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Biomedical subjects

A Sanchez

Publications and source records attributed to A Sanchez.

At least 145 records · Page 8Linked to original sources

Transforming growth factor beta modulates growth and differentiation of fetal hepatocytes in primary culture.

Fetal hepatocytes in primary culture are cells capable to carry out both proliferation and differentiation processes simultaneously. Previous studies have shown that these cells respond to mitogens, such as hepatocyte growth factor (HGF) or epidermal growth factor (EGF), inducing the expression of early genes, such as fos and myc. The transforming growth factor-beta (TGF-beta) family is one of the most influential groups of growth and differentiation factors. In this report, we show that TGF-beta 1 inhibits fetal hepatocyte proliferation, arresting these cells at G1 phase of the cell cycle. In addition, TGF-beta down-regulates the mitogen-induced myc early expression. However, TGF-beta has no effect on the expression of other protooncogenes, such as fos and H-ras. In addition to its inhibitory role on fetal hepatocyte growth, TGF-beta increases the mRNA levels of fibronectin, an extracellular matrix protein, and maintains the expression of some liver specific genes, such as albumin and alfafetoprotein, above control values. The analysis of the expression of some hepatocyte transcriptional factors has shown that TGF-beta increases HNF1 alpha and HNF1 beta mRNA levels. We conclude that TGF-beta may modulate liver growth and differentiation throughout fetal development.

Animals↗

An angiotensin-independent, hypotension-induced, sodium appetite in the rat.

We have investigated the role of peripheral angiotensin II in the generation of hypotension (without hypovolaemia)-induced salt intake in rats treated with an IV infusion of the quaternary ammonium peripheral ganglion blocker Penthonium. At a dose of 15.4 mg/ml this compound induces a significant decrease in blood pressure from the beginning of the infusion. Intake of 3% NaCl was significantly increased only on the first day (1.7 +/- 0.3 ml, n = 7, p < 0.01 vs. the day before) and this induced salt appetite was not altered (2.3 +/- 0.9 ml, n = 7, p < 0.05 vs. the day before) by intracerebroventricular administration of a nonpeptide AII-type 1 receptor specific antagonist, Losartan, at a dose known to block AII-induced drinking (250 ng). We conclude from these results that AII liberated in response to the hypovolaemia is probably not responsible for the subsequent induced intake of NaCl which may be the result of a direct barosensitive input to the salt appetite centers of the brain.

Angiotensin II↗

Identification of a new North American hantavirus that causes acute pulmonary insufficiency.

In May 1993, a pulmonary disease syndrome with novel clinical and epidemiologic features was identified in the southwestern United States. Healthy young adults developed a febrile prodrome followed by the rapid onset of often lethal acute respiratory distress. Although an infectious disease was suspected, intensive investigations initially failed to identify the causative agent. Multiple specialized microbiology laboratories at the National Center for Infectious Diseases (Centers for Disease Control and Prevention) applied classic serologic and culture methods as well as recently developed molecular biological techniques to samples collected from field investigations of the patients. Serologic tests detected the presence of an active immune response to a hantavirus. Reverse transcription and polymerase chain reaction amplification of RNA extracted from human tissues used primers designed from sequences of known hantaviruses to demonstrate genomic sequences of a novel hantavirus. Immunohistochemistry showed the presence of hantavirus antigens in the endothelium of lung tissues from patients and provided the final pathogenetic link to this group of viruses. These methods were concordantly positive in virtually all samples available from 18 patients with compatible clinical histories identified between January and July 1993. Test results of control subjects and searches for other agents in identified cases were negative. This newly recognized hantavirus causes a novel syndrome of acute pulmonary edema and shock; the pathogenesis is related to the presence of virus antigens in the pulmonary capillaries. The virus may be an important cause of severe and fatal disease presenting as adult respiratory distress syndrome in otherwise healthy persons.

Antibodies, Viral↗

Molecular phylogeny of Guanarito virus, an emerging arenavirus affecting humans.

