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Biomedical subjects

B Polack

Publications and source records attributed to B Polack.

At least 37 records · Page 2Linked to original sources

Natural transmission of Pneumocystis carinii in nonimmunosuppressed animals: early contagiousness of experimentally infected rabbits (Oryctolagus cuniculus).

Airborne transmission of Pneumocystis carinii has been established, but the infective form and the sources of infection remain unknown. Animal models for studies of P. carinii have previously been limited to immunosuppressed rodents; however, this study was performed with nonimmunodepressed P. carinii-free rabbits. This study was aimed at determining (i) the delay between inoculation of animals (day zero [D0]) and the onset of contagiousness and (ii) the end of contagiousness of these animals (donors). Five-week-old rabbits were used as contact animals and were housed with the donors. The cohabitation periods were for 4 or 5 days from D0 to D4, D4 to D8, D8 to D13, D13 to D18, and D18 to D22. The highest parasite burdens were observed in contact animals housed with donors from D8 to D13 or D13 to D18. This period (8th to 18th day following the day of inoculation of donors) might correspond to the highest phase of contagiousness of donors.

Animals↗

The Pseudomonas aeruginosa fumc and soda genes belong to an iron-responsive operon.

Pseudomonas aeruginosa contains two superoxide dismutases (SOD), a Mn-containing SOD (Mn-SOD) and a Fe-SOD, which are encoded by sodA and sodB, respectively. We have cloned and sequenced a DNA fragment from P. aeruginosa, strain CHA, which contains the sodA gene and three other open reading frames (ORF). We report here that one of the ORFs upstream from sodA is fumC, which encodes the O2.- resistant isoform of fumarase (or fumarate hydratase). It is shown that fumC and sodA belong to the same operon. By primer extension experiments, the transcription initiation site has been located at -413 from the ATG codon of the fumC gene. The fumC-sodA operon was found to be negatively regulated in presence of iron and the E. coli FUR protein was shown to bind to the 19-bp FUR consensus sequence present at the transcription start site of the operon.

Bacterial Proteins↗

Influence of cytokines on Toxoplasma gondii growth in human astrocytoma-derived cells.

The effects of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), interleukin 1-alpha (IL-1-alpha), and interleukin 6 (IL-6) on the growth of the Toxoplasma gondii RH strain were studied in vitro using a human astrocytoma-derived cell line. Cells were treated with cytokines at different concentrations at 24 h prior to infection with T. gondii tachyzoites. IFN-gamma did not induce any modification in T. gondii growth, whatever the concentration used. TNF-alpha induced a significant decrease in the total number of tachyzoites, whereas IL-1-alpha surprisingly induced an increase in the number of tachyzoites. Our results show that the effects of cytokines on T. gondii growth may be of great importance in the control of cerebral toxoplasmosis but that they can vary, depending on the cell type considered.

Animals↗

Serologic evidence for bovine immunodeficiency virus infection in France.

We report herein on the first serologic detection of antibodies to bovine immunodeficiency virus (BIV) in France. Serum samples from dairy and beef cattle from southwestern and western France (Landes and Vendée) were tested using a western blot assay with a recombinant 53 kDa gag precursor derived from the Louisiana BIV R29 isolate. We performed our study on the oldest animals from 37 different herds that were under serologic follow up for previous bovine leukemia virus infection. Overall, 398 selected bovine sera were assayed and 15 serum samples from 8 herds reacted with the recombinant 53 kDa BIV R29 gag. Interestingly, reactions obtained with French sera were weaker than with positive Louisiana sera, a finding that may indicate the occurrence of distinct French and Louisiana BIV variants.

Animals↗

Role of manganese superoxide dismutase in a mucoid isolate of Pseudomonas aeruginosa: adaptation to oxidative stress.

