Use of critically important antimicrobial classes early in life may adversely impact bacterial resistance profiles during adult years: potential co-selection for plasmid-borne fluoroquinolone and macrolide resistance via extended-spectrum beta-lactam use in dairy cattle.

The transfer of antimicrobial resistance genes commonly occurs via vertical and horizontal gene transfer, as such genes are often found on the same mobile genetic element. This occurrence can lead to the co-selection of resistance to antimicrobials without their application. Dairy cattle located in the south-western United States were enrolled in a matched-pair longitudinal study to evaluate the effects of a two-dose ceftiofur treatment for metritis on levels of third-generation cephalosporin resistance among faecal Escherichia coli temporally. Escherichia coli chosen for further investigation were isolated on selective media, harboured extended-spectrum beta-lactam, fluoroquinolone and macrolide resistance genes. This combination has previously been unreported; importantly, it included genes encoding for resistance to antibiotics that can only be used in dairy cattle less than 20 months of age. Fluoroquinolones, macrolides and third and higher generation cephalosporins are considered critically important and highest priority for human medicine by the World Health Organization.

Altered neuronal network and rescue in a human MECP2 duplication model.

Increased dosage of methyl-CpG-binding protein-2 (MeCP2) results in a dramatic neurodevelopmental phenotype with onset at birth. We generated induced pluripotent stem cells (iPSCs) from patients with the MECP2 duplication syndrome (MECP2dup), carrying different duplication sizes, to study the impact of increased MeCP2 dosage in human neurons. We show that cortical neurons derived from these different MECP2dup iPSC lines have increased synaptogenesis and dendritic complexity. In addition, using multi-electrodes arrays, we show that neuronal network synchronization was altered in MECP2dup-derived neurons. Given MeCP2 functions at the epigenetic level, we tested whether these alterations were reversible using a library of compounds with defined activity on epigenetic pathways. One histone deacetylase inhibitor, NCH-51, was validated as a potential clinical candidate. Interestingly, this compound has never been considered before as a therapeutic alternative for neurological disorders. Our model recapitulates early stages of the human MECP2 duplication syndrome and represents a promising cellular tool to facilitate therapeutic drug screening for severe neurodevelopmental disorders.

Absence of CD38 delays arrival of neutrophils to the liver and innate immune response development during hepatic amoebiasis by Entamoeba histolytica.

To define the role of CD38 in the migration of neutrophils to the liver and consequently in the induction of an innate immune response during murine hepatic amoebiasis by Entamoeba histolytica, we examined amoebic liver abscess development (ALA), presence of amoebae and neutrophils, and expression levels of cytokines and other inflammation mediators mRNA, in infected wild-type and CD38 Knockout (CD38KO) C57BL/6J mice. Results showed that CD38KO mice undergo a delay in ALA development in comparison with the wild-type strain. The presence of amoebae lasted longer in CD38(-/-), and although neutrophils arrived to the liver in both strains, there was a clear difference in the time between the two strains; whereas in the wild-type strain, neutrophils arrived at early times (6-12 h), in the CD38KO strain, neutrophils arrived later (48-72 h). Cytokines profile during the innate immune response development (TNF-α, IL-1ß, IL-6) was, for WT mice concomitant with, and preceded, for CD38KO mice, the time in which neutrophils were present in the liver lesion. In conclusion, CD38 is important for neutrophils migration during hepatic amoebiasis, and in turn, these cells play an important role in the innate immune response.

Quantitative and functional evaluation of innate immune responses in patients with common variable immunodeficiency.

