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Biomedical subjects

M E Collins

Publications and source records attributed to M E Collins.

At least 55 records · Page 3Linked to original sources

Detection of bovine virus diarrhoea virus RNA by in situ hybridisation with digoxigenin-labelled riboprobes.

A non-isotopic in situ hybridisation (ISH) method has been developed for the detection of bovine virus diarrhoea virus (BVDV) RNA. The technique is highly specific and allows the detection of BVDV in formalin-fixed paraffin-wax-embedded sections. Digoxigenin-labelled riboprobes were generated to the coding region for the p125 non-structural protein and consequently it was possible to locate the virus positive-sense RNA in BVDV-infected tissues. Formalin-fixed tissues are ideal for ISH, combining the possibility of a retrospective analysis of tissues for virus distribution with excellent morphological preservation. In situ hybridisation has facilitated a precise examination, at the cellular level, of viral nucleic acid distribution during pestiviral infections, with an increased sensitivity and wider applicability than immunocytochemistry.

Animals↗

Weight control practices of U.S. adolescents and adults.

OBJECTIVE: To estimate the prevalence of various weight-loss practices in U.S. adolescents and adults. DESIGN: The Youth Risk Behavior Survey, a self-administered survey of a random sample of high school students in 1990 and the Behavioral Risk Factor Surveillance System, a random-digit dial survey in 1989. SETTING: Thirty-eight states and the District of Columbia. PARTICIPANTS: High school students (n = 11,467) and adults 18 years and older (n = 60,861). RESULTS: Among high school students, 44% of female students and 15% of male students reported that they were trying to lose weight. An additional 26% of female students and 15% of male students reported that they were trying to keep from gaining more weight. Students reported that they had used the following weight control methods in the 7 days preceding the survey: exercise (51% of female students and 30% of male students); skipping meals (49% and 18%, respectively); taking diet pills (4% and 2%, respectively); and vomiting (3% and 1%, respectively). Among adults, 38% of women and 24% of men reported that they were trying to lose weight, whereas 28% of each sex reported that they were trying to maintain their weight. CONCLUSIONS: Attempts to lose or maintain weight are very prevalent among both adolescents and adults, especially among females.

Adolescent↗

Nucleotide sequence and expression in Escherichia coli of cDNAs encoding papaya proteinase omega from Carica papaya.

We have cloned and sequenced two similar, but distinct, cDNAs from both fruit and leaf tissues of Carica papaya. The C-terminal portion of the predicted amino acid (aa) sequence of one of the clones has complete identity with the mature enzyme sequence of the cysteine proteinase papaya proteinase omega (Pp omega). The second clone contains ten individual bp changes compared with the first and encodes a protein with three single-aa substitutions, only one of which is located in the mature sequence, but most noticeably carries an additional 19-aa C-terminal extension. The clones encode pre-pro precursor isoforms of Pp omega. The former of these clones has been expressed in Escherichia coli using a T7 polymerase expression system to produce insoluble pro-enzyme which has been solubilized and refolded to yield auto-activable pro-Pp omega.

Amino Acid Sequence↗

Characteristics of a retrovirus associated with Jembrana disease in Bali cattle.

A virus causing Jembrana disease in Bali cattle (Bos javanicus) was demonstrated to have characteristics of a retrovirus. Reverse transcriptase activity was detected in virus purified by sucrose gradient centrifugation. Electron microscopic examination of tissue from the affected cattle indicated that the virus matured by C-type budding through the plasma membrane and into intracytoplasmic vacuoles of cells in lymphoid tissue, with the formation of circular enveloped virus particles ranging in diameter from 96 to 124 nm with an eccentric nucleoid. Western immunoblotting using sera from recovered animals demonstrated virus proteins of M(r) 100K, 45K, 42K, 33K, 26K, 16K and 14K. The 26K protein of Jembrana disease virus cross-reacted in Western blots with the 26K capsid protein of bovine immunodeficiency virus (BIV). The apparent morphogenesis, protein structure and antigenic relationship with BIV suggested the virus was a lentivirus.

