Assessing the effects of PMM2 variants on protein stability.

Phosphomannomutase 2 deficiency, PMM2-CDG, is the most frequent disorder of protein N-glycosylation. It is an autosomal recessive disease with a broad clinical and biochemical phenotype. Trying to predict the impact of novel variants is often a challenge due to the high number of variants and the difficulty to establish solid genotype-phenotype correlations. A potential useful strategy is to use computational chemistry calculations as a tool from which relevant information on the structural impact of novel variants may be deduced. Here we present our analyses based on four well-known PMM2 deleterious variants (p.(Leu32Arg), p.(Asp65Tyr), p.(Phe119Leu), p.(Arg141His)) and the polymorphic p.(Glu197Ala) for which we have predicted the effect on protein stability. Our work predicts the effect of different amino acid residues on the conformation and stability of PMM2. These computational simulations are, therefore, an extremely useful methodology which, in combination with routinely used in silico methods of pathogenicity prediction, may help to reveal the structural impact of novel variants at the protein level, potentially leading to a better understanding of target biological molecules.

Carcinoma espinocelular da base do chifre, bilateral e simétrico, em vaca com distúrbio hormonal ˗ relato de caso / [Horn-based squamous cell carcinoma, bilateral and symmetrical in cow with hormonal disorder ˗ case report]

Este trabalho tem por objetivo descrever um caso de carcinoma espinocelular da base do chifre, bilateral e simétrico, em uma vaca e discutir a questão hormonal, possivelmente envolvida na patogênese da doença. Tratava-se de uma fêmea bovina, 11 anos, com histórico de emagrecimento progressivo, presença de massas exofíticas na base de ambos os chifres e em anestro por muitos anos. Foi realizada biopsia incisional de ambas as lesões para exame histopatológico e dosagens hormonais. Devido à progressão do quadro clínico, optou-se pela eutanásia, seguida dos exames necroscópico e histopatológico. O exame histopatológico revelou tratar-se de um carcinoma espinocelular infiltrativo bem diferenciado, e as dosagens hormonais apresentaram alterações nos níveis do hormônio luteinizante, folículo estimulante e estrógenos totais. Apesar de existirem descrições de carcinomas espinocelulares da base do chifre, ainda não havia relatos da ocorrência do mesmo bilateral e em uma vaca com distúrbios hormonais.(AU)

The objective of this study was to describe a case of bilateral and symmetrical squamous cell carcinoma from the horn base in a cow and to discuss the hormonal question, possibly involved in its pathogenesis. A 11-year-old beef cow presenting a history of progressive thinning, presence of exophytic masses at the base of both horns and anestrous for many years was assisted. An incisional biopsy of both lesions was performed for histopathological examination and hormonal dosages. Due to the clinical progression, euthanasia followed by necroscopic and histopathological examination was carried out. Histopathological examination revealed a well differentiated infiltrative squamous cell carcinoma and the hormonal dosages presented changes in luteinizing hormone, follicle stimulating and total estrogen levels. Although there are descriptions of basal squamous cell carcinoma of the horn, there were still no reports of the occurrence of the same bilateral in a cow with hormonal disorders.(AU)

Long-term efficacy of opicapone in fluctuating Parkinson's disease patients: a pooled analysis of data from two phase 3 clinical trials and their open-label extensions.

BACKGROUND AND PURPOSE: The aim was to evaluate the efficacy of the catechol-O-methyltransferase inhibitor opicapone (25 and 50 mg) as adjunct therapy to levodopa in a pooled population of Parkinson's disease patients who participated in the pivotal double-blind trials of opicapone and their 1-year open-label extensions. METHODS: Data (placebo, opicapone 25 mg and opicapone 50 mg) from the BIPARK-1 and BIPARK-2 double-blind and open-label studies were combined. The studies had similar designs, eligibility criteria and assessment methods. The primary efficacy variable in both double-blind studies was the change from baseline in absolute OFF time based on patient diaries. RESULTS: Double-blind treatment with opicapone (25 and 50 mg) significantly reduced absolute daily OFF time from a baseline of 6.1-6.6 h. The mean (and 95% confidence interval) treatment effect versus placebo was -35.1 (-62.1, -8.2) min (P = 0.0106) for the 25 mg dose and -58.1 (-84.5, -31.7) min (P < 0.0001) for the 50 mg dose. Reductions in OFF time were mirrored by significant increases in ON time without troublesome dyskinesia (P < 0.05 and P < 0.0001 for the 25 and 50 mg doses, respectively). No significant differences were observed for ON time with troublesome dyskinesia. Patient diary results from the open-label phase indicated a maintenance of effect for patients previously treated with opicapone 50 mg. The group previously treated with the 25 mg dose benefitted with further optimization of therapy during the open-label phase, whilst switching from placebo to opicapone led to significant reductions in OFF time and increased ON time. CONCLUSIONS: Over at least 1 year of open-label therapy, opicapone consistently reduced OFF time and increased ON time without increasing the frequency of troublesome dyskinesia.