OBJECTIVES: We evaluate the frequency and functional response of innate immune cells in peripheral blood (PB) from patients with common variable immunodeficiency (CVID) and healthy controls upon activation with agonists of the Toll-like receptors (TLR) TLR2, TLR4, and TLR9. In addition, several nonsynonymous single nucleotide polymorphisms (SNPs) within these TLR genes were examined. METHODS: Flow cytometry was used to perform immunophenotyping and evaluate the expression of cell surface markers. Levels of cytokines in the culture supernatants were evaluated using cytometric bead array technology. SNPs in the TLR genes were evaluated from genomic DNA using different sequencing techniques. RESULTS: Our results demonstrate that the frequency of CD1d-restricted TCR invariant natural killer T cells in PB was significantly reduced in the patients with CVID. A marked, though not significant, reduction in absolute numbers of plasmacytoid dendritic cells and natural killer cells was also observed in these patients. Interestingly, CD80 and CD86 expression on innate cells upon stimulation with TLR ligands was not altered in the patients although 3 of them exhibited low baseline levels of these surface molecules on monocytes compared to healthy controls. We also observed a significant increase in TNF-alpha levels in supernatants of PB mononuclear cells from CVID patients after stimulation with lipopolysaccharide. Finally, no association was found between the presence of nonsynonymous SNPs within the TLR genes and the clinical presentation of CVID. CONCLUSIONS: Taken together, our study demonstrates than innate immune responses are disturbed in some CVID patients and prompts the evaluation of innate immunity genes as candidates to explain the CVID clinical phenotype.

Seasonal differences in ventricular proliferation of adult Gallotia galloti lizards.

Lizards present neuronal production throughout the telencephalon in their adult state, both naturally and after experimentally induced brain lesions. As in birds, lizards present seasonal behavioural variations. In birds, such variations have been shown to alter neuronal production. In birds and mammals, lack of stimuli or exposure to stress interferes with adult neurogenetic capacity. The effect of this type of study has not been performed with lizards. In the present study we used bromodeoxyuridine to label dividing cells in the ventricular walls of Gallotia galloti lizards during all four seasons and we investigated the effect of captivity on such proliferation. We found that G. galloti presented a particular distribution that differed from that previously described in other reptiles with respect to regions of greater or lesser proliferative rate. In addition, proliferative rate varied seasonally, with greater production of cells in Spring and low production in Autumn and Winter. Proliferative rate was significantly lower throughout the telencephalon and during all seasons in those lizards kept in captivity as compared with wild animals, even though photoperiod and temperature were similar to natural conditions. Our results indicate that cell production in lizards is species-dependent, varies with seasons and is significantly reduced in captive animals.

Cell-Mediated Immunity (CMI) as the Instrument to Assess the Response Against the Allograft: Present and Future.

The concept of Cell-Mediated Immunity (CMI) monitoring in transplantation has gained popularity over time and is now a reality. Significant technological advances have enabled us to test for multiple molecules and cells implicated in inflammatory or suppressive reactions to the graft. The main challenge nowadays is whether clinicians can use the information provided by the measurement of such markers to predict post-transplant outcome. To date a wide range of markers have been identified as promising biomarkers in the monitoring of individual responses to immunosuppression or in the determination of patient alloreactivity to the graft, which could prove helpful in the assessment of the occurrence of an adverse/side effect. Before these biomarkers are deemed suitable, standardisation of the methodology and validation of its feasibility in clinical outcome remains an ongoing challenge. The research community is currently facing a large effort towards the implementation of a standard methodology that is both highly reproducible and can reduce inter-laboratory variability, therefore generating consistency with data. The aim of this manuscript is to review the current literature regarding CMI monitoring in the field of solid organ transplantation (SOT), undertaking a comprehensive study of the latest findings. In addition, based upon current literature, we aim to propose a comprehensive classification of biomarkers to further aid our current understanding, taking in to account the type of transplantation, when its measurement should be applied and which would be the most suitable biomarker to assess.

[Acute myocardial infarction associated to the Sildenafil consumption. A case report and review of the literature]. / Infarto agudo de miocardio asociado al consumo de sildenafil. Aportación de caso y revisión de la literatura.

Erectile dysfunction affects more than 30 million men in The United States. Since the FDA approved the use of Sildenafil, prescription of this medication has been raising. Adverse events of Sildenafil includes: fatigue, dyspnea, and hypotension. Reported adverse cardiac events associated with the medication use include myocardial infarction, ventricular tachycardia, angina and death, raising concerns about the safety of this agent in patients with coronary artery disease. Published guidelines regarding the management of cardiac patients with erectile dysfunction suggest that Sildenafil may be hazardous in patients with ischemic heart disease. In patients using Sildenafil, myocardial infarctions have been reported to the Food and Drug Administration. Now, we report a patient with myocardial infarction after taking 100 mg of Sildenafil without sexual activity.

