (P less than 0.001). These three concentrations represent the in vivo concentrations of deficiency, normal, and overload levels in serum, respectively. The enhancement was dose dependent. However, the growth of C. trachomatis was inhibited at vitamin C concentrations of 120 and 1,200 mg/dl. The inhibitory effect of erythromycin against C. trachomatis was shown to be reduced in the presence of vitamin C at the three concentrations tested (P less than 0.025-0.001), and MICs were four times greater (1.6 versus 0.4 micrograms/ml). Human amniotic cells were tolerant to vitamin C concentrations of up to 1,200 mg/dl. The results show that vitamin C may be an important nutrient for C. trachomatis and that incorporation of vitamin C in the culture medium may enhance the isolation and propagation of C. trachomatis in cell cultures.

PMID: 1400952 [PubMed - indexed for MEDLINE]

Amnioinfusion with preterm, premature rupture of membranes.

• Strong TH Jr.

Phoenix Perinatal Associates, Arizona.

Excluding labor, the greatest risks to the fetus from preterm PROM are umbilical cord accidents and infection. Heretofore, the clinical options for the PROM patient have been limited. With the advent and refinement of amnioinfusion, the utility of expectant management may be greatly increased. Through the use of amnioinfusion or its permutations, a number of diagnostic and therapeutic procedures previously unavailable to the PROM patient may become routine, including amniotic fluid volume expansion and direct in utero prophylaxis/treatment of amnionitis.

PMID: 1617883 [PubMed - indexed for MEDLINE]

Characterization of oxytocin receptors in rabbit amnion involved in the production of prostaglandin E2.

• Hinko A,
• Soloff MS.

Department of Biochemistry and Molecular Biology, Medical College of Ohio, Toledo 43699-0008.

We characterized oxytocin (OT) receptors in purified plasma membranes from amnion, decidua, and myometrium of late pregnant rabbits using an iodinated OT antagonist (OTA). Saturation studies showed similar Kd values for specific binding sites in the 100- to 250-pM range in all three tissues. OT receptor concentrations in decidua and myometrium did not change until the day of labor (day 31), when they rose about 2.5- and 18-fold, respectively. Increases in amnion receptors were first apparent on day 28 and continued to maximal levels on day 31. There was an increase of about 230-fold from day 26 to labor, reaching 9.5 pmol/mg protein. Competition studies using analogs showed that ligand specificities of amnion and decidual membranes were indistinguishable. Those of myometrial membranes were somewhat different, possibly owing to the presence of both AVP receptors and OT receptors in the myometrium. Binding of OTA corresponded to the OT-induced release of prostaglandin E2 (PGE2) by amnion cells in culture. The effects of OT were dose dependent, agonist specific, and selectively inhibited by OTA. Amnion cells from days 22 and 28 did not respond significantly to either OT or phorbol 12-myristate 13-acetate (PMA), but cells from day 30 pregnant rabbits responded strongly to both. In contrast, calcium ionophore stimulated comparable amounts of PGE2 release from cells cultured on day 22, 28, or 30. These studies show that specific, high affinity OT receptors are associated with the release of PGE2 from rabbit amnion cells. Increases in amnion OT receptor and protein kinase-C activity precede by several days the increases in receptor concentrations in decidua and myometrium, suggesting important roles for the amnion and OT in the initiation of labor in rabbits.

PMID: 1317789 [PubMed - indexed for MEDLINE]

Prostaglandin synthesis regulation in human amnion tissue: involvement of protein kinase C and dependence on ribonucleic acid and protein synthesis.

• Zakar T,
• Olson DM.

Department of Paediatrics, University of Western Ontario, Lawson Research Institute, London, Canada.

The role of protein kinase C (PKC) in the control of prostaglandin production by the human amnion was studied. Amnion membranes delivered spontaneously at term were minced and treated with phorbol esters, protein kinase inhibitors, cycloheximide, and actinomycin D; prostaglandin E2 (PGE2) output then was determined. Untreated tissue produced 3.97 +/- 1.13 ng PGE2/micrograms DNA/14 h (mean +/- SEM, n = 19). Phorbol dibutyrate and 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulated PGE2 output up to 20-fold in a concentration-dependent manner with potencies corresponding to their efficacy as PKC activators. Four-beta-phorbol and 4-methoxy-TPA, which do not stimulate PKC, did not affect PGE2 output. Stimulation by TPA was blocked by staurosporine (IC50 = 57 nM) and H7; however, these PKC inhibitors did not decrease basal prostaglandin production. Cycloheximide inhibited basal and TPA-promoted PGE2 production and amino acid incorporation. Actinomycin D abolished TPA stimulation without decreasing unstimulated prostaglandin synthesis. These results show that amnion PGE2 production after labor is not maintained by PKC action, but PKC activation in this tissue causes a protein synthesis-dependent and RNA synthesis-dependent increase of PGE2 output. However, basal PGE2 production is dependent upon protein synthesis which, presumably, utilizes pre-existing mRNAs.

PMID: 1375515 [PubMed - indexed for MEDLINE]

[Study of embryotoxic and teratogenic effects of amphotericin B and a methyl derivative of amphotericin B in rats after their intravenous and intra-amniotic administration]

• Moguchenok EA.

The embryotoxic action of amphotericin B and its methyl derivative was compared in rats after their intravenous and intraamniotic administration. The concentrations of amphotericin B and its methyl derivative in the amniotic cavity on days 13, 14 and 15 of pregnancy were 1.5 and 36 micrograms/ml, respectively. When administered intravenously during the preimplantation period the antibiotics had no embryotoxic action. Intravenous administration of amphotericin B in a dose of 500 micrograms/kg and its derivative in a dose of 2000 micrograms/kg during organ genesis induced a decrease in the craniocaudal size. In a dose of 3000 micrograms/kg administered intravenously the methyl derivative of amphotericin B induced an increase in postimplantation death rates. Administration of amphotericin B to the amniotic cavity had no damaging action. Administration of the methyl derivative on day 15 of pregnancy led to anomalous development of the lower extremities and slower ossification. The threshold doses by the embryotoxic action for intravenous administration are 500 micrograms/kg for amphotericin B and 2000 micrograms/kg for the methyl derivative. Administration of the antibiotics to the amniotic cavity revealed potential teratogenic properties of the amphotericin B methyl derivative.

PMID: 1514864 [PubMed - indexed for MEDLINE]

Protein kinase-C activation is required for oxytocin-induced prostaglandin production in human amnion cells.

• Moore JJ,
• Moore RM,
• Vander Kooy D.

Department of Pediatrics, Case Western Reserve University School of Medicine, Cleveland, Ohio 44109.