Animals↗

A novel method for the purification of porcine phospholipase A2 expressed in E. coli.

Porcine phospholipaseA2 expressed in E. coli as a fusion protein was isolated, renatured and specifically cleaved by trypsin as described in (1). Active phospholipaseA2, was purified to homogeneity on a column of PBE-94 over a pH region 7.4-4.5. Using this method, several phospholipase A2 mutant enzymes have now been purified in a single step and all behaved identically during chromatofocusing. The method will therefore be extremely useful not only for those interested in understanding the structure-function relationships of phospholipaseA2 but also for preparing the enzyme in large quantities for industrial and pharmaceutical purposes.

Animals↗

Modification of the stability of phospholipase A2 by charge engineering.

Electrostatic interactions play an important role in stabilizing the folded conformation of globular proteins. Here we predict the change in stability of charge engineered mutants, construct these mutants and compare the predicted change in stability with that observed. The change in stability was correctly predicted for two of the three mutants and the factors responsible for the discrepancy between observation and prediction for the third mutant are discussed.

Enzyme Stability↗

An inducible gene expression system for Neurospora crassa.

Neurospora crassa acetyl CoA synthetase is highly induced when the growing mycelium is transferred from sucrose- to acetate-based medium. The inducible promoter of this gene has been isolated and used to control the expression of glutamate dehydrogenase. Transformants containing this expression cassette show gdh levels up to 25 times higher than the nontransformed host strain. This expression cassette will form the basis of a system of heterologous gene expression.

Acetate-CoA Ligase↗

Changes in activity of porcine phospholipase A2 brought about by charge engineering of a major structural element to alter stability.

We have modified the stability of porcine phospholipase A2 by charge engineering. The mutations are situated at the N-terminal of a major helix and are N89D and N89D/E92Q. This engineering has significantly altered the activity of the enzyme to aggregated and monomeric substrates. A N89D/E92K mutant is more stable but considerably less active than wild type. An N89D mutant is more stable and of similar activity to wild type. The substantial change in activity may be due to direct interaction of residue 92 with aggregated substrate or may be via second calcium binding. Second calcium binding may be more probable as activity against monomers is also affected. Additional calcium binding may therefore be an important way of manipulating the activity of phospholipase A2.

Animals↗

Inflammatory disease of the anterior epitympanum.

The sinus epitympani, an anterior extension of the epitympanum, juxtaposes the attic with the protympanum and the eustachian tube. A review of 94 horizontally sectioned temporal bones revealed this space consistently present but variable in size. In 48 temporal bones with acute or chronic inflammatory disease, the pathologic findings in the anterior epitympanum coexisted with those in the middle ear and mastoid. In temporal bones with inflammatory disease, the size of the pediatric sinus was significantly smaller than the sinus in diseased adult bones. Surgical exploration of this area is proposed for more complete removal of diseased tissue and improved ventilation of the attic.

Aged↗

Cloning and characterization of the gene for the '19 kDa' antigen of Mycobacterium bovis.

Monoclonal antibody CMA134.1 reacted with a protein antigen of apparent molecular mass 22 kDa from Mycobacterium bovis and Mycobacterium tuberculosis, and with an apparently 24 kDa antigen of Mycobacterium kansasii, but not with other mycobacteria or related species. This antibody was used to screen a gene library of M. bovis in lambda gt11 and identified a recombinant clone that expressed a protein with an apparent molecular mass of 19-20 kDa. Gene expression occurred from the lac promoter in lambda gt11, but used an unidentified vector promoter, possibly that of the replication primer RNA, in the final plasmid construct. The sequence of an 840 bp fragment was determined and shown to code for a product of 15 kDa. This sequence is identical to that, independently determined, of a gene from M. tuberculosis, usually referred to as the 19 kDa antigen. The reasons for the apparent size discrepancies are discussed.

