ABSTRACT
The aim of this study was to produce high yield of a local bacterial alkaline protease in the yeast system because the scientific involvement of microorganisms in enzyme production is still not given enough attention in Saudi Arabia. Soil samples were collected from the rhizosphere of some desert plants in Saudi Arabia. Ninety-three alkaline protease producing bacterial isolates were recovered on skimmed-milk agar at pH 9.4 and 45°C for 48 hr. Isolate D9 obtained from the rhizosphere of Heliotropium digynum at Dhahran City was the most potent isolate in respect to enzyme productivity (184.6 U/ml). The full gene of alkaline protease was amplified and showed the expected size (1300 bp). Restriction enzymes analysis also verified the integrity of the PCR product. The sequence of the protease gene revealed an open reading frame of 1329 nt correspond to the full length of the protease gene of isolate D9 encoding a 443 aa protein. After ligation of the amplified gene by the TA cloning method, digestion with appropriate restriction enzymes confirmed the integrity of the cloned gene. The insert was prepared by two PCRs that were conducted with a pair of primers specifically designed for this purpose. The digested and purified cloning vector pRS426/GAL1p-207-Glu-MS was ligated with the insert then transformed into various strains of Saccharomyces cerevisiae via the electroporation method. Maximum protease expression was done by recombinant OS303 in galactose containing media (145.5 U/ml) with an approximately 2-fold increase when compared with the wild OS303 strain., this may be due to ability to activate gal operon.
O objetivo deste estudo foi produzir alto rendimento de uma protease alcalina bacteriana local no sistema de leveduras, já que o envolvimento científico de microrganismos na produção de enzimas ainda não recebe atenção suficiente na Arábia Saudita. Amostras de solo foram coletadas da rizosfera de algumas plantas do deserto na Arábia Saudita. Noventa e três isolados bacterianos produtores de protease alcalina foram recuperados em ágar de leite desnatado a pH 9,4 e 45°C por 48 horas. O isolado D9 obtido da rizosfera de Heliotropium digynum na cidade de Dhahran foi o mais potente em relação à produtividade da enzima (184,6 U/ml). O gene completo da protease alcalina foi amplificado e apresentou o tamanho esperado (1300 pb). A análise de enzimas de restrição também verificou a integridade do produto de PCR. A sequência do gene da protease revelou uma fase de leitura aberta de 1329 nt, correspondendo ao comprimento total do gene da protease do isolado D9 que codifica uma proteína 443 aa. Após a ligação do gene amplificado pelo método de clonagem TA, a digestão com enzimas de restrição apropriadas confirmou a integridade do gene clonado. A inserção foi preparada por dois PCRs que foram conduzidos com um par de primers projetados especificamente para esta finalidade. O vetor de clonagem digerido e purificado pRS426/GAL1p-207-Glu-MS foi ligado com a inserção e então transformado em várias cepas de Saccharomyces cerevisiae por meio do método de eletroporação. A expressão máxima da protease foi feita por OS303 recombinante em meio contendo galactose (145,5 U/ml) com um aumento de aproximadamente duas vezes quando comparado com a cepa OS303 selvagem, e isso pode ser por causa da capacidade de ativar o operon gal.
Subject(s)
Animals , Peptide Hydrolases , Saccharomyces cerevisiae , Yeasts , Enzyme Activators , Enzymes/biosynthesis , RhizosphereABSTRACT
BACKGROUND: Carcinoid heart disease (CaHD) develops from vasoactive substances released by neuroendocrine tumors, which can cause significant patient morbidity and mortality without surgical intervention. We performed a systematic review and meta-analysis to elucidate granular perioperative details and long-term outcomes in these patients. METHODS: Electronic search of Ovid, Scopus, Cumulative Index of Nursing and Allied Health Literature, and Cochrane Controlled Trials Register was performed to examine surgical treatment of carcinoid disease. Nine articles comprising 416 patients were selected. Study-level data were extracted and pooled for meta-analysis. RESULTS: Mean patient age was 63 years (95% confidence interval, 57-70) with 53% (95% confidence interval, 46-61) of patients being male. In addition, 75% (95% confidence interval, 54-96) of neuroendocrine tumors originated from the small bowel or colon and 98% (95% confidence interval, 93-100) had liver metastases. Right heart failure was present in 48% (95% confidence interval, 14-81). Moderate or severe regurgitation was present in 97% (95% confidence interval, 95-99) of tricuspid and 72% (95% confidence interval, 58-83) of pulmonary valves. In addition, 99% (95% confidence interval, 98-100) of tricuspid and 59% (95% confidence interval, 38-79) of pulmonary valves were replaced. Bioprosthetic valves were used in 80% (95% confidence interval, 68-93) of tricuspid positions. Mean hospital duration of stay was 16 days (95% confidence interval, 7-25). Thirty-day mortality was 9% (95% confidence interval, 6-12). Mean follow-up was 25 months (95% confidence interval, 11-39). Median survival was 3 years (95% confidence interval, 2.5-3.5). CONCLUSION: For patients >18 years of age, surgical treatment of carcinoid heart disease can be performed with a reasonable safety profile. However, overall survival appears to have ongoing effects of the primary disease.
