Search PubMed⌕ Search

Biomedical subjects

A Sanchez

Publications and source records attributed to A Sanchez.

At least 217 records · Page 12Linked to original sources

Leupeptin does not affect the normal signal transduction mechanism in platelets.

Calpains are Ca2+-dependent serine proteases that can regulate protein kinase C-mediated cellular events by cleaving the membrane-bound native enzyme to yield an activated cytosolic fragment. Inhibition of calpain by leupeptin may cause enhancement or inhibition of cellular functions depending on the nature of the protein kinase C reaction involved. We have studied the effects of leupeptin on platelet responses (aggregation, secretion, thromboxane B2 formation and intracellular Ca2+ and pH changes) induced by either thrombin, collagen or phorbol 12-myristate 13-acetate (TPA), which are known to activate protein kinase C by different mechanisms. Only thrombin-induced responses were inhibited by leupeptin. This suggests that the inhibitory effect of leupeptin is not due to antagonism of calpain in this system, but to direct interference with the proteolytic effect of thrombin.

Adenosine Triphosphate↗

Effect of dietary protein on serum insulin and glucagon levels in hyper- and normocholesterolemic men.

This study was designed to test the effect of dietary protein on blood levels of insulin and glucagon. Twelve normocholesterolemic (less than 200 mg/dl) and 11 hypercholesterolemic greater than 240 mg/dl) healthy male subjects, 31-62 years of age, were randomly given 3 liquid test meals 1 week apart. Meals were identical except for the protein source (soybean, casein, or protein free). Blood was drawn at fasting, and 0.5 and 2 h postprandially. Insulin and glucagon levels were measured by radioimmunoassay. Hypercholesterolemic subjects had a higher (P less than 0.05) insulin/glucagon ratio (1.5) than normocholesterolemic subjects (0.7) 2 h post-prandially when fed the casein test meal. There was no significant difference following the soybean test meal. This implies that the post-prandial insulin/glucagon ratio was affected by the amino acid composition of the diet. There was a consistently higher insulin response to all test meals among hyper- versus normocholesterolemic subjects. These results are consistent with our hypothesis that the hypocholesterolemic effects of soybean protein and the hypercholesterolemic effects of casein were mediated by altered levels of insulin and glucagon.

Adult↗

The nucleoprotein gene of Ebola virus: cloning, sequencing, and in vitro expression.

Genomic and messenger RNAs of a Zaire strain of Ebola virus were cloned, and inserts specific for the nucleoprotein gene were isolated and sequenced. The nucleoprotein gene is located proximal to the 3' end of the genome and is preceeded by a putative leader sequence. The gene begins with the transcriptional start site sequence 3'-UACUCCUUCUAAUU..., and ends with the polyadenylation site sequence 3'-... UAAUUCUUUUUU. The predicted coding region is 2217 bases in length and encodes a protein that contains 739 amino acids, with a calculated molecular weight of 83.3 kDa. The protein has an approximate net charge of -30 and can be divided into a hydrophobic N-terminal half and a hydrophilic and highly acidic C-terminal half. An in vitro transcript, generated from plasmid DNA containing the entire coding region, directs the synthesis of authentic nucleoprotein in a rabbit reticulocyte lysate system. The genomic organization and transcriptional signals of Ebola are similar to those of other nonsegmented, negative-strand RNA viruses, but nucleic acid or amino acid sequence comparisons indicate a lack of similarity.

Amino Acid Sequence↗

Identification of the 5' end of the rubella virus subgenomic RNA.

The 5' end of the subgenomic RNA of rubella virus was determined by primer extension. The Maxam-Gilbert sequence ladder of the primer extension product contained a determinable sequence which was colinear with the complement of the sequence of the genomic RNA through nucleotide 3325 from the 3' end of the genomic RNA and a pair of bands in every lane above the determinable sequence. These results indicated that synthesis of the subgenomic RNA is initiated internally on the minus-polarity genome-length RNA template and that the length of subgenomic RNA is 3327 nucleotides excluding the poly(A) tail. There are thus 77 nucleotides between the 5' end of the subgenomic RNA and the first AUG, which is the initiation codon for the structural protein open reading frame. The initiation site of rubella virus subgenomic RNA synthesis is 20 nucleotides downstream from a block of 28 nucleotides which shares homology with the nucleotide sequence which is conserved at the subgenomic RNA initiation site in the Alphaviruses, the other genus of Togaviruses.

