Kinetics of ascorbate and dithiothreitol oxidation by soluble copper, iron, and manganese, and 1,4-naphthoquinone: Influence of the species concentration and the type of fluid.

An alternative metric to account for particulate matter (PM) composition-based toxicity is the ability of PM-species to generate reactive oxygen species (ROS) and deplete antioxidants, the so-called oxidative potential (OP). Acellular OP assays are the most used worldwide, mainly those based on ascorbic acid (AA) and dithiothreitol (DTT) depletion; OP values are calculated from AA/DTT concentration over time kinetic curves. Since a great variability in OP-DTT and OP-AA values can be found in the literature, the understanding of those factors affecting the kinetic rate of AA and DTT oxidation in the presence of PM-bound species will improve the interpretation of OP values. In this work, a kinetic study of the oxidation rate of AA and DTT driven by species usually found in PM (transition metals and naphthoquinone (NQ)) was carried out. In particular, the influence of the concentration of Cu(II), Fe(II), Fe(III), Mn(II), Mn(III), and 1,4-NQ, and the type of fluid used in the assay (phosphate buffer (PB), phosphate buffer saline (PBS) and artificial lysosomal fluid (ALF)) is analysed and discussed. The reaction orders with respect to the AA/DTT and the active compound, and the kinetic rate constants were also determined. The results show great variability in OP values among the studied species depending on the fluid used; the OP values were mostly higher in PB0.05 M, followed by PBS1x and ALF. Moreover, different species concentration-responses for OP-DTT/OP-AA were obtained. These differences were explained by the different reaction orders and kinetic rate constants obtained for each active compound in each fluid.

Frequencies of the precision ID ancestry panel markers in Basques using the Ion Torrent PGMTM platform.

The Precision ID Ancestry Panel for the Ion Torrent PGM™ platform is able to genotype 165 autosomal SNPs by massively parallel sequencing (MPS). In the present study, we evaluated the depth of coverage, the locus strand balance and the heterozygote balance of the above panel. The SNPs rs1569175, rs2306040, rs9845457, rs1407434, and rs10007810 showed a low performance, due either to a low coverage, locus strand imbalance or heterozygote imbalance. To further to assess this panel, we analyzed 108 Basques. All loci proved to be in Hardy-Weinberg equilibrium and we did not detect any association between them with the exception of the pair rs3916235-rs4891825 in chromosome 18. The forensic parameters combined match probability and combined power of exclusion were estimated to be 3.13×10-35 and 99.9972%, respectively. In conclusion, the Precision ID Ancestry panel along with this new MPS technology constitute a very promising tool for human identification and biogeographical ancestry inference in routine casework in the forensic field.

Splicing regulator SLU7 preserves survival of hepatocellular carcinoma cells and other solid tumors via oncogenic miR-17-92 cluster expression.

Resisting death is a central hallmark of cancer cells. Tumors rely on a number of genetic mechanisms to avoid apoptosis, and alterations in mRNA alternative splicing are increasingly recognized to have a role in tumorigenesis. In this study, we identify the splicing regulator SLU7 as an essential factor for the preservation of hepatocellular carcinoma (HCC) cells viability. Compared with hepatocytes, SLU7 expression is reduced in HCC cells; however, further SLU7 depletion triggered autophagy-related cellular apoptosis in association with the overproduction of reactive oxygen species. Remarkably, these responses were not observed in primary human hepatocytes or in the well-differentiated HepaRG cell line. Mechanistically, we demonstrate that SLU7 binds the C13orf25 primary transcript in which the polycistronic oncomir miR-17-92 cluster is encompassed, and is necessary for its processing and expression. SLU7 knockdown altered the splicing of the C13orf25 primary transcript, and markedly reduced the expression of its miR-17, miR-20 and miR-92a constituents. This led to the upregulation of CDKN1A (P21) and BCL2L11 (BIM) expression, two bona fide targets of the miR-17-92 cluster and recognized mediators of its pro-survival and tumorigenic activity. Interestingly, altered splicing of miR-17-92 and downregulation of miR-17 and miR-20 were not observed upon SLU7 knockdown in non-transformed hepatocytes, but was found in other (HeLa, H358) but not in all (Caco2) non-hepatic tumor cells. The functional relevance of miR-17-92 dysregulation upon SLU7 knockdown was established when oxidative stress, autophagy and apoptosis were reversed by co-transfection of HCC cells with a miR-17 mimic. Together, these findings indicate that SLU7 is co-opted by HCC cells and other tumor cell types to maintain survival, and identify this splicing regulator as a new determinant for the expression of the oncogenic miR-17-92 cluster. This novel mechanism may be exploited for the development of antitumoral strategies in cancers displaying such SLU7-miR-17-92 crosstalk.

