Association of SOD1 gene variants (50 bp Ins/Del, rs4817415, rs2070424, rs1041740, rs17880135) with plasma protein levels in vitiligo patients and their analysis in silico.

OBJECTIVE: Vitiligo is a common systemic, idiopathic autoimmune disease. The aim of this study was to analyze the frequency of variants of the superoxide dismutase 1 (SOD1) gene (50 bp Ins/Del, rs4817415, rs2070424, rs1041740, rs17880135) and circulating plasma protein levels through in-silico analysis. PATIENTS AND METHODS: Blood samples were collected from adult patients of both sexes with a clinical diagnosis of vitiligo. ELISA tests for SOD and analysis of gene variants by qPCR were compared to a disease-free reference group. RESULTS: The population analyzed was young people between 29 and 37 years old, with a higher percentage of women. The population was found in the Hardy-Weinberg equilibrium (HWE). The 50 bp Ins/Del, rs4817415, and rs2070424 variants showed no significant difference between groups (p > 0.05). Although, in the dominant model, the CT and CTTT genotypes of the rs1041740 and rs17880135 variants showed an association with susceptibility to vitiligo compared to the control. Plasma SOD levels showed significant differences between the groups, and when stratified according to the genotypes of each variant, there was a significant difference, except with the rs17880135 variant. The haplotypes InsCGTC and InsAGCC are shown to be risk factors for susceptibility to vitiligo. The in-silico analysis demonstrated that the rs4817415, rs2070424, rs1041740, and rs17880135 variants of the SOD1 gene participate in the modification of selected regulatory elements for differentiating the protein, transcription factors, and long non-coding RNA. CONCLUSIONS: Information regarding the pathogenesis of vitiligo helps recognize risk factors and identify the relationship of diagnostic markers of cell damage inherent to the disease. This will help improve aspects of prevention and the choice of treatment alternatives appropriate to each case.

Variants rs2758346, rs5746094, and rs2758331 of SOD2 gene: association study with breast cancer in a Mexican population and their analysis in silico.

OBJECTIVE: The aim of the study was to analyze the association between the superoxide dismutase 2 (SOD2) gene variants rs2758346, rs5746094, and rs2758331 and breast cancer (BC) in the Mexican population as well as to perform in silico assessments of the variants' potential impact. PATIENTS AND METHODS: We performed in silico analysis and analyzed 489 healthy women and 467 BC patients using TaqMan assays and Real-Time PCR. RESULTS: The TT genotype, the T allele of the rs2758346 variant, and the CC genotype of both rs5746094 and rs2758331 were identified as BC risk factors (p < 0.05). The TT and CTTT genotype of the rs2758346 variant stratified by the presence of ki-67 (> 20%), TCCC, and estrogen receptor (ER)-positive of the rs5746094 variant, and the CC and CT genotypes of rs2758331 stratified by menopause status and non-chemotherapy response were risk factors. The TTC and TTA haplotypes are risk factors for BC. In silico analysis revealed that the rs2758346, rs5746094, and rs2758331 variants could influence SOD2 gene regulation by transcription factors and circulating RNAs (circRNAs). CONCLUSIONS: The rs2758346, rs5746094, and rs2758331 variants of the SOD2 gene were associated with BC risk and could influence SOD2 regulation by transcription factors and circRNAs.

[Comparison of clinical outcomes of anterior cruciate ligament repair in the active military using allograft versus autograft, 2-year follow-up]. / Comparación de resultados clínicos de reparación de ligamento cruzado anterior en militares en el activo con uso de aloinjerto versus autoinjerto, seguimiento de 2 años.