Subject(s)
Carcinoid Heart Disease/surgery , Carcinoid Heart Disease/diagnosis , Carcinoid Heart Disease/mortality , HumansABSTRACT
El síndrome autoinmune tirogástrico (SAT) fue descrito en pacientes en quienes el suero presentaba reacciones cruzadas de anticuerpos dirigidos contra los antígenos de células parietales gástricas y tiroideas. A través de dos casos describimos el espectro patológico de este síndrome. El primero asocia una tiroiditis de Hashimoto y una anemia perniciosa, desarrollando durante el seguimiento un tumor neuroendocrino gástrico. El segundo caso presenta una enfermedad de Graves y una gastritis autoinmune, secundaria a Helicobacter Pylori: esta última es reversible luego de tratamiento. Se considera que la poliendocrinopatía autoinmune de tipo III (dentro de la cual puede inscribirse el síndrome tirogástrico) es rara, pero no lo es en nuestra experiencia. Un total de 13 % (32/240) de los pacientes con tiroiditis que hemos seguido prospectivamente, tienen también una gastritis autoinmune. Helicobacter pylori está claramente implicado en el 16 % de estos casos con gastritis autoinmune. Infección, malabsorción y gastritis son potencialmente reversibles después del tratamiento de erradicación bacteriana. En el 84 % restante de los pacientes con gastritis y tiroiditis, no se encuentran pruebas serológicas o histológicas de Helicobacter pylori. La autoinmunidad gástrica es entonces irreversible, y conduce a una hipergastrinemia, hipoclorhidria y atrofia gástrica severa. La hipergastrinemia estimula la hiperplasia de las células enterocromafines, con riesgo de progresión a un tumor neuroendocrino. Proponemos un esquema diagnóstico novedoso para mejor caracterización del síndrome tirogástrico. Exponemos la literatura sobre el tema y discutimos a partir de algunos modelos animales pertinentes sobre la autoinmunidad gástrica infecciosa. Rev Argent Endocrinol Metab 51:37-43, 2014 Los autores declaran no poseer conflictos de interés.
The thyrogastric autoimmune syndrome (TAS) was described in patients in whom the serum cross-reacted both with gastric parietal cells antigens and thyroid antigens. We report two cases illustrating the spectrum of pathological features of TAS. The first one is a case of Hashimoto’s Thyroiditis associated with pernicious anemia, further developing a gastric neuroendocrine tumor during follow up. The second one is a case of Graves’ disease and autoimmune reversible gastritis, While type III autoimmune polyendocrinopathy (which includes TAS) is considered to be rare, this was not the case in our experience. A total of 13 % (32/240) of the patients with thyroiditis that we have prospectively followed have also autoimmune gastritis. Helicobacter pylori is clearly implicated in 16 % of the cases of autoimmune gastritis. Infection, malabsorption and gastritis are potentially reversible after bacterial eradication treatment. In the remaining 84 % of patients with gastritis, no histological or serological evidence of Helicobacter pyloriwas found. Gastric autoimmunity is then irreversible, leading to gastric severe atrophy, hypochlorhydria and hypergastrinemia. Hypergastrinemia stimulates enterochromaffin cell hyperplasia, progressing eventually to neuroendocrine tumors. We propose a diagnostic approach to improve the characterization of TAS, including a literature review and discussing some relevant animal models of infectious gastric autoimmunity. Rev Argent Endocrinol Metab 51:37-43, 2014 No financial conflicts of interest exists.
ABSTRACT
Approximately 12.5% of all 9,920 extant bird species in the world are threatened with extinction, and yet conservation efforts through natural breeding of captive species continue to encounter difficulties. However, sperm cryopreservation and artificial insemination offer potential benefits over natural breeding, but their applicability is still limited in nondomestic species. In this study, we aimed to exploit the potential of germ cell xenotransplantation as an alternative tool for preserving germplasm of endangered birds. The study was designed to investigate whether transfer of either spermatogonia-enriched cell fraction (SEF) or crude testicular cell fraction (CTF) from adult Japanese quails (as a model for wild species) would result in recolonization of gamma-irradiated gonads of adult recipient chickens. One month after transplantation, 75% of recipients injected with SEF and 25% of recipients injected with CTF resumed spermatogenesis. However, it took more than 3 months for 33% of the negative controls to resume marginal production of sperm. Some SEF recipients produced more spermatozoa bearing head morphology compared with donor controls. DNA analysis using quail-specific primers did not detect donor's DNA in these recipients' semen. However, 6 months after xenotransplantation, presence of quail germ cells was demonstrated by polymerase chain reaction and by immunohistochemistry in 1 rooster injected with SEF. These findings indicate that spermatogonia from adult quails were capable of colonizing immunocompetent testis of adult chickens but failed to produce sufficient sperm. Despite this limitation, the present approach represents a potential conservation tool that may be used to rescue germ cells of endangered adult male birds.
Subject(s)
Chickens , Coturnix , Spermatogenesis , Spermatogonia/transplantation , Spermatozoa/transplantation , Testis/cytology , Transplantation, Heterologous/veterinary , Animals , Breeding , Chickens/physiology , Coturnix/physiology , Endangered Species , Female , Insemination, Artificial , Male , Spermatozoa/physiologyABSTRACT
A method based upon the statistical evaluation of the differentiability of the measure along the trajectory is used to identify determinism in high-dimensional systems. The results show that the method is suitable for discriminating stochastic from deterministic systems even if the dimension of the latter is as high as 13. The method is shown to succeed in identifying determinism in electroencephalogram signals simulated by means of a high-dimensional system.
ABSTRACT
Statistical differentiability of the measure along the reconstructed trajectory is a good candidate to quantify determinism in time series. The procedure is based upon a formula that explicitly shows the sensitivity of the measure to stochasticity. Numerical results for partially surrogated time series and series derived from several stochastic models, illustrate the usefulness of the method proposed here. The method is shown to work also for high-dimensional systems and experimental time series.
ABSTRACT
Genetic and karyotypic studies of some Saccharomyces sensu stricto yeasts from Brazil revealed a genetically isolated population which apparently represents a new sibling species of S. cerevisiae.