Peripheral blood monocyte and T cell subsets in children with specific polysaccharide antibody deficiency (SPAD).

Specific polysaccharide antibody deficiency (SPAD) is a well reported immunodeficiency characterized by a failure to produce antibodies against polyvalent polysaccharide antigens, expressed by encapsulated microorganisms. The clinical presentation of these patients involves recurrent bacterial infections, being the most frequent agent Streptococcus (S.) pneumoniae. In SPAD patients few reports refer to cells other than B cells. Since the immune response to S. pneumoniae and other encapsulated bacteria was historically considered restricted to B cells, the antibody deficiency seemed enough to justify the repetitive infections in SPAD patients. Our purpose is to determine if the B cell defects reported in SPAD patients are accompanied by defects in other leukocyte subpopulations necessary for the development of a proper adaptive immune response against S. pneumoniae. We here report that age related changes observed in healthy children involving increased percentages of classical monocytes (CD14++ CD16- cells) and decreased intermediate monocytes (CD14++ CD16+ cells), are absent in SPAD patients. Alterations can also be observed in T cells, supporting that the immune deficiency in SPAD patients is more complex than what has been described up to now.

Utilization of advanced-age donors in renal transplantation.

The shortage of organ availability in recent years has made it necessary to use grafts from advanced-aged donors to maintain the rate of renal transplantation in our country. The objective of this study was to evaluate the graft function and patient survival using kidneys from deceased donors of over 65 year of age. From 2005 until 2010, we compared the outcomes of patients who received grafts from donors over 65 years old vs less than 65 years. We observed no significant difference in sex, time on dialysis, or cold ischemia time between the groups. As expected the recipient age was significantly different. For the analysis of survival, we used the Tablecloth-Haenzel test and the Kaplan-Meier survival estimator. Actuarial survivals at 3 years after transplantation showed 84.8% among patients transplanted with kidneys from donors over 65 years old versus 97.5% in the control group. The graft survival was 78.8% among expanded criteria versus 86.85% in the control group. When we analyzed graft survival using an "exitus-censured" analysis, we obtained graft survivals of 89.1% in the expanded criteria kidney group versus 88.6% among the controls. We concluded that the use of kidney from donors over 65 years of age allows us to increase the rate of renal transplantation to about 15 to 20 per million population, with good graft and patient survivals provided that the protocol for expanded criteria organs ensured proper macroscopic and microscopic evaluation of the organ for transplantation.

The effect of operational parameters of the process of sludge ozonation on the solubilisation of organic and nitrogenous compounds.

An evaluation of various operational parameters on the process of sludge ozonation was carried out based on semi-batch experiments. Particular reference has been given to examine the main parameters affecting the solubilisation of organic matter and nitrogenous compounds. Various sets of experiments were undertaken using real sewage sludge to feed a semi-industrial ozonation plant. Applying ozone dosages between 25 and 35 mg O(3)/gTSS, the organic matter solubilisation obtained through ozonation increases proportionally to ozone dosage until a maximum value of 430 mg COD/L. Concerning the nitrogenous compounds, no variation in nitrite concentration and a low increase in nitrate concentration were attained, regardless of the applied ozone dosage. Little increase in ammonia concentration was achieved for low ozone dosages, whilst applying dosages higher than 20 mg O(3)/gTSS, the variation of ammonia increased proportionally with ozone dosage. Experiments using hydraulic retention time (HRT) between 10 and 60 min resulted in a similar COD solubilisation, confirming a rapid rate of cell lysis during ozonation of sludge.

[Impact of conservative oncoplastic techniques in a surgery program for women with breast cancer]. / Impacto asistencial de las técnicas oncoplásticas conservadoras en un programa quirúrgico para enfermas con cáncer de mama.

OBJECTIVES: To evaluate the impact of conservative oncoplastic techniques in a surgery program for women with breast cancer. PATIENTS AND METHOD: The study group was composed of women who underwent a conservative oncoplastic technique and the control group consisted of women who underwent conservative (tumorectomy/quadrantectomy) and radical (mastectomy) techniques. Women with tumors smaller than 3 cm, axillary clinical stages N0-N1a-b and multifocal processes (infiltrating and/or in situ) were eligible for inclusion. Women with T3-4 tumors and reduced breast volume, and those in whom postoperative radiotherapy and disease-free margins during intraoperative study were not feasible were excluded. RESULTS: One hundred sixty women underwent surgery during the study period. Fifty oncoplastic techniques (29 to avoid mastectomy and 21 to improve the result of a conservative technique), 57 conservative techniques and 53 mastectomies were performed. The techniques used for breast reconstruction were superior mammoplasty with transversal scar (23 patients), mammoplasty with superior pedicle (10 patients), mammoplasty with inferior pedicle (10 patients), J-shapped mammoplasty (four patients) and oblique mammoplasty (three patients). The mean operating time in the group undergoing oncoplastic techniques (131 minutes) was higher than that in the group undergoing conservative (56 minutes) and radical (93 minutes) techniques. The incidence of postoperative complications was higher with radical techniques (35%), mainly due to axillary seromas after lymphadenectomy, than with conservative (25%) and oncoplastic (24%) techniques. CONCLUSIONS: Oncoplastic techniques are an effective and efficient alternative to conventional surgical techniques in women with breast cancer.

