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Biomedical subjects

M J Kessler

Publications and source records attributed to M J Kessler.

At least 73 records · Page 4Linked to original sources
PubMed1982

High performance liquid chromatography of steroid metabolites in the pregnenolone and progesterone pathways.

Rapid separation of gonadal steroids in the progesterone (delta 4) and pregnenolone pathway (delta 5) has been accomplished by the use of high performance liquid chromatography (HPLC). Two HPLC systems are utilized. THe first requires the use of two separate radial compression columns (C-18 and C-8), with steroids being eluted with a methanol-water gradient. The second employs a stainless steel C-18 (reversed phase) column with a 12% octadecylsilane coating. The latter system separates seven of the eight steroids in the delta 4 and delta 5 pathways in thirty-five minutes. For the quantitation of steroids directly, integration of the peak areas, using 254 nm absorption for the delta 4 pathway steroids (5 ng minimum limit), and 210 nm absorption for the delta 5 pathway steroids (25 ng minimum limit) is used. For the quantitation of radiolabeled metabolites resulting from incubation of gonadal tissue with radiolabeled steroid precursors, either one of two methods is used: (1) the eluent can be recovered from the HPLC using a fraction collector, and counted in liquid scintillation counter or (2) the entire eluent (or a portion of it) can be counted immediately by directing the flow through a radioactivity detector.

Biopsy↗
PubMed1982

Induction of allergic neuritis in rhesus monkeys.

Experimental allergic neuritis (EAN) was induced in rhesus monkey (Macaca mulatta) following sensitization with rabbit nerve (PNS) myelin in complete Freund's adjuvant (CFA) or with bovine P2 protein complexed with phosphatidyl serine (P2-lipid) in CFA. The response of monkeys receiving PNS myelin in CFA differed from the previous studies where monkeys developed clinical signs of fatal EAN within 15-20 days following sensitization. The monkeys in this study (6) showed a much longer delay (40-114 days) before the appearance of severe clinical signs, and 4 of the 6 animals survived without further attack (1 year). Monkeys (4) injected with P2-lipid (2:1 ratio; w/w) developed severe clinical signs of EAN which was fetal in 3 cases. Peripheral lymphocytes from monkeys sensitized to the P2-lipid showed a much stronger mitogeneic response to P2 protein than those from the PNS myelin-sensitized monkeys. on quantitation of the circulating anti-P2 antibodies, the P2-sensitized monkeys generally had much titers than those sensitized with PNS myelin.

Animals↗
PubMed1982

Quantitation of radiolabeled compounds eluting from the HPLC system.

Three techniques are compared for the quantitation of various radiolabeled compounds eluting in the high performance liquid chromatography system. The first technique requires fraction-collecting the effluent from the HPLC, removing an aliquot to scintillation vials, and counting each fraction in a liquid scintillation counter. The second uses direct interface of the HPLC effluent to a flow-through radioactivity detector. The third involves quantitation of various radiolabeled compounds (proteins, steroids, and nucleotides) by splitting the effluent from the HPLC with an electronic steam splitter, thus diverting a present portion to the fraction collector for further chemical characterization and the remainder to the radioactivity flow detector for direct quantitation. A direct comparison of the chromatograms and the radioactivity counting efficiencies of these three techniques is presented.

Adrenal Glands↗
PubMed1982

Characterization of Sertoli cell RNA synthetic activities in vitro at selected times during sexual maturation.

In this study, Sertoli cell RNA synthetic activity in vitro was characterized at selected times during sexual maturation. Sertoli cells, isolated from rat testes undergoing the first wave of spermatogenesis and placed in culture for 4 days, exhibited 2-fold increases in soluble ribonucleotide pools and in total RNA concentrations over the age span of 18-35 days. High performance liquid chromatographic analysis of the ribonucleotide pools in Sertoli cells cultured from 18- and 33- to 34-day-old rats revealed that, in addition to the overall age-related doubling of concentrations, uridine triphosphate (UTP) and cytidine triphosphate (CTP) pools were disproportionately increased 4- and 6-fold, respectively. In general, Sertoli cell contained relatively small amounts of UTP in comparison to several other cell types, but exhibited a high ADP:ATP ratio. A uniform 2-fold increase in the base composition of Sertoli cell RNA per mg DNA was observed over the age span of 18-35 days, with no preferential increase in any one specific nucleotide. There was no change in [3H]uridine incorporation (2 h) into RNA per cell (pmol/mg DNA), but decreased specific activity of the RNA (pmol/mg RNA) in Sertoli cells cultured from 35-day-old rats as compared to those from 18- to 19-day-old rats. Similar differences were noted in the specific activity of label incorporated into specific RNA bases. In contrast, the specific activity of the UTP-CTP soluble pool/mg DNA was only slightly increased. These data indicate that processes related to RNA synthesis in the Sertoli cell undergo a number of changes during the period of sexual maturation.

Animals↗
PubMed1981

Lack of greater seroconversion of rhesus monkeys after subcutaneous inoculation of dengue type 2 live-virus vaccine combined with infection-enhancing antibodies.

Four groups of six nonimmune male rhesus monkeys were inoculated subcutaneously with formulations of dengue type 2 vaccine virus DEN-2/S-1. Group A received 1.9 x 10(4) plaque-forming units of vaccine in normal human serum albumin diluent. Group B received the same dose combined with a dengue type 2-immune human serum diluted 1:1,600, beyond its neutralization endpoint of 1:300, but having an immune enhancement titer of 250,000. Groups C and D received 10-fold dilutions of these respective formulations. No migration-inhibitory factor was found when peripheral blood mononuclear leukocytes obtained on day 68 post-immunization from monkeys of all experimental groups were tested. No viremia was detected in any of the monkeys when sera taken on postvaccination days 1 through 12 were inoculated into adult Toxorhynchites amboinensis mosquitoes and LLC-MK2 cells. By day 89, four of the six monkeys had seroconverted by the neutralization test in each of groups A, B, and C, and three of five monkeys in group D (one monkey died from cardiac collapse after anesthesia) had seroconverted. Immune enhancement of dengue virus infection is known to occur in humans and monkeys circulating heterologous flavivirus antibodies. In this study, there was no enhancing effect when antibody was mixed with dengue type 2 vaccine virus and injected subcutaneously.

Animals↗
PubMed1979

Selection and pairing of 'normal' rhesus monkeys (Macaca mulatta) for research: procedures, techniques, and observations.

A selection process has been developed by the authors to insure investigators that animal subjects used in their experiments will accurately reflect biological changes associated with the project design. The procedure involved the following: complete physical examination; complete hemogram, serum electrolytes, serum biochemistry, body weight; and anthropometric determinations; aging by dentition; electrocardiography; total-body radiography; fecal culture; fecal ova, and parasite examinations; and examination of other conditions. Arithmetic means and standard deviations were calculated and used to eliminate monkeys whose values varied from the mean by more than two standard deviations.

Animals↗
PubMed1979

Clinical description of tetanus in squirrel monkeys (Saimiri sciureus).

Nineteen cases of clinical tetanus developed in Bolivian squirrel monkeys (Saimiri sciureus) which were housed in outdoor corrals with soil-based floors. The disease was initially characterized by stiffness in gait followed by extensor rigidity, trismus and opisthotonus. Eleven of the 19 monkeys (58%) had evidence of external wounds. The case fatality rate was 100%. Tetanus accounted for 12% of adult female and 19% of adult male mortalities in the colony. Immunization with tetanus toxoid was effective in reducing the incidence of tetanus.

Animals↗