Transcranial static magnetic stimulation reduces seizures in a mouse model of Dravet syndrome.

Dravet syndrome is a rare form of severe genetic epilepsy characterized by recurrent and long-lasting seizures. It appears around the first year of life, with a quick evolution toward an increase in the frequency of the seizures, accompanied by a delay in motor and cognitive development, and does not respond well to antiepileptic medication. Most patients carry a mutation in the gene SCN1A encoding the α subunit of the voltage-gated sodium channel Nav1.1, resulting in hyperexcitability of neural circuits and seizure onset. In this work, we applied transcranial static magnetic stimulation (tSMS), a non-invasive, safe, easy-to-use and affordable neuromodulatory tool that reduces neural excitability in a mouse model of Dravet syndrome. We demonstrate that tSMS dramatically reduced the number of crises. Furthermore, crises recorded in the presence of the tSMS were shorter and less intense than in the sham condition. Since tSMS has demonstrated its efficacy at reducing cortical excitability in humans without showing unwanted side effects, in an attempt to anticipate a possible use of tSMS for Dravet Syndrome patients, we performed a numerical simulation in which the magnetic field generated by the magnet was modeled to estimate the magnetic field intensity reached in the cerebral cortex, which could help to design stimulation strategies in these patients. Our results provide a proof of concept for nonpharmacological treatment of Dravet syndrome, which opens the door to the design of new protocols for treatment.

[Appendicitis versus nonspecific acute abdominal pain: diagnostic accuracy of ultrasound]. / Apendicitis versus dolor abdominal agudo inespecífico: rendimiento diagnóstico de la ecografía.

PURPOUSE: The aim of this study is to assess the diagnostic accuracy of ultrasound to differentiate appendicitis from nonspecific acute abdominal pain, that is the most common process requiring differential diagnosis in clinical practice. MATERIAL AND METHODS: Patients admitted for suspicion of appendicitis were prospectively evaluated in our hospital during two years (2013-2014). Cases of nonspecific acute abdominal pain and appendicitis assessed by ultrasound were enrolled in the study. The different variables collected were statistically analyzed by descriptive, univariate and diagnostic accuracy studies. RESULTS: A total of 275 patients were studied, 143 cases of nonspecific acute abdominal pain and 132 cases of appendicitis. Ultrasound sensitivity and specificity to differentiate appendicitis were 94.7% and 87.4% respectively, with a 12.6% rate of false positives and a 5.3% rate of false negatives. The rate of false negatives in perforated group was 17.4% and analysis according to Pediatric Appendicitis Score risk groups showed a 12.2% rate of false positives in low-risk group and a 6.3% rate of false negatives in high-risk group. CONCLUSIONS: The use of ultrasound in low clinical probability cases of appendicitis could rise unnecessary surgery rate, due to the significant number of false positives in this group of patients. In high probability clinical cases, ultrasound does not contribute too much to diagnosis and it could be a confusion factor by the significant number of false negative associated to perforated appendicitis.

OBJETIVO: El objetivo del estudio es evaluar el rendimiento diagnóstico de la ecografía para diferenciar la apendicitis del dolor abdominal agudo inespecífico, principal proceso con el que requiere diagnóstico diferencial en la práctica clínica. MATERIAL Y METODOS: Se evaluaron los pacientes atendidos por sospecha de apendicitis en nuestro centro durante 2 años (2013-2014), incorporando al estudio los casos de dolor abdominal agudo inespecífico y apendicitis en los que se realizó ecografía. Las diferentes variables recogidas se analizaron estadísticamente de manera descriptiva, univariante y con estudios de rendimiento diagnóstico. RESULTADOS: Se estudiaron 275 casos; 143 casos de dolor abdominal agudo inespecífico y 132 casos de apendicitis. La sensibilidad y especificidad de la ecografía para diferenciar apendicitis fue del 94,7% y 87,4%, respectivamente, con un porcentaje de falsos positivos del 12,6% y de falsos negativos del 5,3%. El porcentaje de falsos negativos en el grupo de apendicitis perforada alcanzó el 17,4% y el análisis según los grupos de riesgo establecidos por el Pediatric Appendicitis Score mostró un porcentaje de falsos positivos del 12,2% en el grupo de bajo riesgo y de falsos negativos del 6,3% en el grupo de alto riesgo. CONCLUSIONES: El uso de la ecografía en casos de baja probabilidad clínica de apendicitis podría incrementar la tasa de cirugía innecesaria, debido al significativo número de falsos positivos en este grupo de pacientes. En casos de alta probabilidad clínica de apendicitis, la ecografía aporta poco al diagnóstico, e incluso podría ser un factor de confusión por el significativo número de falsos negativos asociados a la apendicitis perforada.