[Irritable bowel syndrome in patients under age 55 with dyspeptic syndrome]. / Síndrome de intestino irritable en pacientes con síndrome dispéptico menores de 55 años.

The presence of IBS was studied in 289 patients under 55 years of age with dyspeptic syndrome. The subjects were divided into two groups: group I (240 patients with DS) and group II (49 patients with DS and IBS). The mean duration of follow%shy;up was 10.8 years (130 months). IBS was diagnosed in 49 patients (17%). Females accounted for 64% and 77% of groups I and II, respectively (p < 0.05). There were no differences in mean age or the type of FDS. The patients in group II consulted more often and earlier (92% returned for consultation, and took 27 months in doing so) than those in group I (52% and 40 months)(p < 0.05). The number of diagnostic tests requested was greater in group II. Psychiatric pathology was in turn more common in group II than in group I (20 versus 7%)(p < 0.05).

[Prognosis in dyspeptic patients under the age of 55]. / Evolución de los pacientes con dispepsia menores de 55 años.

BACKGROUND: We studied the evolution and diagnostic tests performed in patients with dyspepsia aged less than 55 years. PATIENTS AND METHOD: We included 289 patients under age 55 with clinically manifest dyspepsia who were referred for specialized digestive evaluation from the primary care setting. RESULTS: One half of patients consulted again in the course of the study for the same symptoms. In 131 patients (45%), an endoscopy was proposed at some stage during the study. Gastroduodenal pathology was identified in 37% of the patients who underwent an endoscopy. CONCLUSIONS: In these patients, the approach adopted by the gastroenterologist must comprise assessment of the symptoms with endoscopy.

[Perfusion SPECT with (99m)Tc-HMPAO in type I diabetics with no background of central neurologic symptoms. A study of activation with acetazolamide]. / La SPECT de perfusión con HMPAO-(99m)Tc en los diabéticos tipo I sin antecedentes de clínica neurológica central. Un estudio de activación con acetazolamida.

OBJECTIVE: This study aimed to assess if activation with acetazolamide increases the diagnostic capacity of baseline SPECT with (99m)Tc-HMPAO in the study of brain perfusion in type I diabetic patients with no history of neurological symptoms. MATERIAL AND METHODS: A baseline SPECT was carried out in 11 diabetes mellitus type I patients with no neurological symptoms with 555 MBq of (99m)Tc-HMPAO; 1 g of acetazolamide was administered during the examination and a second SPECT was obtained 20' later with the same methodology used in the baseline SPECT. The images were visually analyzed. The post-acetazolamide studies were analyzed with (CBS) and without (WBS) baseline image subtraction and both methods were compared. RESULTS: The baseline SPECT showed 48 hypoperfused cortical areas. The post-acetazolamide SPECT analyzed without baseline image subtraction detected 14 new hypoperfused areas and those analyzed with it detected 26 areas. 69% of the baseline hypoperfused areas were hyporeactive in the WBS analysis and 54% in the CBS analysis. CONCLUSION: The perfusion SPECT with acetazolamide improves the diagnostic capacity of the baseline perfusion (99m)Tc-HMPAO SPECT, and makes it possible to classify the abnormalities as metabolic or vascular, with a preference for the post-acetazolamide CBS imaging analysis.

Population data on 6 short tandem repeat loci in a sample of Caucasian-Mestizos from Colombia.

Blood samples from 409-452 unrelated Colombian Caucasian-Mestizo individuals were amplified and typed for six short tandem repeat (STR) markers (HUMF13A01, HUMFES/FPS, HUMVWA, HUMCSF1PO, HUMTPOX, HUMTH01). The allele frequencies, genotype frequencies, heterozygosity, mean paternity exclusion chance, polymorphism information content, discrimination power, assumption of independence within and between loci and Hardy Weinberg equilibrium were determined. The results demonstrate that all markers conform to Hardy-Weinberg equilibrium expectations. In addition, the results demonstrate the assumption of independence within and between the loci analysed. The mean exclusion chance (MEC) was 0.9851 for all six STR loci analysed and the discrimination power (DP) was 0.9999973. Therefore, this Colombian population database can be used in identity testing to estimate the frequency of a multiple PCR-based locus DNA profile in forensic cases as well as in paternity testing.

