Heterologous expression of phaC2 gene and poly-3-hydroxyalkanoate production by recombinant Cupriavidus necator strains using canola oil as carbon source.

Many heterologous transformation studies have been carried out using the Cupriavidus necator PHB-4 strain to investigate the expression characteristics of various polyhydroxyalkanoate (PHA) synthase enzymes. In this study, we generated a recombinant C. necator PHB-4 strain by transforming a plasmid (pMRC03) harbouring the synthetic phaC2 gene of Pseudomonas putida CA-3. Under conditions favourable for expression of the phaC2 P.putCA-3 gene, canola oil was used as carbon source for the synthesis of PHAs. The expressed synthase polymerised monomers of 3-hydroxybutyrate (3-HB), 3-hydroxyvalerate (3-HV) and 3-hydroxyhexanoate (3-HHx) in the recombinant C. necator PHB-4 (pMRC03) strain. We then co-expressed the phaC2P.putCA-3 gene with the native phaC1C.ne gene in wild type Cupriavidus necator H16 (C. necator H16 (pMRC03)). This co-expression produced a PHA blend of 3-HB, 3-HV, 3-HHx and 3-hydroxyoctanoate (3-HO) monomers in the presence of canola oil. Gas chromatography analysis revealed the presence of 94mol% 3-HB, 1mol% 3-HV, 4mol% 3-HHx and 1mol% 3-HO in a tetra-polymer. Thus, we confirmed that a synthetic phaC2 gene encoding the synthase enzyme is functionally active with substrates ranging from short to medium chain length PHAs.

Incidence of osmophilic yeasts and Zygosaccharomyces rouxii during the production of concentrate grape juices.

Zygosaccharomyces rouxii is the main spoilage yeast of grape juice concentrates. Detection and identification of Z. rouxii during the production of grape juice concentrate is critical to prevent spoilage in the final product. In this work, three grape juice concentrate processing plants were assessed by identifying osmophilic yeasts in juices and surfaces during different stages of a complete production line. Subsequently, molecular typing of Z. rouxii isolates was done to determine the strain distribution of this spoilage yeast. Osmotolerant yeast species, other than Z. rouxii, were mainly recovered from processing plant environments. Z. rouxii was only isolated from surface samples with grape juice remains. Z. rouxii was largely isolated from grape juice samples with some degree of concentration. Storage of grape juice pre-concentrate and concentrate allowed an increase in the Z. rouxii population. A widely distributed dominant molecular Z. rouxii pattern was found in samples from all three processing plants, suggesting resident microbes inside the plant.

[Primary hypospadias]. / Hipospadias primario.

Hypospadias is one of the most frequent male congenital anomalies. Its surgical correction is under permanent review and it is always controversial. The best surgical technique can only be chosen intraoperatively, since it is the level of corpus spongiosum division what marks the severity of the case, although it is essential to evaluate position of the meatus, penile curvature, quality of the preputial skin and penile size. It is recommended treatment age between 6-12 months. Nowadays, The most frequently used technique for hypospadias repair is the Snodgrass tubularized incised urethral plate (TIP). Moreover, distal and medial hypospadias may be treated with urethral advance or flap techniques and proximal hypospadias with modifications of these in one-step or two-step procedures. Nevertheless, there are other controversies about hypospadias, such as to preserve or not the prepuce, the use of central or peripheral anesthesia blockade, or the use of wound dressings.

Hipospadias primario / Primary hypospadias surgery

El hipospadias es una de las anomalías congénitas más frecuentes en el hombre. Su corrección quirúrgica se encuentra en permanente revisión y es siempre controvertida. La mejor técnica quirúrgica solo puede elegirse intraoperatoriamente, ya que es el nivel de división del cuerpo esponjoso el que marca la gravedad del cuadro aunque es fundamental la valoración de la posición del meato, la curvatura peneana, la calidad de la piel prepucial y el tamaño del pene. Se recomienda que la edad del tratamiento se sitúe entre 6 y los 12 meses como máximo. La técnica de reparación del hipospadias más utilizada en la actualidad es la uretroplastia con tubulo-incisión de la placa uretral de Snodgrass (TIP). Además los hipospadias distales y medios pueden tratarse con técnicas de avance uretral o técnicas de colgajos y los proximales en un tiempo con modificaciones de estas mismas técnicas o en dos tiempos. Pero en torno al hipospadias existen también otros temas controvertidos, como la conservación o no del prepucio, el uso de bloqueos centrales o periféricos en la anestesia o el uso de apósitos