Advanced cell-based modeling of the royal disease: characterization of the mutated F9 mRNA.

Essentials The Royal disease (RD) is a form of hemophilia B predicted to be caused by a splicing mutation. We generated an iPSC-based model of the disease allowing mechanistic studies at the RNA level. F9 mRNA analysis in iPSC-derived hepatocyte-like cells showed the predicted abnormal splicing. Mutated F9 mRNA level was very low but we also found traces of wild type transcripts. SUMMARY: Background The royal disease is a form of hemophilia B (HB) that affected many descendants of Queen Victoria in the 19th and 20th centuries. It was found to be caused by the mutation F9 c.278-3A>G. Objective To generate a physiological cell model of the disease and to study F9 expression at the RNA level. Methods Using fibroblasts from skin biopsies of a previously identified hemophilic patient bearing the F9 c.278-3A>G mutation and his mother, we generated induced pluripotent stem cells (iPSCs). Both the patient's and mother's iPSCs were differentiated into hepatocyte-like cells (HLCs) and their F9 mRNA was analyzed using next-generation sequencing (NGS). Results and Conclusion We demonstrated the previously predicted aberrant splicing of the F9 transcript as a result of an intronic nucleotide substitution leading to a frameshift and the generation of a premature termination codon (PTC). The F9 mRNA level in the patient's HLCs was significantly reduced compared with that of his mother, suggesting that mutated transcripts undergo nonsense-mediated decay (NMD), a cellular mechanism that degrades PTC-containing mRNAs. We also detected small proportions of correctly spliced transcripts in the patient's HLCs, which, combined with genetic variability in splicing and NMD machineries, could partially explain some clinical variability among affected members of the European royal families who had lifespans above the average. This work allowed the demonstration of the pathologic consequences of an intronic mutation in the F9 gene and represents the first bona fide cellular model of HB allowing the study of rare mutations at the RNA level.

Factors of importance when selecting sows as embryo donors.

The improvement in porcine embryo preservation and non-surgical embryo transfer (ET) procedures achieved in recent years represents essential progress for the practical use of ET in the pig industry. This study aimed to evaluate the effects of parity, weaning-to-estrus interval (WEI) and season on reproductive and embryonic parameters at day 6 after insemination of donor sows superovulated after weaning. The selection of donor sows was based on their reproductive history, body condition and parity. The effects of parity at weaning (2 to 3, 4 to 5 or 6 to 7 litters), season (fall, winter and spring), and WEI (estrus within 3 to 4 days), and their interactions on the number of corpus luteum, cysts in sows with cysts, number and quality of viable and transferable embryos, embryo developmental stage and recovery and fertilization rates were evaluated using linear mixed effects models. The analyses showed a lack of significant effects of parity, season, WEI or their interactions on any of the reproductive and embryonic parameters examined. In conclusion, these results demonstrate that fertilization rates and numbers of viable and transferable embryos collected at day 6 of the cycle from superovulated donor sows are not affected by their parity, regardless of the time of the year (from fall to spring) and WEI (3 or 4 days).

Surgical embryo collection but not nonsurgical embryo transfer compromises postintervention prolificacy in sows.

