Biomedical subjects
Publications and source records attributed to J L Moore.
The diagnosis of cancer rarely complicates pregnancy. Despite the fact that advanced disease is often encountered, there is no scientific evidence that the pregnant state alters the neoplastic process. Only through careful attention to the patient can the clinician detect cancer at an early stage and offer the patient reasonable hope for a cure.
Vascular patterns to thoracic limbs, thorax, and neck muscles were studied in 10 dogs (20 limbs) to identify muscles most suitable for transposition in the treatment of large wounds. Gross dissection of injected specimens and angiography were used to identify vascular pedicles. Size and location of pedicles were generally consistent, and any variations would not interfere with most muscle transfers. The cutaneous trunci, latissimus dorsi, sternothyroideus, sternohyoideus, deep pectoral, anconeus, ulnaris lateralis, and ulnar head of flexor carpi ulnaris muscles were identified as suitable for transfer. The cranial trapezius, caudal omotransversarius, cleidobrachialis, and caudal sternocephalicus muscles also had potential for use. Other muscles, because of inaccessibility or unfavorable vascular pattern, were not suitable candidates for transfer.
Malignant hyperthermia is a rare syndrome that occurs in genetically susceptible individuals who are exposed to frequently used inhalation anesthetics. The disorder is most common in children and young adults. It is triggered through a defect in the ability of skeletal muscles to concentrate and release calcium. Signs of malignant hyperthermia include hypercarbia, muscle rigidity and tachycardia. Temperature elevation is often a late sign of the syndrome. Treatment begins with stopping all inhaled anesthetics at the earliest sign of the syndrome. The use of dantrolene has significantly reduced mortality from malignant hyperthermia. No simple screening test exists. Family members or those with a suspicious history need to be counseled and should consider muscle biopsy and testing prior to surgery.
Twenty-nine samples from eighteen patients with chronic lymphocytic leukaemia (CLL) were used in a direct comparison of in vitro response to chlorambucil measured in a metabolic (MTT) and dye exclusion (D.Ex) assay. Reduced ability to produce formazan corresponded to a reduced number of dye-excluding viable cells and a significant correlation was found between dose-response measured in the two assays. Initial low absorbance values obtained with untreated control cells in the MTT assay were effectively overcome by increasing both the cell seeding density and MTT exposure time with the consequent increase in assay sensitivity. The MTT assay provided qualitatively similar dose-response data to that obtained in the D.Ex assay. A wide range in in vitro response was seen for both pretreatment and treatment patient groups. In vitro dose-responses were seen to coincide with decreasing or stable white cell counts, taken around the time of sampling, in samples from 4 patients. Less marked dose-responses were observed for 2 patients considered clinically resistant to chlorambucil. The MTT assay would seem, therefore, to be applicable to in vitro assay of cell response in CLL.
4-Hydroxythiazoles have been identified as potent inhibitors of 5-lipoxygenase in vitro exhibiting IC50's of less than 1 microM. An investigation of structure-activity relationships showed that the most potent inhibitors of this series are the 5-phenyl derivatives. The corresponding thiazolidin-4-one analogues were found to be relatively inactive. The 4-hydroxythiazoles were active inhibitors against 5-lipoxygenase in both intact rat polymorphonuclear leukocytes and human whole blood. The compounds were also selective inhibitors of 5-lipoxygenase, displaying only weak activity against other related enzymes, cyclooxygenase and 12- and 15-lipoxygenase.
Hyperandrogenemia in postmenopausal women requires an evaluation to exclude pathologic ovarian or adrenal causes. Our patient exhibited no signs of hypercortisolism, congenital adrenal hyperplasia, or adrenal or ovarian neoplasia based on biochemical testing and pelvic sonography. We hypothesized that unexplained androgen excess in our patient was due to the development of gonadotropin-dependent excess ovarian stromal androgen production. This syndrome may be comparable to gestational hyperreactio luteinalis where elevated gonadotropins stimulate ovarian stromal androgen production. If tumor can confidently be excluded, such women may benefit from gonadotropin suppression with long-acting GnRH-a.