In previous work we reported that oxytocin activates phospholipase-C (PLC) and increases prostaglandin E2 (PGE2) release in amnion. Whether either of the consequences of activation of PLC by oxytocin, activation of protein kinase-C (PKC) or increases in intracellular calcium, directly results in the production of PGE2 is unknown. Phorbol esters (PMA) and epidermal growth factor (EGF) are also known to increase PGE2 release from amnion. In some tissues these agents are capable of activating the PLC postreceptor cascade system. This study was undertaken primarily to explore the mechanism of oxytocin-induced PGE2 production in amnion and secondarily to determine whether common aspects of PGE2 production by oxytocin, PMA, and EGF include activation of PLC or subsequent steps in this cascade followed by new mRNA/protein production. Involvement of PLC was assessed by inositol phosphate (IP1) turnover. IP1 turnover was increased by oxytocin (2.99 +/- 0.31-fold; P less than 0.01), but not by EGF or PMA. PMA inhibited oxytocin-provoked IP1 turnover (P less than 0.05). PKC involvement was initially evaluated with two PKC inhibitors, H7 and staurosporine. Each inhibited PGE2 production by oxytocin as well as that by PMA and EGF in a dose-dependent fashion. With H7, the IC50 for all agents was 5 microM; the IC50 for staurosporine was 2 nM for PMA and oxytocin and 5 nM for EGF. Agonist-induced PGE2 production was also assessed in cells in which PKC activity had been tachyphylaxed with a high concentration of PMA (400 ng/mL for 48 h). In such cells oxytocin and PMA no longer stimulated (P less than 0.001) PGE2 production, but EGF-stimulated PGE2 production was only slightly reduced. PKC involvement is, thus, implicated for oxytocin and PMA. Other enzymes that are inhibited by H7 and staurosporine are implicated in the production of PGE2 caused by EGF. Although tachyphylaxed cells produced no PGE2 with oxytocin, oxytocin increased intracellular calcium to levels higher than those seen in control cells (435 +/- 102 vs. 286 +/- 1.2) Actinomycin-D (P less than 0.001) and cycloheximide (P less than 0.05) inhibited PGE2 production caused by oxytocin, PMA, and EGF. PGE2 production by oxytocin in human amnion cells proceeds by activation of PKC, followed by new protein and mRNA production. Further, in cells without PKC, oxytocin-induced calcium transients do not increase PGE2. The ability of EGF to stimulate PGE2 in cells with no PKC activity also establishes that PKC activation is not a common intracellular step in the induction of PGE2 production by all agents.

PMID: 2022708 [PubMed - indexed for MEDLINE]

Amniotic membranes in the treatment of necrotizing fasciitis complicating vulvar herpes virus infection.

• Rothman PA,
• Wiskind AK,
• Dudley AG.

Department of Gynecology and Obstetrics, Emory University School of Medicine, Atlanta, Georgia.

Necrotizing fasciitis of the vulva developed in an immunocompromised patient with chronic myelogenous leukemia, apparently from secondarily infected herpes simplex lesions. In addition to surgical debridements and broad-spectrum intravenous antibiotic therapy, the wound was treated using specially prepared amniotic membranes as a wound dressing. The patient died on hospital day 65 because of complications of her immunocompromised state, with autopsy findings of disseminated cytomegalovirus. However, use of amniotic membranes as a wound dressing appeared to be beneficial.

PMID: 2166265 [PubMed - indexed for MEDLINE]

Effects of glucocorticoids on prostaglandin formation by human amnion.

• Gibb W,
• Lavoie JC.

Department of Obstetrics and Gynecology, University of Ottawa, Ottawa General Hospital, Ont., Canada.

The human amnion may be an important source of prostaglandins involved in the onset of human labor and therefore it is important to define the factors that regulate their formation in this tissue. In the present study we demonstrate that glucocorticoids inhibit prostaglandin production by freshly isolated amnion cells. The inhibitory action of the glucocorticoids, however, changes to a stimulatory action when the cells are maintained in primary culture for a few days. For both inhibition and stimulation, concentrations of 10(-8) M dexamethasone or greater were required to give significant effects, and estradiol and progesterone had no effect on the prostaglandin output of the cells. Epidermal growth factor (EGF), which has previously been found to stimulate prostaglandin output by confluent amnion cells, did not alter prostaglandin output of cells initially placed in culture. Furthermore, the stimulatory action of EGF and dexamethasone appeared additive. The calcium ionophore A23187 stimulated prostaglandin output in freshly isolated cells and accentuated the inhibitory effect of dexamethasone. These studies indicate that prostaglandin formation by human amnion during pregnancy could be regulated by glucocorticoids. These steroids are easily available to the amnion by way of cortisone conversion to cortisol by the maternal decidua. The results also indicate that amnion is capable of responding to glucocorticoids in both a stimulatory and inhibitory fashion and whether one or both actions are of importance in vivo is a question that is as yet unresolved.

PMID: 2115393 [PubMed - indexed for MEDLINE]

Arachidonic acid release from cultured human amnion cells: the effect of dexamethasone.

• Potestio FA,
• Olson DM.

Department of Pediatrics, Lawson Research Institute, St. Joseph's Health Centre of London, University of Western Ontario, Canada.

Glucocorticoids inhibit prostaglandin (PG) synthesis in several cell types, presumably by inhibiting arachidonic acid (AA) deacylation from phospholipids. We studied the effects of glucocorticoids on cultured term human amnion cell AA release. Confluent monolayer cultures of amnion cells were adapted to serum-free medium, and phospholipids were labeled for 18 h with [14C]AA. The calcium ionophore A23187 (0.2-5.0 mumol/L) stimulated [14C]AA release (up to 2.2-fold) in a dose- and time-dependent manner. The apparent sources of the liberated [14C]AA were phosphatidylcholine and phosphatidylethanolamine. Pretreatment for 24 h with the synthetic glucocorticoid dexamethasone (0.1-1000 nmol/L) significantly inhibited (P less than 0.01) basal (unstimulated) [14C]AA release by 69% in subsequent 1-h experiments. The sole apparent source of free [14C]AA during this inhibitory state was phosphatidylethanolamine. Dexamethasone pretreatment slightly inhibited (13%; P less than 0.05) calcium ionophore-stimulated [14C]AA release; however, it was still 3.8-fold greater than basal release, suggesting that the glucocorticoid effect on stimulated AA release was not biologically relevant. Further characterization of the glucocorticoid effect revealed that preincubation of the cultures with dexamethasone for as little as 20 min inhibited basal [14C]AA release. Furthermore, studies involving actinomycin-D and cycloheximide demonstrated that inhibition of RNA and protein synthesis failed to block the glucocorticoid inhibition of basal AA liberation. The glucocorticoid receptor antagonist RU 38486, alone or in the presence of dexamethasone, also inhibited unstimulated [14C]AA release. Cortisol, dehydroisoandrosterone sulfate, 17 beta-estradiol, and progesterone all inhibited basal [14C]AA liberation. We conclude that glucocorticoids inhibit unstimulated AA release from cultured amnion cells, but do not prevent calcium ionophore from stimulating a large increase in AA release.(ABSTRACT TRUNCATED AT 250 WORDS)

PMID: 1689736 [PubMed - indexed for MEDLINE]

Application of freeze-dried amniotic membrane: a control trial at the donor site of split-thickness skin grafting.

• Waikakul S,
• Chumniprasas K,
• Setasubun S,
• Vajaradul Y.

Faculty of Medicine, Mahidol University, Siriraj Hospital, Bangkok, Thailand.