INTRODUCTION: Due to the inherent activities of the active duty personnel of the Mexican Navy, ligament injuries are constant, particularly the anterior cruciate ligament of the knee (ACL). Currently, we have various techniques and resources for its repair. OBJECTIVE: identify the clinical results obtained in anterior cruciate ligament reconstruction in active military. MATERIAL AND METHODS: retrospective observational study comparing clinical outcomes of anterior cruciate ligament repair in active military with the use of allograft and autograft in 23 patients who met inclusion criteria for the surgical procedure from 2017 to 2019 at the Naval Medical Center. 23 patients (46 in total) were considered for each category of anterior cruciate ligament repair with autologous graft (contralateral patellar) and heterologous graft (cadaveric anterior cruciate ligament). With an average age of autologous (35.6 years), heterologous (35 years). BMI average: autologous (26.5), heterologous (26.5). Male gender in its entirety. The IKDC and Lysholm scales were applied to all patients for the evolution of subjective results of clinical improvement in a 2-year follow-up; where a significant difference (p = 0.0001) could be observed when comparing both anterior cruciate ligament reconstruction techniques. RESULTS: we included 46 patients who underwent anterior cruciate ligament reconstruction for indication of complete injury respectively by the Joint Surgery Service of the Naval Medical Center. A better level of clinical benefit was observed in patients with heterologous graft, both in evaluation by IKDC scale (median 95.52 ± 1.85) as in Lysholm scale (median 94.91 ± 1.62) compared to autologous grafts, IKDC (median 89.92 ± 2.55) and Lysholm (median 86.04 ± 5.58), with value of p = 0.0001 for both cases. CONCLUSIONS: The results our study suggests that a superiority of functionality is obtained as reported by patients in whom heterograft was used.

INTRODUCCIÓN: Debido a las actividades inherentes del personal del servicio activo de la Armada de México, son constantes las lesiones ligamentarias, en particular la lesión del ligamento cruzado anterior de la rodilla (LCA). Actualmente, contamos con diversas técnicas y recursos para su reparación. OBJETIVO: identificar los resultados clínicos obtenidos en reconstrucción de ligamento cruzado anterior en militares en el activo. MATERIAL Y MÉTODOS: estudio observacional retrospectivo donde se compararon los resultados clínicos de reparación de ligamento cruzado anterior en militares en el activo con uso de aloinjerto y autoinjerto en 46 pacientes que cumplieron criterios de inclusión para el procedimiento quirúrgico del año 2017 al 2019 en el Centro Médico Naval. Se incluyeron 23 pacientes para cada rubro de reparación de ligamento cruzado anterior con injerto autólogo (patelar contralateral) e injerto heterólogo (ligamento cruzado anterior cadavérico). A todos los pacientes se les aplicaron las escalas IKDC y Lysholm, para la evaluación de resultados subjetivos de mejoría clínica a un seguimiento de dos años. RESULTADOS: se reclutaron 46 pacientes que fueron sometidos a reconstrucción de ligamento cruzado anterior con aloinjerto o con autoinjerto. La media de edad por grupo fue: autólogo 35.6 años, heterólogo 35 años. Índice de masa corporal (IMC) promedio: autólogo 26.5, heterólogo 26.5. Sexo masculino en su totalidad. Se observó un mejor nivel de beneficio clínico en los pacientes con injerto heterólogo, tanto en evaluación por escala de IKDC (media 95.52 ± 1.85) como en escala de Lysholm (media 94.91 ± 1.62), en comparación con los injertos autólogos, IKDC (media 89.92 ± 2.55) y Lysholm (media 86.04 ± 5.58), con diferencia significativa (p = 0.0001) al comparar ambas técnicas de reconstrucción de ligamento cruzado anterior. CONCLUSIÓN: Los resultados de nuestro estudio sugieren que se obtiene una superioridad de funcionalidad referida por los pacientes en los que se utilizó heteroinjerto.

Comparación de resultados clínicos de reparación de ligamento cruzado anterior en militares en el activo con uso de aloinjerto versus autoinjerto, seguimiento de 2 años / Comparison of clinical outcomes of anterior cruciate ligament repair in the active military using allograft versus autograft, 2-year follow-up

