Biomedical subjects

J B Weiss

Publications and source records attributed to J B Weiss.

At least 37 records · Page 2Linked to original sources

PubMed1996

Endothelial stimulating angiogenic factor in early fracture healing.

The early vascular response is essential for the normal progress of fracture healing and fracture site blood flow has been shown to reach a peak in the first two weeks after injury. Angiogenesis is an important step in this response as new vessel formation is necessary to vascularize the fracture haematoma and the fracture gap. Changes in serum levels of a low molecular weight endothelial stimulating angiogenic factor (ESAF) have been previously reported in a group of four patients with tibial fractures. In this group, ESAF levels were measured on three occasions only and at different time intervals. We present a more detailed profile of serum ESAF level changes in the first 14 days after the fracture.

Adolescent↗

PubMed1996

Tract savers.

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Enteral Nutrition↗

PubMed1996

Simultaneous PCR detection of Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus types 1 and 2 from genital ulcers.

A multiplex PCR (M-PCR) assay with colorimetric detection was devised for the simultaneous amplification of DNA targets from Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus (HSV) types 1 and 2. By using target-specific oligonucleotides in a microwell format, 298 genital ulcer swab specimens collected in New Orleans during three intervals from 1992 through 1994 were evaluated. The results of the M-PCR assay were compared with the results of dark-field microscopy and H. ducreyi culture on two different culture media. HSV culture results were available for 99 specimens collected during the third interval. Confirmatory PCR assays targeting different gene sequences for each of the three organisms were used to validate the M-PCR results. Specimens were resolved as positive for the determination of sensitivity if the reference diagnostic test was positive or if the results of both the M-PCR and the confirmatory PCR were positive. The resolved sensitivities of M-PCR for HSV, H. ducreyi, and T. pallidum were 100, 98.4, and 91%, respectively. The resolved sensitivities of HSV culture, H. ducreyi culture, and dark-field microscopy were 71.8, 74.2, and 81%, respectively. These results indicate that the M-PCR assay is more sensitive than standard diagnostic tests for the detection of HSV, H. ducreyi, and T. pallidum from genital ulcers.

Bacteriological Techniques↗

PubMed1995

Measurement of serum angiogenic factor in devascularized experimental tibial fractures.

Early recovery of the vasculature is of profound importance in healing fractures, but the exact role of the regenerating endothelium remains controversial. Serum activity of endothelial cell-stimulating angiogenesis factor (ESAF) was measured after tibial osteotomy in two groups of sheep over a period of 42 days. Each osteotomy was held in an instrumented external fixator, and in one group the musculoperiosteal vasculature was excluded from the osteotomy site. In the well-vascularized group (n = 4), ESAF activity was significantly higher 2-5 days after osteotomy, with a peak at 4 days (21.1 +/- 5.4, p < 0.05), and displayed a biphasic pattern of secretion with a second peak at 7 days (24.3 +/- 5.5). In the devascularized group (n = 4) overall activity was lower, with only a single peak occurring at 9 days (17.5 +/- 3.2). These findings were correlated with significant differences in structural properties and on histological examination. The early detection of such regional vascular responses may prove to be of significant clinical value in high-energy trauma, enabling prediction of those fractures at risk of delayed or nonunion.

Angiogenesis Inducing Agents↗

PubMed1995

DNA probes and PCR for diagnosis of parasitic infections.

DNA probe and PCR-based assays to identify and detect parasites are technically complex; however, they have high sensitivity, directly detect parasites independent of the immunocompetence or previous clinical history of the patient, and can distinguish between organisms that are morphologically similar. Diagnosis of parasites is often based on direct detection by microscopy, which is insensitive and laborious and can lack specificity. Most PCR-based assays were more sensitive than DNA probe assays. The development of PCR-based diagnostic assays requires multiple steps following the initial selection of oligonucleotide primers and reporter probe. Generally, the ability to detect the DNA of one parasite was attained by PCR; however, advances in the preparation of samples for PCR (extraction of DNA while removing PCR inhibitors) will be required to achieve that sensitivity with human specimens. Preliminary PCR systems have been developed for many different parasites, yet few have been evaluated with a large number of clinical specimens and/or under field conditions. Those evaluations are essential for determination of clinical and field utility and performance and of the most appropriate application of the assay. Several situations in which PCR-based diagnosis will result in epidemiologic, medical, or public health advances have been identified.

Animals↗

PubMed1995

Multiplex PCR assay and simple preparation method for stool specimens detect enterotoxigenic Escherichia coli DNA during course of infection.

