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Stevens-Johnson syndrome (toxic epidermal necrolysis) has similar pathophysiologic characteristics to extensive partial-thickness burns. Successful treatment of a 6-year-old with this syndrome denuding 95% of her body surface, with 6.5 square feet of amnion is reported.
AA3 is a novel antibody raised against human amnion, which reacts with the basement membrane of various epithelia of ectodermal origin. We used AA3 to examine the epidermal basement membrane zone in normal skin and different genetically determined types of epidermolysis bullosa (EB), by indirect immunofluorescence. AA3 staining was normal in dystrophic and simplex EB, but was markedly reduced in lesional and non-blistered skin in severe forms of junctional EB. In non-lethal junctional EB, the intensity of staining was variable and appeared to be inversely associated with disease severity, but did not correlate with demonstrable abnormalities of hemidesmosomes. AA3 binding was not reduced in pemphigoid lesions or normal suction blisters. It appeared to localize to the lamina lucida, but with different characteristics compared with antibodies to laminin and bullous pemphigoid antigen. These finding suggest that AA3 recognizes an antigen (or antigens) which may be involved in a primary biochemical defect in junctional EB. Moreover, this antibody may act as a new probe for this potentially lethal mechano-bullous disease.
The pectoralis major myocutaneous flap provides an excellent method for reconstruction of oral cavity defects. This flap has been used since 1978. However, the flap is not without problems. The major problem that has been seen is necrosis of the cutaneous and subcutaneous aspects of the flap. However, in all such instances the muscular portion of the flap survived. This complication was found to occur more commonly in the elderly, diabetics, patients with peripheral vascular disease and patients with excessive adipose or breast tissue superimposed between the cutaneous and muscular components of the flap. In this study the technique of oral cavity reconstruction in the high risk patient group has been modified to avoid using the skin component of the pectoralis major myocutaneous flap. Instead, pectoralis major muscle covered with amnion is used for reconstructive purposes with rapid healing and with very satisfactory functional and cosmetic results.
The surface morphology of normal term amnion was studied by scanning electron microscopy. Four patterns were observed. Two of the patterns showed distinctive minority populations of cells. In one, there were cells surrounded by intracellular channels; in the other, there were large flat cells. The microvilli in all four patterns showed variable morphology in different samples. Possible functional implications of the findings are discussed.
Vaginoplasty using human amnion has been performed in 27 patients aged from 13 to 28 years. Excellent epithelialization occurred with good functional results; the technique appears to be preferable to other surgical methods.
Discs of human amnion prepared from nine women delivered by elective caesarean section at term were incubated with and without purified human amniotic fluid surfactant (9 micrograms lipid P/ml), and the output of prostaglandin E (PGE) was estimated by radioimmunoassay. Surfactant stimulated the release of PGE from 3.8 (SD 2.9) to 5.4 (SD 2.5) pmol/mg dry weight/3 h (P less than 0.01). Arachidonic acid accounted for 2% of the fatty acids of surfactant lecithin. These results show that surfactant has a stimulatory effect on amniotic PGE production and that it is an important source of arachidonic acid in amniotic fluid.
Two cases of amnion rupture are presented in 'low-risk' pregnancies. One case ended in fetal death in utero at 21 weeks. The other resulted in the preterm delivery of an otherwise normal male with mild amniotic band syndrome.
PROBLEM: In hematopoietic cells, interleukin (IL)-13 shares many actions with IL-4. The effects of IL-13 in gestational tissues have yet to be reported, however. We compared the effects of IL-4 and IL-13 on the production of cytokines and prostaglandin E2 (PGE2) in epithelial amnion-derived WISH cells. METHOD OF STUDY: WISH cells were treated with IL-4 or IL-13 (0.08-10 ng/ml) with/without cotreatment with IL-1 beta (0.2 ng/ml), tumor necrosis factor-alpha (10 ng/ml) or epidermal growth factor (5 ng/ml). The production of IL-6, IL-8, and PGE2 was measured by immunoassay after 16 hr. RESULTS: Both IL-4 and IL-13 inhibited PGE2 production with indistinguishable concentration-response curves, under basal or stimulated conditions. The maximal inhibition of IL-1 beta-stimulated PGE2 production (to 28% +/- 10% of control) was seen at 10 ng/ml of IL-4 or IL-13. Basal IL-6 production was stimulated approximately twofold by IL-4 and IL-13, whereas IL-4 and IL-13 both inhibited cytokine-stimulated (but not basal) IL-8 production by approximately 50%. In the presence of 1 ng/ml of IL-4, IL-13 was unable to further inhibit PGE2 production. CONCLUSIONS: The inhibition of PGE2 and IL-8 production by IL-4 in WISH cells is mimicked by IL-13. Both cytokines, probably through binding to a common receptor complex, may share a role in suppressing inflammatory reactions within intrauterine tissues.