Recent advances in nonsurgical deep uterine (NsDU) embryo transfer (ET) technology allow the noninvasive transfer of porcine embryos into recipients, overcoming the most important impediment for commercial ET in this species. Although many factors in the porcine ET-field have been recently evaluated, many others remain to be explored. We investigated here the future reproductive performance of donors and recipients after artificial insemination subsequent to the default surgical embryo recovery approach and to the NsDU-ET procedure, respectively. Although surgical embryo collection did not influence subsequent farrowing rates (90.5%), litter size decreased severely (8.9 ± 0.8 piglets) compared to presurgery (10.8 ± 0.3 piglets) and control group (10.7 ± 0.3 piglets). In contrast, NsDU-ETs did neither affect fertility nor prolificacy of recipients in the cycle subsequent to ET, regardless of whether they were pregnant after NsDU-ET or not. These results indicate that while the surgical embryo collection procedure compromises the reproductive future of donor sows, the NsDU-ET approach does not affect the reproductive potential of the recipients after reintroduction to the breeding stock of the farm. Further research is thus needed to improve surgical embryo collection.

The Recipients' Parity Does Not Influence Their Reproductive Performance Following Non-Surgical Deep Uterine Porcine Embryo Transfer.

With the development of the non-surgical deep uterine (NsDU) embryo transfer (ET) technology, the commercial applicability of ET in pigs is now possible. There are, nevertheless, many factors that influence NsDU-ET effectiveness that need to be addressed. The aim of this study was to evaluate the effects of the weaned recipients' parity on fertility and prolificacy following NsDU-ET. The recipients (n = 120) were selected based on their reproductive history and body condition and grouped into three categories according to their parity: primiparous sows, sows of parity 2 and sows of parities from 3 to 5. Thirty fresh embryos (morulae and unhatched blastocysts) were non-surgically transferred into one uterine horn of each recipient. It was possible to insert the NsDU-ET catheter through the cervix along a uterine horn in 98.3% of the recipients. The parity had no influence on the difficulty grade of the insertions or on the percentage of correct insertions. The cervix and uterine wall were not perforated during the insertions, and vaginal discharge was not observed after transfer in any of the recipients. There were no differences in the pregnancy rates (74.8%), farrowing rates (71.2%) or litter sizes (9.6 ± 3.3) between groups. Also, there were no differences between groups regarding to the piglets' birthweights or piglet production efficiency. In conclusion, these results demonstrate that weaned sows from parity 1 to 5 are appropriate to be used as recipients in NsDU-ET programs, which increase the possibilities for the utilization of ET in the recipient farms.

The effects of superovulation of donor sows on ovarian response and embryo development after nonsurgical deep-uterine embryo transfer.

This study aimed to evaluate the effectiveness of superovulation protocols in improving the efficiency of embryo donors for porcine nonsurgical deep-uterine (NsDU) embryo transfer (ET) programs. After weaning (24 hours), purebred Duroc sows (2-6 parity) were treated with 1000 IU (n = 27) or 1500 IU (n = 27) of eCG. Only sows with clear signs of estrus 4 to 72 hours after eCG administration were treated with 750 IU hCG at the onset of estrus. Nonhormonally treated postweaning estrus sows (n = 36) were used as a control. Sows were inseminated and subjected to laparotomy on Days 5 to 6 (Day 0 = onset of estrus). Three sows (11.1%) treated with the highest dosage of eCG presented with polycystic ovaries without signs of ovulation. The remaining sows from nonsuperovulated and superovulated groups were all pregnant, with no differences in fertilization rates among groups. The number of CLs and viable embryos was higher (P < 0.05) in the superovulated groups compared with the controls and increased (P < 0.05) with increasing doses of eCG. There were no differences among groups in the number of oocytes and/or degenerated embryos. The number of transferable embryos (morulae and unhatched blastocysts) obtained in pregnant sows was higher (P < 0.05) in the superovulated groups than in the control group. In all groups, there was a significant correlation between the number of CLs and the number of viable and transferable embryos, but the number of CLs and the number of oocytes and/or degenerated embryos were not correlated. A total of 46 NsDU ETs were performed in nonhormonally treated recipient sows, with embryos (30 embryos per transfer) recovered from the 1000-IU eCG, 1500-IU eCG, and control groups. In total, pregnancy and farrowing rates were 75.1% and 73.2%, respectively, with a litter size of 9.4 ± 0.6 piglets born, of which 8.8 ± 0.5 were born alive. There were no differences for any of the reproductive parameters evaluated among groups. In conclusion, our results demonstrated the efficiency of eCG superovulation treatments in decreasing the donor-to-recipient ratio. Compared with nonsuperovulated sows, the number of transferable embryos was increased in superovulated sows without affecting their quality and in vivo capacity to develop to term after transfer. The results from this study also demonstrate the effectiveness of the NsDU ET procedure used, making possible the commercial use of ET technology by the pig industry.