Infection with enterotoxigenic Escherichia coli (ETEC) is a common cause of diarrhea among travelers and residents of developing countries. ETEC produces either a heat-stable toxin or a heat-labile toxin, or both, encoded by plasmid-borne ST and LT genes, respectively. Diagnosis of infection with this subclass of E. coli can be performed with oligonucleotide hybridization probes; however, the sensitivity and specificity of this method are insufficient. A nonradioactive multiplex PCR assay that provides a sensitive and specific method for detecting the presence of either or both toxin genes has been developed. A simple procedure that removed inhibitors of the PCR while efficiently releasing ETEC DNA from stool specimens for subsequent amplification was used. The results for samples from a human volunteer study of ETEC infection indicated that this method of sample preparation results in greater clinical sensitivity than conventional total nucleic acid extraction and ethanol precipitation. Detection of ETEC by a multiplex PCR assay in stool specimens directly processed with a glass matrix and chaotropic solution had greater sensitivity than culture.

Adult↗

PubMed1994

Specific detection of Clostridium difficile toxin A gene sequences in clinical isolates.

The polymerase chain reaction (PCR) was used to specifically detect toxin A gene sequences of Clostridium difficile in DNA isolated from human faeces. A set of oligonucleotide primers derived from the non-repetitive region of the toxin A gene was developed to amplify a 634-bp DNA fragment. All 28 cytotoxic strains of C. difficile, previously characterized by a toxin B-PCR assay, were positive for the presence of toxin A gene sequences. No amplification products were obtained from DNAs extracted from non-toxigenic strains, strains of C. sordellii, or C. bifermentans. In addition, amplification of DNA extracted from C. difficile 8864, a strain which does not produce toxin A, resulted in multiple bands which probed negative for toxin A gene sequences. DNAs extracted from nine stool specimens which were positive for toxin B by the cytotoxicity assay and by the toxin B-PCR assay were also positive in this assay. Toxin A gene sequences were detected in DNAs obtained from 4/11 stool specimens which were negative by the toxin B cytotoxicity assay. These four specimens were from patients who had a history of relapses due to C. difficile-associated colitis, and whose stools had previously been found to be positive by the toxin B-PCR test despite no detectable toxin B in the specimens. These data indicate a comparable degree of clinical sensitivity between these two toxin-gene PCR-based assays. This rapid, sensitive and specific assay may be useful not only in the diagnosis of C. difficile infections, but also in molecular studies of the toxin A gene in C. difficile strains.

Bacterial Proteins↗

PubMed1994

Endothelial cell stimulating angiogenesis factor--a new biological marker for disease activity in ankylosing spondylitis?

Endothelial cell stimulating angiogenesis factor (ESAF) is raised in conditions of neovascularization, such as may occur at the endochondral surface of the bone growth plate or in association with syndesmophyte formation. Syndesmophytes are formed in response to chronic enthesitis. Twenty unselected patients with AS were studied and enthesitis was measured clinically using a previously validated enthesitis index. Biochemical and haematological markers of disease activity were measured along with ESAF. Radiographs were scored for the degree of sacroiliitis and syndesmophyte formation and all these measurements were correlated to the enthesitis index. ESAF was raised in AS compared with normal controls. ESAF correlated positively with the enthesitis index but not with any of the biochemical or haematological indices. Higher levels of ESAF were found in those with more advanced sacroiliitis. High ESAF activity may be reflecting the angiogenic process involved in new bone formation associated with sacroiliitis and syndesmophyte formation. ESAF may be a biochemical marker for disease activity in AS.

Adult↗

PubMed1993

The origin of HIV-1 isolate HTLV-IIIB.

The striking similarity between the first two human immunodeficiency virus type 1 (HIV-1) isolates Lai/LAV (formerly LAV, isolated at the Pasteur Institute) and Lai/IIIB (formerly HTLV-IIIB, reported to be isolated from a pooled culture at the Laboratory of Tumor Cell Biology (LTCB) of the National Cancer Institute) provoked considerable controversy in light of the high level of variability found among subsequent HIV-1 isolates. In November 1990, the Office of Scientific Integrity at the National Institutes of Health commissioned our group to analyse archival samples established at the Pasteur Institute and LTCB between 1983 and 1985. Retrospective analyses have shown that contamination of a culture derived from patient BRU by one from patient LAI was responsible for the provenance of HIV-1 Lai/LAV; the contaminated culture (M2T-/B) was sent to LTCB in September 1983. Our goals were to determine which HIV-1 variants were present in the samples and the sequence diversity among HIV-1 isolates from the earliest stages of the AIDS epidemic. We examined archival specimens and report here the detection of six novel HIV-1 sequences in the cultures used to establish the pool: none is closely related to HIV-1 Lai/IIIB. A sample derived from patient LAI contained variants of both HIV-1 Lai/IIIB and HIV-1 Lai/LAV, and a sequence identical to a variant of HIV-1 Lai/IIIB was detected in the contaminated M2T-/B culture. We conclude that the pool, and probably another LTCB culture, MoV, were contaminated between October 1983 and early 1984 by variants of HIV-1 Lai from the M2T-/B culture. Therefore, the origin of the HIV-1 Lai/IIIB isolate also was patient LAI.