Chronic infection by alginate-producing (mucoid) Pseudomonas aeruginosa is a leading cause of morbidity among cystic fibrosis (CF) patients. In the lungs of CF patients, the bacteria are exposed to activated oxygen species produced by the phagocytes of the host or resulting from the metabolism of oxygen. Two isoforms of superoxide dismutase are synthesized by P. aeruginosa; they differ by the metal present at their active site, which is either iron or manganese. To evaluate the role of manganese-containing superoxide dismutase (MnSOD), encoded by sodA, we have isolated a sodA mutant of the mucoid P. aeruginosa strain CHA isolated from the bronchopulmonary tract of a CF patient. The sodA mutant exhibited an increased sensitivity to oxidative stress generated by paraquat and was less resistant to oxidative stress in the stationary phase of growth compared with its parental strain. It was observed that MnSOD was expressed in the parental strain solely during the stationary phase of growth and that cells of the sodA mutant taken at the stationary phase resumed growth with a longer delay than the sodA+ cells when reinoculated in a new medium, especially in the presence of paraquat. These results suggest that MnSOD may participate in the adaptation of mucoid strains of P. aeruginosa to the stationary phase of growth in the lungs of CF patients.

Adaptation, Physiological↗

Retinoids induced t-PA synthesis by C6 glioma cells--role in tumoral haemorrhagic necrosis.

Treatment of rat C6 glioma with high doses of 13 cis-retinoic acid (cRA) was responsible for death related to haemorrhagic necrosis localized to the tumor. Our aim was to explore this adverse effect of retinoid treatment. We show that cRA-treated C6 glioma at 25 mg/kg/day for 18 days exhibits in vivo an increase T-PA activity, which is responsible for a localized tumor fibrinolytic activity. Production of t-PA is supported by specific enhancement of gene expression, as was shown by the increase in t-PA mRNA (x 2.3). This production is a direct effect of cRA when treating the tumor, since tumor cells themselves do not produce enough t-PA and treatment of control rats does not increase the t-PA level. T-PA production by rat C6 glioma is in vivo related to the specific synthesis of t-PA by the C6 cell-line. The stimulation of C6 cell-line by cRA in vitro is dose-dependent and reached a maximum for 3 and 30 microM at the 72nd h. So cRA-treated C6 glioma cells produce t-PA which appears to be the major species associated with the fibrinolytic activity-induced intra-tumoral haemorrhage after exposure to retinoid treatment.

Animals↗

Bovine immunodeficiency virus: facts and questions.

Bovine immunodeficiency virus (BIV) is a lentivirus whose serologic prevalence is worldwide. Little is known about its impact on animal health status, pathogenesis and mode of transmission. Understanding BIV biology implies isolation of new viral strains and long-term studies on experimentally-infected cows and surrogate hosts such as rabbits.

Animals↗

Combined hereditary disorders of haemophilia B Leyden (-6 G-->A) and type 1 von Willebrand disease.

Multiple coagulation disorders are unusual. We report here a combination of haemophilia B Leyden with type 1 von Willebrand disease (vWD) affecting different members of the same family. Haemophilia B Leyden was due to a -6 G-->A mutation within the promoter of the factor IX gene and was responsible for a mild haemophilia in the father of the proband. The proband and her sister (age 4 and 6) exhibited a twofold lower level of factor IX activity (0.4 IU/ml) than the paternal grandmother (0.95 IU/ml). The differences in F IX levels in the three carriers of the same -6 G-->A mutation suggest the implication of an age-related mechanism responsible for the increase in factor IX plasma level. Haemophilia B Leyden patient and carriers suffered also from a mild von Willebrand disease. The diagnosis of this associated type 1 vWD was performed by assaying plasma von Willebrand factor together with multimer electrophoretic studies and DDAVP test. The inheritance of this vWD was investigated by haplotype analysis of the vWF gene. Individuals affected by such an association are actually asymptomatic, but per- and post-operative bleeding risk remains to be evaluated.

Adenine↗

The bovine immunodeficiency-like virus (BIV) is transcriptionally active in experimentally infected calves.

We have studied the infection by the bovine immunodeficiency-like virus (BIV) in three experimentally infected calves, by polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR), from the peripheral blood mononuclear cells (PBMC). Two primer pairs located in the gag and pol regions of the viral genome allowed to detect the viral genomic DNA by PCR, as well as the unspliced genomic viral RNA transcript, by RT-PCR. We also present the evidence of the presence in peripheral blood mononuclear cells (PBMCs) of a mRNA transcript of the regulatory trans-activator tat gene, according to the splicing pattern of the viral genome, by use of reverse transcription followed by nested PCR. The active expression of the virus in these animals was further assessed by the sequential rescue of the virus from unstimulated PBMCs in cell culture, from 4 weeks until 15 months following the infection.