It was found that a human hepatoma-associated ALP (orthophosphoric monoester phosphohydrolase, E.C. 3.1.3.1) shared electrophoretic mobility, inactivation by urea, inhibition by inorganic phosphate, ethylenediaminetetraacetate, and amino acids (L-phenylalanine, L-leucine and L-homoarginine), heat stability, sensitivity to neuraminidase, pH optimum, Km value, and antigen site with fast moving ALP isozymes of FL cell strain derived from human amniotic membrane. However, 40-week-old fresh amniotic membrane lacked this isozyme. Instead, it had a placental type ALP consisting of minor components. The other ALP isozyme of FL cells had properties common to hepatoma ALP with regard to L-phenylalanine sensitivity, inhibition by ethylenediaminetetraacetate, inactivation by urea, and antigen site, but differed from it in electrophoretic mobility, sensitivity to L-leucine and L-homoarginine, and the presence of another antigen site. It was more heat stable and more sensitive to inhibition by inorganic phosphate than Hepatoma AP. The possible regulatory mechanism between the hepatoma-type ALP and the placental type ALP in the amnion cells is considered.
15-Deoxy delta(12,14)-prostaglandin J(2) (15d-PGJ(2)), an activator of peroxisome proliferator-activated receptor (PPAR)-gamma and -delta, is a prostanoid metabolite with anti-inflammatory actions. In intrauterine tissues, proinflammatory cytokines and prostaglandins have been identified as playing key roles in the maintenance of pregnancy and the onset of labor. We investigated and compared the early (<3 h) effects of 15d-PGJ(2) with rosiglitazone (PPAR-gamma ligand) and 2-methyl-4-((4-methyl-2-(4-trifluoromethylphenyl)-1,3-thiazol-5-yl)-methylsulfanyl)phenoxy-acetic acid (GW501516) (PPAR-delta ligand) on interleukin (IL)-1beta-induced prostaglandin and cytokine production by amnion-derived WISH cells. We show that 15d-PGJ(2) exerts differential effects depending on concentration. At low concentrations (<0.1 microM), 15d-PGJ(2) inhibited IL-1beta-stimulated prostaglandin E(2) (PGE(2)) but not cytokine (IL-6/IL-8) production or cyclooxygenase-2 (COX-2) expression. This effect was attenuated by a PPAR-gamma inhibitor [2-chloro-5-nitro-N-phenyl-benzamide (GW9662)], by transfection with a dominant-negative PPAR construct, and was reproduced by the PPAR-gamma ligand rosiglitazone. At higher concentrations (1-10 microM), 15d-PGJ(2) inhibited IL-1beta-stimulated PGE(2) and cytokine production and COX-2 expression, and this effect was not blocked by GW9662. Rosiglitazone at high concentrations (1-10 microM) stimulated PGE(2) production in the absence or presence of the dominant-negative PPAR. The PPAR-delta ligand GW501516 also inhibited IL-1beta-stimulated PGE(2) production but only at high concentrations (1 microM). IL-1beta-induced nuclear factor-kappaB (NF-kappaB) DNA binding activity was significantly inhibited by 15d-PGJ(2) (10 microM) and GW501516 (1 microM) but increased with 10 microM rosiglitazone. We conclude that 1) at low concentrations, 15d-PGJ(2) acts through a PPAR-gamma signaling pathway; b) at higher concentrations, its actions are mediated most likely through other pathways such as activation of PPAR-delta and/or inhibition of NF-kappaB; and 3) rosiglitazone exerts PPAR-independent effects at high concentrations (>1 microM).
The ultrastructure of the surface of primary human amnion monolayer cells undergoing cytopathology induced by Clostridium difficile toxin was examined by scanning electron microscopy. Our observations indicated that the type and distribution of cell surface projections were altered dramatically by this toxin. The patterns of such surface changes were specific for the two different types of cells found in this cell culture. Cells with demarcated borders showed rearrangement of microvilli into globular chains or ridges which lined up with the branching membrane. Cells without demarcated borders exhibited studlike microvilli, all arranged into ridges or globular chains. These changes were noted after 1 h of toxin exposure and persisted without further progression, in spite of continued toxin exposure, up to 48 h. These data indicate that C. difficile produces a cytolytic toxin and that scanning electron microscopy may be useful in determining toxin-cell interactions.
The genome of the human polyomavirus JC (Mad-1 strain) was molecularly cloned in Escherichia coli by using the plasmid vector pBR322. Recombinant DNA molecules were constructed with the entire JC genome inserted either at its unique EcoRI site at 0.0 map units or at its unique BamHI site at 0.51 map units. Viral DNA from each of these recombinant plasmids was capable of transforming human amnion cells, and cell lines established from transformed foci were positive for JC tumor antigen as assayed by indirect immunofluorescence.