Sixty five patients (51 males and 14 females) underwent a control trial of freeze-dried amniotic membrane applied in the form of split-thickness skin grafts over an area from which skin had been surgically removed. The average size of the grafts was 15 x 20 cm. The patients were randomly divided into two groups. In the first group (36 patients), the membrane was applied at the upper half of the wound; in the second group (29 patients), the membrane was applied at the lower half of the wound. The remaining areas of the wounds were covered by antibiotic-impregnated fine-mesh greased gauze. Pain, infection, and the progress of healing were monitored. Covering the wound with the amniotic membrane had good results in terms of pain reduction, but the promotion of healing and the prevention of infection were not demonstrated. Hyperemia and hypertrophic scar responses to the membrane in some cases were considered as disadvantages.

PMID: 2163701 [PubMed - indexed for MEDLINE]

Phospholipase C and phospholipase A2 are involved in the antiviral activity of human interferon-alpha.

• Premecz G,
• Markovits A,
• Bagi G,
• Farkas T,
• Foldes I.

Microbiological Research Group, National Institute of Hygiene, Budapest, Hungary.

Treatment of human amniotic cells (UAC) with human interferon-alpha (Hu-IFN alpha) or phorbol myristate acetate (PMA) resulted in translocation of protein kinase C (PK-C) activity from the cytosol fraction to that of the membranes. Analysis of 32P incorporation into phospholipid fractions and studies of alterations in fatty acid content for the major phospholipids of IFN-treated cells suggest that phospholipases C and A2 are activated by Hu-IFN alpha. Addition of neomycin (an inhibitor of phospholipase C), as well as mepacrine (an inhibitor of phospholipase A2) to IFN-treated cells inhibited the antiviral activity of Hu-IFN alpha in the vesicular stomatitis virus (VSV)-UAC system used. These observations indicate that (i) activation of PK-C and (ii) diacylglycerol formation, arachidonic acid and/or lysophosphatidylcholine release are important steps in the mechanism of action of IFN.

PMID: 2544450 [PubMed - indexed for MEDLINE]

Recent advances in burn wound management in China.

• Ding YL,
• Han CM.

The latest advances in burn wound management in China may be summarized as follows: 1. Escharectomy, tangential excision and skin grafting is being performed widely in moderate and small deep burns especially in functional sites; immediate flap transfer after debridement for deep third degree burns; tangential excision, escharectomy and large sheet skin grafting along the division lines of the face for deep facial burns. 2. Micro-skin grafting--a new operative method used in China was introduced to manage extensive deep burns. 3. Softened freeze-dried glutaraldehyde preserved skin, chlorhexidine-alcohol refrigerated porcine skin, frozen amniotic membrane--all are effective as burn dressings. 4. The new methods of prolonging the survival time of allo- and xeno-skin grafts are used with ultraviolet rays, medicinal herbs and in vitro treatment with triamcinolone acetonide. 5. No remarkable rejection occurred after application of composite skin grafts to burn wounds. 6. The time of culturing epidermal cells has been shortened to 5-13 days. Preliminary successful results were obtained in patients after transplantation of cultured epidermal cell plate or cell emulsion dropped into the holes of allo-skin grafts. No rejection occurred for 50 days up to 16 months after allo-transplantation of epidermal cell plate. 7. Silver norfloxacin, zinc sulphadiazine and cerium nitrate/silver sulphadiazine had a strong bacteria inhibiting action and promoted wound healing.

PMID: 2479205 [PubMed - indexed for MEDLINE]

Glucocorticoids stimulate prostaglandin synthesis in human amnion cells by a receptor-mediated mechanism.

• Potestio FA,
• Zakar T,
• Olson DM.

Department of Pediatrics, St. Joseph's Health Centre of London, University of Western Ontario, Canada.

Prostaglandin E2 (PGE2) synthesis by human amnion increases with the onset of labor and is thought to participate in the initiation and maintenance of parturition. Since cortisol levels increase in amniotic fluid in late pregnancy, we studied the effects of glucocorticoids on cultured term human amnion cell PGE2 output. In 24-h studies, the synthetic glucocorticoid dexamethasone stimulated basal PGE2 output 2-fold over control levels at 16-500 nmol/L. PGE2 output was dramatically stimulated (greater than 10-fold) when, after dexamethasone pretreatment, the cells were incubated with calcium ionophore A23187 or arachidonic acid (AA) for 2 h. Maximum effects were achieved at 31 nmol/L dexamethasone. Basal PGE2 output was stimulated at 12 h of dexamethasone treatment, whereas A23187- or AA-stimulated PGE2 output was enhanced after 3-6 h of dexamethasone pretreatment. Cortisol (50 and 500 nmol/L) also enhanced basal and stimulated PGE2 output, while dehydroepiandrosterone sulfate, 17 beta-estradiol, and progesterone were ineffective. The glucocorticoid receptor antagonist RU 38486 attenuated dexamethasone-enhanced basal and stimulated PGE2 output. Dexamethasone pretreatment had no effect on basal or stimulated PGE2 output from cultured term chorion cells, suggesting tissue specificity. We conclude that glucocorticoids specifically enhance PGE2 output from cultured amnion cells via a receptor-mediated mechanism. We speculate that the action of glucocorticoids is to increase the capacity of the cells to convert AA to PGE2.

PMID: 3142916 [PubMed - indexed for MEDLINE]

Stimulation of human amnion prostaglandin E2 production by activators of protein kinase-C.

• Zakar T,
• Olson DM.

Department of Pediatrics, Lawson Research Institute, St. Joseph's Health Centre, London, Ontario, Canada.

We tested the possibility that the activation of protein kinase-C by the tumor-promoter phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) or diacylglycerol stimulates the production of prostaglandin E2 (PGE2) by the amnion. Confluent primary cultures of human amnion epithelial cells were adapted to serum-free medium and treated with the agonists for up to 8 h. Cumulative PGE2 output in the medium was measured by RIA. TPA, a potent activator of protein kinase-C, stimulated basal PGE2 output from less than 50 pg/well.5 h to 3 ng/well.5 h (P less than 0.01) in a time- and dose-dependent manner. 4-Methoxy-TPA, a weak tumor promoter derivative of TPA, was ineffective when tested in the same concentration range as TPA (1 nmol/L to 1 mumol/L). Neither calcium ionophore A23187 (20 nmol/L) nor arachidonate (1 mumol/L) stimulated PGE2 output alone, but each agonist potentiated the effect of TPA as much as 5-fold (P less than 0.01). 1,2-Dioctanoyl-sn-glycerol stimulated PGE2 output 4- to 7-fold (P less than 0.05), and this effect was potentiated by Ca ionophore and arachidonate. Studies involving actinomycin-D and cycloheximide indicated that the stimulatory effect of TPA was dependent on RNA synthesis during the first 60 min and on protein synthesis during the entire length of the phorbol ester treatment period (300 min). TPA was also able to stimulate PGE2 production after irreversible inactivation of PG endoperoxide synthase activity with acetylsalicylic acid. These results suggest that activation of protein kinase-C in amnion cells increases the de novo synthesis of the PG endoperoxide synthase enzyme in a RNA synthesis-dependent manner. Elevated intracellular calcium levels contribute to the stimulation apparently by increasing the availability of endogenous arachidonate for subsequent conversion to PGE2.