Resumen: Introducción: Debido a las actividades inherentes del personal del servicio activo de la Armada de México, son constantes las lesiones ligamentarias, en particular la lesión del ligamento cruzado anterior de la rodilla (LCA). Actualmente, contamos con diversas técnicas y recursos para su reparación. Objetivo: identificar los resultados clínicos obtenidos en reconstrucción de ligamento cruzado anterior en militares en el activo. Material y métodos: estudio observacional retrospectivo donde se compararon los resultados clínicos de reparación de ligamento cruzado anterior en militares en el activo con uso de aloinjerto y autoinjerto en 46 pacientes que cumplieron criterios de inclusión para el procedimiento quirúrgico del año 2017 al 2019 en el Centro Médico Naval. Se incluyeron 23 pacientes para cada rubro de reparación de ligamento cruzado anterior con injerto autólogo (patelar contralateral) e injerto heterólogo (ligamento cruzado anterior cadavérico). A todos los pacientes se les aplicaron las escalas IKDC y Lysholm, para la evaluación de resultados subjetivos de mejoría clínica a un seguimiento de dos años. Resultados: se reclutaron 46 pacientes que fueron sometidos a reconstrucción de ligamento cruzado anterior con aloinjerto o con autoinjerto. La media de edad por grupo fue: autólogo 35.6 años, heterólogo 35 años. Índice de masa corporal (IMC) promedio: autólogo 26.5, heterólogo 26.5. Sexo masculino en su totalidad. Se observó un mejor nivel de beneficio clínico en los pacientes con injerto heterólogo, tanto en evaluación por escala de IKDC (media 95.52 ± 1.85) como en escala de Lysholm (media 94.91 ± 1.62), en comparación con los injertos autólogos, IKDC (media 89.92 ± 2.55) y Lysholm (media 86.04 ± 5.58), con diferencia significativa (p = 0.0001) al comparar ambas técnicas de reconstrucción de ligamento cruzado anterior. Conclusión: Los resultados de nuestro estudio sugieren que se obtiene una superioridad de funcionalidad referida por los pacientes en los que se utilizó heteroinjerto.

Abstract: Introduction: Due to the inherent activities of the active duty personnel of the Mexican Navy, ligament injuries are constant, particularly the anterior cruciate ligament of the knee (ACL). Currently, we have various techniques and resources for its repair. Objective: identify the clinical results obtained in anterior cruciate ligament reconstruction in active military. Material and methods: retrospective observational study comparing clinical outcomes of anterior cruciate ligament repair in active military with the use of allograft and autograft in 23 patients who met inclusion criteria for the surgical procedure from 2017 to 2019 at the Naval Medical Center. 23 patients (46 in total) were considered for each category of anterior cruciate ligament repair with autologous graft (contralateral patellar) and heterologous graft (cadaveric anterior cruciate ligament). With an average age of autologous (35.6 years), heterologous (35 years). BMI average: autologous (26.5), heterologous (26.5). Male gender in its entirety. The IKDC and Lysholm scales were applied to all patients for the evolution of subjective results of clinical improvement in a 2-year follow-up; where a significant difference (p = 0.0001) could be observed when comparing both anterior cruciate ligament reconstruction techniques. Results: we included 46 patients who underwent anterior cruciate ligament reconstruction for indication of complete injury respectively by the Joint Surgery Service of the Naval Medical Center. A better level of clinical benefit was observed in patients with heterologous graft, both in evaluation by IKDC scale (median 95.52 ± 1.85) as in Lysholm scale (median 94.91 ± 1.62) compared to autologous grafts, IKDC (median 89.92 ± 2.55) and Lysholm (median 86.04 ± 5.58), with value of p = 0.0001 for both cases. Conclusions: The results our study suggests that a superiority of functionality is obtained as reported by patients in whom heterograft was used.

SOD1 gene variants rs4817415, rs2070424, and rs1041740 and their association with breast cancer risk.

OBJECTIVE: The aim of this investigation was to determine the frequency and association of the variants rs4817415, rs2070424, and rs1041740 of the SOD1 gene in healthy women and breast cancer (BC) patients. PATIENTS AND METHODS: Genomic DNA samples from 146 healthy women and 130 patients with BC were analyzed. RESULTS: GG genotype (OR 2.54, 95% CI 1.31-4.91, p = 0.0073) and the G allele (OR 1.37, 95% CI 1.09-1.73, p = 0.007) of the rs2070424 variant and CC genotype (OR 1.67, 95% CI 1.04-0.2.70, p = 0.0444) and allele C (OR 1.58, 95% CI 1.09-2.29, p = 0.0183) of the rs1041740 variant of SOD1 gene were associated as risk factors for BC susceptibility relative to the control group. Study groups comparison of the stratification by menopausal status showed an association of susceptibility to BC risk with carriers of the GG genotype (OR 2.9, 95% CI 1.11-7.81, p = 0.042) of the rs2070424 variant and with the premenopausal status of the study group and the TT (OR 2.89, 95% CI 1.73-4.85, p = 0.001) genotype of the rs1041740 variant. Furthermore, differences were observed in the patients with BC who were carriers of the CC genotype of the rs4817415 variant with elevated Ki-67 (≥ 20%) and who presented lymph node metastasis and stage III-IV BC (p<0.05). Two common haplotypes were identified in the study groups: CAC (protective factor), and CGC (risk factor) (p<0.05). CONCLUSIONS: The rs2070424 and rs1041740 variants of the SOD1 gene and the CGC haplotype were associated as risk susceptibility factors of BC in this sample analyzed.