Hypospadias is one of the most frequent male congenital anomalies. Its surgical correction is under permanent review and it is always controversial. The best surgical technique can only be chosen intraoperatively, since it is the level of corpus spongiosum division what marks the severity of the case, although it is essential to evaluate position of the meatus, penile curvature, quality of the preputial skin and penile size. It is recommended treatment age between 6-12 months. Nowadays, The most frequently used technique for hypospadias repair is the Snodgrass tubularized incised urethral plate (TIP). Moreover, distal and medial hypospadias may be treated with urethral advance or flap techniques and proximal hypospadias with modifications of these in one-step or two-step procedures. Nevertheless, there are other controversies about hypospadias, such as to preserve or not the prepuce, the use of central or peripheral anesthesia blockade, or the use of wound dressings

Incidence and risk factors for infections after liver transplant: single-center experience at the University Hospital Fundación Santa Fe de Bogotá, Colombia.

The incidence of infections in liver transplant patients is higher compared with recipients of other organs, and infections are one of the major complications after transplantation. The aim of our study was to evaluate the incidence, presentation and risk factors of infections in liver transplant recipients in a Latin-American population, and to compare the results with data worldwide. We performed a retrospective analysis of 94 consecutive patients undergoing liver transplantation between 2004 and 2008 at the University Hospital Fundación Santa Fe de Bogotá, Colombia. The patients contributed a total of 64.4 person years (PY). Fifty-two patients (55.3%) developed one or more infections, in total 83 events (128.9 infections/100 PY). Bacterial infections represented the most frequent event (71.1%), followed by viral (19.3%) and fungal infections (8.4%). In 1%, no causative organism was identified. More than one-third of infections (37%) occurred during the first 30 days, whereas 83% of all events were seen during the first 6 months. The most common site of pathogen localization was the bloodstream (25.3%), followed by the urinary tract (15.7%), liver with bile tract (14.5%), abdomen (10.8%), surgical site (7.2%), and lungs (9.6%). The overall mortality after 1 year was 14.9%, and 57.1% of the deaths were attributed to infections. We found that risk factors significantly associated with increased incidence rate ratio for infection were prolonged stay at the intensive care unit, the need for parenteral nutrition, and blood transfusion requirement. Our data provide additional information about etiology and epidemiology of infections after liver transplantation.

Production of polyhydroxyalkanoates (PHAs) with canola oil as carbon source.

Wautersia eutropha was able to synthesize medium chain length polyhydroxyalkanoates (PHAs) when canola oil was used as carbon source. W. eutropha was cultivated using fructose and ammonium sulphate as carbon and nitrogen sources, respectively, for growth and inoculum development. The experiments were done in a laboratory scale bioreactor in three stages. Initially, the biomass was adapted in a batch culture. Secondly, a fed-batch was used to increase the cell dry weight and PHA concentration to 4.36 g L(-1) and 0.36 g L(-1), respectively. Finally, after the addition of canola oil as carbon source a final concentration of 18.27 g L(-1) PHA was obtained after 40 h of fermentation. With canola oil as carbon source, the polymer content of the cell dry matter was 90%. The polymer was purified from dried cells and analyzed by FTIR, NMR and DSC using PHB as reference. The polymer produced by W. eutropha from canola oil had four carbon monomers in the structure of the PHA and identified by 1H and 13C NMR analysis as 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), 3-hydroxyoctanoate (3HO), and 3-hydroxydodecanoate (3HDD).

Endospore dipicolinic acid detection during Bacillus thuringiensis culture.