Animals↗

Comparison of the hypocholesterolemic effects of dietary soybean protein with those of formaldehyde-treated casein in rabbits.

Treatment of casein with formaldehyde changes its tertiary structure and decreases its hypercholesterolemic properties in rabbits. To investigate whether formaldehyde-treated casein exerts this hypocholesterolemic effect in the same manner as soybean protein, rabbits were fed high or low cholesterol diets containing soybean protein, casein, formaldehyde-treated casein or a mixture of casein and formaldehyde-treated casein. Formaldehyde-treated casein was hypocholesterolemic when fed in a low, but not in a high, cholesterol diet. The hypocholesterolemic effect of soybean protein was independent of the amount of cholesterol included in the diet. In contrast to rabbits fed soybean protein, steroid absorption in those fed formaldehyde-treated casein did not differ from that in rabbits fed native casein. Furthermore, the absorption of phosphorus and nitrogen was lower in rabbits fed formaldehyde-treated casein than in those fed native casein, whereas the absorption found in rabbits fed soybean protein resembled that of their casein-fed counterparts. The diets containing soybean protein and formaldehyde-treated casein produced a comparable ratio of lysine to arginine in serum. The results presented in this paper indicate that the hypocholesterolemic action of dietary formaldehyde-treated casein does not resemble that of soybean protein.

Amino Acids↗

Junin virus monoclonal antibodies: characterization and cross-reactivity with other arenaviruses.

Twenty-one monoclonal antibodies reactive with Junin virus structural proteins were produced and characterized. Using radioimmunoprecipitation and Western blot assays, 13 were found to react with the nucleoprotein, seven with the surface glycoprotein and one failed to react, but showed a fluorescent antibody staining pattern consistent with other glycoprotein-specific antibodies. In radioimmunoprecipitation assays, glycoprotein-specific monoclonal antibodies reacted not only with the 35K structural glycoprotein, but also with what is presumed to be the glycoprotein precursor. Four of seven glycoprotein-specific antibodies neutralized Junin virus to high titres. Cross-reactivity with other arenaviruses was found to be restricted to nucleoprotein-specific monoclonal antibodies and occurred only with New World arenaviruses. Cross-reactivity also shows the Junin virus to be most closely related to Machupo and Tacaribe viruses.

Animals↗

Alpha-blockade on blood pressure and on cardiac noradrenaline content in SHR and DOCA-salt rats.

The current study was designed to explore the effects of peripherally administered alpha-adrenoceptor inhibiting drugs (either prazosin or yohimbine) in spontaneously hypertensive (SHR) and DOCA-salt hypertensive rats before the development of established hypertension, as well as to characterize changes induced by prazosin and/or yohimbine in the cardiac noradrenaline (NA) content that might be responsible for the development of hypertension in these strains. The alpha-adrenoceptor blockade can prevent hypertension in DOCA and salt-treated animals in such a way that their blood pressure stabilizes at levels significantly lower than those observed in similarly treated normotensive controls. Significantly lower cardiac NA content was observed in DOCA-salt rats under basal and experimental conditions. The blood pressure of the treated rats and the heart NA content of the SHR were unaltered by treatment. Thus, administration of the alpha-adrenoceptor blocking agents, prazosin and/or yohimbine, throughout the developmental stage of SHR hypertension failed to alter either the progressive rise in blood pressure or in NA content. There may be differences between the cardiac adrenergic mechanisms responsible for the development of hypertension in each of these two models of hypertension.

Adrenergic alpha-Antagonists↗

Tuberculosis of the larynx: clinical aspects in 19 patients.

