Anti-endotoxin monoclonal antibodies.
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Biomedical subjects
Publications and source records attributed to S Rosenfeld.
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The mouse neuroblastoma x rat glioma hybrid NG108-15 was previously shown to express delta opioid receptors. Because neuroblastoma cells display different phenotypes and cloned cell lines are heterogenous, we studied the characteristics and distribution of human 125I-beta-endorphin (125I-beta E) binding sites in cultures of NG108-15 cells with the use of micro-autoradiography and light microscopy. 125I-beta E labeled delta sites in NG108-15 in the presence of the non-opioid blocking peptide, beta-endorphin (6-31) (beta E (6-31)). Silver grains resulting from 125I-beta E binding to the opioid sites occurred in diffuse patches over several cells, with preferential location in dense cell patches. Pretreatment of NG108-15 with the delta agonist DADLE, previously shown to decrease beta E binding to delta sites on intact cells, also reduced silver grain density; however, some cells located in dense cell clusters were resistant to substantial agonist induced loss of labeling. These results suggest that delta opioid binding has a heterogenous cellular distribution in NG108.
B19 parvovirus is pathogenic in humans, causing fifth disease, transient aplastic crisis, some cases of hydrops fetalis, and acquired pure red cell aplasia. Efforts to develop serologic assays and vaccine development have been hampered by the virus's extreme tropism for human bone marrow and the absence of a convenient culture system. We constructed recombinants containing either the major (VP2) or minor (VP1) structural proteins of B19 in a baculovirus-based plasmid, from which the polyhedrin gene had been deleted; these recombinant plasmids were used to generate recombinant infectious baculovirus. Subsequent infection of insect cells in vitro resulted in high-level expression of either B19VP1 or VP2. Parvovirus capsids were obtained by self-assembly in cell cultures coinfected with either VP1- and VP2-containing baculoviruses or, surprisingly, VP2-containing baculoviruses alone. Empty B19 capsids composed of VP1 and VP2 could replace serum virus as a source of antigen in a conventional immunoassay for detection of either IgG or IgM antiparvovirus antibodies in human serum. Immunization of rabbits with capsids composed of VP1 and VP2 resulted in production of antisera that recognized serum parvovirus on immunoblot and neutralized parvovirus infectivity for human erythroid progenitor cells. Baculovirus-derived parvovirus antigen can substitute for scarce viral antigen in immunoassays and should be suitable as a human vaccine.
We used 18 monoclonal antibodies against B19 parvovirus to identify neutralizing epitopes on the viral capsid. Of the 18 antibodies, 9 had in vitro neutralizing activity in a bone marrow colony culture assay. The overlapping polypeptide fragments spanning the B19 structural proteins were produced in a pMAL-c Escherichia coli expression system and used to investigate the binding sites of the neutralizing antibodies. One of the nine neutralizing antibodies reacted with both VP1 and VP2 capsid proteins and a single polypeptide fragment on an immunoblot, identifying a linear neutralizing epitope between amino acids 57 and 77 of the VP2 capsid protein. Eight of nine neutralizing antibodies failed to react with either of the capsid proteins or any polypeptide fragments, despite reactivities with intact virions in a radioimmunoassay, suggesting that additional conformationally dependent neutralizing epitopes exist.
Diseases associated with asbestos are prevalent in the construction trades primarily as the result of the previously widespread use of insulation materials containing asbestos in the building industry. Workers in metal related trades, who are employed at construction sites, but who do not routinely use such materials in their work, may also be at risk for asbestos hazards. To assess such risk, a clinical survey was conducted on 869 ironworkers from the New York metropolitan area. A high prevalence of abnormalities on chest radiographs was found. Three hundred and twenty nine (38%) of the examined workers had pleural abnormalities on their chest radiographs consistent with asbestos induced effects. The prevalence of radiographic abnormalities indicating interstitial lung disease was low (7%) as was the prevalence of restrictive pulmonary function impairment (7%). Association was found between forced vital capacity (FVC) and pleural abnormalities on chest radiographs. Although ex-smokers appeared to have the highest prevalence of abnormalities, regression analyses showed that duration of employment in the ironworkers' trade was the most important factor associated with them.
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A decertification action by the Department of Health and Human Services (formerly the Department of Health, Education and Welfare) substantially impacts on the lives of Medicaid patients who reside in the affected nursing home. Decertification means that the federal government, the state government, or both will no longer pay for the care of Medicaid patients in the decertified home. Thus, a decision to decertify necessitates the transfer of elderly and infirm Medicaid patients out of the decertified home. Since this transfer may threaten the lives and health of these patients, they should be granted the right to participate in pre-decertification proceedings. However, in O'Bannon v. Town Court Nursing Center, the Supreme Court decided that patients do not have the right to participate in predecertification proceedings. The Court rejected the patients' due process arguments, finding: 1) that decertification does not deprive the patients of any constitutionally protected interest in life, liberty, or property; and 2) that any adverse consequences of decertification are only an "indirect and incidental" result of government action. This Comment analyzes the Supreme Court opinion and concludes that the patients have protectable property and life interests that entitle them to participate in some form of hearing prior to the decertification of the nursing home where they reside. In addition, this Comment suggests alternative methods for asserting nursing home patients' legal rights.
The intracranial pressure "reserve" test seems to be the most reliable method of determining when the brain's natural mechanisms for pressure compensation for added intracranial volume have been compromised or exhausted. The test employs a timed sequence of intracranial fluid injections, but as a safety precaution injections are discontinued if intracranial pressure remains elevated more than 10 Torr over baseline. In this case, a linear extrapolation is then calculated to determine the elevation which might have been achieved by a full series of injections. However, this linear extrapolation has been criticised on the expectation that an exponential response should be expected. A series of experimental observations in dogs and baboons and a review of clinical records in humans have been made to determine the observed slope of increase following aliquot injection during performance of the intracranial pressure reserve test. In these species the observed response was actually linear in shape rather than exponential. This held true even for different initial baseline values and with different volumes of "lesion" balloon inflations in experimental animals. A theoretic explanation is proposed.
The hemostatic properties and effect on osteogenesis of gelatin foam paste and bone wax were compared on surgical bone lesions in experimental animals. Thirty rabbits each received four trephine craniotomies and four lumbar laminectomies. Alternate bone incisions in each animal were treated with either gelatin foam paste or bone wax. Blood loss was measured by absorbing the blood into dry surgical cottonoids weighed before and after use. Bone healing sites of three rabbits were examined histologically to assess the effect of each agent on osteogenesis. The trephination sites of eight rabbits were subjected to fracture force testing at 6 weeks postoperatively to compare the effect ot the two agents on bone healing. No significant difference was found between gelatin foam paste and bone wax in either effectiveness of hemostasis or effect on osteogenesis.
An elderly woman of Ashkenazic Jewish descent, who had had adult Gaucher's disease diagnosed seven years previously, was found to have IgA myeloma. The diagnosis of IgA myeloma was confirmed by the presence of lytic lesions in the skull and long bones, atypical plasma cells in bone marrow, and monoclonal elevation of IgA, kappa-type. In addition, the marrow contained Gaucher's cells. A causal relationship between these two pathologic entities remains speculative.
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Limited proteolysis of rabbit skeletal troponin C with bovine thrombin yielded two fragments, TH1 (Mr = 11000) containing Ca2+ binding regions I--III and TH2 (Mr = 6000) containing region IV. Determination of the partial sequences of the fragments established the site of cleavage at Arg120-Ala121. Secondary cleavage by thrombin at other arginyl or lysyl residues in troponin C was ruled out by the sequence data and by the amino acid compositions of the two fragments.