The rs1008562, rs2234671, and rs3138060 polymorphisms of the CXCR1 gene are associated with breast cancer risk in a Mexican population.

OBJECTIVE: The rs1008562, rs2234671 and rs3138060 polymorphisms of the CXCR1 gene have been shown to be associated with many diseases, but in breast cancer (BC) their association has not been detected. The purpose of this study was to determine the frequency and association of the rs1008562, rs2234671 and rs3138060 polymorphisms of CXCR1 gene in BC patients in the Mexican population. PATIENTS AND METHODS: The CXCR1 polymorphisms were determined by Polymerase Chain Reaction (PCR) and real time-PCR in healthy Mexican subjects and BC patients. RESULTS: The prevalent patron in BC patients was observed, the majority were overweight and obesity (72%) with metastatic lymph nodes (48%), luminal A/B subtypes (63%), and advanced stages (60%). Triple negative breast cancer (TNBC) patients: they were younger (58%) than 43 years old, overweight (33%), obesity (42%), ductal type histological (98%), metastasis to lymph nodes (47%), advanced stages III-IV (61%) and metastasis (33%). The rs2234671 polymorphism was associated with BC susceptibility when BC patients and the control group were compared for the CC genotype (p=0.037), CG (heterozygous model: p=0.018), GC/CC (dominant model: p=0.004), and the C allele (p=0.001), as well as the GC/CC genotype with hormone replace therapy (HRT, p=0.016). The rs3138060 polymorphism was associated with BC susceptibility for CG/GG genotype (dominant model: p=0.032) and G allele (p=0.018). Although the association between the dominant model of rs1008562, rs2234671, rs3138060 polymorphisms and BC patients and control was evident for tobacco and alcohol consumption (p<0.05). The rs1008562, rs2234671, and rs3138060 polymorphisms of the CXCR1 gene classified by molecular subtype and stage were also associated with BC patients, indicating that these factors may significantly contribute to BC risk. The CCC (OR 1.75, 95% CI 1.03- 2.97, p=0.046), GGG (OR 3.73, 95% CI 1.61- 8.65, p=0.0018) haplotypes were also associated with BC susceptibility. CONCLUSIONS: Rs2234671 and rs3138060 polymorphisms in the CXCR1 gene were associated with BC susceptibility in the Mexican population. The dominant model of the rs1008562, rs2234671 and rs3138060 polymorphisms could significantly contribute to BC risk in tobacco and alcohol consumption, molecular subtype and stage. The rs1008562, rs2234671 and rs3138060 polymorphisms, and the haplotypes CCC and GGG could significantly contribute to BC risk in the Mexican population analyzed.

Increased homocysteine plasma levels in breast cancer patients of a Mexican population.

AIM: Hyperhomocysteinemia has been associated with different pathologies, including cardiovascular diseases, hypertension, diabetes, and breast cancer (BC). To examine the differences in total homocysteine (tHcy) plasma levels, we compared healthy women to BC patients from a Mexican population. MATERIALS AND METHODS: The tHcy plasma levels were measured using high-performance liquid chromatography with a fluorescence detector in 89 female controls and 261 BC patients. RESULTS: The observed plasma tHcy levels were significantly higher among the BC patients (11.1019 ± 5.9161 µmol/l) compared to the controls (9.1046 ± 1.3213 µmol/l) (p = 0.002), and these differences were evident when stratified by age (≥ 50 years old), menopause status, overweight and obesity, miscarriages, node metastases, progression, subtype classification (luminal, Her2 and triple negative) and nonresponse to chemotherapy. CONCLUSIONS: The tHcy plasma levels could be a good marker for the progression and chemosensitivity of BC in the analyzed sample from a Mexican population.

Total elbow arthroplasty in haemophilia.