Laryngeal tuberculosis continues to exist in spite of extensive tuberculosis eradication campaigns. We present 19 cases of laryngeal tuberculosis seen in our institution between 1982 and 1987. From the clinical findings it is clear that the localization of the lesions has changed since the preantibiotic era. Today the spread appears to be mainly via lymphatic and hematogenic routes, as opposed to the direct spread more common previously. The fibrosis caused by the lesions is incapacitating and frequently requires surgical correction.

Adolescent↗

The effect of a dentifrice containing triclosan and a copolymer on plaque formation and gingivitis: a 14-week clinical study.

One hundred eighteen male and female adult subjects were entered into a 14-week, double-blind clinical study to compare the effect of a dentifrice containing 0.3% triclosan and 2% of a copolymer of methoxyethylene and maleic acid on plaque formation and gingivitis. The subjects were stratified into two balanced groups according to baseline plaque scores. They then received an oral prophylaxis and were assigned to the use of either a placebo dentifrice or the triclosan/copolymer dentifrice for the next 14 weeks. After 6 weeks' use, the triclosan/copolymer dentifrice provided a 21.34% statistically significant (99% level of confidence) reduction in supragingival plaque deposits, as compared to the placebo dentifrice. The triclosan/copolymer dentifrice also provided a 5.49% reduction gingivitis after 6 weeks' use, as compared to the placebo dentifrice. This reduction was not statistically significant. After 14 weeks' use, the triclosan/copolymer dentifrice provided a 38.80% statistically significant (99% level of confidence) reduction in supragingival plaque deposits, as compared to the placebo dentifrice. The dentifrice containing triclosan and copolymer also provided 50.72% statistically significant (99% level of confidence) reduction in gingivitis, as compared to the placebo dentifrice. When tooth surfaces with the highest degree of plaque formation (i.e. Quigley-Hein score greater than or equal to 3) were examined at 6 weeks, the triclosan/copolymer dentifrice provided a 42.92% statistically significant (99% level of confidence) reduction in supragingival plaque deposits, as compared to the placebo dentifrice.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Effect of diet on growth of M. leprae in mouse footpads.

Semipurified diets, with equal amounts of vitamins, minerals and fibre, but varied in protein and fat content from pork, barbel fish or soya beans were tested for their possible effect on the growth of M. leprae in mouse footpads. 105 BALB/c male weanling mice were randomly divided into five diet groups of 21 mice each and fed for six months. Differences between bacterial counts of diet groups were found. The mouse foot pad model is suitable for dietary study in leprosy.

Animals↗

Atherogenic effect of oxidized products of cholesterol.

Cholesterol under certain in vitro and possibly in vivo conditions may be oxidized to oxysterols, which are suspected of being initiators of atherosclerotic plaques. Oxysterols inhibit HMG-CoA reductase activity resulting in a decreased cholesterol concentration in the cell membrane, which leads to endothelial membrane injury and probable premature cell death. Exogenous oxidation of cholesterol in human tissues under certain unusual conditions is highly probable. Dietary oxysterols are absorbed in the gastrointestinal tract and are selectively transported by the athrogenic lipoproteins LDL and VLDL. The oxysterols cholestanetriol and 25-OH cholesterol have been shown to be atherogenic. Oxysterols are commonly found in dried egg products, powdered milk, cheeses and in a variety of high temperature dried animal products.

Animals↗

Effects of dietary composition on growth of M. leprae in mouse footpads.

The number of bacteria per mouse footpad were measured at intervals beginning with the third month in male, weanling BALB/c mice infected with M. leprae and fed for a period of 6 months to test the effects of diet on multiplication of bacteria. The mean bacteria count per footpad in mice remaining at 6 months in the two high fat diets was higher (p = 0.014) than the mean of the two low fat diets. Likewise, the pooled mean bacterial count of mice fed the two diets of animal origin had a tendency to a higher mean bacterial count compared to mice fed the two diets of plant origin. Low level of dietary protein in early life also seemed to predispose to M. leprae multiplication. Our data in mice suggest that the association of diet with human leprosy should be investigated.

Animals↗

Thrombin-induced changes of intracellular [Ca2+] and pH in human platelets. Cytoplasmic alkalinization is not a prerequisite for calcium mobilization.

