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Biomedical subjects

B Jefferson

Publications and source records attributed to B Jefferson.

24 records · Page 2Linked to original sources

Recombinant domain III of perlecan promotes cell attachment through its RGDS sequence.

Perlecan has been previously been shown to support attachment of a wide variety of cells through interactions of its core protein with the cell surface. The core protein domains involved in cell adhesion are, however, unknown. The laminin-like domain III of murine perlecan contains an RGDS sequence and is a likely candidate for supporting integrin-mediated cell attachment. We made a cDNA construct corresponding to domain III and containing an in frame signal peptide at the 5' end as well as in frame a stop codon at the 3' end by using cDNA clones to perlecan. The construct was inserted into the pRC/CMV vector and transfected into HT1080 cells, and the secreted recombinant domain III, a 130-kDa protein, was purified from the medium. The size of proteolytic fragments produced by digestion with V8 protease as well as analysis of the rotary shadowed image of the recombinant protein indicated it was produced in a native conformation. Recombinant domain III coated on tissue culture dishes, supports adhesion of an epithelial-like mouse mammary tumor cell line MMT 060562 in a dose-dependent manner. This interaction was inhibited specifically by the RGDS synthetic peptide and intact perlecan, but not laminin. This domain III RGD-dependent cell attachment activity indicates a role for perlecan in integrin-mediated signaling.

Amino Acid Sequence↗

A serological survey of bovine syncytial virus in Ontario: associations with bovine leukemia and immunodeficiency-like viruses, production records, and management practices.

Of the 920 cows tested, 56.7% showed antiretroviral serological reactivity. Prevalence rates (95% confidence interval) of antiretroviral antibodies among individual dairy cows in Ontario were: BIV 5.5% (4.0-7.0), BLV 25.7% (22.9-28.6), and BSV 39.6% (36.4-42.8). The following percentages of cows showed serological reactivity against the specified retroviruses: BIV 2.3%, BLV 14.0%, BSV 27.5%, BIV and BSV 1.3%, BIV and BLV 0.9%, BLV and BSV 9.9%, BIV and BLV and BSV 0.9%. These rates of sero-positivity are similar to those found in other countries. Serological test results were not adjusted for sensitivity and specificity. The prevalence rates of antibodies to the three retroviruses (BIV, BLV, and BSV) were significantly different, but no associations were observed between specific retroviral serological test results among individual cows. The prevalence rates of BIV and BSV seropositivity were constant across Ontario, whereas, there was a significant trend for the prevalence rate of BLV seropositivity to decrease going from southwestern to eastern Ontario; cows in eastern Ontario had approximately half the prevalence rate of those in southwestern Ontario. Cows that were seropositive for BSV were significantly older than BSV seronegative cows. There was no association between culling rate and BSV serology. Significant negative associations were found with winter or summer housing of calves separate from adults and summer outdoor exercise for dry cows. The use of calf hutches in the summer had a significant positive association with BSV seropositivity. Regression analyses were done to assess the association of retroviral (BIV, BLV, and BSV) seropositivity on calving interval, milk somatic cell count, and milk production. Serological test results for BIV, BLV, and BSV were entered into all models and all models were adjusted for intra-cluster (intraherd) correlation. Herd size and age were found to be important confounding variables. BIV seropositivity was not associated with any changes in production using this approach, however when considered in isolation BIV seropositivity remained associated with decreased milk production. BLV seropositivity was significantly associated with longer calving intervals and higher somatic cell counts in older cows. As well, in older cows, BSV seropositivity was significantly associated with higher milk production.

Age Factors↗

Haematological and lymphocyte subset analyses in sheep inoculated with bovine immunodeficiency-like virus.