Population frequency for the short tandem repeat loci D18S849, D3S1744, and D12S1090 in Caucasian-Mestizo and African descent populations of Colombia.

Blood samples from 489 unrelated Caucasian Mestizo and 252 individuals of African descent in Colombia were amplified and typed for three short tandem repeat (STR) markers (D12S1090, D3S1744, and D18S849). All markers conformed to Hardy-Weinberg equilibrium expectations in both populations studied. In addition, heterozygosity, mean exclusion chance, polymorphism information content, discrimination power, and the assumption of independence within and between loci were determined. The mean exclusion chance for all three STR markers is 0.9750 in the Caucasian Mestizo population and 0.9731 in the African Colombian Population. The discrimination power is 0.999925 and 0.999911 in the Caucasian Mestizo and African Colombian respectively.

[Inhaled corticosteroids and wheezing post-bronchiolitis]. / Glucocorticoides inhalados y sibilancias posbronquiolitis.

OBJECTIVE: To determine whether inhaled corticosteroid therapy given for 3 months after mild bronchiolitis decreases the incidence and/or severity of wheezing in the following 12 months. DESIGN: Multicentric, single-blind, controlled, randomised intervention study. SETTING: Primary Health Care Centers in Lezo, Beraun, Andoain and Irún (Gipuzkoa, Spain). PATIENTS: Infants less than 12 months old (n = 94) diagnosed with mild bronchiolitis. INTERVENTION: We established two groups of patients: group 1 (n = 47) was treated with inhaled beclomethasone (250 pg/12 hours) using a valved holding chamber (Babyhaler); the treatment started eight days after diagnosis of bronchiolitis and lasted 3 months. Group 2 (n = 47) received no treatment. We compared the number of wheezing episodes and their severity during the intervention period (3 months) and the follow-up period (12 months) with the Students t-test and the Chi-squared test. RESULTS: We studied 89 infants (group 1, n = 42; group 2, n = 47), 67% of whom wheezed during the study period (15 months). There were no significant differences between the treatment and the control group in the study periods. CONCLUSIONS: Inhaled beclomethasone given for 3 months does not significantly modify the occurrence of wheezing episodes during the treatment period or during the following 12 months.

[Follow-up in lung transplant by the study of the pulmonary epithelial permeability and the pulmonary scintigraphies of ventilation and perfusion: preliminary results]. / Seguimiento del trasplante pulmonar mediante el estudio de la permeabilidad epitelial pulmonar y las gammagrafías pulmonares de ventilación y de perfusión: resultados preliminares.

INTRODUCTION: Acute rejection is the most common complication in lung transplantation. OBJECTIVE: This work aimed to assess the contribution of pulmonary clearance of radioaerosols and relative pulmonary perfusion to diagnose acute rejection in lung transplantation. MATERIAL AND METHOD: We have designed a prospective study and present the results obtained for the preliminary phase. This work includes 5 patients who have received a lung transplantation and in whom 28 studies of pulmonary clearance of 99mTc-DTPA and relative pulmonary perfusion with 99mTc microspheres were performed. The pulmonary biopsy diagnosed 9 rejection episodes, 2 associated to CMV infection. RESULTS: The mean radioaerosol clearance time increased when the follow-up was favorable and decreased in 6 of the 9 rejection episodes, including 2 associated to CMV infection. There was only one case with decreased mean clearance time that was not associated to rejection. CONCLUSIONS: Calculating relative pulmonary perfusion can be useful in the follow-up of single lung transplantation but not in bipulmonary ones. Our results suggest that measuring 99mTc-DTPA clearance is useful to suspect a rejection episode.

[Effects of treatment with OKT3 on brain perfusion in a heart transplant patient with SPET with 99mTc-HMPAO]. / Efecto sobre la perfusión cerebral del tratamiento con OKT3 en paciente con trasplante cardíaco mediante SPET con HMPAO-99mTc.

A heart transplant patient treated with OKT3 developed a severe headache which worsened and was accompanied by a sudden decrease in the patient's consciousness level and aphasia when the treatment course was completed. CT was performed and was normal. SPET imaging with 99mTc-HMPAO of cerebral blood flow done 16 hours later revealed multiple and clear focal defects in the blood flow. Analysis of cerebral spinal fluid revealed aseptic pleocytosis. Five days after the completion of treatment, the symptoms remitted and a new control SPET 3 weeks later was completely normal. A diagnosis of neurotoxicity secondary to OKT3 administration was established.