Características clínicas y pronóstico de pacientes con enfermedad cerebrovascular crónica (estudio ICBAR) / Clinical features and prognosis of patients with chronic cerebrovascular disease (ICBAR study)

Introducción. Los pacientes con enfermedad cerebrovascular (ECV) son un grupo de muy alto riesgo cardiovascular, ademásde por su propia patología vascular, probablemente por un inadecuado control de factores de riesgo y por la presencia de comorbilidades.Pacientes y métodos. Estudio multicéntrico, con participación de 34 médicos de atención primaria y registro de característicasde 473 pacientes con historia de episodio de ECV que precisó hospitalización. Tras seguimiento clínico de la cohorte, se analizaron reingresos hospitalarios, mortalidad y causas. Resultados. La edad media de los pacientes (el 52% varones) fue de 75 ± 10 años, y los factores de riesgo más prevalentesfueron hipertensión arterial (79%), dislipidemia (66%), obesidad (43%) y diabetes (29%). El 68% de los pacientes tenía diagnóstico de ictus y el 32%, de ataque isquémico transitorio. El tiempo medio transcurrido desde el primer episodio de ECV fue de 6,6 ± 5,5 años. Presentaban situación de dependencia el 29% y sólo un tercio mostraron buen control tensionaly lipídico. Durante un seguimiento de 8,2 ± 2,3 meses, el 7,2% de los pacientes sufrió algún episodio cardiovascular (muerte u hospitalización), del que resultaron determinantes independientes la insuficiencia cardíaca previa (hazard ratio, HR = 2,74; intervalo de confianza del 95%, IC 95% = 1,3-5,9), la miocardiopatía (HR = 3,32; IC 95% = 1,4-8,2), laanemia (HR = 3,09; IC 95% = 1,6-6,2), la insuficiencia renal (HR = 2,4; IC 95% = 1,0-5,6), la situación de dependencia (HR = 2,57; IC 95% = 1,3-5,7) y los ingresos cardiovasculares en el último año (HR = 3,05; IC 95% = 1,5-5,6).Conclusiones. Los pacientes con ECV seguidos en el ámbito de atención primaria presentan una prevalencia elevada y un escaso grado de control de hipertensión arterial. Su pronóstico está condicionado por comorbilidades cardiovasculares ysecuelas de su patología cerebrovascular (AU)

Introduction. Patients with cerebrovascular disease (CVD) are a group with a very high cardiovascular risk, in addition tothat arising from their own vascular pathology, probably due to an inadequate control of risk factors and owing to the presence of comorbidities. Patients and methods. This research consisted in a multi-centre study involving the collaboration of 34 primary carephysicians and recording of the features of 473 patients with a previous history of a CVD event that required hospitalisation.After a clinical follow-up of the cohort, hospital readmissions, mortality and causes were analysed.Results. The mean age of patients (52% males) was 75 ± 10 years and the most prevalent risk factors were arterial hypertension (79%), dyslipidaemia (66%), obesity (43%) and diabetes (29%). Sixty-eight per cent of patients had been diagnosed with stroke and 32% with transient ischaemic attack. The mean amount of time elapsed since the first CVDevent was 6.6 ± 5.5 years. Twenty-nine per cent of patients had a situation of dependence and only one third showed good blood pressure and lipid control. During a follow-up lasting 8.2 ± 2.3 months, 7.2% of patients suffered some kind of cardiovascular event (death or hospitalisation), which independent determinants were found to be previous heart failure (hazard ratio, HR = 2.74; 95% confidence interval, CI 95% = 1.3-5.9); cardiomyopathy (HR = 3.32; CI 95% = 1.4-8.2); anaemia (HR = 3.09; CI 95% = 1.6-6.2); renal failure (HR = 2.4; CI 95% = 1.0-5.6); the situation of dependence (HR = 2.57; CI 95% =1.3-5.7) and cardiovascular admissions over the past year (HR = 3.05; CI 95% = 1.5-5.6).Conclusions. Patients with CVD followed up in the area of primary care present a high prevalence of arterial hypertension and little is done to control it. Their prognosis is conditioned by cardiovascular comorbidities and sequelae of their cerebrovascular disease (AU)