INTRODUCTION: Advanced haemophilic arthropathy of the elbow can be a devastating manifestation of severe haemophilia with significant implications for activities of daily living. AIM: To summarize the currently available data on total elbow arthroplasty (TEA) for haemophilic arthropathy of the elbow, to draw comparisons to TEA for other indications, and to outline some considerations regarding TEA in this unique population. METHODS: A narrative review of the literature has been performed. RESULTS: The scant data regarding outcomes of TEA for haemophilic arthropathy are limited to small case series and case reports. A review of the available literature shows that, while pain relief and patient satisfaction are promising, variable outcomes with significant complications and infection rates may discourage routine use of TEA for haemophilic arthropathy of the elbow. CONCLUSION: While patients with severe haemophilic arthropathy of the elbow are likely to make gains in terms of pain control and range of motion following TEA, there are insufficient data to routinely recommend its use. Complication and infection rates are concerning, and the lack of survival analysis data makes it difficult to quantify the benefit to the patient in light of the risks and resources involved in the procedure.

Relación entre el índice de masa corporal bajo previo al embarazo y el riesgo de parto prematuro / Relationship between low body mass index before pregnancy and the risk of preterm birth

Resumen OBJETIVOS: identificar si existe relación entre el índice de masa corporal bajo previo a la concepción y el riesgo de parto pretérmino y de ruptura prematura de membranas pretérmino. MATERIALES Y MÉTODOS: estudio transversal, analítico, de una muestra al azar de pacientes atendidas entre enero de 2015 y enero de 2016. Variables de estudio: edad, índice de masa corporal, ganancia ponderal durante el embarazo, antecedentes de infección de vías urinarias, cervicovaginitis, amenaza de aborto y ruptura prematura de membranas. El análisis se efectuó con un modelo de regresión lineal generalizada para identificar la influencia del índice de masa corporal y del resto de las variables estudiadas en el riesgo de parto prematuro. RESULTADOS: se analizaron los datos de 120 pacientes y la regresión lineal generalizada mostró una relación estadísticamente significativa entre el índice de masa corporal previo al embarazo y el riesgo de parto pretérmino (r2 = 0.016, p = < 0.001). CONCLUSIONES: el IMC bajo, previo al embarazo, se relaciona con aumento poco importante del riesgo de parto pretérmino; el riesgo de ruptura prematura de membranas se incrementa discretamente en pacientes con antecedente de amenaza de aborto y ruptura prematura de membranas pretérmino.

Abstract OBJECTIVE: To identify if there is a relationship between the low pre-conceptional body mass index and the risk of preterm delivery and / or PROM. MATERIALS AND METHODS: Analytical cross-sectional study was conducted with a random sample of 120 patients treated between January 2015 to January 2016. Patients were analyzed for age, body mass index, and weight gain during pregnancy, history of urinary tract infection, vaginal infections, threat of abortion and premature rupture of membranes. The analysis was performed using a generalized linear regression model to identify the influence of body mass index and the rest of the variables studied on the risk of preterm delivery. RESULTS: The generalized linear regression showed a statistically significant relationship between the pre gestational body mass index and the risk of preterm delivery. (R2 = 0.016, p = <0.001) CONCLUSION: Body mass index influenced the risk of preterm delivery but not premature rupture of preterm membranes.

Trends in use of anti-thrombotic agents and outcomes in patients with non-ST-segment elevation myocardial infarction (NSTEMI) managed with an invasive strategy.

OBJECTIVE: To analyze trends in utilization of anti-thrombotic agents (ATA) and in-hospital clinical outcomes in non-ST-elevation myocardial infarction (NSTEMI) patients managed with an invasive strategy from 2007 to 2010. METHODS & RESULTS: Using ACTION Registry(®)-GWTG™ data, we analyzed trends in use of ATA and in-hospital clinical outcomes among 64,199 NSTEMI patients managed invasively between 2007 and 2010. ATA included unfractionated heparin (UFH), low molecular weight heparin (LMWH), glycoprotein IIb/IIIa inhibitors (GPI) and bivalirudin. Although the proportion of NSTEMI patients treated with PCI within 48h of hospital arrival was similar in 2007 and 2010, percentage use of bivalirudin (13.4-27.3%; p<0.01) and UFH increased (60.0-67.5%, p<0.01), and that of GPI (62.3-41.0%; p<0.01) and LMWH (41.5-36.8%; p<0.01) declined. Excess dosing of UFH (75.9-59.3%, p<0.01), LMWH (9.6-5.2%; p<0.01) and GPI (8.9-5.9%, p<0.01) was also significantly lower in 2010 compared with 2007. Though in-hospital mortality rates were similar in 2007 and 2010 (2.3-1.9%, p=0.08), the rates of in-hospital major bleeding (8.7-6.6%, p<0.01) and non-CABG related RBC transfusion (6.3-4.6%, p<0.01) were significantly lower in 2010 compared with 2007. CONCLUSION: Compared with 2007, patients with NSTEMI, who were managed invasively in 2010 received GPI and LMWH less often and bivalirudin and UFH more frequently. There were sizeable reductions in the rates of excess dosing of UFH (though still occurred in 67% of patients), GPI and LMWH. In-hospital major bleeding complications and post-procedural RBC transfusion were lower in 2010 compared with 2007.