AIMS: The aim of this work was to detect Bacillus thuringiensis endospore production during fermentation under conditions hindering endospore detection, i.e. in a complex undefined industrial medium with a high content of solids in suspension. METHODS AND RESULTS: Bacterial endospore production was measured using the photoluminescence of dipicolinate (DPA) with Tb3+. The high temperature and pressure of a conventional autoclave was used to release DPA from the endospores. The endospore was obtained from B. thuringiensis var. kurstaki HD-73 fermentations in industrial-type media with 25.1 and 54.1 g l(-1) glucose, 4.4 and 35.3 g l(-1) soybean meal, 5.8 g l(-1) yeast extract, 9.2 g l(-1) corn steep solids and mineral salts. CONCLUSIONS: In this study, we successfully determined the DPA concentrations during the culture of B. thuringiensis in high-concentration soybean meal media. A good correlation was found between microscope endospore counting and DPA measurements in the cultures. SIGNIFICANCE AND IMPACT OF THE STUDY: Because of synergy between Cry protein and endospore in B. thuringiensis bioinsecticides formulation, it is important to be able to determine endospore development during B. thuringiensis industrial-type fermentation, in order to ascertain the beginning of sporulation.

[Giant renal oncocytoma]. / Oncocitoma renal gigante.

Renal oncocytoma is a benign neoplasms arising from cells of the distal renal tubule. They acount for 3-7% of all renal tumors. most are incidental findings. Differential diagnosis with renal cells carcinoma is often difficult. Here we report a case of big renal oncocytoma as an incidental finding while performing an abdominal ultrasound in a patient with low abdominal pain. We also review the diagnostic an therapeutic approach in this kind of malignancies.

Oncocitoma renal gigante / Giant renal oncocytoma

El oncocitoma renal es un tumor benigno derivado de las células del túbulo renal distal. Aunque no es muy frecuente, representa entre el 3-7 % de las masas renales. La mayoría de las ocasiones se presenta como un hallazgo incidental en pruebas de imagen realizadas por otros motivos. El diagnóstico diferencial preoperatorio con el carcinoma de células renales es complejo y a menudo sólo se puede confirmar después de la cirugía. Presentamos un caso clínico de oncocitoma renal de gran tamaño diagnosticado mediante una ecografía abdominal realizada por molestias en hipogastrio. Revisamos el enfoque diagnóstico y terapéutico de este tipo de tumores (AU)

Renal oncocytoma is a benign neoplasms arising from cells of the distal renal tubule. They acount for 3-7% of all renal tumors. Most are incidental findings. Differential diagnosis with renal cells carcinomais often difficult. Here we report a case of big renal oncocytoma as an incidental finding while performingan abdominal ultrasound in a patient with low abdominal pain. We also review the diagnostican therapeutic approach in this kind of malignancies (AU)

Degradation of the main components of cellulose-paint thinner by the mould Scopulariopsis brevicaulis cultured on rice hulls.

AIMS: Biodegradation of the main components of the cellulose-paint thinner (toluene, acetone, isopropanol and xylenes) by Scopulariopsis brevicaulis, isolated from a thinner biodegradation microbial consortium was investigated. METHODS AND RESULTS: Our results showed that 90% of S. brevicaulis conidia survived after 4 weeks in a cellulose-paint thinner saturated atmosphere. The mould was able to grow under these environmental conditions with a low development of conidia. The biodegradation potential of S. brevicaulis was established with and without support material (rice hulls). Biodegradation without support was very limited, <10% for all the components quantified. There was notable thinner biodegradation when the fungus was grown on rice hulls. CONCLUSIONS: Our results suggest the potential use of fungi in biofiltration systems employed in biodegradation of the main components of the cellulose-paint thinner. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of volatile organic compounds biodegradation by this fungal species.

[Spinal angiolipoma: case report]. / Angiolipoma espinal: presentación de un caso.

Spinal extradural angiolipomas are rare benign tumours, accounting for only 0.14-1.2% of all spinal neoplasms. They are usually localized within the thoracic extradural space of the spinal canal and their common clinical presentation is myelopathy, mainly in the way of a slowly progressive paraparesis and sometimes in an acute form. We report the case of a 85 year old man who complained of bilateral lower limb weakness with sphincter disturbance of acute instauration. The MRI showed a posterior extradural lesion at L1-L2 level that compressed conus medularis and cauda equina. He underwent urgent surgical treatment and we achieved total resection of the lesion via a L1-L2 laminectomy. The pathological examination confirmed the tumour as an angiolipoma.

A randomized, controlled trial of weekly administration of lymphoblastoid interferon in patients with chronic hepatitis C.