Sharp rates of decay of solutions to the nonlinear fast diffusion equation via functional inequalities.

The goal of this paper is to state the optimal decay rate for solutions of the nonlinear fast diffusion equation and, in self-similar variables, the optimal convergence rates to Barenblatt self-similar profiles and their generalizations. It relies on the identification of the optimal constants in some related Hardy-Poincaré inequalities and concludes a long series of papers devoted to generalized entropies, functional inequalities, and rates for nonlinear diffusion equations.

Decision-making, behavioral supervision and learning: an executive role for the ventral premotor cortex?

In order to adjust the behavioral performance in a changing environment, subjects have to monitor their evolving actions and to know whether their responses were correct or incorrect. This requires self-awareness, cognitive flexibility, working memory (WM), and decision making that frequently are impaired in psychosis. What is the neural substrate of these processes and where are these substrates located? Dysfunction of prefrontal, parietal, temporal cortices, and associated subcortical structures are known to be involved in some of these symptoms. The prefrontal-subcortical circuits have been the main focus of study while other cortical areas such as the premotor cortex have received less attention. The main focus of this review is about the evidence that the ventral premotor cortex processes both recent sensory information and that from long-term memory to decide and evaluate the behavior of previous decisions. This process may serve for learning and thus adapting future behavior to environmental demands. Therefore, dysfunction of this cortical area could be related to some cognitive neuropsychiatric disorders.

[Familial spontaneous intracranial hypotension]. / Síndrome de hipotensión intracraneal espontánea de presentación familiar.

INTRODUCTION: Spontaneous intracranial hypotension (SIH) is characterized by orthostatic headache and diffuse dural gadolinium enhancement on magnetic resonance imaging. Spontaneous intracranial hypotension is caused by spinal cerebrospinal fluid leaks and evidence for an underlying generalized connective tissue disorder is found in about two thirds of patients. There had been no familial cases reported to date. CASE REPORT: We describe the case report of three healthy female patients with spontaneous intracranial hypotension who belong to the same family. They have no cutaneous or articular stigmas of connective tissue disease. DISCUSSION: These familial cases of spontaneous SIH suggest that certain underlying genetic susceptibility, probably linked to lower resistance of the spinal meninges, could favor the development of spontaneous intracranial hypotension.

WT1 gene silencing by aerosol delivery of PEI-RNAi complexes inhibits B16-F10 lung metastases growth.

The Wilms' tumor gene 1 (WT1) is a universal tumor antigen and consequently a good therapeutic target for the development of gene therapy strategies. Earlier, we reported the in vitro efficacy of WT1 RNAi in the inhibition of B16F10 murine melanoma cell line growth. In this study, we used an aerosol system to deliver WT1 RNAi complexes, polyethyleneimine (PEI)-WT1-1 or PEI-WT1-2, to the lungs of mice with B16F10 lung metastasis. This treatment produced a statistically significant (P=0.020) reduction in the number and size of lung tumor foci, resulting in decreased lung weight and tumor index in treated mice compared with controls. The WT1 RNAi treatment also reduced the number and size of tumor blood vessels, suggesting decreased angiogenesis. Furthermore, the treated lung tissue showed cells in the tumor infiltrations undergoing apoptosis and elevated expression of the proapoptotic genes Bcl-xS and Bax, suggesting an activation of the intrinsic apoptotic pathway. Overall, WT1-1 treatment prolonged the mean survival time of tumor-bearing mice in comparison with the control and WT1-2-treated mice. Our data show that WT1 gene silencing in vivo by aerosol delivery of PEI-WT1 RNAi complexes is an effective therapeutic strategy for the treatment of lung metastases.