Association between TNF-α-308G>A and -238G>A gene polymorphisms and TNF-α serum levels in Mexican colorectal cancer patients.

The objective of this study was to examine the association between TNF-α serum levels and -308G>A and -238G>A polymorphisms in the corresponding gene by comparing healthy subjects to colorectal cancer (CRC) patients from a Mexican population. Serum levels of TNF-α were found to significantly differ between CRC patients and controls (P = 0.001), but no relationship between the -308G>A and -238G>A polymorphisms and increased CRC risk was established (P > 0.05). However, an association between the -308G>A variant and disease became evident when the distribution of AA-GA genotypes was examined in patients with hematologic toxicity (neutropenia) and those without (odds ratio = 3.356, 95% confidence interval = 1.295- 8.698, P = 0.013). The GG haplotype was more common in controls than CRC patients, with a frequency of 0.85 among the former, but this difference was not significant (P > 0.05). In conclusion, TNF-α serum levels and AA-AG genotypes of the TNF-α-308G>A polymorphism may significantly contribute to CRC susceptibility in the population examined in this investigation.

Invasive lobular carcinoma of the breast with extracellular mucin: A case report.

BACKGROUND: Invasive lobular carcinoma is the second most common histological type of breast carcinoma, accounting for approximately 5%-15% of all invasive breast cancers. The extracellular mucin secretion is by default a feature of ductal carcinoma. Only four cases of infiltrative lobular carcinoma with extracellular mucin have been report. CASE SUMMARY: A 60 year old female asymptomatic patient with palpable breast mass and architectural distortion by mammography on external upper quadrant of the right breast was diagnosed as invasive lobular carcinoma with extracellular mucin in the resection, confirmed with immunohistochemistry markers. DISCUSSION: Previous report in the literature of four cases of Invasive lobular carcinoma of breast with extracellular mucin, all of them sharing the same histologic features: the presence of extracellular and intracellular mucin with appearance of infiltrates lobular carcinoma with signet ring cells and "Indian files". CONCLUSION: It is important to know that extracellular mucin production is not exclusive of ductal lesions and keep in mind the lobular carcinomas with extracellular mucin as a differential diagnosis.

Safety and efficacy of plasmid DNA expressing two isoforms of hepatocyte growth factor in patients with critical limb ischemia.

VM202, a plasmid DNA that expresses two isoforms of hepatocyte growth factor, may elicit angiogenic effects that could benefit patients with critical limb ischemia (CLI). In a phase 2, double-blind trial in 52 CLI patients, we examined the safety and potential efficacy of intramuscular injections of low-dose (n=21) or high-dose (n=20) VM202 or placebo (n=11) in the affected limb (days 0, 14, 28 and 42). Adverse events and serious adverse events were similar among the groups; no malignancy or proliferative retinopathy was seen. In exploratory efficacy analyses, we found no differences in ankle or toe-brachial index, VAS, VascuQuol or amputation rate among the groups. Complete ulcer healing was significantly better in high-dose (8/13 ulcers; P<0.01) versus placebo (1/9) patients. Clinically meaningful reductions (>50%) in ulcer area occurred in high-dose (9/13 ulcers) and low-dose (19/27) groups versus placebo (1/9; P<0.05 and P<0.005, respectively). At 12 months, significant differences were seen in TcPO2 between the high-dose and placebo groups (47.5 ± 17.8 versus 36.6 ± 24.0 mm Hg, respectively; P<0.05) and in the change from baseline among the groups (P<0.05). These data suggest that VM202 is safe and may provide therapeutic bioactivity in CLI patients.

Fertilization capacity of cryopreserved Iberian ibex epididymal sperm in a heterologous in vitro fertilization assay.