Animals↗

Validation of electronic cell counts by quantitative buffy coat analysis (QBC).

A total of 458 eight blood cell counts, accompanied by blood film reviews of the same samples, were performed with an electronic cell counter and with the QBC. In the great majority of cases, the QBC gave fast and accurate control of the flags from the electronic counter, thus avoiding the necessity for manual validation with its associated risks of infection and contaminations. Furthermore, QBC non readability could be related to microcytosis and hypochromia and hence point to possible cases of congenital or acquired haemoglobinopathy.

Blood Cell Count↗

In vivo leukocyte tropism of bovine leukemia virus in sheep and cattle.

Bovine leukemia virus (BLV), an oncovirus related to human T-cell leukemia virus type I, causes a B-cell lymphoproliferative syndrome in cattle, leading to an inversion of the T-cell/B-cell ratio and, more rarely, to a B-cell lymphosarcoma. Sheep are highly sensitive to BLV experimental infection and develop B-cell pathologies similar to those in cattle in 90% of the cases. BLV tropism for B cells has been well documented, but the infection of other cell populations may also be involved in the BLV-induced lymphoproliferative syndrome. We thus looked for BLV provirus in other leukocyte populations in sheep and cattle by using PCR. We found that while B cells harbor the highest proviral load, CD8+ T cells, monocytes, and granulocytes, but not CD4+ T cells, also bear BLV provirus. As previously described, we found that persistent lymphocytosis in cows is characterized by an expansion of the CD5+ B-cell subpopulation but we did not confirm this observation in sheep in which the expanded B-cell population expressed the CD11b marker. Nevertheless, BLV could be detected both in bovine CD5+ and CD5- B cells and in sheep CD11b+ and CD11b- B cells, indicating that the restricted BLV tropism for a specific B-cell subpopulation cannot explain its expansion encountered in BLV infection. Altogether, this work shows that BLV tropism in leukocytes is wider than previously thought. These results lead the way to further studies of cellular interactions among B cells and other leukocytes that may intervene in the development of the lymphoproliferative syndrome induced by BLV infection.

Animals↗

A simultaneous evaluation of three multiparametric coagulation instruments: BFA, HEMOLAB and STA.

In the present paper, three multiparametric coagulation instruments were evaluated with regard to chronometric tests for aPTT (CK-Prest and automated APTT), PT (Recombiplastin and Thromborel), fibrinogen and factors of the prothrombin complex. Analysis of within-run precision and linearity and comparative studies showed the analytical performances of the instruments to differ according to the reagents used and emphasized the difficulty of finding the best compromise between instrument and reagent. On the basis of this study, the mechanical instrument appeared to be more versatile than the optical machines. This conclusion could however be modified after further evaluation of the recent new generation coagulation instruments.

Blood Coagulation Tests↗

[Comparative and simultaneous evaluation of three automated counters in hematology: Coulter STKS, Sysmex NE 8000, Technicon H-2].

We evaluated and compared three automated blood cell counters, Coulter STKS, Sysmex NE 8000 and Technicon H-2. These perform both a complete blood cell count and a full white cell differential count. Carry-over was found to be acceptable, except for the leucocyte count by the Sysmex NE 8000 (1.5%; p < 0.001). Linearity over a wide concentration range for all of the measured parameters, haemoglobin, red blood cells, white blood cells and platelets, was excellent (r > 0.98). Within-run precision was verified with 22 samples covering a wide range of cell counts by repeated analysis (n = 20) of the same sample. Coefficients of variation (CV) were acceptable, < 4% for values within the normal range. The CVs for eosinophils and basophils were less good but without clinical impact. Analysis of 270 samples showed that the Coulter counter indicated the presence of atypical cell populations more frequently (19%; p = 0.02). Message of qualitative abnormalities displayed by the three analysers were often discordant. These three blood cell counters differed in their sensitivity and positive predictive value for detecting abnormal blood cells rather than in their specificity and negative predictive value. Other operating aspects of three instruments that are important in haematological practice are documented.

Equipment Contamination↗