Prevalence of metabolic syndrome and its association with lower urinary tract symptoms and sexual function. / Prevalencia del síndrome metabólico y su asociación con síntomas del tracto urinario inferior y función sexual.

OBJECTIVES: To estimate the frequency of metabolic syndrome (MetS) in a daily urology practice and to determine its association with lower urinary tract symptoms (LUTS) and erectile dysfunction (ED). MATERIAL AND METHODS: A retrospective study was conducted. Data from all male patients aged ≥40 years who attended our outpatient urology clinic from 2010 to 2011 was collected. Prevalence of MetS was determined, and LUTS and ED were assessed. A logistic model was used to determine possible associations, controlling for confounders and interaction factors. RESULTS: A total of 616 patients were included. MetS was observed in 43.8% (95% CI 39.6-48.3). The bivariate model showed an association between MetS and LUTS (p<0.01), but not between MetS and ED. The logistic model showed an association between MetS and the International Prostate Symptom Score (IPSS), while controlling for other variables. Patients exhibiting moderate LUTS had a greater risk for MetS than patients with mild LUTS (OR 1.83, 95% CI 1.14-2.94). After analyzing for individual components of MetS, positive associations were found between diabetes and severe LUTS (OR 1.3, 95% CI 1.24-7.1), and between diabetes and ED (OR 2.57, 95% CI 1.12-5.8). CONCLUSION: This study was able to confirm an association between MetS and LUTS, but not for ED. Specific components such as diabetes were associated to both. Geographical differences previously reported in the literature might account for these findings. Given that MetS is frequent among urological patients, it is advisable that urologists actively screen for it.

Modeling anorexia nervosa: transcriptional insights from human iPSC-derived neurons.

Anorexia nervosa (AN) is a complex and multifactorial disorder occurring predominantly in women. Despite having the highest mortality among psychiatric conditions, it still lacks robust and effective treatment. Disorders such as AN are most likely syndromes with multiple genetic contributions, however, genome-wide studies have been underpowered to reveal associations with this uncommon illness. Here, we generated induced pluripotent stem cells (iPSCs) from adolescent females with AN and unaffected controls. These iPSCs were differentiated into neural cultures and subjected to extensive transcriptome analysis. Within a small cohort of patients who presented for treatment, we identified a novel gene that appears to contribute to AN pathophysiology, TACR1 (tachykinin 1 receptor). The participation of tachykinins in a variety of biological processes and their interactions with other neurotransmitters suggest novel mechanisms for how a disrupted tachykinin system might contribute to AN symptoms. Although TACR1 has been associated with psychiatric conditions, especially anxiety disorders, we believe this report is its first association with AN. Moreover, our human iPSC approach is a proof-of-concept that AN can be modeled in vitro with a full human genetic complement, and represents a new tool for understanding the elusive molecular and cellular mechanisms underlying the disease.

Trojan horse or proton force: finding the right partner(s) for toxin translocation.

Much is known about the structure function relationships of a large number of bacterial protein toxins, the nature of their cell surface receptors, and their enzymatic activities which lead to the inactivation of their respective cytosolic targets. Despite this wealth of knowledge a detailed understanding of the mechanisms which underlie translocation of the catalytic domain across the eukaryotic cell membrane to the cytosol, the penultimate event in the intoxication process, have been slow in developing. In the case of diphtheria toxin, two prominent hypotheses have been advanced to explain how the catalytic domain is translocated from the lumen of endocytic vesicles to the target cell cytosol. We discuss each of these hypotheses and provide an overview of recent observations that tend to favor a mechanism employing a Cytosolic Translocation Factor complex in the entry process. This facilitated mechanism of translocation appears to rely upon protein-protein interactions between conserved domains within the transmembrane domain of diphtheria toxin with host cell factors to effect delivery of the enzymatic moiety. We have recently identified a 10 amino acid motif in the transmembrane domain of diphtheria toxin that is conserved in anthrax Lethal and Edema Factors, as well as in botulinum neurotoxins A, C and D. Stable eukaryotic cell transfectants that express a peptide containing this motif become resistant to the toxin, and sensitivity is completely restored by co-expression of siRNA which inhibits peptide expression. Data obtained from use of the protein fusion toxin DAB(389)IL-2 in cytotoxicity assays using susceptible Hut 102/6TG and resistant transfectant Hut102/6TG-T1 cells, as well as pull down assays have led to the formulation of a working model of facilitated delivery of the diphtheria toxin catalytic domain to the cytosol of target cells which is discussed in detail.