To evaluate the efficacy of a treatment of weekly interferon administration in patients with chronic hepatitis C, 36 patients were randomly assigned to two groups. In one group lymphoblastoid interferon was given at a dose of 6 million units, intramuscularly, once per week for 24 weeks, and no treatment was given to the other. Serum alanine aminotransferase levels in the treated group were significantly lower during therapy than in the control group, although there was no significant difference between these two groups before therapy. The normalization of serum alanine aminotransferase levels at the end of therapy was observed in 50% of the treated group, and in 11.1% of the control group. This difference was statistically significant (p < 0.03). Response to interferon was better in patients with chronic persistent hepatitis or with chronic active hepatitis than in patients with chronic active hepatitis with cirrhosis. Relapse after the end of therapy was observed in 83.3% of the responders. These results indicate that the weekly administration of 6 million units of lymphoblastoid interferon is effective in decreasing serum alanine aminotransferase levels in patients with type C chronic persistent hepatitis or chronic active hepatitis.

Effect of dexamethasone, dimethylsulfoxide and sodium butyrate on a human hepatoma cell line PLC/PRF/5.

The effects of agents which are known to be differentiation inducers on a human hepatoma cell line PLC/PRF/5 were investigated. Dexamethasone (DEX), sodium butyrate (SB) or dimethylsulfoxide (DMSO) were examined. They all reduced cell proliferation but differ from each other in effect on the secretion of alphafetoprotein (AFP) and hepatitis B surface antigen (HBsAg), changes in morphology and RNA transcription. SB changed the cell from polygonal into a fibroblast-like type and decreased AFP secretion. DMSO decreased the cell size and changed AFP secretion in the same manner as SB. DEX changed the cell into a larger size, as well as increased AFP secretion. HBsAg secretion and also HB virus DNA transcription was enhanced by 3 agents. AFP and myc gene transcriptions were reduced by SB but DMSO reduced only AFP. Albumin gene transcription was enhanced by SB and DEX. These results indicate that the decrease of PLC/PRF/5 proliferation is induced through different mechanisms by these 3 agents.

Treatment of chronic hepatitis C with weekly administration of interferon-alpha.

To evaluate the efficacy of weekly administration of interferon (IFN)-alpha, we studied 23 anti-HCV positive patients with chronic hepatitis diagnosed by liver needle biopsy. Thirteen patients received weekly intramuscular injections of 6 MU human lymphoblastoid interferon (HLBI) for 24 weeks, and the other 10 patients were given no treatment. We examined liver-specific idiotype-bearing antibody (LSIA) in the patients' sera. This HLBI treatment was easily tolerated by all the treated patients. In the treated group serum alanine aminotransferase (ALT) level significantly decreased during HLBI treatment. Normalization of serum ALT level by the end of treatment was observed in 7/13 (54%) of the treated patients but in 0% of the non-treated patients. Anti-HCV was detectable in all the patients during the treatment. Those who had high LSIA levels did not respond to HLBI treatment. These results demonstrate that weekly administration of IFN is sufficient to suppress disease activity in patients with chronic hepatitis C and that patients with high LSIA levels are unlikely to respond to IFN therapy.

[Levels of L-carnitine in the Venezuelan infant population]. / Niveles de L-carnitina en la población infantil venezolana.

The L-carnitine blood serum and amniotic fluid levels were measured in 133 samples obtained from clinically healthy patients: 39 pregnant women with a fetal gestational age ranging from 14 to 40 weeks, 13 newborn children less than a day old, 19 newborn children between the ages of 1 and 30 days, 8 breast-feed babies and 19 children over two years of age. No significant statistical differences were seen in the maternal blood serum and amniotic fluid samples for the different gestational ages considered in the study. The average values were found to be 22.6 +/- 5.1 nmol/mL for maternal blood serum and 25.3 +/- 9.4 nmol/mL for amniotic fluid. The blood serum levels were found to be greater in the group between the ages of 1 to 30 days, reaching levels of 43.1 +/- 7.4 nmol/mL. In the group aged 1 month-18 years, the serum levels were on the average 34.3 +/- 6.7 nmol/mL. The variations found among these groups reflect characteristics specific of our population. These values should be further researched since they differ from those values reported by the Anglo-Saxons. L-carnitine concentration; blood serum level; amniotic fluid level.