PMID: 2460486 [PubMed - indexed for MEDLINE]

Clinical effectiveness of a new cervical indwelling catheter in the management of premature rupture of the membranes: a Japanese collaborative study.

• Ogita S,
• Mizuno M,
• Takeda Y,
• Arai M,
• Sugawa T,
• Kuwabara Y,
• Hashimoto T,
• Nishijima M,
• Imanaka M.

Osaka City Perinatal Center, Japan.

The aim of this multiinstitutional study was to evaluate a new cervical indwelling catheter in 84 patients with premature rupture of the membranes at less than 33 weeks' gestation. The average time of insertion of the catheter was 29.1 weeks' gestation, at an average of 2.0 days after rupture, and was left in place for an average of 6.5 days. The mean birth weight was 1417 gm. The mortality rate and the incidence of respiratory distress syndrome were 5.7% and 11.9%, respectively. Amniotic fluid culture was positive in 39.1% of patients before catheter insertion and only 4.3% at the time of delivery. Infection was noted in only four of 84 infants. The incidence of infection was very low in those treated for 3 days or more after catheter insertion.

PMID: 3407690 [PubMed - indexed for MEDLINE]

The action of epidermal growth factor on human amnion prostaglandin E2 output.

• Zakar T,
• Olson DM.

Department of Paediatrics, St Joseph's Health Centre of London, University of Western Ontario, Canada.

The mechanism of stimulatory action of epidermal growth factor on term human amnion prostaglandin E2 production was studied. Monolayer cultures of amnion epithelial cells from spontaneous vaginal deliveries were preincubated for 24 h with serum-free media and treated with epidermal growth factor, calcium ionophore A23187 (4.5 microM), and arachidonate. Cumulative prostaglandin E2 output was not stimulated by epidermal growth factor (less than or equal to 200 ng/mL) or A23187 alone or the two added together. Pretreating the cells with epidermal growth factor for at least 2 h followed by A23187 or arachidonate (in the continuing presence of epidermal growth factor), however, stimulated prostaglandin E2 output up to 14-fold. The maximum effect of epidermal growth factor was attained at 1-10 ng/mL, while the EC50 was 0.2-0.32 ng/mL. Ionophore- or arachidonate-promoted prostaglandin E2 output was not stimulated by pretreatment with platelet-derived growth factor, fibroblast growth factor, and beta-transforming growth factor. Cycloheximide added before, at the same time as, or up to 30-60 min after epidermal growth factor completely abolished the stimulation. Epidermal growth factor did not affect [14C]arachidonate incorporation into cells or cell lipids. These results suggest that epidermal growth factor promotes, specifically and in a protein synthesis dependent manner, the conversion of arachidonate to prostaglandin E2. The provision of exogenous or endogenously liberated arachidonate is also necessary for enhanced amnion prostaglandin E2 production.

PMID: 3139271 [PubMed - indexed for MEDLINE]

Transcervical amnioinfusion of antibiotics: a basic study for managing premature rupture of membranes.

• Ogita S,
• Imanaka M,
• Matsumoto M,
• Oka T,
• Sugawa T.

Department of Obstetrics and Gynecology, Osaka City Perinatal Center, Japan.

To determine the best method of preventing ascending infection in the management of premature rupture of membranes, antibiotics such as latamoxef sodium, cefoperazone sodium, and cefotaxime sodium were infused directly into the amniotic cavity in 64 patients undergoing induction of labor at term. A single infusion of 100 or 500 mg of each drug resulted in a concentration of 200 to 1000 micrograms/ml immediately after infusion, and the concentration remained above 10 micrograms/ml for about 24 hours without significant increase in fetal or maternal blood levels. Consequently, a daily single dose of 100 mg or more is probably effective prophylaxis in cases of premature rupture of membranes. When intrauterine infection is suspected, the dose can be increased to 500 mg or more, and transplacental administration may be added to achieve a higher concentration in fetal blood. The present study simulates well premature rupture of membranes, and an amnioinfusion of antibiotics will be reliable and effective in managing premature rupture of membranes.

PMID: 3337177 [PubMed - indexed for MEDLINE]

Intra-amniotic injection of oxytetracycline hydrochloride for termination of mid-trimester pregnancy.

• Faktor JH,
• Frenkel Y,
• Mashiach S,
• Serr DM.

Department of Obstetrics and Gynecology, Sheba Medical Center, Tel-Aviv University, Israel.

Seventy-eight women underwent induced mid-trimester abortion. Fifty-two women aborted after an intra-amniotic injection of 1 g oxytetracycline hydrochloride. The control group comprised 16 women who received an intra-amniotic injection of hypertonic saline and 9 women with prostaglandin F2 alpha. All but 2 women aborted after one injection. Thirty-five women of the oxytetracycline hydrochloride group received intravenous oxytocin after the appearance of uterine contractions, 17 did not. The mean injection abortion interval in the women who received intravenous oxytocin was 38.6 +/- 2.7 h, whereas in the group without oxytocin it was 31.3 +/- 2.3 h. There is no statistically significant difference between these two groups (p less than 0.2). The mean injection abortion interval in the hypertonic saline group was 18.4 +/- 2.2 h. In the F2 alpha group it was 13.2 +/- 1.3 h. There is no statistically significant difference between hypertonic saline and prostaglandins (p less than 0.2). The mean injection abortion interval is significantly shorter in the F2 alpha and hypertonic saline groups as compared to the oxytetracycline hydrochloride group (p less than 0.001). It is advisable therefore to use oxytetracycline hydrochloride only in cases when the use of F2 alpha or hypertonic saline is contraindicated.

PMID: 3240887 [PubMed - indexed for MEDLINE]

Mechanism of interferon action. Interferon alpha inhibits vesicular stomatitis virus primary transcript accumulation in P1/eIF-2 alpha protein kinase-deficient human fibroblast cells.

• Zhang XT,
• Samuel CE.

Department of Biological Sciences, University of California, Santa Barbara.

The molecular basis of the inhibition of vesicular stomatitis virus (VSV) replication by purified recombinant alpha interferon (IFN-alpha A/D) in human fibroblast GM2767A cells was examined. A saturating concentration of IFN-alpha A/D inhibited infectious VSV yield by about four to five log10. By use of the VSV mutant tsG41, which is competent in RNA transcription but defective in RNA replication at 40 degrees C, it was shown that IFN-alpha A/D treatment significantly inhibited primary viral protein synthesis. However, the apparent IFN-induced inhibition of VSV protein synthesis was due primarily to a reduction in the accumulation of VSV primary transcripts in IFN-alpha A/D treated GM2767A cells rather than to a direct effect on translation per se. The IFN-induced reduction in VSV primary genome expression was detectable after only 1 hour of IFN treatment; actinomycin D treatment of GM2767A cells prior to IFN-alpha A/D treatment blocked the establishment of the IFN-induced inhibition of VSV. In contrast to the results obtained with GM2767A cells, IFN-alpha A/D produced no detectable effect on the accumulation of VSV primary transcripts in human amnion U cells even though VSV primary protein synthesis and infectious virus yield were significantly reduced. In summary, the principal cause of the IFN-alpha induced inhibition of VSV replication in protein P1/eIF-2 alpha kinase-deficient human fibroblast GM2767A cells appears to be at or prior to primary transcript accumulation; thus, the antiviral mechanisms of IFN-alpha in GM2767A cells is fundamentally different from the IFN-alpha induced translation inhibition observed in kinase-sufficient human amnion U cells.

