Biomedical subjects
Publications and source records attributed to P Ng.
Intraventricular neurocytoma is a rare cerebral tumour which is usually associated with a good prognosis. It has imaging features which help differentiate it from other cerebral tumours. We report three cases including the first to originate from the pineal gland. The report emphasizes the radiological appearance and reviews the literature.
The aim of this study was to compare the efficacy of plain films and computed tomography (CT) in defining inferior orbital fractures and any muscle involvement. Forty-four patients with final diagnosis of orbital floor fractures in the period 1990-94 were retrospectively studied. Computed tomography was performed in 28 patients, 20 being direct coronal acquisitions and eight being fine axial acquisitions with coronal reconstructions. Water's view radiographs were performed in 34 patients. Fourteen fractures on plain films were associated with soft tissue opacities to suggest inferior rectus (IR) muscle involvement, but only two required surgical elevation of the orbital floor. The remaining patients were successfully treated conservatively. Three patients had IR entrapment on direct coronal CT, all requiring surgical elevation of the orbital floor. Seven patients had IR muscle displacement on direct coronal CT and all had conservative management. In four patients with axial acquisition and coronal reconstructions, the CT images were of inadequate quality to determine the presence or absence of a fracture. One patient who had no IR muscle involvement identified on reconstructed coronal CT required surgical elevation of the orbital floor on clinical grounds. We conclude that: (i) soft tissue opacities on plain films are not an accurate indicator of clinically significant IR involvement; (ii) axial CT is not efficacious in detection of fractures or IR involvement; and (iii) direct coronal CT is the most efficacious imaging modality.
Ectopic recombination occurs between DNA sequences that are not in equivalent positions on homologous chromosomes and has beneficial as well as potentially deleterious consequences for the eukaryotic genome. In the present study, we have examined ectopic recombination in mammalian somatic (murine hybridoma) cells in which a deletion in the mu gene constant (Cmu) region of the endogenous chromosomal immunoglobulin mu gene is corrected by using as a donor an ectopic wild-type Cmu region. Ectopic recombination restores normal immunoglobulin M production in hybridomas. We show that (i) chromosomal mu gene deletions of 600 bp and 4 kb are corrected less efficiently than a deletion of only 2 bp, (ii) the minimum amount of homology required to mediate ectopic recombination is between 1.9 and 4.3 kb, (iii) the frequency of ectopic recombination does not depend on donor copy number, and (iv) the frequency of ectopic recombination in hybridoma lines in which the donor and recipient Cmu regions are physically connected to each other on the same chromosome can be as much as 4 orders of magnitude higher than it is for the same sequences located on homologous or nonhomologous chromosomes. The results are discussed in terms of a model for ectopic recombination in mammalian somatic cells in which the scanning mechanism that is used to locate a homologous partner operates preferentially in cis.
OBJECTIVE: To define the incidence, characteristics, and survival of children with perinatally acquired human immunodeficiency virus (HIV) infection and encephalopathy. DESIGN: Cross-sectional and longitudinal data collected from 1811 HIV-infected children in a multicenter active surveillance study. SETTING: Health departments and medical centers in six areas of the United States. RESULTS: HIV encephalopathy was diagnosed in 178 (23%) of 766 children with perinatally acquired immunodeficiency syndrome (AIDS). The median age at diagnosis of encephalopathy was 19 months. Among infected children, the estimated risk of having HIV encephalopathy by age 12 months was 4.0% (95% confidence interval, 2.6% to 6.0%). Children with HIV encephalopathy had more hospitalizations (median, 4) than children with other AIDS-defining conditions (median, 2; p = 0.002) and lower CD4+ T-lymphocyte counts in the first year of life (median, 444 cells/mm3). Estimated median survival after diagnosis was 22 months, similar to the 20 months for children with Pneumocystis carinii pneumonia. CONCLUSION: HIV encephalopathy in children with perinatally acquired AIDS is a common condition and is associated with severe morbidity evidenced by frequent hospitalizations, severe immunodeficiency, and short survival.
