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.

Papel de la biopsia de ganglio centinela en el manejo diagnóstico-terapéutico del melanoma de cabeza y cuello / The role of the sentinel node biopsy in the diagnosis and therapeutic management of melanoma of the head and neck

Objetivo. La creciente incidencia del melanoma facial constituye un auténtico reto en muchas regiones del mundo, a pesar de medidas como la educación sanitaria de la población y realización de screenings poblacionales. Aunque también se emplean terapias no quirúrgicas, lamayoría de los melanomas se tratan en la actualidad mediante excisión quirúrgica. En los últimos años, la biopsia del ganglio centinela se ha incorporadocomo una técnica diagnóstica y terapéutica adicional, y ha permitido una reducción significativa de la morbilidad asociada al manejo quirúrgico del cuello. En el presente artículo pretendemos revisar la experienciade nuestro equipo con el melanoma de cabeza y cuello en los últimos años, así como analizar retrospectivamente la incidencia de metástasis locoregionales, y la predictibilidad de la biopsia de ganglio centinela para su estadiaje. Diseño del estudio. Se ha revisado retrospectivamente nuestra experiencia en los dos últimos años con 12 casos de melanoma de cabeza y cuello en que se realizó linfoescintigrafía y biopsia de ganglio centinela. Resultados. La identificación del ganglio centinela mediante sonda de captacióngamma se consiguió en 11 de los casos (91,6%). Se identificaron un total de 21 ganglios centinela, es decir, 1,75 ganglios por paciente, siendoel nivel II cervical la localización más frecuente. Dos de ellos (9,52%) resultaron afectados por melanoma. La morbilidad residual como consecuencia de la biopsia de los ganglios centinelas ha sido mínima. Conclusiones.A pesar del reducido tamaño de la muestra y de la falta de seguimiento a largo plazo, nuestros resultados son similares a los obtenidos por otros autores, lo que nos induce a considerar la linfoescintigrafía y biopsiade ganglio centinela como una técnica fiable, incluso ya desde fases iniciales de su implantación

Objective. The ongoing incidence of malignant cutaneousmelanoma of the head and neck has become a challenge in many regions of the world, in spite of prophylactic trials such as popular education and screening. Although non-surgical therapies are performed, most melanomas are actually treated by surgical excision. In the last few years, sentinel node biopsy has evolved as a diagnosticand therapeutic tool, and it has permitted a significant minimizing of the morbidity associated with the surgical management of the neck. This article is aimed at providing a thorough review of our experience in head and neck melanoma, as well as a retrospectiveanalysis of locoregional metastases and the predictability of sentinel node biopsy for the staging of melanoma. Design. A retrospective review of our experience over the last two years has been carriedout that includes 12 cases of head and neck melanoma where lymphoscintigraphy and sentinel node biopsy were performed. Results. Sentinel lymph nodes were identified using preoperative lymphoscintigraphy and intraoperative gamma probe in 11 cases (91.6%). A total of 21 nodes were identified in the 12 patients, with an average number of 1.75 nodes per patient. The most frequentsite where nodes were located was level II of the neck. Two nodes (9.52%) were affected by melanoma. There was minimal morbidity related to this procedure. Conclusions. In spite of the reduced size of the sample and the short-term follow-up, our results are similarto those obtained by other authors, which leads us to believe that sentinel lymph node mapping with biopsy is a reliable technique for the diagnosis of regional spread in head and neck cutaneous melanoma, even in early development stages

