RESUMO
The expert system FLORIDA (Fuzzy Logic Orientated Rule Interpreter for Diagnostic Applications) is equipped with a knowledge base applying linguistic rules of clinical experts according to the pathophysiologic conception of the sepsis-3 definition and the mathematics of fuzzy logic. It works independently of any subjective factors. FLORIDA detects sepsis by an increase of a second incoming valueâ¯≥ 25% compared with the reference value. It requires dynamics of the parameters used in knowledge base; thus, if the dynamics are absent, the system is unable to detect sepsis. In a retrospective clinical validation study, FLORIDA was used to scan 498 consecutive patients in a medical intensive care unit, which corresponded to 1700 patient-days. During the study period, the prevalence of sepsis was 10%. In all, 423 patients were identified as not having sepsis, which was confirmed by clinical experts. Among the 48 patients identified as having sepsis, 26 were confirmed (true positive). In 22 patients, sepsis could not be confirmed by the clinician (false positive). FLORIDA did not detect sepsis in 4 patients, but was diagnosed by the clinician (false negative). In the 22 false-positive patients, a life-threatening disease existed requiring intensive care. Because of the system philosophy, FLORIDA is unable to recognize patients with full blown sepsis at admission. With this in mind, the sensitivity was 1.0 and specificity was 0.95. Thus, FLORIDA is qualified for the early detection of a developing sepsis. Sepsis is detected on average 12.5⯱ 8.6â¯h after the start of sepsis has been determined by a clinical expert. FLORIDA should be used on normal wards and should contribute to early detection of sepsis and potentially earlier therapeutic intervention in order to decrease hospital mortality. However, prospective validation is needed.
Assuntos
Sistemas Inteligentes , Sepse , Diagnóstico Precoce , Florida , Humanos , Estudos Prospectivos , Estudos Retrospectivos , Sepse/diagnósticoRESUMO
Bone loss with aging may at least in part be due to inadequate bone formation. In this study, we examined whether the proliferation of osteoblast-like cells in vitro in response to local and systemic factors might be attenuated with age. A total of 36 cultures of osteoblast-like cells were obtained from outgrowths of human trabecular bone. Parathyroid hormone, growth hormone, calcitonin, transforming growth factor beta, insulin-like growth factor I, and platelet-derived growth factor BB dose dependently increased DNA synthesis in all cultures. Increases in DNA synthesis with each of these factors were significantly negatively correlated with donor age in cultures obtained from the iliac crest bone of 50- to 70-year-old women. Cells from 61- to 70-year-old donors required approximately 10-fold higher concentrations of growth factors and hormones to yield comparable increases in DNA synthesis than cells from 51- to 60-year-old donors. A significant negative correlation between age and mitogenic responsiveness to platelet-derived growth factor and growth hormone, but not toward the other factors, was also observed in cultures from the femoral head trabecular bone of 60- to 90-year-old women. Our findings suggest that bone loss with aging may be partially due to a decreased capacity of osteoblasts to proliferate in response to systemic or locally released osteotropic factors.
Assuntos
Envelhecimento/metabolismo , Substâncias de Crescimento/farmacologia , Osteoblastos/efeitos dos fármacos , Idoso , Idoso de 80 Anos ou mais , Calcitonina/farmacologia , Divisão Celular/efeitos dos fármacos , Células Cultivadas , DNA/biossíntese , Feminino , Hormônio do Crescimento/farmacologia , Humanos , Fator de Crescimento Insulin-Like I/farmacologia , Pessoa de Meia-Idade , Osteoblastos/citologia , Hormônio Paratireóideo/farmacologia , Fenótipo , Fator de Crescimento Derivado de Plaquetas/farmacologia , Fator de Crescimento Transformador beta/farmacologiaRESUMO
Reports from numerous laboratories have shown that the gene coding for the bacterial enzyme chloramphenicol-3-O-acetyltransferase can be used as a reporter gene (cat) in mammalian and plant systems to analyze gene activity at the transcriptional level when combined with endogenous regulatory signals; the enzyme activity can be quantified by a chromatographic or a photometric assay. To adapt this simple and highly sensitive test for the yeast system, we constructed a series of yeast vectors containing the cat gene together with selectable markers for Escherichia coli and yeast; integrating, autonomously replicating and centromere-carrying plasmids were used. We show that the cat gene lacking the endogenous promoter is expressed at low levels in yeast transformants. To demonstrate functional expression of the cat gene placed under the control of a yeast promoter, we chose the PHO5 regulatory region. We found that cat expression was induced via the PHO5 promoter in a manner as observed for the endogenous PHO5 gene, whereas in the repressed state cat expression remained low. Using these vectors, it should be feasible to analyze other sequences conferring promoter activity or other control functions in yeast.