PMID: 2845723 [PubMed - indexed for MEDLINE]

Identification of calmodulin-like activity in term human amnion: effect of calmodulin inhibitors on prostaglandin biosynthesis.

• Olson DM,
• Kramar D,
• Smieja Z.

Human amnion prostaglandin E2 (PGE2) synthesis increases with the onset of labour, and this synthesis is Ca2+-dependent. To understand better the mechanism of Ca2+-stimulated PGE2 biosynthesis, studies were performed to identify the presence of the intracellular Ca2+-mediator, calmodulin, in human amnion and to examine its role in PGE2 synthesis. Calmodulin-like activity was identified by the ability of the microsomal and cytosolic fractions of the 105,000g centrifugation of amnion homogenate to stimulate cyclic AMP-dependent phosphodiesterase activity. Cytosolic fractions consistently stimulated phosphodiesterase activity more than microsomal fractions (P less than 0.001) in paired samples from term human amnions. This activity was calcium-dependent. The cytosolic and microsomal factors increased the Vmax but not the Km of phosphodiesterase. There were no differences in these parameters with the onset of labour. The distribution of calmodulin-like activity between microsomes and cytosol was similar to the distribution of calmodulin mass as determined by radioimmunoassay. Three structurally different inhibitors of calmodulin activity, calmidazolium, trifluoperazine and W7, were tested for their ability to inhibit cytosolic factor-stimulated phosphodiesterase activity and to inhibit PGE2 output from dispersed amnion cells obtained before the onset of labour at term (cesarean section cells) or after spontaneous labour and vaginal delivery (spontaneous labour cells). The 50% inhibitory concentrations of the calmodulin antagonists in the phosphodiesterase assay were: trifluoperazine (6.7 microM), calmidazolium (0.11 microM), and W7 (24 microM). Trifluoperazine inhibited both basal and calcium ionophore (A23187)-stimulated PGE2 output from cesarean section cells and spontaneous labour amnion cells. Calmidazolium inhibited basal PGE2 output in cesarean section cells and spontaneous labour cells, but had no effect on A23187-stimulated output. W7 inhibited only the ionophore-stimulated PGE2 output in cesarean section amnion cells. The rank order of inhibition of both phosphodiesterase activation and basal PGE2 output was: calmidazolium greater than trifluoperazine greater than W7. These results suggest that human amnion contains calmodulin and that its distribution, concentration and activity remain unchanged with the onset of labour. The data suggest, although not conclusively, that calmodulin may, in part, play a role in amnion cell PGE2 production. Further investigation of calmodulin effects upon specific enzymes in the PGE2 synthetic pathway will be necessary to elucidate a role for calmodulin in PGE2 production.

PMID: 3038990 [PubMed - indexed for MEDLINE]

Infections of the diabetic foot.

• DeValentine S,
• Fredenburg M,
• Loretz L.

Diabetic foot infections are frequently much worse than the initial clinical appearance would indicate. Altered immune response, peripheral vascular disease, and neuropathy are key factors in the production of infection. Treatment must be based upon careful bacteriologic analysis and individualized antibiotic therapy. Surgical debridement, when indicated, is the mainstay of treatment. Partial foot amputation and salvage are always preferred over higher amputation when possible.

PMID: 2952249 [PubMed - indexed for MEDLINE]

[Cytopathogenic action of intact and antibiotic-altered populations of Bordetella pertussis on a human amnion cell culture]

• Bakulina NA,
• Efimova OG,
• Vysotskii VV.

Cytopathogenic and adhesive action of intact B. pertussis population and population changed under the effect of antibiotics was studied comparatively. It was shown that the level and character of the changes in the cell culture of line FL human amnion depended on the antibiotic used in the experiments. Populations of B. pertussis changed under the effect of tetracycline had the highest cytopathogenic and adhesive activity, while the activity of the population changed under the effect of erythromycin was the least. The decrease in the cytopathogenic and adhesive activity of B. pertussis changed under the effect of erythromycin correlated with lowering of the toxicity and the loss of the main agglutinogenic factors of the surface of the bacteria cell wall. The increase in the toxicity of the population changed under the effect of tetracycline was accompanied by changes in the cytopathogenic and adhesive properties of the culture.

PMID: 2888430 [PubMed - indexed for MEDLINE]

[Electron microscopic study of the interaction of cultures of Bordetella pertussis altered by antibiotic action with a human amniotic cell culture]

• Efimova OG,
• Vysotskii VV,
• Bakulina NA.

The adhesive properties of B. pertussis subjected to long-term subcultivation on nutrient media with antibiotics were studied with electron microscopy and it was shown that under the action of erythromycin the adhesive capacity of B. pertussis markedly decreased. Subcultivation on media with tetracycline did not lower the adhesive activity of B. pertussis. Changes in the adhesive properties of B. pertussis under the action of the antibiotics correlated with changes in toxicity, serovars and ultrastructure of the bacteria in the population.

PMID: 2878640 [PubMed - indexed for MEDLINE]

The functions of oligosaccharide chains associated with influenza C viral glycoproteins. I. The formation of influenza C virus particles in the absence of glycosylation.

• Hongo S,
• Sugawara K,
• Homma M,
• Nakamura K.

The effect of a glycosylation inhibitor, tunicamycin (TM) on the replication of influenza C virus was investigated. Incorporation of [3H]-glucosamine into the gp88 glycoproteins of this virus was completely inhibited by TM at the concentrations higher than 0.25 microgram/ml. Under these conditions, the synthesis of internal proteins NP and M was shown in TM-treated cells but the synthesis of gp88 was not. The disappearance of gp88 was however accompanied with the appearance of two new polypeptides with molecular weights of 80,000 (T80) and 76,000 (T76). While T80 was identified by peptide mapping as a host cell protein whose synthesis was enhanced by TM, T76 was shown to correspond to a nonglycosylated form of gp88. Pulse-chase experiments revealed that there was no significant difference in the intracellular stability of T76 and gp88. Although TM depressed the production of infectious progeny virus greater than 100-fold, only a five-fold decrease was observed in the release of noninfectious physical particles, suggesting that glycosylation is not essential for the formation of influenza C virus particles. However, the virions from TM-treated cells had a lower buoyant density in isopycnic sucrose gradients and lacked surface proteins in either glycosylated or nonglycosylated form.

PMID: 3718234 [PubMed - indexed for MEDLINE]

Platelet-activating factor metabolism in human amnion and the responses of this tissue to extracellular platelet-activating factor.

• Billah MM,
• Di Renzo GC,
• Ban C,
• Truong CT,
• Hoffman DR,
• Anceschi MM,
• Bleasdale JE,
• Johnston JM.

