Computer infrastructure.
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Computer Communication Networks↗
Biomedical subjects
C Smith
Publications and source records attributed to C Smith.
An evaluation of one and two airflow filters in preventing the movement of bacteria through the anesthesia circle system.
A review of the literature demonstrated that controversy exists as to whether anesthesia circle systems can be a source of transmission of bacteria to patients. The purpose of this study was to evaluate the use of one airflow filter (on the expiratory limb) versus two airflow filters (one on the inspiratory limb and one on the expiratory limb) in the prevention of bacterial migration in the anesthesia circle system, during general anesthesia, using semiquantitative analysis of the number of bacterial colony forming units (CFUs). This study consisted of two randomized groups receiving one or two airflow filters. Thirty-five volunteers participated in the study. At the conclusion of the surgical cases, a culture was taken of the anesthesia circle system. The cultures were read at 24 and 48 hours for the presence of CFUs. No CFUs were reported in either group. The results of the study were inconclusive. Therefore, the use of one airflow filter on the expiratory limb should remain the standard of care, pending additional research. Some practitioners may use a second airflow filter when there is a greater risk of infection, such as in the case of an immunocompromised patient. However, this study can neither support nor question the effectiveness of this practice.
Air Microbiology↗
Transfer of marker genes into hemopoietic progenitor cells.
Ex vivo gene marking of normal and malignant hemopoietic cells allows the cells to be subsequently tracked in vivo. Marking has shown that even when marrow is in remission, it may contain malignant cells that contribute to relapse. These studies have also shown that it is possible to obtain long-term gene expression in human long-lived hemopoietic progenitor cells and T lymphocytes in vivo. Current marker studies use two distinguishable vectors to track two distinctively treated cell populations in a single individual. This modification greatly increases the power of the technique. It is now possible to study the effects of purging on residual malignant cells in marrow, to determine the action of growth-promoting agents (such as cytokines and stroma) on short- and long-term repopulation by transduced marrow, and to discover which phenotypic subsets of hemopoietic progenitor cells have long-term repopulating potential. The information gained will be invaluable for improving therapeutic gene transfer protocols in which marrow-derived cells are the targets.
Bone Marrow Cells↗
The strange case of David Acer: myth and reality.
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Acquired Immunodeficiency Syndrome↗
Prourokinase-annexin V chimeras. Construction, expression, and characterization of recombinant proteins.
Annexin V is a human protein that binds with high affinity to the abundant phosphatidylserine molecules exposed on activated platelets and accumulates selectively in thrombi after intravenous administration in animal models of arterial thrombosis. We designed two chimeras that use annexin V as a means to target thrombolytic agents to platelet-containing thrombi: prourokinase (1-411)-annexin V (1-320); and prourokinase (144-411)-annexin V (1-320) (amino acid numbers of parent proteins given in parentheses). Chimeras were produced by cytoplasmic expression in Escherichia coli, refolded, and purified in single-chain form. Both chimeras had the same specific activity as annexin V in binding to cell membranes containing exposed phosphatidylserine. After activation with plasmin, both chimeras had specific amidolytic activity similar to that of urokinase. Both chimeras activated plasminogen in vitro with kinetic parameters similar to those for urokinase, and both showed full activity compared to urokinase in an assay of clot lysis in vitro. This study shows the feasibility of producing chimeric plasminogen activators in which annexin V provides the thrombus-targeting component; although not yet tested in vivo, such chimeras may have advantages over antibody-based targeting agents.
Amino Acid Sequence↗
Spine rehabilitation. Secondary and tertiary nonoperative care.
To achieve desirable behavioral outcomes, physicians treating spinal pain patients should be aware of appropriate algorithms for conservative care. Lower cost secondary rehabilitation can be effective if deconditioning, severity of physical symptoms, surgical equivocation, or psychosocial barriers to recovery are not present. Patients who have extended disability in excess of 6 months, recognized psychosocial barriers (depression, substance abuse, personality disorders, secondary gain), or severe deconditioning have a better prognosis with tertiary care.
Humans↗
In-utero percutaneous cystoscopy in the management of fetal lower obstructive uropathy.