Assuntos
Cloranfenicol O-Acetiltransferase/genética , Vetores Genéticos , Regulação da Expressão Gênica , Plasmídeos , Regiões Promotoras Genéticas , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/genética , Transformação GenéticaRESUMO
TYR1, the gene from Saccharomyces cerevisiae, which encodes prephenate dehydrogenase, one of the tyrosine biosynthetic enzymes, has been cloned by complementing a yeast tyr1 mutant strain. The DNA fragment containing the gene is part of a 45-kb cosmid clone which represents a region of chromosome II covering the genetically mapped tyr1 locus. The nucleotide sequence of a 3.1-kb region carrying the TYR1 gene and adjacent regions has been determined. The open reading frame contains 441 codons, corresponding to about 52.2 kDa for the encoded protein. The canonical NAD-binding domain is located within the first 45 amino acids of the protein. By primer extension, we show that there is one transcription start point. Presumably, the expression of TYR1 is not under the general GCN4 control. Instead, we find a dependence on the presence or absence of phenylalanine. These data were obtained by analysing CAT activity in constructs containing promoter fragments of TYR1 and the cat reporter gene.
Assuntos
Genes Fúngicos , Oxirredutases/genética , Prefenato Desidrogenase/genética , Saccharomyces cerevisiae/genética , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Dados de Sequência Molecular , Mutação , Regiões Promotoras Genéticas , RNA Mensageiro/genética , Mapeamento por Restrição , Saccharomyces cerevisiae/enzimologiaRESUMO
A new procedure of extracting data from measurements by single-probe radionuclide ventriculography is presented which reduces the phase differences during summation of single-beat actions and permits the use of a greater number of single actions for summation. Sum curves so determined are better suited for measurements of ejection fractions, compliance and ventricular blood flow rates.
Assuntos
Imagem do Acúmulo Cardíaco de Comporta/métodos , Função Ventricular Esquerda/fisiologia , Angina Instável/fisiopatologia , Velocidade do Fluxo Sanguíneo , Diástole/fisiologia , HumanosRESUMO
Genes for gentamicin-3-acetyltransferases [ACC(3)] of types III and IV have been cloned from various R-plasmids. In two R-plasmids, pWP14a (AAC(3)-III) and pWP7b [AAC(3)-IV], resistance genes have been found directly adjacent to a single copy of an IS element, IS140. Nucleotide sequence determination of the AAC(3)-IV gene from plasmid pWP7b and of part of IS140 from three different sources suggested that the -35 region of the AAC(3)-IV promoter was part of the IS element. A similarly built-up promoter was found in pWP14a. It was found also, that a hygromycin B phosphotransferase was expressed from a locus neighbouring the AAC(3)-IV gene in pWP7b which was under the control of the same promoter. In two other R-plasmids, pWP113a and pWP116a, the AAC(3)-III gene was found in different genetic environments, namely close to Tn3-like structures.
Assuntos
Acetiltransferases/genética , Elementos de DNA Transponíveis , Escherichia coli/enzimologia , Genes Bacterianos , Genes , Sequência de Bases , Enzimas de Restrição do DNA , Resistência Microbiana a Medicamentos , Escherichia coli/genética , Hibridização de Ácido Nucleico , Plasmídeos , Recombinação Genética , Transcrição GênicaRESUMO
In the yeast Saccharomyces cerevisiae, the majority of the tRNA genes are found associated with transposable elements one of which is the Ty element. We noticed that the transcriptional activity of several of these genes was rather different depending on the type of the accompanying element(s) [Nelböck et al. (1985) Biol. Chem. Hoppe-Seyler 366, 1041-1051]. In order to study the influence of a Ty element on tRNA gene expression, we used the method of micro-injection into Xenopus oocytes taking advantage of a Ty+/Ty- allelic pair of a tRNALys1 gene recently isolated in our laboratory. A direct comparison showed that the expression of the plus allele was c. sixfold higher than that of the minus allele. In order to determine sequences of the Ty responsible for this effect, we constructed a number of variants in which distinct segments of the Ty were deleted or rearranged. The activating ability was localized to a 590-bp segment of the Ty containing the 'promoter delta', independent of its orientation or location towards the tRNA gene thus resembling the effects described for transcriptional enhancers. This is the first time that a long-range effect has been observed on the expression of a gene transcribed by RNA polymerase III.