Distribution of selenium in sheep treated with dipheny diselenide / Distribuição do selênio em ovinos tratados com disseleneto de difenila

The aim of the present study was to report the in vivo distribution of selenium in sheep. For this, animals were allocated into two groups (control group and treated group) and kept in metabolic cages for a period of 37 days. The treated group received a single dose (6µmol/kg) of Diphenyl Diselenide, intravenously. Plasma and erythrocytes samples were collected at different times. Adipose tissue, muscles (latissimusdorsi, semitendinosus, and supra-scapular) heart, liver, lung, kidney, intestine and brain were sampled at 30 days post-treatment, in order to determine the selenium concentration. The results demonstrated that the selenium, from the Diphenyl Diselenide group, was higher in erythrocytes (4.8mg/L, six hours post-treatment) when compared with the control sheep. The deposition of selenium occurred in the liver (7.01µg/g), brain (3.53µg/g) and kidney (2.02µg/g). After 30 days of a single intravenous injection of Diphenyl Diselenide, liver was the main organ of selenium deposition.(AU)

O objetivo do presente estudo foi investigar a distribuição in vivo do selênio em ovinos. Para isso, os animais foram distribuídos em dois grupos (grupo controle e grupo tratado) e mantidos em gaiolas metabólicas por um período de 37 dias. O grupo tratado recebeu uma dose única (6µmol/kg) de disseleneto de difenila, por via intravenosa. As amostras de plasma e de eritrócitos foram recolhidas em momentos diferentes. Tecido adiposo, músculos (latissimus dorsi, semitendinoso e supraescapular) coração, fígado, pulmão, rim, intestino e cérebro foram amostrados aos 30 dias pós-tratamento, a fim de se determinar a concentração de selênio. Os resultados demonstraram que o selênio, do grupo disseleneto de difenila, foi maior em eritrócitos (4,8mg/L, seis horas após o tratamento) quando comparado com o grupo controle. A deposição de selênio ocorreu no fígado (7,01µg/g), cérebro (3,53µg/g) e rim (2,02µg/g). Após 30 dias de uma única injeção intravenosa de disseleneto de difenila, o fígado foi o principal órgão de deposição de selênio.(AU)

Efeito metafilático do zinco injetável sobre parâmetros metabólicos e oxidativos de ovelhas no pós-parto imediato / Metaphylactic effect of injectable zinc on metabolic and oxidative parameters of sheep in the immediate postpartum

The aim of this study was to evaluate a new source of injectable organic zinc (zinc edetate) on the energy and oxidative profile in sheep during the immediate postpartum period. Twenty-six Texel breed animals were previously identified and divided into two experimental groups: the treated group (TG; n= 13) that comprised the animals that received a subcutaneous (SC) injection of 100 mg of zinc edetate (2 mL) fifteen days before the parturition expected date and the control group (CG; n=13) that comprised the animals that received 2mL of physiological solution at the same date of TG. Blood samples were collected on the parturition day for the assessment of serum fructosamine, cholesterol and triglycerides, insulin-like growth factor type 1 (IGF-1), the oxidative stress index (OSi) and blood zinc concentration. In addition to these parameters, the measurement of zinc was made in food given to the animals. There was no difference in metabolic parameters and OSi between the experimental groups (P>0.05), as well as in blood zinc concentrations (P>0.05). The parenteral zinc edentate does not change the energy and oxidative profile of sheep in immediate postpartum.(AU)

Identification of SNPs in growth-related genes in Colombian creole cattle.