Extended pelvic lymphadenectomy in patients with clinically localised prostate cancer: A prospective observational study. / Linfadenectomía pélvica extendida en pacientes con cáncer de próstata clínicamente localizado: estudio observacional prospectivo.

OBJECTIVE: To determine the frequency of lymph node involvement in patients with clinically localised prostate adenocarcinoma who had radical prostatectomy and extended pelvic lymphadenectomy. MATERIAL AND METHODS: A prospective observational study was conducted on 137 patients with clinically localised prostate cancer of low, intermediate or high risk according to the D'Amico classification. All participants underwent radical prostatectomy plus extended pelvic lymphadenectomy in 3 reference centres in Bogota, Colombia, between 2013 and 2014. The following variables were assessed: age, prostate specific antigen levels, Gleason score of the biopsy, probability of lymph node involvement calculated with Partin tables and the histopathology result of the surgical specimen, with the definitive Gleason pattern and the total number of resected and involved lymph nodes per tumour, according to the territory of the dissection. RESULTS: A total of 2,876 lymph nodes were extracted (an average of 20.99 lymph nodes per patient). There was lymph node involvement in 14 (10.22%) patients. The high-risk and intermediate-risk group presented lymph node metastases in 28.57% and 5.25%, respectively. There was no lymph node involvement in the low-risk group. Of the patients at risk of lymph node involvement (≥2% according to the Partin tables), 19.40% had lymph node metastases. CONCLUSION: Lymph node involvement in our population is similar to that reported in the worldwide literature. Extended pelvic lymphadenectomy increased the probability of detecting lymph node metastases in our community.

[Vein of Galen arteriovenous malformations and neonatal convulsions]. / Malformacion arteriovenosa de la vena de Galeno y convulsiones neonatales.

INTRODUCTION: Vein of Galen arteriovenous malformation (VGAM) appears during the embryonic period and gives rise to a complex network of arterial and venous vessels that generates a blood shift, from the brain parenchyma towards the malformation, with haemodynamic repercussions. Heart failure is the most frequent presenting symptom during the neonatal period, yet, convulsions or other neurological signs have occasionally been reported in this stage of life. CASE REPORT: A term infant with symptoms of heart failure and convulsions that began during the first 12 hours of life. CAT and magnetic resonance angiography scans revealed a vascular malformation and areas of cerebral ischaemia. CONCLUSIONS: In the case of our patient, detecting areas of ischaemia in the cerebral hemispheres suggested that the damage could be caused by a "steal" syndrome leading the blood flow away from these areas towards the malformation. This situation can occur either before or after birth and the self-limiting nature of the seizures in cerebral infarcts could lead to them going noticed because they take place inside the uterus or when the patient is not being observed directly by his or her health care providers. We suspect that the convulsions in newborn infants with this malformation may well be more frequent than is currently believed.

Metformin's effects on glucose and lipid metabolism in patients with secondary failure to sulfonylureas.

OBJECTIVE: To compare results obtained with metformin versus those obtained with DNA-recombinant insulin in obese patients with NIDDM suffering from secondary failure to sulfonylureas. RESEARCH DESIGN AND METHODS: We conducted an open, prospective, randomized, and comparative study comprising a total of 60 patients selected and placed in two parallel groups. We had previously confirmed that the subjects had secondary failure to high doses of sulfonylureas. The initial metformin dosage was a single 850 mg tablet, and the dosage was increased to two or three tablets depending on the patient's metabolic changes. The initial dosage of DNA-recombinant insulin was 24 U, subcutaneously administered and divided into two portions: two-thirds at around 8:00 A.M., before breakfast, and the remaining third at 8:00 P.M., before dinner. The dosage was adjusted based on the patient's clinical and metabolic response. RESULTS: The initial average glucose value for the metformin group was 269.1 +/- 32.2 mg/dl, decreasing by the end of the study to 159.7 +/- 30.5 mg/dl. For the insulin group, these figures went from 270.7 +/- 24.0 mg/dl at the beginning of the study to 134.8 +/- 26.7 mg/dl. This decrease correlates with the reduction in glycosylated hemoglobin from 12.8 to 8.9% for the first group and from 12.3 to 8.2% for the second, as well as with the reduction in triglyceride values from 230.3 to 183.1 mg/dl and from 218.4 to 186.3 mg/dl, respectively. The BMI (27.5-26.4), blood pressure (systolic from 145.7-132.1 mmHg, diastolic from 90.3-84.8 mmHg), and total cholesterol levels (235-202 mg/dl) decreased in only the metformin group. CONCLUSIONS: Metformin is an effective, safe, and well-tolerated treatment that improves metabolic control and favorably modifies secondary clinical alterations due to insulin resistance, such as arterial hypertension, overweight, and hyperlipidemia, in obese patients with NIDDM suffering from secondary failure to sulfonylureas.