Mammalian cell culture-derived biotechnical products for therapeutic use have risks associated with cell substrate DNA. Regulatory authorities in the USA and Europe have set stringent limits for this contaminant, requiring orders of magnitude reduction from fermenter harvests to purified products. This paper addresses the relevant unit processes (e.g., cell separation, affinity chromatography and ion-exchange chromatography) utilized for purification of mammalian cell culture-derived products as they pertain to removal of DNA contaminants.
We have examined the stability of trinitrophenyl (TNP) -specific IgM production in hybridoma cell lines in which homologous recombination was used to change the variable region of an endogenous chromosomal immunoglobulin kappa gene to one specific for TNP. Mutant hybridomas that have lost TNP-specific IgM production are detected with a frequency of approximately 1%. Characterization of the mutant cells reveals a variety of gross rearrangements in the recombinant kappa TNP gene as well as in the endogenous kappa and muTNP genes.
The potential use of vaccines for treatment of chronic or persistent virus infections is an area of great interest and controversy. Previous experiments have shown that the incidence and severity of spontaneous recurrent genital herpes in latently infected guinea pigs could be significantly reduced by vaccination with herpes simplex virus glycoprotein subunit vaccines. The current study shows the critical role of adjuvant in an effective formulation. Immunization of previously infected guinea pigs with a subunit vaccine containing a muramyl peptide derivative, MTP-PE, in a low-oil emulsion as adjuvant reduced the incidence of recurrent disease up to 80% compared with formulations lacking MPT-PE. Vaccines containing adjuvant alone failed to modify recurrences. Alum, the traditional adjuvant, was not effective. Glycoprotein subunit vaccines elicited high-titer ELISA and neutralizing antibody responses far greater than those generated by virus infection. However, neither antibody titers nor lymphoproliferative responses reproducibly correlated with the pattern of recurrent disease.
PURPOSE: We studied oncologists' attitudes and behavior with regard to their participation in randomized clinical trials. METHODS: We surveyed the 1,737 physician members of the Eastern Cooperative Oncology Group (ECOG) using the Physician Orientation Profile (POP), a self-administered mailed questionnaire. A response rate of 86% was achieved (1,485 of 1,737); each physician's actual patient accrual was recorded. RESULTS: All respondents indicated that they had a systematic pattern of patient preselection for entry onto trials beyond the formal inclusion/exclusion trial criteria. Eighty-nine percent stated that improving patient quality of life rather than prolonging survival was more personally satisfying. Sixty-two percent did not enter a single patient during the 12-month period following the survey, while 10% entered 80% of all patients during that time. Physicians overestimated their accrual rate by a factor of 6. Eighty-three percent defined randomization and adherence to trial protocol as a serious challenge to their ability to make individualized treatment decisions. CONCLUSION: This study raises questions regarding the following: (1) the perceived generalizability of trial findings, (2) the role of end points other than survival for clinical trials, (3) the consequences of physician overestimation of patient accrual, and (4) the impact of randomized trials on the behavior of clinicians. Further investigation into these critical issues will provide meaningful recommendations to enhance the future design, implementation, and conduct of randomized clinical trials in cancer.
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To evaluate factors that may affect the timely diagnosis of children with human immunodeficiency virus (HIV) infection, we compared data derived from two population-based pediatric HIV studies. Data from anonymous newborn HIV serosurveys were used to estimate the number of children born to HIV-seropositive mothers. A statewide active surveillance project determined the number of HIV-exposed children who had been clinically recognized. Of 88,732 Massachusetts newborn specimens tested anonymously for HIV antibodies during a 12-month period (November, 1987, to October, 1988), 223 were positive. As of October, 1991, 78 of these children (35%) had been identified by a statewide network of infectious disease physicians. HIV-exposed children born in inner city hospitals were more likely to have come to medical attention than those born in suburban hospitals (47% vs. 17%). Among the 29 children with confirmed HIV infection (13% of 223), the initial evaluation for HIV occurred at an earlier age among children born in inner city hospitals than among children born in other areas. HIV testing practices that rely heavily on risk assessment may result in delayed diagnosis of HIV infection in children whose mothers are not perceived to be at risk.