Colombian creole cattle have important adaptation traits related to heat tolerance and reproductive and productive efficiency. Romosinuano (ROMO) and Blanco Orejinegro (BON) are the most common breeds used by Colombian cattle breeders. Growth traits are of prime importance in these animals, which are mainly raised for beef production. Genes encoding growth hormone, growth hormone receptor, homeobox protein, insulin growth factor binding protein 3, leptin, and myostatin have been associated with physiological growth pathways in cattle and other species. We therefore aimed to identify single nucleotide polymorphisms (SNPs) within these genes in ROMO, BON, and Zebu cattle. DNA regions of these genes were sequenced in 386 animals; 47 new SNPs were found, of which 14 were located in the exonic regions, thereby changing the protein sequence. An association of SNPs with weaning weight (WW), daily weight gain at weaning (DWG), and weight at 16 months (W16M) traits was deduced. The genetic analysis revealed several SNPs related to these traits. The SNP GhRE06.2 had a significant association with WW and the SNP Lep03.4 was highly associated with DWG and W16M. Other polymorphisms were significantly associated with WW and DWG, although they did not surpass the Bonferroni significance threshold. The new mutations identified may indicate important points of genetic control in the DNA that could be responsible for changes in the expression of the analyzed traits. These SNPs might be used in future breeding programs to improve the productive performance of cattle in beef farms.

Effect of opicapone on levodopa pharmacokinetics, catechol-O-methyltransferase activity and motor fluctuations in patients with Parkinson's disease.

BACKGROUND AND PURPOSE: Opicapone (OPC) is a novel third generation catechol-O-methyltransferase (COMT) inhibitor that enhances levodopa availability. This study investigated the effects of OPC in comparison with placebo on levodopa pharmacokinetics, tolerability and safety, COMT activity and motor response to levodopa in Parkinson's disease (PD) patients with motor fluctuations. METHODS: This was a randomized, multicentre, double-blind and placebo-controlled study in four parallel groups of PD patients treated with standard-release 100/25 mg levodopa/carbidopa or levodopa/benserazide and with motor fluctuations (wearing-OFF phenomenon). Subjects were sequentially assigned to be administered, once-daily, up to 28 days (maintenance phase), placebo (n = 10) or 5 (n = 10), 15 (n = 10) and 30 mg (n = 10) OPC. Two levodopa tests were performed, one at baseline and another following the maintenance phase. Subjects kept a diary to record motor fluctuations (ON/OFF periods) throughout the study. RESULTS: In relation to placebo, levodopa exposure (AUC0-6) increased 24.7%, 53.9% and 65.6% following 5, 15 and 30 mg OPC, respectively. Maximum COMT inhibition (Emax) ranged from 52% (5 mg OPC) to 80% (30 mg OPC). The study was not designed to detect any significant differences in motor performance, but the exploratory analysis performed shows improvement in various motor outcomes, including a dose-dependent change in absolute OFF time corresponding to a percentage decrease of 4.16% (P > 0.05), 29.55% (P > 0.05) and 32.71% (P < 0.05) with 5, 15 and 30 mg OPC, respectively. Treatments were generally well tolerated and safe. CONCLUSIONS: OPC is a promising new COMT inhibitor that significantly decreased COMT activity, increased systemic exposure to levodopa and improved motor response.

Androecia in two Clusia species: development, structure and resin secretion.

Clusia fluminensis and C. lanceolata are dioecious shrubs having resiniferous flowers with strongly distinct androecia. The aim of this study was to investigate the development and anatomy of their androecia and the ultrastructure, histochemistry and secretory process of their androecium resin glands, examining whether the cellular aspects of resin secretion differed between these two morphologically distinct androecia. Stamens differ, being free in C. fluminensis and clustered in a synandrium in C. lanceolata. Staminode sterility is due to the undifferentiated nature of the anthers in C. lanceolata and degeneration of meiocytes and anther indehiscence in C. fluminensis. Resin is produced in subepidermal cavities and canals with wide lumens. In the secretory stage, epithelial cells present sinuous walls, voluminous nuclei, polymorphic plastids associated with periplastidial reticulum, mitochondria, oil bodies, multivesicular bodies, endoplasmic reticulum and dictyosomes. The resin is released through rupture points on the distal surface of stamens and staminodes, associated with disrupted cavities and canals. Our results show morphological diversity associated with functional similarity. Also, a secretion pattern shared by the two species includes initiation of the secretory process in young floral buds, compartmentalisation of the secretion in pre-anthesis buds and release of secretions at anthesis. Cellular aspects of resin secretion in these species are quite similar, as are the chemical identities of the main components of the floral resins of the genus.