The nucleotide sequence of a portion of the nucleocapsid (N) gene of the Guanarito virus prototype strain (INH-95551) has been determined. It was obtained by direct RNA and polymerase chain reaction (PCR) fragment sequencing of the 3' end of the small (S) RNA fragment. A comparison of this 782-nucleotide segment was done with the known homologous gene sequences of five other arenaviruses: Junin, Machupo, Tacaribe, Pichinde, and lymphocytic choriomeningitis (LCM). Phylogenetic analysis of the N gene open reading frame showed that Guanarito virus is genetically distinct from other members of the Arenavirus family, with 32% nucleotide sequence divergence from Junin, 30% from Machupo, 32% from Tacaribe, 41% from Pichinde, and 45% from LCM. Comparison of amino acids encoded by this sequence region indicated a probable antigenic domain (amino acids 55-63) shared among all arenaviruses studied to date. Along with its host restriction and focal distribution, our data support the hypothesis that this virus has been evolving independently in its endemic focus, for some time.

Amino Acid Sequence↗

Genome structure and variability of a virus causing hantavirus pulmonary syndrome.

A previously unrecognized hantavirus (family Bunyaviridae) has recently been detected and shown to be associated with a severe respiratory illness with high mortality, termed hantavirus pulmonary syndrome (HPS). This disease has now been identified throughout the western United States. We present nucleotide sequence characterization of the three RNA segments composing the HPS virus genome and address the question of the apparent emergence of this highly lethal virus. No evidence of genetic reassortment with previously recognized hantaviruses was found, each RNA segment being unique and approximately 30% different at the nucleotide level to the segments of the closest relative, Prospect Hill virus. These findings, together with the observed extensive genetic diversity of HPS viruses and examples of geographic clustering of distinct virus genotypes, suggest that HPS and associated virus have likely existed undetected for many years. The virus genome M segment was determined to be 3696 nucleotides in length and encode G1 and G2 proteins, 652 and 488 amino acids in length. The S segment was found to be 2059 nucleotides in length and to encode a nucleocapsid protein, 428 amino acids in length. S segment analysis also revealed an unusually long noncoding region with numerous repeats and evidence for a potential NSS protein encoded in an overlapping frame.

Amino Acid Sequence↗

Increased P-type ATPase activity in Leishmania tropica resistant to methotrexate.

In the present report, we show evidence that the membrane from the protozoan parasite Leishmania tropica (LRC-L39), in vitro resistant to 1 mM of methotrexate (MTX), has a significative increased ATPase activity with respect to wild-type line. This ATPase activity is vanadate sensitive, a characteristic of the P-type ATPases included in the ATP-binding casette (ABC) superfamily of transporters, such as P-glycoprotein involved in the multidrug resistance in mammalian cells. Also, this ATPase activity is not modified by MTX, ammonium molybdate and other detergents such as Triton X-100, Brij 58 and lysophosphatidylcholine. However, unlike the P-glycoprotein, we have observed that the ATPase activity is not stimulated by the drugs verapamil and puromycin. This significative ATPase activity could be related to the overexpressed putative P-glycoprotein, with unknown function in these MTX-resistant parasites.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Effects of extremely-low-frequency electromagnetic fields on ion transport in several mammalian cells.

We have investigated the effects of sinusoidal electromagnetic fields (EMF) on ion transport (Ca2+, Na+, K+, and H+) in several cell types (red blood cells, thymocytes, Ehrlich ascites tumor cells, and HL60 and U937 human leukemia cells). The effects on the uptake of radioactive tracers as well as on the cytosolic Ca2+ concentration ([Ca2+]i), the intracellular pH (pHi), and the transmembrane potentsial (TMP) were studied. Exposure to EMF at 50 Hz and 100-2000 microT (rms) had no significant effects on any of these parameters. Exposure to EMF of 20-1200 microT (rms) at the estimated cyclotron magnetic resonance frequencies for the respective ions had no significant effects except for a 12-32% increase of the uptake of 42K within a window at 14.5-15.5 Hz and 100-200 microT (rms), which was found in U937 and Ehrlich cells but not in the other cell types.

Animals↗

Characterization of filoviruses based on differences in structure and antigenicity of the virion glycoprotein.