Concentrations of polychlorinated biphenyls (PCBs), dibenzo-p-dioxins, and dibenzofurans, organochlorine pesticides, and their metabolites were measured in eggs of the common snapping turtle (Chelydra s.serpentina) collected from four wetlands on the shorelines of Lakes Ontario, and Erie, and one control location in central Ontario, Canada. Snapping turtle eggs from these sites were also artificially incubated to determine hatching success, and incidence of deformities in embryo and hatchling turtles. The hypothesis that elevated incidences of egg death and/or deformities of hatchling turtles would occur in populations with high concentrations of organochlorine contaminants in eggs was tested. The results were elevated using epidemiological criteria. Unhatched eggs and deformities occurred at significantly higher rates in eggs from Lake Ontario wetlands. Two of three sites from Lake Ontario had substantially higher levels of PCBs, dioxins, and furans compared to eggs from Lake Erie and the control site. It could not be shown that contamination of eggs preceded the occurrence of poor development of eggs, although excellent hatching success and low numbers of deformities in eggs from the control site were considered representative of development in healthy eggs. The statistical association between contaminant levels in eggs and poor development of these eggs supported the hypothesis that eggs from sites with the greatest contamination had the highest rates of abnormalities. PCBs were the most strongly associated chemicals, although possible effects due to the presence of other chemicals in eggs was a confounding factor. The deformities and rates of unhatched eggs were similar to those occurring in other vertebrates collected from highly contaminated areas of the Great Lakes. There were several chemicals present in the eggs that can cause similar reproductive effects in other species; therefore a specific chemical effect was not identified. Results were coherent with known statistical and biological information. Theoretical and factual evidence of PCB contamination in wild-caught snapping turtles supported and hypothesis. However, lack of controlled studies of reproductive effects of polychlorinated hydrocarbons upon this species hindered the agreement of all factual and theoretical evidence with the hypothesis.
Oligodeoxypurine- and oligodeoxypyrimidine-containing strands were mixed under conditions conducive to the formation of triple stranded assemblies. The mixtures were characterized both by their UV absorbance change with increasing temperature and by their mobility in non-denaturing polyacrylamide gels. Duplexes 34 bp long containing 15 central purines on one strand and 15 complementary pyrimidines on the other strand yielded new melting transitions and showed different gel mobilities upon combination with oligopyrimidine 15-mers. The dependence of the thermal denaturation profiles on pH, salt concentration, GC content, strand orientation, base mismatches, and strand length was investigated.
A strategy, termed homolog-scanning mutagenesis, was used to identify the epitopes on human growth hormone (hGH) for binding to its cloned liver receptor and eight different monoclonal antibodies (Mab's). Segments of sequences (7 to 30 residues long) that were derived from homologous hormones known not to bind to the hGH receptor or Mab's, were systematically substituted throughout the hGH gene to produce a set of 17 chimeric hormones. Each Mab or receptor was categorized by a particular subset of mutant hormones was categorized by a particular subset of mutant hormones that disrupted binding. Each subset of the disruptive mutations mapped within close proximity on a three-dimensional model of hGH, even though the residues changed within each subset were usually distant in the primary sequence. The mapping analysis correctly predicted those Mab's which could or could not block binding of the receptor to hGH and further suggested (along with other data) that the folding of these chimeric hormones is like that of HGH. By this analysis, three discontinuous polypeptide determinants in hGH--the loop between residues 54 and 74, the central portion of helix 4 to the carboxyl terminus, and to a lesser extent the amino-terminal region of helix 1--modulate binding to the liver receptor. Homolog-scanning mutagenesis should be of general use in identifying sequences that cause functional variation among homologous proteins.