It was found previously that platelet-activating factor (PAF, 1-0-alkyl-2-acetyl-sn-glycero-3-phosphocholine) is undetectable in human amniotic fluid obtained before labor but is present in the majority of samples of amniotic fluid obtained after labor. In the present investigation, the amount of PAF in amnion tissue and the ability of this tissue to produce PAF and respond to PAF were investigated. Amounts of PAF in amnion obtained either during the second trimester of gestation or at term (before labor) were similar. After labor, however, the amount of PAF in amnion increased to 2.5-times that in amnion before labor without any discernible changes in the amounts of two related glycerophospholipids viz., 1-0-alkyl-sn-glycero-3-phosphocholine and 1-0-alkyl-2-acyl-sn-glycero-3-phosphocholine. The Ca2+-ionophore A23187, in the presence of Ca2+, caused an increase in the amount of PAF in amnion tissue disks but PAF did not appear to be released into the incubation medium. The stimulation of PAF formation by A23187 and Ca2+ was not affected by the addition of indomethacin. Addition of PAF to disks of amnion tissue resulted in an increase in the concentration of prostaglandin E2 in the incubation medium. An increase in prostaglandin E2 formation of similar magnitude was induced by A23187. Based on these results it is concluded that PAF can be synthesized in amnion tissue and net production is stimulated by Ca2+. In addition, amnion is receptive to extracellular PAF and exhibits, as one response, an increased production of prostaglandin E2.

PMID: 3936121 [PubMed - indexed for MEDLINE]

Relation between cyclic adenosine monophosphate and prostaglandin output by dispersed cells from human amnion and decidua.

• Warrick C,
• Skinner K,
• Mitchell BF,
• Challis JR.

We have examined the ability of activators of adenylate cyclase and cyclic adenosine monophosphate to affect the output of prostaglandins E and F by dispersed cells of amnion and decidua collected from women following spontaneous labor. Cyclic adenosine monophosphate production by amnion and decidua cells was stimulated in a dose-dependent fashion by cholera toxin and by forskolin in the absence or presence of the phosphodiesterase inhibitor 3-isobutyl-1-methyl xanthine. Forskolin and cholera toxin also stimulated prostaglandin E and F output from amnion and decidua cells. Similar effects were seen with cells incubated with dibutyryl cyclic adenosine monophosphate +/- 3-isobutyl-1-methyl xanthine. The beta-adrenergic receptor agonists salbutamol, isoproterenol, and epinephrine all stimulated prostaglandin E and F output from dispersed cells of both tissues. The stimulatory effect of 3-isobutyl-1-methyl xanthine was partially additive with the Ca2+ ionophore A23187. Basal outputs of prostaglandin and outputs stimulated by A23187 and by N6, O2'-dibutyryl adenosine 3':5'-cyclic monophosphate were attenuated by the calmodulin antagonist trifluoperazine in a dose-dependent fashion. We conclude that mechanisms exist for stimulation of adenylate cyclase in human amnion and decidua resulting in enhanced prostaglandin output. This pathway requires basal interaction with Ca2+-calmodulin and may be additive with cyclic adenosine monophosphate-independent mechanisms for prostaglandin stimulation.

PMID: 2412440 [PubMed - indexed for MEDLINE]

Monovalent cations transfer through isolated human amnion: a new pharmacological model.

• Bara M,
• Guiet-Bara A,
• Durlach J.

Transfer of monovalent cation through the isolated human amnion consists of different factors: paracellular, coupling, ATPase dependent cellular transfer, leak cellular transfer. Understanding this transfer permits testing of the action of various substances. Physiological substances (Mg, taurine) increase ionic transfer and there is a vicarious effect between Mg and taurine. The tocolytic agents MgSO4 and ethanol do not exhibit a good effect on the transfer: decrease with ethanol; equality between entry and exit fluxes with MgSO4. On the other hand, amphotericin B increases mother-to-fetus transfer. Polluting metals (Pb, Cd, Hg, As) dramatically reduce exchanges and almost completely inhibit amnion permeability. Ingestion of ethanol also exhibits a dramatic effect on the exchange between mother and fetus through the amnion. Study of ionic transfer in vitro can be considered a pharmacological model to investigate the modifications of mother-fetus exchanges by various substances.

PMID: 4021652 [PubMed - indexed for MEDLINE]

Synergism of antiviral activity in cell cultures treated with low concentrations of interferon and interferon-treated lymphocytes.

• Weigent DA,
• Lloyd RE,
• Blalock JE,
• Stanton GJ.

Human T cells treated with low levels of interferon (IFN) (1-10 units/ml), and washed to remove the IFN, transferred the same level of antiviral activity to recipient WISH cells as an equivalent IFN treatment alone could induce in WISH cells. Further, when T cells pretreated with IFN (1-10 units/ml) were cocultivated with WISH cells in the presence of IFN (1-10 units/ml), a 2.5- to 5-fold greater level of protection developed than could be expected from the additive effect of each. Antibody to leukocyte, fibroblast, or immune IFN blocked the antiviral effect of the respective IFN types but had no effect on the transfer of antiviral activity initiated by leukocyte, fibroblast, or immune IFN. Also, treatment of T cells with actinomycin D blocked the transfer of antiviral activity of IFN-treated T cells. Taken together, the data suggest that the increased antiviral activity is not merely an additive effect of the IFN, but represents a synergistic amplification of protection most likely due to the combination of the separate effects of IFN and IFN-induced transfer. Such interactions would be expected to play a major role in early protection against virus infections in vivo when low levels of interferon are present and lymphocytes are migrating into the area.

PMID: 6333028 [PubMed - indexed for MEDLINE]

[Distribution of ampicillin, gentamicin and ceporin in fetal organs after intra-amniotic and parenteral administration]

• Gurtovoi BL,
• Krasil'nikova AIa,
• Amiraslanova LA,
• Fursova SA.

PMID: 6507773 [PubMed - indexed for MEDLINE]

Preparation of amniotic membranes for surgical use with antibiotic solutions.

• Gannaway WL,
• Barry AL,
• Trelford JD.

A comparative study was conducted to investigate the effectiveness of antibiotic solutions for disinfecting vaginally delivered amniotic membranes. The microbial flora of eight amniotic membranes was defined and six antibiotic solutions were tested for their effectiveness in eliminating aerobic and anaerobic organisms. A solution containing 600 micrograms lincomycin HCl/ml, 500 micrograms neomycin sulfate/ml, 25 micrograms polymyxin B sulfate/ml, and 25 micrograms amphotericin B sulfate/ml in 0.5N saline was found to be capable of disinfecting amniotic membranes stored at 4 degrees C for 24 hours. This solution is now being used to disinfect surgically used amniotic membranes at our institution.

PMID: 6710355 [PubMed - indexed for MEDLINE]

Perinatal bacterial infection after prolonged rupture of amniotic membranes: an analysis of risk and management.

• St Geme JW Jr,
• Murray DL,
• Carter J,
• Hobel CJ,
• Leake RD,
• Anthony BF,
• Thibeault DC,
• Ross IB,
• Drage JS.