The lacertidian reticular thalamic nucleus topographically upon the dorsal thalamus: experimental study in Gallotia galloti.

The projection pattern of the ventral thalamic reticular nucleus onto the dorsal thalamus was studied in the lizard Gallotia galloti using in vitro horseradish peroxidase and fluorescent carbocyanine labelling techniques. Localized label deposits at three dorsoventrally spaced sites in the dorsal thalamus elicited retrograde transport into separate, though partly overlapping, medial, dorsolateral and ventrolateral sectors within an extended cytoarchitectonic complex which may be globally identifiable as the reticular nucleus. Neurons found in the dorsolateral and ventrolateral sectors mainly corresponded to the cell group named nucleus ventromedialis (or nucleus of the dorsal supraoptic decussation) in the literature, whereas neurons labelled in the medial sector corresponded to the so-called dorsal hypothalamic nucleus. Sparser cells appear labelled in the superficially placed nucleus suprapeduncularis. Thalamotelencephalic fibers arising from the injected dorsal thalamic nuclei also project to the corresponding retrogradely labeled sectors within the reticular nucleus. These findings reveal a rough topographic organization in the connections of the extended reticular nucleus complex with the whole dorsal thalamus. This supports the hypothesis of hodological homology between this ventral thalamic formation in Gallotia and the mammalian thalamic reticular nucleus.

Reelin expression during embryonic brain development in lacertilian lizards.

The expression of reelin mRNA and protein was studied during embryonic brain development in the lacertilian lizards L. viridis and L. galloti, by using radioactive in situ hybridization and immunohistochemistry. At all stages studied, high reelin expression was consistently found in the olfactory bulb, in the lateral cortex, and in neurons of the marginal zone and subplate of medial and dorsal cortical sectors. In the dorsal ventricular ridge (DVR), reelin expression was confined to deeply located, large cells which were more abundant in the caudal than the rostral part of the DVR. In the diencephalon, the ventral lateral geniculate complex and the perirotundal were strongly positive, whereas other nuclei were mostly negative. High reelin signal was associated with some layers in the tectum, with the torus semicircularis, cerebellar granule cell layers, and the ventral horn of the spinal cord. A more moderate signal was detected in the septal nuclei, striatum, retina, habenular nuclei, preoptic and periventricular hypothalamic components, and in reticular nuclei of the mid- and hindbrain. The medial and dorsal cortical plate and Purkinje cells were reelin-negative but expressed disabled-1 (Dab1) mRNA. When they are compared with reelin expression during mammalian brain development, our data reveal an evolutionarily conserved canvas of reelin expression, as well as significant differences, particularly in developing cortical fields. The developing lizard cortex differs from that of turtles, birds, crocodiles, and mammals in that it displays heavy reelin expression not only in neurons of the marginal zone that might be homologous to mammalian Cajal-Retzius cells, but also in subplate neurons. This difference in the pattern of reelin expression suggests that the elaborate radial organization of the lacertilian cortical plate, somewhat reminiscent of its mammalian counterpart, results from evolutionary convergence. Our data lend support to the hypothesis that the reelin signaling pathway played a significant role during cortical evolution.

The humoral immune response to the kinetoplastid membrane protein-11 in patients with American leishmaniasis and Chagas disease: prevalence of IgG subclasses and mapping of epitopes.