The deoxyoligonucleotide 5' AATCGGGCATGGATT (15-mer) was synthesized containing 12 phosphoramidate linkages derived from 2 primary and 2 secondary amines. The oligonucleotides were purified by reverse phase HPLC and characterized by PAGE. The thermal stability of the duplexes derived from these compounds, when hybridized to the complementary diester linked oligomer, were determined and compared to the diester and methanephosphonate oligomer. The results indicated that all analogue oligomers form less stable duplexes then the diester oligomer. A large difference was observed between primary and secondary amine derived phosphoramidates.
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Infusion studies of recombinant factor VIII were performed in hemophilic (factor VIII-deficient) animals. Functional activity was determined using a standardized model of bleeding and kinetic characteristics charted by performing assays of factor VIII functional and antigenic activities over time. To obtain an adequate comparison with plasma-derived factor VIII, a crossover study in two animals was performed in which each animal received either recombinant factor VIII or a highly purified plasma-derived factor VIII on day 1 and the alternative three days later (day 4). Both factor VIII preparations were functionally effective with complete correction of the cuticle bleeding time occurring one hour after infusion. The observed recovery was full and close to predicted for both preparations. The survival curves obtained for both functional and antigenic activities for both preparations were virtually identical and within the anticipated range determined from previous experiments using infusions of conventional factor VIII preparations. The plasma half-disappearance time (T 1/2) for recombinant factor VIII and plasma-derived factor VIII was 9.2 and 7.9 hours, respectively. Plasma samples obtained following infusion were subjected to chromatography on Sepharose 4B. The elution profile of factor VIII antigen activity was compared with that obtained with the infusates. A clear shift in profile was observed with the plasma samples, suggesting complexing of the infused factor VIII material with circulating canine von Willebrand factor (vWF). The elution profile of vWF antigen was superimposable, thus providing further evidence in support of this assumption. The study provided evidence that recombinant factor VIII possesses full functional activity in vivo, binds to circulating vWF, and exhibits normal recovery and survival characteristics.
A gene for bovine pancreatic trypsin inhibitor (BPTI) was fused to the coding sequence for the Escherichia coli alkaline phosphatase signal peptide and expressed in E. coli under the control of the alkaline phosphatase promoter. When induced in phosphate-depleted medium such cells produced a trypsin inhibitor that was indistinguishable from native, properly folded BPTI. In particular, the BPTI produced by E. coli had three disulfide bonds that appeared to be identical to those found in native BPTI, as assayed by sensitivity to iodoacetate, dithiothreitol, and urea. This expression/secretion system will make possible the production of variant BPTI molecules, thus allowing the perturbing effects of amino acid substitutions on BPTI folding, structure, and function to be assessed.
The prominent role of plasma fibronectin (pFN) in the host defense system as an opsonin for gelatin (collagen)-coated colloids has been established. In the present study we investigated the interaction of pFN and membrane isolates from cells devoid of collagen, as well as several tissues. In a liver slice assay system it was shown that subcellular membrane fractions from lung macrophages, peritoneal exudate cells, spleen, tests, and liver were able to competitively inhibit the pFN-mediated uptake of 125I-gelatin coated latex beads (gLtx) at low concentrations. Endocytosis of 125I-labeled membrane isolates by macrophage monolayers was also promoted by addition of pFN. In an attempt to characterize the membrane component(s) interacting with pFN, it was found that mild extraction procedure with 1 M KBr could release a significant amount of this inhibitory activity. Further studies demonstrated that the agent(s) responsible for inhibition of gLtx uptake was heat sensitive, not altered by trypsin treatment, and did not contain actin, a protein known to interact with pFN. This work indicates that pFN interacts specifically with an as yet unknown membrane component(s) and that such interaction will promote clearance of cellular debris by macrophages. This suggests that pFN may be an important opsonin for the reticuloendothelial system in clearance of collagenous and noncollagenous cellular debris once they are exposed to interact with it.