Assuntos
Elementos Facilitadores Genéticos , Genes Reguladores , RNA Fúngico/genética , RNA de Transferência/genética , Saccharomyces cerevisiae/genética , Animais , Elementos de DNA Transponíveis , Microinjeções , Oócitos , Plasmídeos , Transcrição Gênica , XenopusRESUMO
From histogram analysis the value of computed tomography for early detection of breast carcinoma or differentiation of benign and malign processes of the breast was evaluated. The results show, that CT is not applicable and gives no improvement of diagnostics.
Assuntos
Neoplasias da Mama/diagnóstico por imagem , Tomografia Computadorizada por Raios X , Neoplasias da Mama/patologia , Feminino , Humanos , Estadiamento de Neoplasias , Fatores de TempoRESUMO
THR1, the gene from Saccharomyces cerevisiae, encoding homoserine kinase, one of the threonine biosynthetic enzymes, has been cloned by complementation. The nucleotide sequence of a 3.1-kb region carrying this gene reveals an open reading frame of 356 codons, corresponding to about 40 kDa for the encoded protein. The presence of three canonical GCN4 regulatory sequences in the upstream flanking region suggests that the expression of THR1 is under the general amino acid control. In parallel, the enzyme was purified by four consecutive column chromatographies, monitoring homoserine kinase activity. In SDS gel electrophoresis, homoserine kinase migrates like a 40-kDa protein; the native enzyme appears to be a homodimer. The sequence of the first 15 NH2-terminal amino acids, as determined by automated Edman degradation, is in accordance with the amino acid sequence deduced from the nucleotide sequence. Computer-assisted comparison of the yeast enzyme with the corresponding activities from bacterial sources showed that several segments among these proteins are highly conserved. Furthermore, the observed homology patterns suggest that the ancestral sequences might have been composed from separate (functional) domains. A block of very similar amino acids is found in the homoserine kinases towards the carboxy terminus that is also present in many other proteins involved in threonine (or serine) metabolism; this motif, therefore, may represent the binding site for the hydroxyamino acids. Limited similarity was detected between a motif conserved among the homoserine kinases and consensus sequences found in other mono- or dinucleotide-binding proteins.
Assuntos
Regulação Enzimológica da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Genes Fúngicos , Fosfotransferases (Aceptor do Grupo Álcool) , Fosfotransferases/genética , Saccharomyces cerevisiae/enzimologia , Treonina/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Evolução Biológica , Clonagem Molecular , Dados de Sequência Molecular , Peso Molecular , Fosfotransferases/isolamento & purificação , Mapeamento por Restrição , Saccharomyces cerevisiae/genética , Homologia de Sequência do Ácido Nucleico , Transcrição GênicaRESUMO
The gene encoding threonine synthase (THR4) from the yeast Saccharomyces cerevisiae was cloned by complementation of a thr4 mutant. This gene was also found on a lambda clone (5239) consisting of a fragment of chromosome III inserted in the vector lambdaMG3. The THR4 gene encodes a protein of 514 amino acids (M.W. 58 kDa), which has extensive homologies with E. coli threonine synthase (thrC) and B subtilis threonine synthase. The 5' flanking region of the gene contains three regulatory sequences [TGACT(C)] for the general amino acid control (GCN). About 130 bp downstream of the THR4 gene another large open reading frame (563 amino acids) is found in the opposite orientation. This may imply that this open reading frame, called CTR86, shares a terminator region with THR4. The function of the protein encoded by CTR86 is not yet clear, but the fact that the upstream region contains a GCN4 responsive site suggests that the gene product may also be involved in amino acid biosynthesis.