The kinetoplastid membrane protein-11 (KMP-11) is a major target of the humoral immune response during Leishmania-infections. The majority of sera from visceral leishmaniasis, mucocutaneous leishmaniasis and even some cutaneous leishmaniasis patients contain detectable IgG antibodies against KMP-11. We also provide evidence that this protein may act as a potent antigen in T. cruzi infections, since most Chagas sera show immunological cross-reactivity. Therefore, KMP-11 cannot be used as a specific diagnostical tool for the serodiagnosis of leishmaniasis in those regions where both, Leishmania and T. cruzi infections overlap geographically. When analyzing the subclass specificity of the antibody response to KMP-11 we observed the following order of reactivity: IgG1 > > IgG3 > IgG2 > IgG4, which is similiar to that seen in crude parasite extract. The mapping of antigenic determinants by using synthetic 20-mer peptides revealed the existence of predominantly conformational epitopes in leishmaniasis, while 50% of sera from Chagas patients reacted with a particular KMP-11 peptide. These results therefore suggest the presence of disease-specific B-cell epitopes.

Assessment of the efficacy and safety of nimesulide vs naproxen in paediatric patients with respiratory tract infections. A comparative single-blind study.

A total of 99 paediatric patients (57 male, 42 female) aged 1 to 12 years, weighing 10 to 40kg and with acute pharyngo-amygdalitis were enrolled in a single-blind study to assess the efficacy and tolerability of nimesulide in comparison with naproxen when both drugs were administered over an 8-day treatment period. Among the 2 treatment groups comprising 99 evaluable patients, demographic analysis of age, weight and height did not reveal statistically significant differences. Evaluation of fever, pain, inflammation and nasal obstruction over the 8-day treatment period showed a significant improvement in these parameters for those patients treated with nimesulide when compared with naproxen from day 1, with remission of symptoms starting from day 3. These findings were complemented by a superior tolerability profile reported for nimesulide-treated patients. In conclusion, nimesulide appears to be a safe and effective treatment for paediatric patients with pharyngo-amygdalitis and it has shown superior efficacy and tolerability when compared with naproxen.

Prospective study of amniocentesis performed between weeks 9 and 16 of gestation: its feasibility, risks, complications and use in early genetic prenatal diagnosis.

This paper demonstrates that the outcome of amniocenteses performed between the 9th and the 14th weeks is similar to that of amniocenteses performed between the 15th and 20th weeks. We have performed and prospectively followed 615 amniocenteses between the 9th and 16th weeks of gestation. The outcome, risks, and complications are similar to those of amniocenteses at the usual time (after 15 weeks) and to the other groups of early amniocentesis (before 15 weeks). Early amniocentesis differs from amniocentesis at the usual time in that it carries higher rates of fetal losses and of amniotic fluid leakage, more confined cytogenetic abnormalities, and an increased number of patients who have the procedure postponed. Two cultures (0.32%) failed to produce results, 595 (96.7%) samples were obtained at the first tapping, 20 (3.3%) at the second attempt. alpha-Fetoprotein levels reach their maximum at 13 weeks. Amniocenteses between 15 and 16 weeks (293, or 47%) constitute the control group, those between 9 and 14 weeks (322) the experimental group. Early amniocentesis appears to be a safe early genetic prenatal diagnosis technique, an alternative to chorionic villi sampling.

[Influence of epilepsy-related factors on scholastic achievement in an epileptic child]. / Influencia de algunos factores relacionados con la epilepsia sobre los resultados escolares del niño epiléptico.

We studied 30 children aged between 5 and 12 years (mean = 8.7 years: standard deviation = (+)-1 9678) who suffered from partial epileptic crises, with or without secondary generalization, cryptogenic, who had had this disorder for more than one year, and who attended ordinary schools. The objective was to ascertain the educational results obtained, and to evaluate the effect of some of the factors related to epilepsy, in these results. The scholar's family was interviewed in all cases to record: the age of onset and duration of the illness, the frequency of crises, attendance at school and results obtained in the previous school year. The school results were: 5 children had results E (16.67%). 10 (33.33%) were classed as MB, 6 (20%) as B, 5 had R (16.67%) and 4 (13.33%) were evaluated as M. It is concluded that the factors most closely related to poor school results in the group studied were: a frequency of more than three crises in the previous year, partial crises with secondary generalization and failure to attend school because of crises.