Genome-wide association study on growth traits in Colombian creole breeds and crossbreeds with Zebu cattle.

Whole genome selection represents an important tool for improving parameters related to the production of livestock. In order to build genomic selection indexes within a particular breed, it is important to identify polymorphisms that have the most significant association with a desired trait. A genome-wide marker association approach based on the Illumina BovineSNP50 BeadChip(TM) was used to identify genomic regions affecting birth weight (BW), weaning weight (WW), and daily weight gain (DWG) in purebred and crossbred creole cattle populations. We genotyped 654 individuals of Blanco Orejinegro (BON), Romosinuano (ROMO) and Cebú breeds and the crossbreeds BON x Cebú and ROMO x Cebú, and tested 5 genetic control models. In total, 85 single nucleotide polymorphisms (SNPs) were related (P < 0.05) to the 3 evaluated traits; BW was associated with the highest number of SNPs. For statistical false-positive correction, Bonferroni correction was used. From the results, we identified 7, 6, and 4 SNPs with strong associations with BW, WW, and DWG, respectively. Many of these SNPs were located on important coding regions of the bovine genome; their ontology and interactions are discussed herein. The results could contribute to the identification of genes involved in the physiology of beef cattle growth and the development of new strategies for breeding management via genomic selection to improve the productivity of creole cattle herds.

Genetic variability in Blanco Orejinegro breed cattle populations in Colombia.

We analyzed population structure and genetic diversity in Blanco Orejinegro Creole cattle with 12 microsatellite markers, genotyping 138 individuals belonging to 7 commercial and 3 conservation herds. These markers showed a high level of polymorphism; 171 alleles were identified. The mean number of alleles per locus was 5.63 (3.82-6.58). The total number of alleles per marker was 14.2 and ranged from 16 (TGLA126) to 22 (TGLA227). The mean expected heterozygosity (0.73) was higher than the observed heterozygosity (0.65), with a significant excess of heterozygosity in almost all populations (FIS = 0.09; P < 0.05). This may be due to crossing between different lines of this breed, affecting the inbreeding levels. Analysis of relationships among populations, assessed by principal component analysis and Nei's genetic distances, indicated a close relationship between some herds. Furthermore, analysis of population structure demonstrated a low probability of admixture with Zebu breeds, as it shows the cluster assignment and the FST values obtained. We conclude that there is high allelic diversity in this breed, even though a low effective population size has been maintained and the level of inbreeding has not been monitored. Therefore, appropriate conservation efforts should be undertaken, such as adopting strategies aimed at minimizing inbreeding, to avoid losing genetic variability.

Pharmacokinetic interaction study between eslicarbazepine acetate and lamotrigine in healthy subjects.

OBJECTIVE: Anti-epileptic drugs are often used in combination. Both eslicarbazepine (main metabolite of eslicarbazepine acetate, ESL) and lamotrigine undergo conjugation with glucuronic acid, and both eslicarbazepine and its glucuronide and lamotrigine glucuronide undergo extensive renal elimination; therefore, there is a potential for interaction. This study investigated the interaction between ESL and lamotrigine in healthy subjects. METHODS: Open-label study in two parallel groups of 16 healthy volunteers each. After an 8-day treatment with ESL or lamotrigine, ESL (1200 mg once-daily) and lamotrigine (150 mg once-daily) were co-administered for 19 days. Geometric mean ratios (GMR) and 90% confidence intervals (90% CI) for maximum plasma concentration (C(max)) and area under the plasma concentration-time curve in the dosing interval (AUC(0-24)) were calculated for eslicarbazepine (ESL active metabolite) and lamotrigine. RESULTS: The C(max) and AUC(0-24) GMR (90% CI) were, respectively, 95% (87-102%) and 96% (91-102%) for eslicarbazepine, and 88% (82-94%) and 86% (81-92%) for lamotrigine. The 90% CI of the C(max) and AUC(0-24) GMR fell within the prespecified acceptance interval (80-125%) both for eslicarbazepine and lamotrigine. CONCLUSION: There was no significant pharmacokinetic interaction between ESL and lamotrigine in healthy subjects. Therefore, no dosage adjustment appears to be usually required in either lamotrigine or ESL when the drugs are co-administered.

