Cultured keratinocytes and keratinocyte grafts.
Explore the source record for details and available documents.
Burns↗
Biomedical subjects
K Hancock
Publications and source records attributed to K Hancock.
Urethral placement through a bilateral myocutaneous gracilis flap neovagina.
Malignant melanoma of the vagina is an uncommon genital tumor. When the lesion arises in the distal vagina, radical surgery has been the only treatment to produce long-term survivors. A total vaginectomy, vulvectomy, and bilateral groin node dissection is recommended to assure adequate margins. Sexual function is lost unless reconstruction is performed. The bilateral myocutaneous gracilis flap neovagina has been a successful plastic procedure after exenterative procedures that result in loss of bladder and/or rectum. The authors present a technique in which bladder function was preserved after creation of a gracilis flap neovagina in a patient with Stage I melanoma of the distal vagina. Placement of the distal urethra through the reconstructed gracilis flap neovagina resulted in preservation of urinary function and continence, primary closure of the vulvar defect, and satisfactory sexual function. Two and one-half years after surgery the patient has good bladder and vaginal function without significant sequel.
Female↗
Modification of the FAST-ELISA for field diagnosis of schistosomiasis mansoni with serum or blood samples.
Using well-characterized serum pools from patients (with schistosomiasis mansoni), we modified the FAST-ELISA system for detecting serum antibodies to Schistosoma mansoni for field use. We found little or no change in test function or results when the test was run 1) with blood instead of serum, 2) after addition of sodium azide preservation to diluted and lysed blood specimens, 3) using impure water in wash steps, and 4) without a spectrophotometer (i.e., read with the naked eye). Our evaluation of the modified FAST-ELISA showed that it can be successfully used under the restraints of minimally equipped field laboratories.
Animals↗
Cisplatin, doxorubicin, and cyclophosphamide (PAC) in the treatment of mixed mesodermal tumor of the ovary.
Explore the source record for details and available documents.
Antineoplastic Combined Chemotherapy Protocols↗
Development and optimization of the FAST-ELISA for detecting antibodies to Schistosoma mansoni.
The Falcon assay screening test (F.A.S.T.) system was used to develop a rapid, sensitive, and quantitative kinetic-based enzyme-linked immunosorbent assay (k-ELISA) for detecting antibodies against Schistosoma mansoni adult microsomal antigens (MAMAs). The FAST-ELISA uses polystyrene beads on sticks molded to the lid of a microtitration plate. The beads are coated with antigen. Reagents and sera are placed in microtitration plates and the beads exposed to reagents by immersion. The exposure time required for a single dilution of serum or other antibody source, conjugate, and substrate is 5 min each. Excluding preparation time, two plates can easily be assayed in 30 min. The optima for assay conditions, reproducibility, quantitative linearity, and sensitivity are delineated. A battery of sera from patients with both homologous and heterologous infections was tested, and a dilution series of a standard reference serum pool was included with each test. Results were expressed in number of units as calibrated against the standard reference sera pool. Antigen-coated bead storage studies were performed with untreated and three chemically treated antigens. The storage stability of MAMA, ability to perform the assay with minimal equipment, sensitivity, short assay time, and ease of operation make the FAST-ELISA ideal for field studies.
Antibodies↗
Quantitative capacities of glutaraldehyde and sodium m-periodate coupled peroxidase-anti-human IgG conjugates in enzyme-linked immunoassays.
Covalently linked peroxidase-anti-human IgG conjugates were prepared by either glutaraldehyde or NaIO4 coupling techniques. Sodium dodecyl sulfate polyacrylamide gel electrophoresis shows that the glutaraldehyde coupled conjugate is composed of generally lower molecular weight components than the NaIO4 coupled product. The NaIO4 conjugate, when used to quantitate human immunoglobulin (Ig) in enzyme-linked immunoassays, appears to be highly sensitive in that small amounts of Ig elicited relatively high reactivities. The quantitative range of this type of conjugates, where reactivities are linearly proportional to the amount of human Ig present, is, however, extremely narrow (0.01-0.10 micrograms/ml of human IgG). Conversely, the glutaraldehyde coupled type conjugate is capable of sustaining a much wider range of linearity (0.01-0.6 micrograms/ml), but with a more gradual rise of reactivity which corresponds well to the amount of human Ig present. Conjugates prepared with glutaraldehyde are thus more useful in quantitative assays where wide quantitative ranges are desirable. NaIO4 conjugates on the other hand, are more suited to qualitative assays where sensitivity is more important.