In vitro fertilization (IVF) can be used to assess the fertilization capacity of sperm. Heterologous IVF may be useful when assessing that of wild animals as it is often difficult to obtain adequate numbers of naturally corresponding oocytes. The aim of the present study was to assess the fertilization capacity of frozen-thawed ibex epididymal spermatozoa via heterologous IVF involving the oocytes of prepubertal domestic goats. The effect on fertilization and embryo development of adding oestrous sheep serum (ESS) to the fertilization medium was also examined. Cumulus-oocyte complexes (COCs) were matured in TCM-199 for 24-27 h at 38.5°C in a 5% CO2 in air atmosphere. Frozen-thawed epididymal spermatozoa were selected by density gradient centrifugation. After maturation, the oocytes were co-incubated with spermatozoa in synthetic oviductal fluid (SOF) with different concentrations of ESS: SOF-C (0%), SOF-2 (2%) and SOF-20 (20%). At 17 h post-insemination (hpi), zygotes with one female and one male pronucleus (2PN) were categorised as normal; zygotes with 3PN were recorded as polyspermic, and oocytes with 1PN as asynchronous. Cleavage and blastocyst development were assessed at 48 and 168 hpi respectively. The percentage of zygotes with 2PN was higher in the SOF-2 than in the SOF-20 treatment group (27.7% versus 2.9% P < 0.05). The percentage of blastocysts formed with the SOF-C, SOF-2 and SOF-20 treatments were 1.1%, 7.5% and 0% respectively. These results show that the presence of 2% ESS achieves better results than the use of no serum or the standard 20% concentration. Heterologous IVF may be an effective method for predicting the fertilization capacity of ibex spermatozoa, and therefore perhaps that of other wild mountain ungulates.

²¹°Po, Cd and Pb distribution and biomagnification in the yellowfin tuna Thunnus albacares and skipjack tuna Katsuwonus pelamis from the Eastern Pacific.

We measured Cd and Pb in the muscle and stomach contents of Thunnus albacares and Katsuwonus pelamis to define the distribution of the elements in the tissues and their degrees of biomagnification. (210)Po was measured in the livers of both species and compared to the results of similar studies. The trophic position of the tuna species was determined by N isotope measurements. The average activity of (210)Po in the liver ranged from 119 to 157 (Bq kg(-1) wet weight) in K. pelamis and T. albacares. The trophic position of T. albacares (4.60) was higher than that of K. pelamis (3.94). The Cd content of the muscle increased significantly with the trophic position of the tuna. δ(13)C in T. albacares and K. pelamis varied, with values of 3.13 and 1.88‰, respectively. The δ(15)N values in yellowfin tuna were higher than in skipjack tuna. The trophic position of T. albacares (4.60 ± 0.67) was therefore more elevated than that of K. pelamis (3.94 ± 1.06). Pb was biomagnified in T. albacares (transfer factor=1.46).

Transferring from clopidogrel loading dose to prasugrel loading dose in acute coronary syndrome patients. High on-treatment platelet reactivity analysis of the TRIPLET trial.

High on-treatment platelet reactivity (HPR) has been identified as an independent risk factor for ischaemic events. The randomised, double-blind, TRIPLET trial included a pre-defined comparison of HPR in acute coronary syndrome (ACS) patients undergoing percutaneous coronary intervention (PCI) following a placebo/600-mg clopidogrel loading dose (LD) immediately before a subsequent prasugrel 60-mg or 30-mg LD. Platelet reactivity was assessed using the VerifyNow® P2Y12 assay (P2Y12 Reaction Units, PRU) within 24 hours (h) following the placebo/clopidogrel LD (immediately prior to prasugrel LD), and at 2, 6, 24, 72 h following prasugrel LDs. The impact of CYP2C19 predicted metaboliser phenotype (extensive metabolisers [EM] and reduced metabolisers [RM]) on HPR status was also assessed. HPR (PRU ≥240) following the clopidogrel LD (prior to the prasugrel LD) was 58.5% in the combined clopidogrel LD groups. No significant difference was noted when stratified by time between the clopidogrel and prasugrel LDs (≤6 hs vs>6 h). At 6 h following the 2nd loading dose in the combined prasugrel LD groups, HPR was 7.1%, with 0% HPR by 72 h. There was no significant effect of CYP2C19 genotype on pharmacodynamic (PD) response following either prasugrel LD treatments at any time point, regardless of whether it was preceded by a clopidogrel 600-mg LD. In conclusion, in this study, patients with ACS intended for PCI showed a high prevalence of HPR after clopidogrel 600-mg LD regardless of metaboliser status. When prasugrel LD was added, HPR decreased substantially by 6 h, and was not seen by 72 h.