Effect of eslicarbazepine acetate on the pharmacokinetics of metformin in healthy subjects.

PURPOSE: Eslicarbazepine acetate (ESL) is a new voltage-gated sodium channel blocker currently in development for the treatment of neuropathic pain, including that of diabetic origin. The primary objective was to investigate the effect of ESL on the pharmacokinetics of metformin, a commonly used oral antidiabetic drug. METHODS: Randomized, open-label, two-way crossover study in 20 healthy subjects. The volunteers received an 850 mg single-dose of metformin hydrochloride on two occasions - once as such and once after pre-treatment with an oral once-daily dose of ESL 1200 mg for 6 days - separated by a washout period of at least 2 weeks. The bioequivalence approach was used for assessing the effect of ESL on the pharmacokinetics of metformin. Test/Reference geometric mean ratios (GMR) and 90% confidence intervals (90% CI) were calculated for AUC0- yen, AUC0-12 and Cmax of metformin. RESULTS: Test/Reference metformin GMR (90% CI) was 0.95 (0.86; 1.05) for AUC0- yen, 0.95 (0.88; 1.06) for AUC0-12, and 0.88 (0.77; 1.00) for Cmax. Formal bioequivalence could not be demonstrated for metformin Cmax. However, the extent of exposure to metformin, as reflected by AUC0-12 and AUC0- yen, allows the claim of bioequivalence since the 90% CI of the GMR fall within the pre-specified bioequivalence acceptance interval (0.80; 1.25). CONCLUSION: Once-daily administration of ESL 1,200 mg had no relevant effect on the systemic exposure to metformin pharmacokinetics in healthy subjects.

Effect of food on the pharmacokinetic profile of trans-resveratrol.

OBJECTIVE: It has been postulated that trans-resveratrol may act as an antioxidant, cardioprotective, neuroprotective and cancer chemopreventive agent. The objective of this study was to investigate the effect of food on the bioavailability of trans-resveratrol following oral administration. MATERIAL AND METHODS: Single-centre, open-label, randomized, 2-way crossover study on 24 healthy subjects. The study consisted of two consecutive treatment periods separated by a washout of 7 days or more. On each of the study periods subjects were administered a single-dose of 400 mg of trans-resveratrol following either a standard high fat content meal or 8 hs of fasting. RESULTS: There was a large interindividual variability in the trans-resveratrol pharmacokinetic parameters. Mean +/- SD maximum plasma concentration (Cmax) was 42.2 +/- 36.6 ng/ml in fed and 47.3 +/- 30.0 ng/ml in fasting conditions. Median time to Cmax (tmax) was 2.0 h in fed and 0.5 h in fasting (p < 0.0001). The fed/fasting geometric mean ratio (GMR) and 90% confidence interval (90% CI) were 79.4 and 53.8, 117.0% for Cmax, and 106.0 and 86.8, 128.0% for the area under the plasma concentration-time curve (AUC0- yen). The 90% CI for the GMR of AUC0- yen and Cmax fall outside the usual bioequivalence acceptance range of 80, 125%, but that of AUC0- yen was close to the bioequivalence standard. CONCLUSION: The rate of absorption of trans-resveratrol following an oral 400 mg single-dose was significantly delayed by the presence of food, as reflected by Cmax and tmax. However, the extent of absorption, as reflected by AUC- yen, was not affected in a relevant way.

Simultaneous supratentorial and brainstem abscesses due to Listeria monocytogenes.

Multiple supratentorial abscesses caused by Listeria monocytogenes are rare. We report the simultaneous occurrence of multiple supratentorial and brainstem abscesses due to Listeria, in a patient under corticotherapy for an exacerbation of ulcerative colitis. MR imaging features before and after successful conservative treatments are depicted. In immunocompromised patients with supratentorial listerial abscesses, the coexistence of brainstem abscedation is exceptional. Despite high mortality associated with listerial abscesses, this case illustrates the possibility of a good clinical outcome, if the appropriate antibiotic regimen is instituted and the immunosuppressant agent is discontinued.

