Biomedical subjects

L A Rivas

Publications and source records attributed to L A Rivas.

16 recordsLinked to original sources

PubMed2003

Essential Bacillus subtilis genes.

To estimate the minimal gene set required to sustain bacterial life in nutritious conditions, we carried out a systematic inactivation of Bacillus subtilis genes. Among approximately 4,100 genes of the organism, only 192 were shown to be indispensable by this or previous work. Another 79 genes were predicted to be essential. The vast majority of essential genes were categorized in relatively few domains of cell metabolism, with about half involved in information processing, one-fifth involved in the synthesis of cell envelope and the determination of cell shape and division, and one-tenth related to cell energetics. Only 4% of essential genes encode unknown functions. Most essential genes are present throughout a wide range of Bacteria, and almost 70% can also be found in Archaea and Eucarya. However, essential genes related to cell envelope, shape, division, and respiration tend to be lost from bacteria with small genomes. Unexpectedly, most genes involved in the Embden-Meyerhof-Parnas pathway are essential. Identification of unknown and unexpected essential genes opens research avenues to better understanding of processes that sustain bacterial life.

Bacillus subtilis↗

PubMed2001

Functional analysis of the Streptomyces lividans type I signal peptidases.

Type I signal peptidases are responsible for the proteolytic cleavage of the signal peptide of secreted proteins. In the gram-positive bacterium Streptomyces lividans, four adjacent genes (sipW, sipX, sipY and sipZ) were isolated encoding putative type I signal peptidases. In this work, the different sip genes were cloned and expressed. Subsequently, the Sip proteins were purified to raise antibodies. Although the four Sip proteins share a low degree of sequence similarity and differ significantly in size and pI, anti-Sip antibodies cross-reacted intensively. Functional signal peptidase processing activity for each of these Sip proteins was shown both in vitro and in vivo. The different Sip proteins did not exhibit the same cleavage efficiency on the Bacillus subtilis pre-chitosanase.

Antibodies, Bacterial↗

PubMed1999

Direct versus indirect signs of traumatic aortic injury revealed by helical CT: performance characteristics and interobserver agreement.

OBJECTIVE: The purpose of this study was to evaluate the relative value of and interobserver agreement on direct versus indirect (hematoma) signs of traumatic aortic injury using helical CT. MATERIALS AND METHODS: From April 1994 through January 1997, 40 patients who were suspected to have traumatic aortic injury and who underwent contrast-enhanced helical CT had subsequent proof or exclusion of aortic injury. All available CT scans of these patients were combined with CT scans of 13 randomly chosen patients that had been initially interpreted as negative, and clinical follow-up showed no evidence of aortic injury. Two emergency radiologists and a nonemergency radiologist who were unaware of clinical outcome performed independent review of these cases to evaluate for mediastinal hematoma, periaortic hematoma, and direct signs of aortic injury. RESULTS: Direct signs of injury were seen on helical CT by both emergency radiologists in all 17 cases of aortic injury with no false-positive interpretations. The nonemergency radiologist failed to observe subtle direct signs in two cases of aortic injury, but patient management would not have been adversely affected. All observers had more false-negative interpretations for both mediastinal hematoma and periaortic hematoma than for direct signs. Interobserver agreement was higher for direct signs (kappa = .93) than for either mediastinal hematoma (kappa = .65) or periaortic hematoma (kappa = .71). CONCLUSION: In this study, helical CT revealed direct signs of traumatic aortic injury that were more accurate and more often observed than were indirect signs. Emphasis on direct signs should improve confidence in using helical CT to evaluate traumatic aortic injury.

Adolescent↗

PubMed1996

Erythropoietin gene expression in human, monkey and murine brain.

The haematopoietic growth factor erythropoietin is the primary regulator of mammalian erythropoiesis and is produced by the kidney and the liver in an oxygen-dependent manner. We and others have recently demonstrated erythropoietin gene expression in the rodent brain. In this work, we show that cerebral erythropoietin gene expression is not restricted to rodents but occurs also in the primate brain. Erythropoietin mRNA was detected in biopsies from the human hippocampus, amygdala and temporal cortex and in various brain areas of the monkey Macaca mulatta. Exposure to a low level of oxygen led to elevated erythropoietin mRNA levels in the monkey brain, as did anaemia in the mouse brain. In addition, erythropoietin receptor mRNA was detected in all brain biopsies tested from man, monkey and mouse. Analysis of primary cerebral cells isolated from newborn mice revealed that astrocytes, but not microglia cells, expressed erythropoietin. When incubated at 1% oxygen, astrocytes showed >100-fold time-dependent erythropoietin mRNA accumulation, as measured with the quantitative reverse transcription-polymerase chain reaction. The specificity of hypoxic gene induction in these cells was confirmed by quantitative Northern blot analysis showing hypoxic up-regulation of mRNA encoding the vascular endothelial growth factor, but not of other genes. These findings demonstrate that erythropoietin and its receptor are expressed in the brain of primates as they are in rodents, and that, at least in mice, primary astrocytes are a source of cerebral erythropoietin expression which can be up-regulated by reduced oxygenation.

Adult↗

PubMed1996

Cervical spine trauma: how much more do we learn by routinely using helical CT?