The hypothesis of an intrinsic defect of germ plasma put forward by Streeter to explain the abnormalities observed in 'amniotic bands sequence' (ABS) was not supported by Torpin, who suggested that the bands derived from early amnion rupture, with formation of mesodermal strings and naked chorion, which then constricted or adhered to fetal parts. Recently, several authors have recorded and discussed possible mechanisms for 'non-band related' malformations in patients with otherwise typical limb constrictions and amputations. The mouse mutant disorganisation (Ds) is a semidominant with 72% of heterozygotes manifesting abnormalities, which include cranioschisis, limb duplications and deficiencies, gastro/thoracoschisis, and papillae protruding from other parts of the body. We report similar abnormalities including papillae in five fetuses and one newborn with ABS and, based on these observations and published reports, we suggest that a human homologue for Ds may be the cause of at least some examples of ABS.
A first-trimester transabdominal ultrasound (US) study was performed on twin pregnancies to determine the utility of US in predicting chorionicity and amnionicity. Among 85 dichorionic-diamniotic (DC-DA) twin pairs, a thick membrane was present in 78 (92%). Four of the DC-DA cases without a thick membrane had two distinct placental sites, allowing 82 DC-DA pregnancies (96%) to be predicted. Among 16 monochorionic-diamniotic (MC-DA) twin pairs, a thin membrane was present in 14 (88%). None of the four monochorionic-monoamniotic (MC-MA) cases had an identifiable membrane. The lambda sign had no value in this evaluation and was actually misleading, while a thick membrane or the identification of two separate placentas was always predictive of DC-DA twinning. However, a thin membrane, while usually predictive of an MC-DA pregnancy, did not exclude a DC-DA gestation. When no membrane is present, an MC-MA gestation is probable; however, a diamniotic pregnancy may still be present, and further evaluation is suggested.
Previously we grew differentiated primary epithelial tissue cultures from rabbit renal medullary thick ascending limbs but were unable to subculture them into lines. Now, following the use of amnion as a support during the initial passages, two cell lines have grown from single fragments of medullary thick ascending limbs. Cells have now been in culture past 12 passages over more than 2 yr. On confluence they formed morphologically differentiated epithelial monolayers with polarization of the cells visible on electron microscopy. They had apical zonula occludens and microvilli, lateral cellular interdigitations, and basal membranes flat against the support. "Domes" often were visible when the epithelia formed on dishes, indicative of salt and water transport. Other functional differentiation in some passages of one line or the other included presence of Tamm-Horsfall protein (demonstrated by immunofluorescence) or transepithelial voltage oriented apical surface positive. Both the Tamm-Horsfall protein and the voltage are normally expressed by intact medullary thick ascending limbs and are characteristic of this particular nephron segment.
A method is described for the sequential detergent and high ionic strength extraction of human amnion with the progressive enrichment of the intermediate filament (IF) cytoskeleton and its associated structures including hemidesmosomes (HD). TEM of the extracted epithelium in situ reveals IF bundles beneath the apical cell surface, around the nucleus and at the lateral edges of the cells where association with desmosomes occurs. IF bundles are also very prominent within basal cell processes where they loop through the cytoplasm adjacent to the HDs. A novel connecting filament network is observed running between the IFs and the hemidesmosomal dense plaque. The adjacent IF network contains both cytokeratin and vimentin, the latter revealed much more fully as a result of the extraction protocol. The hemidesmosomal plasma membrane contains integrin subunits alpha 6 and beta 4 and these are quantitatively retained as the basal cell surface during extraction, while nonjunctional plasma membrane is solubilised. Integrin beta 1 is found at the basolateral cell surface but, like actin, is extracted quantitatively and is not present in HDs. The extracted epithelial cells may be recovered by scraping and the IF network depolymerised to produce a particulate fraction containing short residual IFs, associated thin filaments and plaque material. This fraction contains immunoreactive cytokeratin and vimentin. Integrin alpha 6 beta 4 has been used as a biochemical criterion of the presence of HD material in the fraction. Both subunits are highly enriched. The fraction also contains the hemidesmosomal components HD1, BP230 and BP180. This method is likely to be useful in further characterisation of the HD.
The sensitivity of several group C arboviruses (Bunyaviridae) to human amnion interferon was compared to that of vesicular stomatitis virus (VSV) and Sindbis virus. In CPE inhibition assays. Apeu virus was the most sensitive of the group C arboviruses tested; it was significantly more inhibited than VSV, but was in the same range as Sindbis virus. In plaque reduction assays, the increasing order of sensitivity was Apeu, Marituba, VSV and Sindbis viruses. Single-cycle yields of VSV and Apeu virus were reduced to the same extent with 1-10 interferon units; with 230 units, VSV growth was inhibited to a much greater extent (100-fold) than Apeu virus.
FL-amnion cells have mainly two alkaline phosphatase (AP) isozymes, of which the faster migrating one (FL-APF) on 5% polyacrylamide gel electrophoresis has proved to be identical to the Kasahara isozyme, a tumor-associated AP of intestinal type. But the other slower migrating one (FL-APs) remains to be characterized. Immunological and enzymic examination of purified FL-APs revealed that it is a hybrid form of AP consisting of one subunit with a molecular weight of 66,200 coming from the same subunit as that of FL-APF and another one with a molecular weight of 58,000 from placental AP subunit with modified glycosylation.