Acquired Immunodeficiency Syndrome↗

PubMed1993

Collagenase and gelatinase production by calcifying growth plate chondrocytes.

The regulation of cartilage resorption is important both in cartilage pathophysiology and angiogenesis. Previous work has identified a link between calcification and activation of collagenolysis. The aim here was to test whether production of collagenase itself is also calcification-dependent, using a high-density growth plate chondrocyte culture model of calcification. Ultrastructural studies indicated that calcification occurred around large hypertrophic cells. There was no indication of phagocytosis of crystals even by cells lying next to mineral aggregates, although remodeling of the organic matrix by cell processes was evident. Release of collagenase activity increased dramatically between 24 and 48 h postcalcification, from low or undetectable basal levels. In contrast, gelatinase production was not calcification-dependent. Collagenase was released almost entirely in the latent form, being a consequence of increased protein synthesis rather than activation of existing enzyme. This linkage of calcification with latent collagenase production represents part of a coordinated remodeling of both collagenous and mineral components of the matrix which may also extend, in vivo, to the control of microvascular invasion and resorption.

Animals↗

PubMed1993

Specific detection of toxigenic strains of Clostridium difficile in stool specimens.

Clostridium difficile is the infectious agent responsible for antibiotic-associated colitis. We report the use of the polymerase chain reaction technique to identify toxigenic strains of C. difficile in human stool specimens. A set of primers based on the nucleotide sequence of the toxin B gene, which amplified a 399-bp fragment from isolates producing toxin B, was designed. We examined 28 known toxigenic strains, which were all positive by this assay. DNAs from the nontoxigenic strains examined and from strains of Clostridium sordellii and C. bifermentans were not amplified with these primers. The sensitivity of this assay allowed us to identify as little as 10% toxigenic C. difficile cells in the presence of 90% nontoxigenic cells and to detect the toxin B gene in 1 pg of DNA from a toxigenic strain. DNAs extracted from 18 clinical stool specimens that were positive for toxin B by the tissue culture cytotoxicity assay were also positive by this assay. In addition, we detected toxin B sequences in DNA from 2 of 18 stool specimens that were negative for toxin B by the cytotoxicity assay. These two stool specimens were from patients who had a clinical pattern of colitis that was compatible with C. difficile causation. This rapid, sensitive assay will be useful for specific identification of toxigenic C. difficile and for revealing cases that are undetected by analysis of fecal samples for toxin B alone.

Bacterial Proteins↗

PubMed1992

Characterization of a series of novel fucose-containing glycosphingolipid immunogens from eggs of Schistosoma mansoni.

Lipid extracts of eggs, worms, and cercariae of the parasitic trematode Schistosoma mansoni have been shown to contain a large number of highly immunogenic glycolipids (Weiss, J. B., Magnani, J. L., and Strand, M. (1986) J. Immunol. 136, 4275-4282). Three fractions of schistosome egg glycolipids were selected on the basis of their reactivity with an anti-schistosome monoclonal antibody (128C3/3), which recognizes a developmentally regulated carbohydrate epitope present on both glycolipid and glycoprotein antigens from S. mansoni. These fractions were purified by silica gel chromatography and preparative high performance thin layer chromatography and characterized by monosaccharide, fatty acid, and linkage analysis with gas chromatography-mass spectrometry, as well as by positive and negative ion fast atom bombardment-mass spectrometry. The immunogens were shown to be glycosphingolipids having homologous structures based on a highly novel extension of glucosylceramide. Monosaccharide inhibition studies indicated that the epitope recognized by 128C3/3 residues in an outer region of the immunogens consisting of Fuc2GlcNAc (where Fuc is fucose) repeating units. The largest antigen characterized may have the following structure, based on the evidence presented in this paper. [sequence: see text] The evidence indicated the existence of a series of glycan structures created by deletions of one or more Fuc1----3 side chains from the above structure.

Animals↗

PubMed1992

Classification of subgroups of Giardia lamblia based upon ribosomal RNA gene sequence using the polymerase chain reaction.