In fetuses with lower obstructive uropathy, sonography cannot establish the cause of obstruction. We assessed whether percutaneous fetal cystoscopy could be useful in the evaluation and treatment of obstructive defects in utero. We inserted a fibreoptic endoscope through the lumen of the needle or trocar into the fetal bladder at the time of vesicocentesis or vesicoamniotic-shunt placement and looked at the the urethra, bladder neck, and ureteral orifices. Urethral vesicoamniotic shunting was considered in suitable cases; otherwise a percutaneous shunt was inserted. Fetal cystoscopy was possible in 11 of 13 patients referred. The bladder mucosa appeared haemorrhagic or oedematous in three. The ureteral orifices were seen in 9/11 fetuses, dilation was seen in five, but was only suspected in two by ultrasound. Ureteral webs were noted in two other fetuses. Two of seven fetuses underwent urethral vesicoamniotic shunting; urethral patency was achieved with urethral probing alone in one fetus. The remaining four fetuses were shunted with a standard technique. Fetal cystoscopy helps define the underlying conditions responsible for sonographic findings of lower obstructive uropathy, and allows the introduction of new treatments.
Amniotic Fluid↗
The phosphorylation state of the microtubule-associated protein tau as affected by glutamate, colchicine and beta-amyloid in primary rat cortical neuronal cultures.
The effects of the excitatory amino acid glutamate, the microtubule destabilizing agent colchicine, and beta 25-35-amyloid peptide on the phosphorylation state of tau were studied in rat cortical neurons in primary culture. Using immunocytochemistry and Western-blot analysis, we demonstrated that a proportion of tau in these cultures is normally highly phosphorylated, but most of this tau fraction is dephosphorylated after treatment of the cultures with glutamate or colchicine, but not with beta-amyloid; the glutamate- and colchicine-induced changes in tau phosphorylation commenced before cell death, as assessed by release of lactate dehydrogenase. Dephosphorylation of tau was readily revealed by using the monoclonal antibodies Tau.1 and AT8, which have phosphate-sensitive epitopes that both centre around serine-199 and -202 (numbering of the largest tau isoform). On Western blots and by immunocytochemistry, AT8 labelling strongly decreased after glutamate and colchicine treatments, whereas Tau.1 staining was more intense. Neurofilament monoclonal antibodies, including RT97, 8D8, SMI31 and SMI310, all additionally known to recognize tau in a phosphorylation-dependent manner, also demonstrated that glutamate and colchicine treatments of the cultures induced a dephosphorylation of tau. We also showed immunocytochemically that there is an increase in tau immunoreactivity in neuronal perikarya in response to glutamate and colchicine treatment, and this occurs concomitantly with the dephosphorylation of tau. Treatment of the primary rat cortical neuronal cultures with beta 25-35-amyloid peptide, under conditions which induce neuronal degeneration, did not induce a change in tau phosphorylation, and failed to act synergistically with glutamate to produce an increase in dephosphorylation of tau over that produced by glutamate treatment alone. These findings demonstrate that glutamate and colchicine induce tau dephosphorylation, as opposed to increased tau phosphorylation, which would be more indicative of Alzheimer-type neurodegeneration.
Amyloid beta-Peptides↗
Efficient gene transfer with adeno-associated virus-based plasmids complexed to cationic liposomes for gene therapy of human prostate cancer.
We have shown previously that treatment of rats bearing the Dunning R3327 MatLyLu prostatic tumor with human interleukin 2 (IL-2) gene-modified tumor cell preparations induces potent antitumor immunity in the animal. To test the clinical feasibility of using genetically modified tumor vaccines for the treatment of prostate cancer, we have explored the use of a simplified gene delivery system based on liposomes to introduce and express the IL-2 gene in the Dunning rat R3327 MatLyLu prostatic tumor cell line (MatLyLu) and in short-term cultures of primary human prostatic tumor cells. Liposome-DNA complexes containing the adeno-associated virus inverted terminal repeats exhibited 3-10-fold higher levels of gene transfer and IL-2 expression than did liposome complexes with non-adeno-associated virus containing plasmids. Single transfections resulted in IL-2 expression for an extended period of time that exceeded severalfold the amount of IL-2 secreted from retrovirally transduced MatLyLu cells. X-irradiation of cells (4000 rads) prior to transfection did not affect cytokine secretion, indicating that liposome-mediated gene transfer does not depend on cell proliferation. High levels of gene transfer and IL-2 expression were also achieved in short-term cultures of primary human prostatic tumor cells established from tumor specimens obtained following radical prostatectomy of cancer patients. Depending on the type of liposome used, IL-2 levels secreted from the human prostatic tumor cells were comparable to or exceeded the levels of IL-2 secreted from retrovirally transduced MatLyLu cells, which induced antitumor immunity in the rat model. The ability to culture and expand ex vivo human prostatic tumor cells, and the use of a simple and highly efficient gene transfer method to generate genetically modified tumor vaccines, set the stage for clinical exploration of gene-based immunotherapy of prostate cancer.