UV irradiation provides a simple and efficient way to minimize contamination or false positivity which often occurs in laboratories performing routine PCR tests. Here, we characterize several parameters of the effect of UV irradiation on DNA template, primers, deoxynucleoside triphosphate and Taq polymerase. UV irradiation of DNA results in the formation of pyrimidine dimers and thus prevents them from being effective templates in subsequent PCR. Reduction of the HIV DNA templates in polypropylene microcentrifuge tubes by more than 1000-fold can be achieved by UV irradiation. The sensitivity of the primers is sequence- and concentration-dependent. Oligonucleotides with neighboring thymine bases are more susceptible to UV than those without. Taq polymerase is highly UV sensitive, whereas deoxynucleotide triphosphate is relatively UV resistant.
A method has been developed to estimate the post-irradiation survival of cytochalasin B-induced polyploidization of adherent Chinese hamster ovary cell using the flow cytometer. After exposure to radiation, surviving cells are allowed to become polyploid in the presence of cytochalasin, and are detached using trypsin, fixed by the addition of glutaraldehyde and stained using mithramycin. DNA content distributions are polymodal, and the absolute number of cells per culture in any given ploidy class is estimated by reference to a non-fluorescent bead internal standard, detected using forward scatter. Post-irradiation survival is defined as the ability to reach a given DNA content, and is reduced exponentially with dose. A bioassay to determine optimum cytochalasin concentrations can be derived from the relative size of the 2C (G0/G1) peak in the DNA content distribution. At culture densities greater than about 8 x 10(4) cell/cm2 the relative number of cells reaching at least 16C is reduced, but this inhibition is partially reversible by an increase in the medium glucose concentration, but not by the use of cytochalasin D or dihydro B.
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To evaluate the correlation of detection of human immunodeficiency virus (HIV) by polymerase chain reaction (PCR) with detection of HIV antibody, 271 simultaneous serum and peripheral blood mononuclear cell samples were examined from 242 persons whose activities placed them at increased risk for HIV infection: 142 from homosexual men, 86 from hemophilic men, and 43 from heterosexual partners of HIV-infected persons. PCR was performed using the gag region primer pair SK38/39 and the env region primer pairs SK68/69 and CO71/72. Amplified HIV DNA was detected using specific oligomer probes. Of 63 HIV antibody-positive samples, 58 (92%) had HIV DNA by PCR. Of 208 HIV antibody-negative samples, 7 (3.4%) had HIV DNA by PCR. On follow-up, 4 of the latter persons were seropositive when next tested; 2 were well and antibody- and PCR-negative; 1 had died of a stroke before retesting. Thus, PCR detects HIV in most antibody-positive persons; detection is increased by use of multiple primer pairs. PCR-positive antibody-negative specimens may indicate HIV infection in which antibody has not yet developed or may be false-positive PCR results. When PCR is discordant with HIV antibody, testing of additional specimens and clinical follow-up are necessary to assess HIV infection status.
The human immunodeficiency virus (HIV) infects and depletes or alters the function of cells involved in immune responsiveness. While both T helper lymphocytes and monocyte/macrophages can be infected via cell-surface CD4 in vitro, previous studies showed that few blood cells express HIV RNA in vivo. This study used DNA amplification to determine the levels of HIV DNA in purified lymphocytes, monocytes, and neutrophils from HIV-infected asymptomatic individuals and persons with AIDS. The average numbers of HIV DNA copies in lymphocytes from AIDS patients and asymptomatic individuals were similar (approximately 100-140 copies/150,000 cells). However, when expressed on the basis of numbers of CD4+ T cells, AIDS patients' cells contained approximately 2.5 times more HIV DNA. While HIV DNA was present in lymphocytes from all 27 subjects, little or no HIV DNA was observed in monocytes or neutrophils. Only 1 asymptomatic person contained levels of HIV DNA in monocytes (125 proviral copies/150,000 cells) that were comparable to levels expressed in lymphocytes (160/150,000). While expression of monocyte HIV DNA in this person was persistent over at least 8 months, it was not observed in neutrophils, suggesting that monocyte HIV DNA did not originate in myeloid precursors. This study shows that in AIDS or asymptomatic HIV infection, lymphocytes are the predominant infected cell found in blood.