Sex-sorting sperm by flow cytometry in pigs: issues and perspectives.

Several hundred thousand offspring of preselected sex of various species have been born since sperm sexing technology based on flow cytometric sorting of X- and Y-chromosome-bearing sperm and DNA was first demonstrated in 1989. The advantages derived from application of sexing technology to commercial dairy cattle production have been demonstrated worldwide. Utilizing sex-sorting technology for pig production systems offers many similar advantages. However, several factors currently limit implementation of sexing technology in pigs. Anatomical and physiological features inherent to the female pig, together with the relatively low sperm output of a flow sorter, are the main limitations to widespread use of this technology in pig production systems. This review analyzes the factors that limit the efficiency of sperm sorting technology for commercial swine production. In addition, this review discusses recent innovations in technical instrumentation and applied reproductive techniques that may help to overcome some of these limitations.

Improving the efficiency of insemination with sex-sorted spermatozoa.

The sorting of X- and Y-chromosome-bearing spermatozoa by flow cytometry is nowadays one of the most apt assisted-reproduction technologies in livestock production. Potential economic and biological benefits, as well as those related to easier management of herds, have been reported arising out of the application of this technique, especially in cattle. Yet, the sex-sorting procedure induces damage to spermatozoa, affecting their function and fertilizing ability. Different species present varying degrees of susceptibility to damage from the sorting process and each has its own requirements for sex-sorted insemination procedures. Thus, several new protocols and strategies have been designed for the handling of sorted spermatozoa, with the main objective of optimizing their fertilizing ability and the consequent application of flow-cytometric sex-sorting technology. This article reviews current advances in this technology, pointing out the components to be improved before this technology may be widely applied in different domestic species.

New developments in low-dose insemination technology.

New nonsurgical procedures for inseminating swine with a low number of spermatozoa have been developed and/or evaluated over the last few years. These procedures allow the deposition of the insemination dose into the uterine body (post-cervical insemination) or directly into the uterine horn (deep intrauterine insemination). With the use of the post-cervical insemination, a threefold reduction in the number of fresh sperm has been successfully used to achieve pregnancy. Using deep intrauterine insemination (DUI), up to a 20-fold reduction in the number of fresh spermatozoa or a sixfold reduction in the number of frozen/thawed spermatozoa can be achieved, with reproductive performance very similar to that obtained after standard AI. Complementing these nonsurgical insemination techniques, a new procedure for depositing spermatozoa into the oviduct by laparoscopy has been recently described. This laparoscopic technique has proven to be applicable to diluted and sex-sorted spermatozoa. The development of new insemination procedures will help achieve more efficient application of currently available sperm technologies. Using appropriate insemination procedures, it is now feasible to achieve high fertility rates with cooled, frozen-thawed, or sex-sorted semen.

Low-dose insemination in pigs: problems and possibilities.

Low-dose AI procedures are required by the pig industry to efficiently utilize emerging sperm technologies, such as cryopreservation and sex-sorting. Currently, several different procedures for inseminating with a low or very low number of spermatozoa have been described. Deep intrauterine insemination allows the deposition of the spermatozoa in the depth of the uterine horn, allowing a significant reduction in the number of spermatozoa inseminated with maintenance of optimal reproductive performance. Intra-oviductal laparoscopic insemination has been recently applied in pigs. This technique has proved to be applicable with diluted and sex-sorted spermatozoa. This review discusses several problems encountered during the development of deep intrauterine insemination and intra-oviductal laparoscopic insemination of pigs and provides potential solutions for the practical application of both the technologies.

Effect of Breathwalk on body composition, metabolic and mood state in chronic hepatitis C patients with insulin resistance syndrome.