Animals↗
Core RNA polymerase and promoter DNA interactions of purified domains of sigma N: bipartite functions.
The sigma N class of sigma factors confer upon RNA polymerase the requirement for enhancer-binding activator proteins. The sigma-N (sigma N) protein of Klebsiella pneumoniae was analysed by the assay of purified peptides comprising domains or regions of sigma N defined by proteolysis or by homology alignment, respectively. The NH2-terminal Region I is required for the correct interaction of holoenzyme with the promoter, and promoter complexes forming with a truncated sigma N lacking Region I are not activatable. The complexes lack the DNA structure believed to represent nucleated strand separation but still make close contacts with this promoter part. Determinants of specific DNA recognition by sigma N were shown to reside in a C-terminal 16 kDa peptide, and core RNA polymerase binding determinants in an adjacent peptide. The latter contacts and appears to pack against the DNA-binding domain. Thus the DNA-binding and core-binding domains are bipartite in function, consistent with core functioning as an allosteric effector of the sigma DNA-binding activity. The DNA-binding and core-binding domains together include Region III of sigma N. Although not the primary determinant of core or DNA recognition, the acidic Region II of sigma N influenced both activities. Regions I and II in combination with core RNA polymerase thus appear to control the activity of C-terminal DNA contacting surfaces to allow formation of a closed promoter complex that is susceptible to activation.
Allosteric Regulation↗
Possible incorporation of an immunotoxicological functional assay for assessing humoral immunity for hazard identification purposes in rats on standard toxicology study.
The objective of this study was to examine the feasibility of conducting an immunotoxicological assay for assessing humoral immunity in rats on standard toxicology study. Male CD rats were untreated or dosed intraperitoneally daily for 30 or 90 days, excluding weekends, with vehicle or 2 mg/kg cyclophosphamide (CY). Six days prior to sacrifice, selected rats were injected intravenously with sheep red blood cells (SRBC). One day prior to necropsy, blood samples for hematological and clinical chemical measurements were collected from each rat. On the day of necropsy standard protocol tissues were collected, weighed, processed to slides, and examined microscopically. One-half of each spleen was used to prepare a single cell suspension in order to assess spleen cell numbers. Serum was analyzed for anti-SRBC IgM antibody using an enzyme-linked immunosorbent assay. A second set of studies was performed to examine further the effect of SRBC administration on lymphoid organ weights using 30- and 90-day study age-equivalent naive male CD rats. Exposure of animals to 2 mg/kg CY for 30 or 90 days resulted in a 28% and 61% decrease, respectively, in SRBC-specific serum IgM levels. CY treatment also caused mild alterations in some leukocytic parameters, with significant decreases of 35% and 33% in white blood cell and lymphocyte counts, respectively, observed in 30-day CY-treated animals receiving SRBC. Injection of SRBC alone did not alter hematological or clinical chemistry parameters. With the expected exception of the spleen (increased number and size of germinal centers), administration of SRBC did not significantly alter the weights or morphology of routine protocol tissues. Furthermore, administration of SRBC did not mask the immunosuppressive effects of CY treatment under the conditions of this study. Based on our preliminary findings, a functional assay for assessing humoral immunity may be conducted in animals on standard toxicology study.
Animals↗
Developmental expression of a type II collagen/beta-galactosidase fusion gene in transgenic mice.
The correct temporal and spatial expression of the type II collagen gene is believed to be important for normal development and growth of the skeleton and the eye, i.e., tissues where the protein product is predominantly found. To study transcriptional activation of type II collagen gene in skeletal and nonskeletal tissues we produced transgenic mice carrying murine proalpha1(II) collagen/beta-galactosidase fusion gene constructs. The expression of the fusion gene was found to depend on the presence of intron 1 deleted failed to reveal any beta-galactosidase activity confirming the important role of regulatory sequences within intron 1 of the gene. High-level expression of the functional construct was clearly confined to cartilaginous tissues but transient low-level expression was also observed in extraskeletal locations, such as the developing brain and the notochord. The results demonstrate that the regulatory elements in the proalpha1(II) collagen/beta-galactosidase fusion gene construct confer both temporal and spatial specificity indistinguishable from that of the endogenous proalpha1(II) collagen gene as determined by the presence of the corresponding mRNA by in situ hybridization. Furthermore the beta-galactosidase activity correlated well with the progression of chondrogenesis as seen by staining of whole mouse embryos with Alizarin red S and Alcian blue in the hybrid mouse strain used for microinjections. The transgenic mouse line produced should prove useful for studies on various aspects of chondrogenesis. Furthermore, the data shows that the regulatory elements present in the construct are sufficient for targetting the expression of other genes in cartilage.