Bovine immunodeficiency-like virus (BIV) was passaged in vivo by intraperitoneal transfusion of ovine whole blood. Prior to transfusion, the recipient sheep were given sodium thioglycolate intraperitoneally to induce mild non-suppurative inflammation. The anti-BIV antibody response, haematology, and peripheral blood lymphocyte subsets (B, gamma delta, CD2+, CD4+ and CD8+) of recipient sheep were assessed for one year following transfusion. Passaging was successful since serum anti-BIV antibody responses were detected in 5 of the 6 recipient sheep; 1 of the 5 remained seropositive throughout the study. Lentivirus was not isolated from the recipient sheep, but provirus was detected by the polymerase chain reaction in DNA from peripheral blood leukocytes in 3 of the 5 sheep that seroconverted. In the BIV-inoculated sheep, neutrophils and eosinophils were significantly increased (p < or = 0.05) at 3 months and between 6 and 8 months postinoculation, respectively. B, CD2+ and CD4+ cells and the CD4+/CD8+ ratios were significantly increased (p < or = 0.05) 2 months postinoculation. Mild, transient haematological changes occurred in BIV-exposed sheep, but illness was not detected in the year.

Animals↗

Detection of bovine leukemia virus RNA in serum using the polymerase chain reaction.

A method was developed for detecting bovine leukemia virus (BLV) RNA in serum samples using a pair of primers from the BLV polymerase gene in the polymerase chain reaction (PCR). The PCR was able to detect 3800-7600 molecules of BLV RNA. At this level of sensitivity eleven pools of adult and one fetal bovine serum appeared free from BLV contamination.

Animals↗

Proviral detection and serology in bovine leukemia virus-exposed normal cattle and cattle with lymphoma.

Twenty-seven cattle with lymphoma and 46 cows from a known bovine leukemia virus (BLV)-infected herd were tested for anti-BLV antibody by the agar gel immunodiffusion (AGID) test and an enzyme-linked immunosorbent assay (ELISA). The polymerase chain reaction (PCR) and Southern hybridization were used to detect BLV provirus in the tumor DNA of the 27 cattle with lymphoma. The PCR was used to detect BLV provirus in the peripheral blood mononuclear cell DNA of the 46 normal known-exposed cattle. Two presumed false negative AGID test results compared to ELISA were found. Of ten cattle three years of age or less with "sporadic" forms of lymphoma, four had BLV provirus in tumor DNA, detectable by PCR. In two of these four, BLV provirus was clonally integrated based on digestion of tumor DNA with restriction enzymes followed by Southern hybridization. The BLV provirus was not detected by PCR in 5 of 17 cattle with "enzootic" lymphoma and two of these five were seronegative. Among normal BLV-exposed cows, 6.5% (3 of 46) were serologically positive and PCR negative; serologically negative and PCR positive cows occurred with the same frequency. Serological and PCR test results, when considered in all cattle (n = 73), had a concordance rate of 83.6%. Discordant test results occurred with approximately equal frequency between serologically positive and PCR negative (7 of 73, 9.6%) and serologically negative and PCR positive (5 of 73, 6.8%) groups. These data suggest that the role of BLV in some "sporadic" bovine lymphomas, previously unassociated with BLV, should be reexamined. The BLV provirus was not demonstrable in the tumor DNA from five adult cattle with lymphoma, suggesting that BLV may not be the etiological agent in all adult bovine lymphomas. The findings of persistently seronegative PCR positive and seropositive PCR negative cattle indicate that further work is needed to more fully understand the host-virus interaction. Present serological screening methods may not have sufficient sensitivity for determining BLV status in some circumstances.

Animals↗

Adherence to high-carbohydrate, high-fiber diets: long-term studies of non-obese diabetic men.

High-carbohydrate, high-fiber (HCF) diets provide short-term benefits for selected individuals with diabetes. Long-term effects of HCF diets, however, have not previously been documented. The effectiveness and acceptability of HCF diets for 14 men with diabetes followed for 4 years as outpatients were assessed. Hospital HCF diets with 70% of energy as carbohydrate and 65 gm plant fiber daily lowered insulin doses, serum cholesterol, and triglycerides. Home HCF diets with 55% to 60% carbohydrate and 50 gm plant fiber daily sustained improvements throughout the observed period of up to 86 months. Of the 11 patients for whom diet adherence was evaluated, 9 had good to excellent adherence, 1 had fair adherence, and 1 had poor adherence.

Adult↗