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

Objetivos. Evaluar el impacto asistencial de las técnicas oncoplásticas conservadoras en un programa quirúrgico de mujeres con cáncer de mama. Pacientes y método. El grupo a estudio lo constituyeron enfermas sometidas a una técnica oncoplástica conservadora mientras que el grupo control lo formaron mujeres intervenidas con técnicas conservadoras (tumorectomía/cuadrantectomía) y radicales (mastectomía). Los criterios de inclusión para una técnica oncoplástica comprendieron mujeres con tumores de diámetro inferior a 3 cm, tumores T2 tratados con quimioterapia neoadyuvante y reducidos a un tamaño menor de 3 cm, en estadios clínicos axilares N0-N1a-b y procesos multifocales (infiltrante y/o in situ). Se excluyó del estudio a las mujeres con tumores T3-4, imposibilidad para la radioterapia postoperatoria, volumen mamario reducido e imposibilidad de márgenes libres de enfermedad durante el estudio intraoperatorio. Resultados. Durante el período se intervino a 160 mujeres en las que se practicaron 50 técnicas oncoplásticas (29 para evitar una mastectomía y 21 para mejorar el resultado de la técnica conservadora), 57 técnicas conservadoras y 53 mastectomías. Las técnicas utilizadas para la remodelación mamaria fueron la mamoplastia horizontal (23 casos), la mamoplastia vertical de pedículo superior (10 casos), la mamoplastia vertical de pedículo inferior (10 casos), la mamoplastia de rotación (4 casos) y la mamoplastia lateral (3 casos). El grupo de técnicas oncoplásticas presentó un tiempo medio de intervención (131 min) superior a las técnicas conservadoras (56 min) y radicales (93 min). La incidencia de complicaciones postoperatorias fue mayor en las técnicas radicales (35%), fundamentalmente por seromas axilares tras linfadenectomía, respecto a las técnicas conservadoras (25%) y oncoplásticas (24%). Conclusiones. Las técnicas oncoplásticas constituyen una alternativa eficaz y eficiente a las técnicas quirúrgicas convencionales en la mujer con cáncer de mama (AU)

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 (AU)

[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).

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

Se estudió la presencia de síndrome intestino irritable (SII) en 289 pacientes menores de 55 años con síndrome dispéptico (SD). El número de pacientes diagnosticados de SII fue 49 (17 por ciento).El grupo I se consideró a los pacientes con SD (240 pacientes), mientras que el grupo II eran los pacientes con SD y SII (49 pacientes). El grupo II acudió con más frecuencia a las consultas y antes (92 por ciento volvio a las consultas y tardaron 27 meses en volver) que los pacientes del grupo I (52 por ciento y 40 meses) (p<0,05). El número de pruebas diagnósticas solicitadas fue superior en los pacientes del grupo II. La patología psiquiátrica fue más frecuente en el grupo II que en el grupo I (20 por ciento frente al 7 por ciento) (p<0,05). Los pacientes con SD que son diagnosticados de SII, acuden con más frecuencia y antes a la consulta. El número de pruebas diagnósticas solicitadas por paciente es mayor en los pacientes con SII y SD (AU)

[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.

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 / The perfusion SPECT with 99m Tc-HMPAO in type I diabetics with no background of central neurologic symptoms. A study of activation with acetazolamide

Objetivo: El objetivo de este trabajo fue evaluar si la activación con acetazolamida incrementa la capacidad diagnóstica de la SPECT basal con HMPAO-99mTc en el estudio de la perfusión cerebral de los diabéticos tipo I sin clínica neurológica central (DMISCNC). Material y métodos: En 11 pacientes DMISCNC, realizamos una SPECT basal con 555 MBq de HMPAO-99mTc, durante la adquisición de las imágenes se inyectó 1 g de acetazolamida y a los 20' se realizó una segunda SPECT con la misma metodología que la basal. Las imágenes fueron analizadas visualmente. Los estudios post-acetazolamida se evaluaron con y sin substracción de las imágenes basales (CSB/SSB) comparando ambos métodos de evaluación. Resultados: La SPECT basal detectó 48 hipoperfusiones corticales. La SPECT post-acetazolamida analizada SSB reveló 14 nuevas áreas y analizada CSB 26. El 69 por ciento de las hipoperfusiones basales eran hiporreactivas en el análisis SSB y el 54 por ciento en el análisis CSB.Conclusión: la SPECT de perfusión tras estimulación con acetazolamida aumenta la capacidad diagnóstica de la SPECT de perfusión basal con HMPAO-99mTc, y tipifica las alteraciones como metabólicas o vasculares, siendo preferible el análisis de las imágenes post-acetazolamida CSB (AU)

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