Animals↗
Parenteral methotrexate: effective shelf-life following initial puncture of preservative-protected vial.
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Drug Storage↗
Physical training improves body composition of black obese 7- to 11-year-old girls.
We determined the effect of supervised physical training without dietary intervention, on body composition of obese girls. The subjects were 25 obese 7- to 11-year-old black girls, divided into physical training and lifestyle education groups which were comparable on baseline body composition; 22 girls finished all aspects of the study. Twelve girls engaged in aerobic training (10 weeks, 5 days/week) while 10 engaged in weekly lifestyle discussions without formal physical training. Total body and regional body composition were measured with dual energy x-ray absorptiometry, skinfolds and circumferences. Aerobic fitness was measured by heart rate response to submaximal treadmill exercise. The physical training group attended 94% of scheduled sessions and kept their heart rates at an average of 163 bpm for 28 minutes/session. The lifestyle group attended 95% of their sessions; they remained stable in aerobic fitness and most body composition measurements. The physical training group showed a significant improvement in aerobic fitness and a significant decline of 1.4% body fat. Skinfold and circumference indices of fatness also declined significantly in the training group. We conclude that controlled physical training, without dietary intervention, improved the fitness and body composition of obese black girls.
Absorptiometry, Photon↗
Glycoconjugates in normal and abnormal secondary neurulation.
In chick embryos, the anterior greater portion of the neural tube develops by the folding, apposition, and fusion of the neuroectoderm. The smaller caudal portion that forms the secondary neural tube (lumbosacral and coccygeal regions) is derived from the tail bud, an aggregate of mesenchymal cells located at the caudal limit of the body. Tail bud mesenchyme, arranged in a solid cord, undergoes mesenchymal-epithelial transformation to form the secondary neural tube. Previous evidence suggests that this transformation is accompanied by modulation of cell surface glycoconjugates in the differentiating tissues. In this study, we show by lectin histochemistry and lectin blotting of proteins isolated by SDS-PAGE, that Datura stramonium agglutinin (DSA) binds preferentially to differentiating tail bud cells. This lectin is specific for beta 1-4-linked N-acetylglucosamine oligomers, such as the oligosaccharides of the poly-N-acetyllactosamine series that have been previously implicated in cell differentiation. Ultrastructural lectin cytochemistry indicates that at least some of the proteins binding DSA are localized extracellularly. The use of DSA as a teratogen resulted in embryos showing a variety of neural tube and notochord defects. We have also examined the binding of DSA to embryos that were treated with teratogenic doses of retinoic acid by sub-blastodermal injection, and find that the DSA-binding patterns are perturbed. Analysis of DSA-treated embryos using the TUNEL technique indicated that cell death was not a factor in DSA teratogenesis. This strongly suggests that the glycoconjugates of the cell surface have a role in the normal differentiation of tail bud mesenchyme into the neuroepithelium of the secondary neural tube. Perturbations of glycoconjugate activity results in defects of the secondary neural tube and associated tail bud derivatives.
Abnormalities, Drug-Induced↗
AIDS and HIV infection.
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Acquired Immunodeficiency Syndrome↗
Percutaneous fetal cystoscopy and endoscopic fulguration of posterior urethral valves.
Percutaneous fetal cystoscopy was performed in a male fetus with ultrasonographic evidence of lower urinary tract obstruction at 19 weeks of gestation. The diagnosis of posterior urethral valves was confirmed. Percutaneous endoscopic fulguration of the valves was successfully performed at 22 weeks of gestation, and urethral patency was established. This case illustrates the feasibility of performing diagnostic and therapeutic endoscopic procedures within the human fetus for the management of a congenital anomaly. While we believe that fetal cystoscopy may improve our diagnostic, prognostic, and therapeutic capabilities in the management of fetuses with lower obstructive uropathy, studies are needed to establish the actual value, risks, and limitations of this new approach in fetal medicine.
Adult↗