Amino Acid Sequence↗

A professional preparation course on AIDS/HIV infection.

This paper describes the conceptualization, development, planning, and implementation of a new professional preparation course entitled "AIDS/HIV Education: Issues and Strategies." The 15-week, campuswide course was offered through the Dept. of Health Science Education at the University of Florida during spring semester 1990. Though universities have been identified as providers of preservice education for future teachers and health professionals, current activities often have been deemed inadequate to meet the needs of those charged with providing AIDS/HIV education. This course was implemented to address an array of issues related to HIV education, ranging from epidemiological and biomedical information and educational strategies to examination of personal and ethical issues surrounding the disease. Delivery of a successful course and student reaction are discussed, along with recommendations for programs considering similar educational efforts. A detailed planned information packet is available from the authors.

Acquired Immunodeficiency Syndrome↗

SPORT database.

SPORT addresses needs that have recently surfaced, where athletics and its effect on society have grown. Consciousness of drug use among high profile athletes, as well as among those in high school and college athletics, has generated new research into the effects of this phenomenon on the physiology and psychology of the athletes themselves. Other topics, such as public health, exercise physiology, and the physiology and sociology of sport have made this database essential to researchers.

Canada↗

The electrostatic fields in the active-site clefts of actinidin and papain.

The active sites of actinidin (EC 3.4.22.14) and papain (EC 3.4.22.2) display different reactivity characteristics to probes targeted at the active-site cysteine residue despite the close structural similarity of their active sites. The calculated electrostatic fields in the active-site clefts of actinidin and papain differ significantly and may explain the reactivity characteristics of these enzymes. Calculation of electrostatic potential also focuses attention on the electrostatic properties that govern formation of the active-site thiolate-imidazolium ion-pair. These calculations will guide the modification of the pH-activity profile of the cysteine proteinases by site-directed mutagenesis.

Binding Sites↗

Education for healthy body weight: helping adolescents balance the cultural pressure for thinness.

Though education for healthy body weight traditionally has focused on obesity, the increased incidence of anorexia nervosa and bulimia among young women suggests education also is needed to address the opposite end of the spectrum. Anorexia nervosa and bulimia are complex and multidimensional disorders associated with individual, family, and sociocultural factors. This article examines the cultural pressure for dieting and thinness currently experienced in America and its impact as a possible predisposing factor for developing eating disorders among adolescent females. Literature is reviewed related to the changing American standard of attractiveness for females reflected by 20th century mass media and its subsequent influence on adolescent concerns for dieting and thinness. Preventive strategies are recommended to help adolescents balance the cultural pressure for thinness and their own desires for attractiveness within the larger context of overall good health.

Adolescent↗

Identification of restriction fragments from two cryptic Clostridium butyricum plasmids that promote the establishment of a replication-defective plasmid in Bacillus subtilis.

Clostridium butyricum NCIB 7423 carries two cryptic plasmids, pCB101 (6.05 kbp) and pCB102 (7.8 kbp). Sites for the restriction enzymes EcoRI, EcoRV, HindIII, ClaI and PstI have been found in one or both of these plasmids and their relative positions determined. Restriction fragments from both plasmids have been inserted into a vector plasmid (pJAB1) that is able to replicate in Escherichia coli but not in Bacillus subtilis and the recombinant plasmids have been established in E. coli. A 3.3 kbp Sau3A fragment of pCB101 conferred upon the vector the ability to transform both Rec+ and Rec- strains of B. subtilis. Plasmid pRB1, a representative chimaera carrying only the 3.3 kbp Sau3A fragment of pCB101, was successfully transferred from B. subtilis back to E. coli. Plasmid pRB1 was readily lost from B. subtilis in the absence of selection. This evidence, together with the results of hybridization experiments, suggests that pRB1 is present as a weakly replicating autonomous element in B. subtilis. A recombinant plasmid carrying a 2.0 kbp Sau3A fragment of pCB102 underwent integration into the B. subtilis chromosome.

Bacillus subtilis↗