Eight different filovirus isolates, representing major episodes of filovirus hemorrhagic disease, were propagated for structural and antigenetic analyses of their glycoprotein (GP). Carbohydrate analysis revealed that N- and O-glycosylation are features of filovirus GPs. Oligosaccharide side chains differed in their sialylation pattern and seemed to be cell line-dependent. Marburg virus (MBG) isolates are clearly distinguished from Ebola (EBO) and Reston viruses by a lack of terminal sialic acids when propagated in E6 and MA-104 cells. It was also determined that GP-specific antisera failed to show any cross-reactivity between MBG isolates and other filoviruses. These data, together with prior findings, indicate that the genus Filovirus can be divided into a MBG group and EBO group.

Animals↗

An analysis of Xenopus tyrosine kinase genes and their expression in early development.

Xenopus laevis and X. borealis were screened for tyrosine kinase genes using the polymerase chain reaction (PCR) and reverse transcriptase (RT)-PCR and 34 X. laevis and 23 X. borealis tyrosine genes were identified. Eighteen of the genes represented novel tyrosine kinase family members. The rest could be classified into known tyrosine kinase subfamilies, of which however only three have been previously identified in Xenopus. Eight clones, including bFGFR (xFGFR1) and potential Trk, Ins R., Fak, Fyn, and Abl homologs, were used to probe temporal and spatial gene expression in early development. Quantitative RT-PCR and whole-mount and in situ hybridization showed that most of these mRNAs were present throughout development and were broadly distributed, mainly in ectodermal and mesodermal derived tissues. At the blastula stage, bFGFR mRNA was detected within the ectoderm and a gradient of expression was noted within the invaginating mesoderm. The unexpected promiscuous expression of many tyrosine kinase genes in early development is discussed.

Amino Acid Sequence↗

Antibiotic resistance and penicillin tolerance in clinical isolates of group B streptococci.

The aim of this study was to determine the susceptibility patterns of 100 group B streptococcal strains isolated in our hospital and to ascertain tolerance to penicillin by determining quantitative killing curves. We found two strains with intermediate susceptibility to penicillin and eight strains to ampicillin. Seventeen isolates were tolerant to penicillin, with bacterial counts decreasing 2 to 3 log during the first 8 h but still above 10(2) CFU/ml after 24 h. The kinetic study shows that penicillin tolerance is not rare among group B streptococci isolated in our hospital.

Ampicillin Resistance↗

Outbreak of Brucella melitensis among microbiology laboratory workers.

We report on an outbreak of laboratory-acquired brucellosis involving four technicians working at a microbiology laboratory. All cases occurred in a period of 4 months. Blood cultures and the Rose Bengal test were positive for Brucella spp. in all cases. Microagglutination was positive for Brucella spp. at titers of between 1/40 and 1/160. All patients were cured after treatment.

Adult↗

Hormonal, renal, and metabolic alterations during hypertension induced by chronic inhibition of NO in rats.

The evolution of renal excretory function and circulating vasoactive systems was studied during progressive increases in blood pressure (BP) induced in rats by oral administration of NG-nitro-L-arginine methyl ester (L-NAME; 5-30 mg/100 ml) for 5 wk. L-NAME induced a stepped elevation (P < 0.05) in BP levels without changing creatinine clearance, urine flow, or sodium excretion rate along the study. Reductions (P < 0.05) in plasma renin activity and plasma aldosterone concentration were found only during treatment with 30 mg/100 ml of L-NAME. Plasma norepinephrine and epinephrine concentrations were elevated (P < 0.05) in the last week of the study. Plasma concentrations of endothelin-1 and urinary excretion of prostaglandin E2, 6-ketoprostaglandin F1 alpha, and thromboxane B2 were not significantly affected by L-NAME. Similarly, no changes in plasma concentrations of glucose, insulin, total cholesterol, or triglycerides were observed. In summary, during long-term administration of L-NAME, progressive increases in BP levels were observed without changes in either sodium excretion or enhanced circulating vasoconstrictor activity. Thus, it is likely that inhibition of synthesis of nitric oxide (NO) in the vasculature leads to an imbalance between the tonic relaxing action of NO and the influences of vasoconstrictor agents even when the latter remain at normal levels.

Aldosterone↗