La enfermedad meningococica en cuidados intensivos (I): valor de los factores pronósticos / The meningoccocal illness in looking after intensive (I): courage of the factors presages

Se realiza un estudio de los factores pronosticos en los pacientes con enfermedad meningococica ingresados en la unidad de cuidados intensivos del hospital provincial clinico quirurgico docente "Dr. Gustavo Aldereguia Lima", en el periodo comprendido entre los años 1986- 1990. Se determina que estos factores junto a la experiencia práctica del médico de asistencia pueden ayudar a establecer índices pronóstico y realizar una terapéutica mas eficaz, asi como hacer la prevención de aquellas complicaciones que pueden aparecer una vez identificados estos signos. Se estudian 23 pacientes y se analiza la presencia de estos signos de mal pronóstico, tanto en los pacientes engresados vivos, como en los fallecidos encontrándose que la hipocelularidad del líquido cefalorraquídeo y la acidosis metabólica fueron los factores mas frecuentes en los fallecidos

La enfermedad meningococica en cuidados intensivos (II): algunos aspectos clínicos y complicaciones / The meningoccocal illness in looking after intensive (II): some clinical looks and complications

En el número anterior se publicó la primera parte de este artículo, la cual estuvo dedicada la análisis de los factores pronósticos en los pacientes con enfermedad meningococica, en esta segunda parte se aborda la patogenia de las complicaciones fundamentales de esta entidad, haciendo especial enfasis en el choque septico, dificultad respiratoria del adulto, coagulación intravascular diseminada, insuficiencia suprarrenal aguda y se expone el comportamiento de las mismas en una serie estudiada en la unidad de cuidados intensivos del hospital provincial clinico quirurgico de Cienfuegos

Embolismo graso: problemas en su diagnóstico / Fatty embolism: problems in their diagnosis

Se presenta un estudio del embolismo graso y los principales problemas en su diagnóstico, asi como el tratamiento que se debe implantar, preventivo, sintomático y específico. En la provincia esta enfermedad en los dos últimos años se diagnosticó en 10 pacientes, en los que la fractura diafisiaria de femur en combinación con otros huesos fracturados fue la causa mas frecuente, a todos se les aplicó tratamiento con una evolución satisfactoria. Se concluye que el embolismo graso no es una rareza clinica, sino un diagnóstico en el que no se piensa ante un politraumatizado

The anterior tibial artery flap: anatomic study and clinical application.

Satisfactory replacement of skin defects over the lower leg remains a difficult problem. Various forms of coverage, including, local rotation flaps, muscle flaps, and fascial and free flaps, have their specific indications and inherent disadvantages. In this work, a new axial skin flap based on perforating vessels in the territory of the anterior tibial artery is described. A series of 50 lower leg dissections was carried out in 25 fresh cadavers after latex injection into the femoral artery. Detailed studies of the cutaneous distribution of the anterior tibial artery showed that three main arteries perfuse the anterior lateral portion of the lower leg. The superior lateral peroneal artery and the inferior lateral peroneal artery interseptal cutaneous perforators arise at an average of 25.6 and 17.2 cm from the lateral malleolus, respectively. The superior lateral peroneal artery was present in 100 percent of the specimens, whereas the inferior lateral peroneal artery was present in 70 percent of the specimens. In their course, they give several muscular branches to the peroneus longus and brevis prior to perforating the fascia and arborizing in the subcutaneous tissues of the anterolateral portion of the leg. The average external diameter was 1.6 cm for the superior and 1.4 cm for the inferior lateral peroneal artery. The superficial peroneal nerve accessory artery is the third artery which contributes to the skin of the lower leg. It arises from the superior lateral peroneal artery in 30 percent of cases, from the inferior lateral peroneal artery in 40 percent, and from both in 30 percent. The artery runs along with the superficial peroneal nerve and gives several cutaneous perforators along its descending course. Several cutaneous axial flaps can be fashioned around this anatomy. The operative technique along with demonstrative clinical cases is presented followed by pertinent discussion.