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The vascular patterns to pelvic limb muscles were studied in 6 dogs (12 limbs) to identify muscles most suitable for transposition in the treatment of large wounds. Gross dissection of injected specimens and angiography were used to identify the vascular pedicles. The vascular pedicles to several muscles were generally consistent, and any variations would not interfere with most muscle transfers. The cranial part of the sartorius, gracillis, semitendinosus, and rectus femoris muscles were identified as suitable candidates for transfer. The caudal part of the sartorius, cranial tibial, and long digital extensor muscles have segmentalized vascular patterns that would limit its arc of rotation.
A novel dye exclusion assay based on differential staining of fixed cells has been evaluated using the MOLT4 (T-lymphoblastic leukaemia) and DAUDI (Burkitt lymphoma) cell lines. Reproducible differential staining was found with fixed samples kept for up to 2 months. A small increase (less than 13%) in the proportion of stained cells was observed over the first 30 days in fixative. Reproducible dose-response data were obtained with cells treated with radiation or drugs and assayed after 4 days in culture. This ability to fix cells prior to measurement of viability is of general use particularly where time constraints prevent haemocytometer counting. More specifically, it may have potential use in the field of chemosensitivity testing particularly where large sample numbers require rapid processing.
When isolated nuclei of human lymphocytes are challenged with 2 M NaCl a histone-free-DNA-protein (HF-DNA) complex is released. In a linear sucrose gradient (pH 8.0) the sedimentation distance of HF-DNA is reduced when immediately isolated from cells irradiated in vitro. At low doses, if irradiated cells are incubated at 37 degrees C the sedimentation behaviour approaches that of unirradiated cells (i.e. repair). In the present study such repair was usually complete within 1 h. The radiation damage to lymphocytes from a healthy donor group and three patient groups consisting of new patients (before radiotherapy), well patients (2 to 6 years post-radiotherapy) and patients with complications attributed to radiotherapy was similar. The lymphocytes from most healthy donors and new patients demonstrated complete repair of radiation damage following an incubation of 1 h at 37 degrees C. However, 2/29 (6.9 per cent) healthy donors and 2/25 (8 per cent) new patients demonstrated poor repair. Of those patients now attending with 'bowel complications' attributed to radiotherapy 7/16 (44 per cent) demonstrated poor repair. In contrast, all those (11/11) described as 'well and complication-free' showed good repair.
Polymerase chain reaction (PCR) was used to detect human immunodeficiency virus (HIV)-1 DNA in peripheral blood mononuclear cells to assess in hemophilic men whether any were HIV-seropositive but uninfected or seronegative but infected and in seronegative sex partners of seropositive hemophilic men whether any were infected. Of 40 seropositive men, 38 (95%) were PCR-positive; one was PCR-indeterminate and one PCR-negative. None of 41 seronegative men who used only donor-screened, virus-inactivated coagulation factor products were PCR-positive. However, two of six who received noninactivated products were PCR-positive; one had low T-helper cell counts and died of unrelated causes and the other had seroconverted 11 mo later. PCR with a second primer pair also detected HIV-1 DNA in these two men. None of 25 seronegative female sex partners of seropositive men, including six men with AIDS and seven with AIDS-related symptoms, were PCR-positive. These data suggest that most seropositive hemophilic men are HIV-infected; whether some are infected with defective virus remains to be resolved as does the infection status of seropositive PCR-negative men. Identification of two seronegative PCR-positive men supports the possibility that HIV-1 DNA can be detected before seroconversion.
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Rifabutin (ansamycin; LM427), a semisynthetic rifamycin derivative, inhibits the replication of human immunodeficiency virus type 1, the virus etiologically linked to acquired immunodeficiency syndrome. The inhibition of virus production was observed in human immunodeficiency virus type 1-infected peripheral blood mononuclear cells both by reverse transcriptase assay and virus-antigen assay. In addition, rifabutin effectively reduced human immunodeficiency virus type 1-associated cytopathic effect to OKT4-positive cells. Virion-associated reverse transcriptase was also markedly inhibited by rifabutin. The inhibitory dose of rifabutin was nontoxic to lymphoid cells, which further suggests that it might serve as a useful agent in the therapeutic modalities of acquired immunodeficiency syndrome.