Sperm characteristics and heterologous in vitro fertilisation capacity of Iberian ibex (Capra pyrenaica) epididymal sperm, frozen in the presence of the enzymatic antioxidant catalase.

The aim of this work was to evaluate the protective effect of catalase (CAT) on frozen/thawed ibex epididymal sperm recovered post mortem, and to detect any harmful effect this might have on sperm fertilisation capacity. Epididymal spermatozoa were diluted using a Tris-citric acid-glucose medium (TCG) composed of 3.8% Tris (w/v), 2.2% citric acid (w/v), 0.6% glucose (w/v), 5% glycerol (v/v), and 6% egg yolk (v/v). Sperm masses from the right epididymis were diluted with TCG medium, while those from the left were diluted with TCG medium supplemented with 200IU/mL CAT. Heterologous in vitro fertilisation (IVF) was used to assess the fertilisation capacity of this sperm. The addition of CAT to the extender did not improve frozen/thawed sperm variables. Moreover, a reduced fertilisation capacity was detected: sperm diluted with TCG provided 25.5% 2PN zygotes, while just 13.2% was recorded for that diluted with TCG-CAT (P<0.01). The percentage of cleaved embryos at 48hpi was higher (P<0.01) with the TCG sperm than with the TCG-CAT sperm (16.7% vs. 7.6%). The use of 200IU/mL CAT as an additive cannot, therefore, be recommended for the preservation of ibex epididymal sperm. Other antioxidants should, however, be tested in both this and related wild mountain ungulates.

Association of the tumor necrosis factor-alpha -308G>A polymorphism with breast cancer in Mexican women.

The tumor necrosis factor-alpha (TNF-α) gene plays an important role in cell proliferation, differentiation, apoptosis, lipid metabolism, coagulation, insulin resistance, and endothelial function. Polymorphisms of TNF-α have been associated with cancer. We examined the role of the -308G>A polymorphism in this gene by comparing the genotypes of 294 healthy Mexican women with those of 465 Mexican women with breast cancer. The observed genotype frequencies for controls and breast cancer patients were 1 and 14% for AA, 13 and 21% for GA, and 86 and 65% for GG, respectively. We found that the odds ratio (OR) for AA genotype was 2.4, with a 95% confidence interval (95%CI) of 5.9-101.1 (P = 0.0001). The association was also evident when comparing the distribution of the AA-GA genotype in patients in the following categories: 1) premenopause and obesity I (OR = 3.5, 95%CI = 1.3-9.3, P = 0.008), 2) Her-2 neu and tumor stage I-II (OR = 2.5, 95%CI = 1.31-4.8, P = 0.004), 3) premenopause and tumor stage III-IV (OR = 1.7, 95%CI = 1.0-2.9, P = 0.034), 4) chemotherapy non-response and abnormal hematocrit (OR = 2.4, 95%CI = 1.2-4.8, P = 0.015), 5) body mass index and Her-2 neu and III-IV tumor stage (OR = 2.8, 95%CI = 1.2- 6.6, P = 0.016), and 6) nodule metastasis and K-I67 (OR = 4.0, 95%CI = 1.01-15.7, P = 0.038). We concluded that the genotypes AA-GA of the -308G>A polymorphism in TNF-α significantly contribute to breast cancer susceptibility in the analyzed sample from the Mexican population.

Intracytoplasmic sperm injection (ICSI) in small ruminants.

Small ruminants are an important component of the global production systems of meat and wool, and their reproductive biology is well known. However, the incorporation of assisted reproduction techniques (ART) in the production systems of small ruminants is not as well developed as for other domestic species. Normally, production systems that incorporate ARTs are restricted to artificial insemination or in vivo embryo transfer. Intracytoplasmic sperm injection (ICSI) is one of the ARTs techniques reported for small ruminants and consists of the injection of spermatozoa inside an oocyte, bypassing the natural process of sperm-oocyte interaction. In goats and sheep, there are few live births by ICSI reported, with no reports from other species of small ruminants. Currently, there has not been intensive research about ICSI in small ruminants. However, ICSI has potentially important applications in animal production systems, primarily its use with semen of valued animals, with epididymal sperm, in the fertilization of prepubertal or cryopreserved oocytes. Other applications include more advanced techniques, such as transgenic-ICSI or its combination with spermatogonial transplantation. In this article, we review the "state of the art" of this technique in small ruminants including its historical development, research needs for its improvement and future applications.