Assessment of suspected cervical spine injuries remains a major debate in trauma care. It is generally accepted that many fractures are missed or incompletely shown at radiography, mainly because of suboptimal studies obtained in obtunded, uncooperative trauma victims. In a series of 88 severely traumatized patients, the authors retrospectively determined the type, distribution, and significance of such missed lesions. This assessment was made by comparing radiographs and helical computed tomographic (CT) scans of the cervical spine and reviewing medical records in these cases. Of the 88 patients, 32 patients had cervical spine fractures (n = 50) that were not revealed or were incompletely demonstrated at radiography. Most missed fractures occurred at the C-1 to C-2 and C-6 to C-7 levels, and most involved the transverse processes and the posterolateral elements of the vertebrae. One-third of the patients with missed fractures had either clinically significant or unstable injuries, as determined on the basis of mechanistic or imaging criteria. Helical CT can depict significant fractures not shown by plain radiography and should be added routinely to the initial screening for cervical spine fractures in polytrauma victims.

Adolescent↗

PubMed1995

Localization of specific erythropoietin binding sites in defined areas of the mouse brain.

The main physiological regulator of erythropoiesis is the hematopoietic growth factor erythropoietin (EPO), which is induced in response to hypoxia. Binding of EPO to the EPO receptor (EPO-R), a member of the cytokine receptor superfamily, controls the terminal maturation of red blood cells. So far, EPO has been reported to act mainly on erythroid precursor cells. However, we have detected mRNA encoding both EPO and EPO-R in mouse brain by reverse transcription-PCR. Exposure to 0.1% carbon monoxide, a procedure that causes functional anemia, resulted in a 20-fold increase of EPO mRNA in mouse brain as quantified by competitive reverse transcription-PCR, whereas the EPO-R mRNA level was not influenced by hypoxia. Binding studies on mouse brain sections revealed defined binding sites for radioiodinated EPO in distinct brain areas. The specificity of EPO binding was assessed by homologous competition with an excess of unlabeled EPO and by using two monoclonal antibodies against human EPO, one inhibitory and the other noninhibitory for binding of EPO to EPO-R. Major EPO binding sites were observed in the hippocampus, capsula interna, cortex, and midbrain areas. Functional expression of the EPO-R and hypoxic upregulation of EPO suggest a role of EPO in the brain.

Animals↗

PubMed1987

Synthesis of beta-glucans in Prototheca zopfii. Evidence for the existence of a glycoprotein primer.

Membrane preparations from the non-photosynthetic alga Prototheca zopfii incorporate glucose from UDP-[3H]glucose into the trichloroacetic-acid-insoluble fraction and the polysaccharides insoluble in hot alkali. Time course and pulse-chase experiments indicate that the acid-insoluble fraction was a precursor of the alkali-insoluble fraction. Isolation of 3H-labeled membrane or soluble fraction showed that only membrane fractions were able to transfer radioactivity into polysaccharides. Treatment of glucosylated membranes with trypsin or cellulase only partially affect their transfer ability, indicating that the precursor was internalized in vesicles. Analysis of the in vitro synthesized polysaccharides by enzymatic and acid hydrolysis showed that glucose and cellobiose were present as radioactive sugars. Permethylation of the polysaccharide indicates that 80% of the glucose was beta-1,4-bonded with 20% in beta-1,3-linkages. This polysaccharide was found to be identical with the cell-wall beta-glucan obtained in vivo [Rivas, L.A. & Pont Lezica, R. (1978) Planta (Berl.) 165, 348-353].

Carbohydrate Metabolism↗

PubMed1987

Synthesis of beta-glucans in Prototheca zopfii. Isolation and characterization of the glycoprotein primer.

When Prototheca zopfii cells were pulse-labeled with 14C-containing amino acids and homogenized, 14C-labeled membranes were obtained. In vitro incubations with the previously labeled membranes and UDP-[3H]Glc produced a trichloroacetic-acid-insoluble fraction having both isotopes. A double-labeled glucoprotein was isolated and characterized. It has a relative molecular mass of 28,000-30,000 and a carbohydrate content of 10%. The oligosaccharide chain is linked to the protein through an O-glycosidic bond between hydroxyproline and glucose. The oligosaccharide has a polymerization degree ranging over 10-20 hexose units. Glucose is the only monosaccharide found; most of the glucose residues are beta-1,4-linked (90%) but some are beta-1,3-linked (10%).

Cell Wall↗

PubMed1983

Inhibition of beta-1,4-Glucan Biosynthesis by Deoxyglucose : The Effect on the Glucosylation of Lipid Intermediates.

GDP- and UDP-deoxyglucose inhibit the incorporation of glucose from UDP-glucose into dolichyl phosphate glucose and dolichyl pyrophosphate oligosaccharides. GDP-deoxyglucose inhibits by competing with the physiological nucleotide sugars for dolichyl phosphate, and dolichyl phosphate deoxyglucose is formed. This inhibition is reversed by excess of dolichyl phosphate. UDP-deoxyglucose does not give rise to a lipid-linked derivative, and inhibition by this analog is not reversed by dolichyl phosphate. The UDP- and GDP-derivatives of deoxyglucose inhibit the incorporation of glucose into glucose-containing glycoproteins. This effect seems to be the result of the inhibition of lipid intermediates glucosylation and is comparable to the effect produced by coumarin. Cellulose synthetase activity is not affected by UDP- or GDP-deoxyglucose. On the other hand, deoxyglucose inhibits the formation of beta-1,4-glucans in vivo.

Journal Article↗

PubMed1976