A sensitive and specific polymerase chain reaction-based assay has been developed to detect and analyze polymorphism in the Giardia lamblia 18S ribosomal RNA gene. Efficient amplification required the inclusion of cosolvents (glycerol and dimethyl sulfoxide) in the reaction. Following the optimization of conditions for amplification and subsequent hybridization of amplified product with radiolabeled oligonucleotide probe, a detection limit of less than one organism's worth of DNA was achieved. Thirty-five different G. lamblia strains obtained from various human and animal host types and geographic locations were analyzed by this method. The strains could be divided into 3 groups on the basis of defined nucleotide substitutions within the 183-bp amplified DNA fragment of the 18S ribosomal RNA gene. The groupings based upon the 18S ribosomal RNA gene sequence correlated with groupings previously assigned based upon patterns of surface antigens and restriction enzyme analysis. Analysis of the G. lamblia 18S ribosomal RNA gene sequences present in fecal specimens obtained from giardiasis patients revealed the presence of the different sequence types in these specimens. Some specimens contained more than one sequence type. The identification of subgroups of G. lamblia may facilitate studies of virulence, infectivity, and the epidemiology of giardia infection.

Amino Acid Sequence↗

PubMed1992

Concentrations of endothelial-cell-stimulating angiogenesis factor, a major component of human uterine angiogenesis factor, in human and bovine embryonic tissues and decidua.

Embryonic development involves the establishment of new patterns of vascular growth in the fetus and within the lining of the womb. A factor, human uterine angiogenesis factor, has been purified from the decidua and stimulates the growth of blood vessels in collagen sponge implants and in the chick chorioallantoic membrane. Evidence is presented that suggests that a major active component of human uterine angiogenesis factor is an activator of latent matrix metalloproteinases, of low M(r), called endothelial-cell-stimulating angiogenesis factor and that this factor is present in substantial quantities in a number of embryonic tissues.

Angiogenesis Inducing Agents↗

PubMed1991

Activation of the matrix metalloproteinase inhibitor complex by a low molecular weight angiogenic factor.

Tissue inhibitor of matrix metalloproteinases is the major inhibitor of the collagenolytic enzymes and the inhibitory complex has been thought to be irreversible. In this paper we show that a low molecular weight non-protein endothelial cell stimulating angiogenic factor is able to reactivate the enzyme from the inhibitor complex and liberate free inhibitor. The importance of an angiogenic factor able to initiate limited degradation of extra-cellular matrix such that space is created for new capillary growth is discussed.

Angiogenesis Inducing Agents↗

PubMed1991

Increased endothelial cell stimulating angiogenesis factor in patients with tibial fractures.

Angiogenesis is an important step in the normal process of fracture healing, irrespective of the method of fixation. Using a quantitative assay, the amount of endothelial cell stimulating factor (ESAF) has been determined in a small group of patients with tibial fractures. ESAF levels were found to be increased when compared with normal volunteers, although this increase was relatively less in fractures held with intramedullary nails than with external fixation.

Adult↗

PubMed1991

Low molecular weight angiogenesis factors.

A number of substances have been proposed for the role of angiogenesis factors. Many of these are of protein origin and are therefore amenable to the tools of the molecular biologist. However a number of low molecular weight angiogenesis factors are emerging as important initiators and/or cofactors of neovascularization. Of these a number are known to stimulate angiogenesis indirectly, possibly through an inflammatory response. Some putative angiogenic factors stimulate microvessel endothelial cells nonspecifically, also causing migration and proliferation of large vessel cells. Others are specific for microvessel cells either for stimulating migration, proliferation or both. The nature and action of the low molecular weight factors in vivo and in vitro are reviewed.

Angiogenesis Inducing Agents↗

PubMed1991

Raised levels of latent collagenase activating angiogenesis factor (ESAF) are present in actively growing human intracranial tumours.

Endothelial cell stimulating angiogenesis factor (ESAF) is a potent, low molecular mass mitogen, specific for endothelial cells. In common with various protein growth factors, it displays angiogenic activity in a variety of biological test systems. However, it differs from these other factors by virtue of its low molecular mass and its ability to activate latent matrix metalloproteinases in a dose dependent manner. This activity has been used to quantify the factor in both normal and diseased brain tissue. The concentration of ESAF determined in biopsies from different types of intracranial tumours varied: in some tumour types the level was close to that of control samples whereas in others it rose to levels comparable to those encountered in the pineal gland, the richest source of ESAF in mature mammals. Tumours considered to be benign contained significantly less ESAF than those neoplasms classified as being malignant (P = 0.025). There was also a correlation between the mitotic activity of tumour samples, as determined by conventional H & E histochemical staining and the ESAF concentration present. These findings agree with previous studies in which elevated ESAF levels have been found in tissue where proliferation of vascular elements has been observed.

Adult↗