AIM: To identify the anthropometric, metabolic and mood state in hepatitis C virus (HCV)-infected patients from the west of Mexico and to evaluate the effect of Breathwalk (BW), a combination of walking, synchronized breathing and focussed attention, on those patients. METHODS: In an experimental study, 17 patients with serological and molecular diagnosis of HCV, not receiving pharmacological treatment, were studied. One hour sessions of BW were practiced 3 times at week for six months. Body composition was assessed by electric impedance. Biochemical profiles and insulin resistance (IR) risk was assessed by conventional methods. Mood state was evaluated with specific and open questions at the beginning and at the end of the program. RESULTS: Seventy percent of patients were overweight or obese, and 77% of the patients presented with IR at the beginning of the study. Improvements were observed at the 3rd mo, and statistically significant differences were recorded at the 6th mo using the fitness score (76 vs 83, P < 0.01), in alanine aminotransferase (ALT) (106 +/- 93 U/L vs 59 +/- 32 U/L, P < 0.01), total bilirubin (0.09 +/- 1 mg/dL vs 0.62 +/- 0.2 mg/dL, P < 0.01), ALT/AST ratio (1.04 vs 0.70, P < 0.01), triglycerides (165 +/- 86 mg/dL vs 124 +/- 49 mg/dL, P < 0.01) and the IR risk (4.0 vs 2.7). Most patients (88%) indicated to feel better at the end of BW (P < 0.01). CONCLUSION: Breathwalk has an important effect on body composition, lipid profile and liver enzymes. It is also easy, inexpensive and has a beneficial effect on metabolic and mood state in HCV patients.

Improving the fertilizing ability of sex sorted boar spermatozoa.

The sex sorting of spermatozoa by flow cytometry induces damage, since sperm cells are highly diluted, affecting their functionality and fertilizing ability. In this work it was investigated whether the concentration of sex sorted spermatozoa by the sedimentation method, rather than centrifugation, in combination with the presence of the seminal plasma protein PSP-I/PSP-II heterodimer may improve their fertilizing ability. Spermatozoa were sorted by flow cytometry and collected in BTS with 10% of seminal plasma (group C: control) or with 1.5mg/mL of PSP-I/PSP-II heterodimer (group H). Collected spermatozoa from each medium were split into two aliquots. One aliquot of each group was centrifuged (800 x g/5 min) just after sorting and stored 16-18 h at 17 degrees C (groups Cc and Hc) at 6 x 10(6)sperm/mL. The second aliquot was directly stored at 17 degrees C for 16-18 degrees C (group Cs and Hs). After storage the supernatant was discarded and the sedimented pellet adjusted to 6 x 10(6)sperm/mL. Membrane integrity, acrosome status and motility characteristics of spermatozoa from all groups were assessed. Post-weaning pre-ovulatory sows were inseminated by laparoscopy into the oviduct with 0.3 x 10(6) sex sorted spermatozoa to assess their ability to penetrate oocytes in vivo. Putative zygotes were collected 18 h after insemination by washing the oviduct. Penetration and monospermic rates were evaluated. After 16-18 h of storage, centrifuged spermatozoa collected with 10% seminal plasma or 1.5 mg/mL PSP-I/PSP-II heterodimer after sex sorting showed lower (p<0.05) percentages of membrane integrity, motility and fertilization than sedimented spermatozoa. Overall, the presence of 10% seminal plasma or PSP-I/PSP-II heterodimer did not affect the results. However, a positive effect of PSP-I/PSP-II heterodimer (p<0.05) was observed in sedimented spermatozoa. Hence, our results indicate that the sedimentation method in the presence of PSP-I/PSP-II heterodimer improves the in vivo fertilizing ability of sex sorted boar spermatozoa.

[Preoperative diagnostic imaging in stump appendicitis]. / Diagnóstico preoperatorio por métodos de imagen de una apendicitis del muñón.

Stump appendicitis is a rare condition in which the diagnosis is often delayed and is eventually reached during exploratory surgery. Knowledge of stump appendicitis makes it possible to reach the diagnosis using imaging techniques and thereby reduce the morbidity of this condition. We present the case of a patient with appendicitis of the appendicular stump that was diagnosed prior to surgery using plain-film radiography and abdominal CT.