Aldehydes↗
Calibration of prestained protein molecular weight standards for use in the "Western" or enzyme-linked immunoelectrotransfer blot techniques.
Prestained protein molecular weight standards allow easy, direct visual location of electrophoretically transblotted lanes on nitrocellulose. They also provide a simple and accurate means for calibrating the molecular weights of resolved bands. Commercial prestained protein molecular weight standards, however, appear to have significantly different molecular weights from the original unstained proteins. We describe a calibration of these prestained molecular weight standards.
Collodion↗
Demonstration of species-specific and cross-reactive components of the adult microsomal antigens from Schistosoma mansoni and S. japonicum (MAMA and JAMA).
Analysis of human serum reactivities to the Schistosoma mansoni adult microsomal antigens (MAMA) showed that S. japonicum and S. haematobium infection sera, as a rule, did not react as well to MAMA as did the homologous S. mansoni infection sera. The degree of species specificity, although not absolute, was quite pronounced. Purification of the corresponding microsomal antigens from S. japonicum adults (JAMA) and subsequent assays with both homologous and heterologous infection sera show a distinct and reciprocating species specificity between S. mansoni and S. japonicum microsomal antigens. The specificities of these antigens were quantitated by k-ELISA. Qualitative analysis of active antigenic components for both JAMA and MAMA involved assay by the "Western blot" or enzyme-linked immunoelectrotransfer blot (EITB). The EITB patterns of both antigens, after resolution by SDS-PAGE, show species-specific reactive bands at the 16,000 to 35,000 m.w. region. S. japonicum-specific antigens are located at the 18,000 to 35,000 m.w. region whereas S. mansoni-specific antigens were associated with m.w. components of 16,000 to 29,000. High m.w. antigen components (greater than 40,000) of both JAMA and MAMA are recognized by both heterologous and homologous infection sera and are thus not species specific. The demonstration of the clear separation of species-specific antigen bands of JAMA and MAMA by physical size offers a unique opportunity to isolate and characterize the species specificities of antibody-antigen reactions in these parasitic infections.
Animals↗
India ink staining of proteins on nitrocellulose paper.
India ink staining of proteins that have been electrotransfer blotted onto nitrocellulose paper is described. This stain proved to be a useful adjunct to the enzyme-linked immunoelectrotransfer blot technique. It is more sensitive than Coomassie blue, amido black, and fast green stains and is simple to use.
Animals↗
Adaptation of two avian rotaviruses to mammalian cells and characterization by haemagglutination and RNA electrophoresis.
Turkey and chicken rotaviruses were successfully adapted to replicate in a rhesus monkey embryo kidney (MA104) cell line. Trypsin treatment of virus and cells was essential for serial passage of the viruses. Polyacrylamide gel electrophoresis separated the viral RNA into 11 segments with clear resolution of segments 10 and 11. Segment 5 migrated with size class I, and this characteristic appeared to be a unique feature of the avian rotavirus genome. The ability of the avian rotaviruses to haemagglutinate a variety of erythrocytes was demonstrated. A type-specific antigen was detected by haemagglutination-inhibition assays and a group or subgroup antigen by enzyme immunoassays. Treatment of serum with heparin-MnCl2 was shown to be the best method for removing non-specific inhibitors of haemagglutination.
Animals↗
Evidence that the hypothalamus may be a source of a circulating Na+-K+-ATPase inhibitor.
Acetone extracts from a variety of rat tissues were tested for their ability to stimulate renal glucose-6-phosphate dehydrogenase (G6PD) activity at 2 min in an in-vitro cytochemical assay which is a marker of the sodium potassium-dependent adenosine triphosphatase (Na+-K+-ATPase) inhibiting activity. Extracts of the hypothalamus were the only ones found to be active in this system. Acetone extract of hypothalamus also inhibited renal Na+-K+-ATPase activity in vitro. The G6PD-stimulating activity from one hypothalamus was about 10000 to 100000 times greater than that of 1 ml plasma. The G6PD-stimulating activity of hypothalamic extracts from rats which had been on a high sodium intake for 4 weeks were approximately 150 times more active than those obtained from rats which had been on a low sodium diet. The G6PD-stimulating activity of the corresponding plasma was sixfold more active. These findings suggest that a circulating sodium transport inhibitor(s) may be secreted from the hypothalamus.