We have studied the effects of thrombin (0.1 U/ml) on intracellular Ca2+ ([Ca2+]i) and pH (pHi) in human platelets loaded with fluorescent indicators. Thrombin produced a transient decrease of pHi which reached its maximum within 15-25 seconds (s) and was followed by a sustained alkalinization which brought pHi above the resting value. [Ca2+]i increased transiently peaking at 5-10 s. The late alkalinization induced by thrombin was antagonized by ethylisopropylamiloride, an inhibitor of Na+-H+ exchange, and by sphingosine, an inhibitor of protein kinase C, with little effect on the [Ca2+]i transient. The early acidification was not inhibited by these treatments. We conclude tha the thrombin-induced changes of [Ca2+]i and pHi are mediated by different mechanisms. The late alkalinization is due to activation of Na+/H+ exchange mediated by protein kinase C and, contrarily to previous proposals (Siffert, W. and Akkerman, J.W.N. (1987) Nature 325, 456-458), it is not necessary for calcium mobilization from intracellular stores.

Amiloride↗

Testing a mechanism of control in human cholesterol metabolism: relation of arginine and glycine to insulin and glucagon.

Eight men were given 2 casein meals, one with and one without a supplement of arginine and glycine, to measure the effect on plasma amino acids, insulin and glucagon. Supplementation resulted in increased levels of plasma glucagon, glycine and arginine, a tendency to decreased insulin and significantly lower insulin/glucagon ratio, tryptophan and tyrosine. The data suggest that insulin and glucagon, which control cholesterol metabolism, respond to dietary and postprandial plasma amino acid levels of arginine and glycine.

Amino Acids↗

Physicochemical properties of Marburg virus: evidence for three distinct virus strains and their relationship to Ebola virus.

The physicochemical and antigenic properties of three groups of Marburg (MBG) virus isolates, separated temporally and geographically, were compared to each other and to another member of the same family, Ebola (EBO) virus. Each MBG isolate contained seven virion proteins, one of which was a glycosylated surface protein. Peptide mapping of glycoproteins, nucleoproteins (NP) and viral structural protein (VP40) demonstrated extensive sequence conservation in the proteins of viruses isolated over a 13-year period, but homology was not evident in VP24. Some homology between the NPs of MBG and EBO was observed. A close antigenic relationship between MBG strains was found by radioimmunoassay but no evidence was found of antigenic cross-reactivity with EBO viruses. MBG virion proteins are produced from virus-specific monocistronic mRNA species. Five of the seven viral proteins were produced by in vitro translation of these RNAs. MBG virions contained one RNA species with an Mr of 4.2 x 10(6) and virions had a density of 1.14 g/ml in potassium tartrate. Virus isolates from different outbreaks had distinct T1 oligonucleotide maps, but had approximately 95% homology in base sequence. No two geographically distinct virus pairs were more closely related to each other than to a third virus isolate. MBG viruses are thus similar to EBO viruses in morphology and other physicochemical properties and are very similar to each other in RNA and protein composition. Each of the three geographically and temporally distinct MBG virus outbreaks appears to have been due to a genetically distinguishable, but antigenically closely related virus strain. In addition, these studies confirm the belief that MBG and EBO viruses are members of the new virus family, the Filoviridae.

Animals↗

Age-related changes of norepinephrine content in kidneys of spontaneously hypertensive and Wistar-Kyoto rats.

Renal norepinephrine (NE) content was determined during the development of spontaneously hypertensive rats (SHR) and age-matched Wistar-Kyoto (WKY) rats in an attempt to correlate biochemical changes with the reported functional changes occurring in hypertension development in the SHR. In contrast to WKY rats, in which the levels of NE remained relatively constant with age, renal NE content in the SHR was highest at the 4th week of age, decreasing transiently during the 5th, 6th, and 7th weeks, and then again reaching a plateau during the 8th week. The fall in NE content in the kidney is associated with a rise in blood pressure with age in SHR and suggests a relationship between NE levels and hypertension.

Aging↗