Chi-square and logistic stepwise multiple regression analysis of perinatal determinants of infant bacterial infection following prolonged rupture of amniotic membranes for 24 hours or more prior to delivery was applied in 33 infected infants and 66 matched control infants from the NINCDS Collaborative Project. In order of statistical significance, the most important variables were placental inflammation (P = 0.002), gestational age less than 34 weeks (P = 0.008), gestational age 34 to 37 weeks (P = 0.013), male sex (P = 0.015), Apgar score less than 6 at 5 minutes (P = 0.023), and clinical amnionitis (maternal fever, fetal tachycardia, or amniotic or gastric fluid leukocytes or bacteria) (P = 0.044). Duration of labor during PROM, race, and maternal age and parity were insignificant. Using these predictive variables, identification of infected infants for either microbial surveillance (superficial and systemic cultures) or microbial surveillance and anticipatory antibiotic therapy (discontinued after 3 days of negative cultures) was highly significant (P = 0.0001). Incorporating these variables and derived coefficients from multivariate analysis, a mathematical model was used for evaluation and prediction of perinatal bacterial infection with a sensitivity of 82% and specificity of 70%. Analysis of 46 infants prior to and 310 infants after implementation of this process at Harbor-UCLA Medical Center indicated significant improvement in the appropriate management of these infants at risk (from 59% to 87% of the population, P less than 0.05). Inappropriate antibiotic therapy decreased from 35% to 10% (P less than 0.05). In the absence of a shift in the median days of hospitalization of non-PROM infants, determination of the grand median days of PROM infant hospital stay showed a decrease (P less than 0.01) after initiation of this evaluation and management scheme.

PMID: 6707823 [PubMed - indexed for MEDLINE]

Predictive value of amniotic-membrane cultures for the development of postcesarean endometritis.

• Yonekura ML,
• Appleman M,
• Wallace R,
• Boucher M,
• Nakamura R.

This study assesses (1) the relationship between the bacteriology of amniotic-membrane cultures (AMCs) obtained at the time of primary cesarean section and the subsequent postoperative course of afebrile, laboring patients whose membranes ruptured greater than or equal to 4 hr before delivery and (2) the impact of perioperative antibiotics on this relationship. Therapy with perioperative antibiotics was begun after the cord was clamped and the membrane specimen was obtained for culture. Specimens for AMC were obtained from 127 patients; 62 received placebo and 65 received perioperative antibiotics. In spite of the homogeneity of the patients' clinical risk factors, their AMCs demonstrated considerable variation in the degree of bacterial contamination present at the time of cesarean section. The presence of no growth or of low-virulence isolates only in the AMC usually was associated with a benign postoperative course, and therapy with perioperative antibiotics did not significantly decrease the incidence of endometritis associated with these AMC patterns. On the other hand, the presence of a mixture of high- and low-virulence organisms in the AMC or of high-virulence anaerobes only--more specifically, gram-negative anaerobes--was predictive of subsequent postcesarean endometritis only for patients who did not receive perioperative antibiotics, and therapy significantly decreased the incidence of endometritis associated with these AMC patterns. In conclusion, the correlation between specific bacteriologic patterns found in the AMC and the subsequent development of endometritis for patients who did not receive prophylactic antibiotics was better than the correlation based on clinical risk factors alone.

PMID: 6372024 [PubMed - indexed for MEDLINE]

Management of chorioamnionitis.

• Looff JD,
• Hager WD.

Chorioamnionitis presents as a serious threat to both mother and fetus. We have evaluated the diagnostic criteria and method of management by means of a retrospective analysis. Ninety-nine per cent of mothers had two of three major risk factors: 1, rupture of membranes; 2, leukocytosis of greater than 15,000, and 3, temperature greater than 100.4 degrees F. All three risk factors were present in 82.7 per cent. The method of management included culturing the endocervix when premature rupture of the membranes was present, administering parenteral antibiotics once the diagnosis of chorioamnionitis was made and performing cesarean sections for obstetric indications only. This management protocol resulted in a perinatal survival rate of 94.9 per cent in infants weighing more than 1,000 grams compared with 98.3 per cent in the general population and no perinatal deaths in infants weighing more than 1,500 grams. There were no maternal deaths. Our data support the recommendations that endocervical cultures be obtained for all patients with premature rupture of membranes, mothers with ROM and normal temperature and white blood cell count be managed without antibiotics, mothers with a diagnosis of chorioamnionitis have induction of labor and cesarean section be performed for obstetric indications only.

PMID: 6695310 [PubMed - indexed for MEDLINE]

Adhesion of Proteus species to various cell types.

• Savoia D,
• Martinetto P,
• Achino A,
• Pugliese A.

Pathogenic and saprophytic Proteus strains from the urine of patients with urinary tract infections and healthy adults respectively were evaluated with regard to their ability to adhere in vitro to homologous cells (uroepithelial cells from urinary sediment and cultured WISH cells) and heterologous cells (RK-13 and MDCK cells). The effect on attachment of pretreating bacteria with subinhibitory concentrations of piperacillin and sagamicin was also determined for one sensitive and one resistant strain. Fifty percent of the pathogenic Proteus strains demonstrated good adherence to human urinary epithelial cells, whereas saprophytic strains did not adhere. There was a lower rate of attachment to culture cells. Piperacillin and sagamicin in subinhibitory concentrations caused a decrease in the attachment of the sensitive Proteus strain.

PMID: 6667683 [PubMed - indexed for MEDLINE]

Mycoplasma hominis: a placental pathogen?

• Embree JE,
• Krause VW,
• Embil JA.

Current information on the role played by Mycoplasma hominis in placental inflammation was reviewed. M. hominis is associated with chorioamnionitis and funisitis, but the clinical significance of this association in not clear. Further research on the role of the organism in perinatal disease is needed. Meanwhile, in a gravely ill infant with a history of prolonged rupture of the placental membranes, chorioamnionitis, and funisitis, M. hominis should probably be considered a potential pathogen, cultures for this organism should be performed, and therapy with an antibiotic regimen effective against M. hominis should be instituted.

PMID: 6665673 [PubMed - indexed for MEDLINE]

The effects of polyamines and aminoglycosides on phosphatidylinositol-specific phospholipase C from human amnion.

• Sagawa N,
• Bleasdale JE,
• Di Renzo GC.