Animals↗
Schistosoma mansoni adult microsomal antigens, a serologic reagent. II. Specificity of antibody responses to the S. mansoni microsomal antigen (MAMA).
The purified Schistosoma mansoni adult microsomal antigen, MAMA, was used in the quantitative single-tube kinetic dependent enzyme-linked immunosorbent assay (k-ELISA) to measure antibody levels of various human patient sera. The 511 serum specimens tested were from patients with both homologous and heterologous infections. Sera from U.S., Egyptian, Brazilian, and Puerto Rican patients infected with S. mansoni reacted strongly with MAMA. Chinese patients infected with S. japonicum, and Nigerians or Egyptians infected with S. haematobium produced much lower responses to this antigen than those infected with S. mansoni. Sera from patients with echinococcosis, filariasis, paragonimiasis, clonorchiasis, trichinosis, amebiasis, and hepatitis and from healthy uninfected control individuals generally contained no detectable antibodies against this antigen. The S. mansoni adult microsomal antigen, MAMA, therefore, appears to be a highly potent and specific reagent for the serodiagnosis of S. mansoni infections.
Animals↗
Schistosoma mansoni adult microsomal antigens, a serologic reagent. I. Systematic fractionation, quantitation, and characterization of antigenic components.
A systematic and quantitative search was conducted to identify and isolate a serologically pertinent antigen with high specific activity and low cross-reactivity from adult worms of Schistosoma mansoni. Adult worms of S. mansoni quickly thawed from liquid N2 temperatures were disrupted by controlled homogenization in isotonic buffered sucrose. Differential centrifugation of the homogenate yielded three particulate and one soluble fractions: the 480 x G pellet (nuclear), the 7650 x G pellet (mitochondrial), the 360,000 x G pellet (microsomal), and the 360,000 x G supernatant (cytosol). Quantitative analysis indicated a major concentration of specific antigenic activities in the microsomal fraction. Further purifications of the urea-solubilized, n-butanol-treated microsomal particles by gel filtration and ionic-exchange chromatography resulted in a microsomal antigen (MAMA) possessing high specific activity and low cross-reactivity. The final purification post-ionic exchange chromatograph showed a 30-fold increase of specific antigen activity over that of the cytosol fraction. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunotransfer blot by the "Western blot" technique (EITB) indicated specific antigenic activities in association with several different m.w. bands (heterogeneous m.w. by extrapolation = 4.3 to 11.9 x 10(6), 2.0 x 10(5), and 2.0 x 10(4) daltons) of the MAMA fraction. When compared with other reported serologic antigens, MAMA showed substantially higher specific activity and lower cross-reactivity.
Animals↗
Ten-year survival rates in breast cancer using combination chemotherapy.
The outcome in 254 patients with all stages of breast cancer treated by combination chemotherapy is presented. All the patients were treated 10 or more years ago. The 10-year survival rate for Stages I and II combined is 60 per cent, in Stage III 19 per cent and in Stage IV 3 per cent. The combined rate in Stages I and II differed markedly according to hormonal status. In premenopausal patients the rate was 84 per cent compared with 42 per cent in postmenopausal patients.
Adult↗
Attitudes toward the aged as a function of age and education.
Explore the source record for details and available documents.
Age Factors↗
Rapid characterization of S. mansoni expression library clones of potential interest.
A S. mansoni adult worm cDNA expression library was screened with sera from baboons in a early phase after infection. The clones that were positive with the early infection sera were examined for reactivity with pre-infection sera and heterologous infection sera. In order to discriminate a positive antibody reaction from the reactivity due to residual anti-E. coli antibodies, an unrelated cDNA clone was plated with the positive clone. The unrelated clone provided the negative background and the contrast necessary to discern a positive antibody reaction. In this way, we were able to eliminate selected clones that were positive with the pre-infection sera or heterologous infection sera. This characterization of the expression library clones enabled us to quickly target only clones with the desired pattern of antibody reactivity for sequencing, subcloning, and expressing.
Animals↗
A new direction for postnatal infant care.
Explore the source record for details and available documents.
Child Health Services↗