The effects of polyvalent cations (polyamines and aminoglycoside antibiotics) on Ca2+-dependent phosphatidylinositol-specific phospholipase C activity of human amnion tissue were examined. In the presence of 1 mM Ca2+, the hydrolysis of phosphatidylinositol (2 mM) by phospholipase C was increased greatly (240-560% of control) by spermine (0.4 mM), spermidine (1 mM), neomycin (0.1 mM), gentamicin (0.2 mM), kanamycin (0.4 mM) and streptomycin (0.8 mM). Putrescine and cadaverine (0.1-2.0 mM), however, stimulated phospholipase C activity only slightly. The effects of spermidine, spermine and gentamicin on phospholipase C activity were characterized and found to be dependent upon the concentrations of phosphatidylinositol, Ca2+ and the particular polyvalent cation. At low concentrations of phosphatidylinositol and Ca2+ the predominant effect of polyamines and aminoglycosides was to inhibit phospholipase C activity. When the concentrations of phosphatidylinositol and Ca2+ were increased, spermidine, spermine and gentamicin stimulated phospholipase C activity. In the presence of 16 mM Ca2+, however, phospholipase C activity was maximal and was unaffected by either polyamines or aminoglycosides. At all concentrations of Ca2+ examined, the maximal stimulation of phospholipase C activity by a given polyvalent cation occurred at a fixed molar ratio of the particular polyvalent cation to phosphatidylinositol. Polyamines and aminoglycosides appeared to modulate the Ca2+ requirement for phospholipase C activity, but could not substitute completely for Ca2+. The activities of phospholipase A2, diacylglycerol lipase, monoacylglycerol lipase and diacylglycerol kinase in amnion tissue were unaffected by any of the polyvalent cations examined. It is proposed that any in vivo influences (stimulatory or inhibitory) of polyamines and aminoglycosides on amnion phospholipase C activity would depend upon the effective concentrations of Ca2+ and phosphatidylinositol.

PMID: 6849963 [PubMed - indexed for MEDLINE]

[Cefalexin therapy in amnionitis]

• Tork J,
• Bartfai G,
• Benko M.

PMID: 7177632 [PubMed - indexed for MEDLINE]

Chorioamnionitis and possible neonatal infection associated with Lactobacillus species.

• Lorenz RP,
• Appelbaum PC,
• Ward RM,
• Botti JJ.

A patient is described with premature labor at 32 weeks of gestation complicated by chorioamnionitis associated with Lactobacillus species, subsequent premature delivery, and possible neonatal infection. Mother and infant did well with antibiotic therapy. The significance of chorioamnionitis and neonatal pneumonia due to this organism group is discussed.

PMID: 7130367 [PubMed - indexed for MEDLINE]

[Induction of immune interferon by erythrogenic toxins A and B from Streptococcus pyogenes]

• Tonew E,
• Gerlach D,
• Tonew M,
• Kohler W.

By interaction of streptococcal erythrogenic toxins A and B with chick embryo fibroblasts and human amnion (FL) cells an antiviral interferon-like factor was secreted. It inhibited the replication of vesicular stomatitis, vaccinia and Mengo viruses. The streptococcal toxin type B was 50 times more cytotoxic for both cell cultures in comparison with streptococcal toxin A. The maximum tolerated doses of the two types of streptococcal toxins induced approximately the same antiviral protection effect. The production curve of the antiviral factor showed a maximum at the 12th hour after incubation at 37 degrees C with graduate decrease up to the 24th hour using a 6 hours induction time. The interferons induced by the streptococcal erythrogenic toxins A and B were thermostable at 56 degrees C for 30 min and were partially destroyed at pH 2 as tested against Mengo virus in FL cells. The antiviral effect could be reversed by addition of streptococcal erythrogenic toxin A at the maximum tolerated dose simultaneously with the glycosylating inhibitors streptovirudin and D-glucosamine by 90 and 100 per cent, respectively.

PMID: 6817542 [PubMed - indexed for MEDLINE]

Puerperal and perinatal infections with group B streptococci.

• Pass MA,
• Gray BM,
• Dillon HC Jr.

Twenty-one patients were seen with puerperal sepsis owing to group B streptococci (GBS), resulting in an attack rate of 2/1,000 deliveries. Most were young primiparous black women from a population with a known high incidence of GBS carriage. The association among abdominal delivery, endometritis, and puerperal sepsis was striking. Cultures of the birth canal or lochia were commonly positive for the same serotype recovered from the blood. Forty-seven patients with nonbacteremic GBS infections were seen; 27 had endometritis or amnionitis. Twenty patients had GBS urinary tract infection: Eight infections occurred prenatally, seven at delivery, and five post partum. Seven neonates developed serious GBS infections; intrauterine exposure occurred in at least four cases. Fetal exposure to GBS also occurred in three of four cases in which parturients with GBS bacteremia were delivered of their infants by cesarean section. Because of the high incidence of puerperal and perinatal GBS infections in this population, antibiotic prophylaxis regimens may be beneficial.

PMID: 7044126 [PubMed - indexed for MEDLINE]

Synthesis of prolactin by human decidua in vitro.

• Liu TI,
• Minaguchi H,
• Taga M,
• Mori H,
• Sakamoto S.

Human decidua, chorion, amnion and placenta from the 1st., 2nd. and 3rd. trimesters of gestation were investigated for synthesis and secretion of prolactin by in vitro incubation of these tissue fragments in medium 199. Prolactin content in decidua was found to be significantly higher than that in chorion, amnion or placenta at any stages of gestation. During 6 hours of incubation, decidua secreted significantly more prolactin into medium than did chorion, amnion or placenta. The amount of prolactin secreted by decidua was significantly higher than the prolactin content in tissue before incubation. Decidua were also incubated in medium 199 with or without actinomycin-D (20-200 micrograms/ml), puromycin (200 micrograms/ml) or cycloheximide (100-200 micrograms/ml) for 12 hours. Both total prolactin secreted into medium and prolactin content in tissue after incubation were significantly lower than the control without inhibitor. Decidua of 2nd. trimester of gestation was noted to secrete more prolactin into medium than decidua of 1st. or 3rd. trimester of gestation. In further studies, 3H-leucine (200 muCi) was incubated for 12 hours with decidua (10 grams) from 2nd. trimester of gestation. The incorporated protein in medium or tissue was extracted, fractionated by gel filtration through Sephadex G-100 column and by 10% SDS-polyacrylamide gel electrophoresis. Peaks of 3H count and immunoreactive prolactin in gel slices were coincident with the peak of standard pituitary prolactin at gel slices' No. 31 (Rf = 0.62). These results demonstrate that prolactin is synthesized and secreted by human decidua, which is identical to human pituitary prolactin. The synthetic activity is most prominent in the decidua of the 2nd. trimester of pregnancy.

PMID: 7052926 [PubMed - indexed for MEDLINE]

Intraamniotic infection due to group G streptococcus: treatment and antibody response.

• Larsen JW Jr,
• London WT,
• Baker CJ,
• Curfman BL,
• Sever JL.

Penicillin treatment and antibody response were studied using a rhesus monkey model for intraamniotic infection with type III group B streptococci (T3GBS). Acute and convalescent phase sera from mothers and their offspring were tested with a radioactive antigen-binding assay to determine the concentration of antibody to the capsular T3GBS antigen. The frequency of placentitis was significantly lower in penicillin-treated animals (3 of 8) than in controls (10 of 10; P less than .01). The penicillin group also had a significantly lower neonatal mortality (1 of 9) than controls (6 of 10; P less than .05). Both groups of rhesus mothers developed a significant increase in concentration of antibody to T3GBS, but the antibody response was of lesser magnitude in the penicillin-treated group. This experimental model appears to be useful for studying both therapy for intraamniotic infection and the humoral immune response to infection with T3GBS.

PMID: 7019795 [PubMed - indexed for MEDLINE]

Intra-amniotic antibiotic infusion.

• Goodlin RC.

PMID: 7223805 [PubMed - indexed for MEDLINE]