Identification of reference genes for studies of quantitative gene expression in male and female quail tissues.

In industrial poultry, quail production has gained increasing prominence over the years. It is known that the intensification of genetic studies has contributed greatly to this growth, through techniques, such as analysis of gene expression by PCR, for example. This study aimed to evaluate stability and recommend reference genes for quantitative real-time PCR in different tissues from male and female broiler quails. The stability of 10 housekeeping genes (GAPDH, RPL5, MRPS27, MRPS30, TFRC, HMBS, EEF1, LDHA, B2M, and UBC) by means Bestkeeper, NormFinder, GeNorm softwares with ΔCq method. The tissues analyzed were: heart, thigh muscle, brain, and spleen, considering that they are tissues commonly used in nutrigenomic, immunological, and poultry performance research. As expected, the reference genes tested showed varying stability depending on the tissue evaluated. According to the present study, the most stable housekeeping genes were MRPS30, TFRC, and HMBS in heart; MRPS30, EEF1, and HMBS in thigh muscle; B2M, GAPDH, and UBC in brain; and EEF1, LDHA, and HMBS in spleen. Therefore, it is recommended to be used as reference genes for gene expression studies of male and female quails.

Anti-JCV antibody serostatus and longitudinal evaluation in a Portuguese Multiple Sclerosis population.

Multiple Sclerosis (MS) treatment with natalizumab is associated with Progressive Multifocal Leukoencephalopathy (PML). The risk of PML being related to the anti-JCV antibody index is well established, but there is less known about seroconversion rates in natalizumab-treated patients and longitudinal variation in the anti-JCV antibody index. Our objective was to assess anti-JCV antibody prevalence in an MS population and to evaluate the evolution of the anti-JCV antibody index in natalizumab-treated patients. To assess anti-JCV antibody prevalence, we included all patients who had the anti-JCV antibody test in our consultation, regardless of the treatment. To evaluate the evolution of the anti-JCV antibody index and seroconversion, only natalizumab-treated patients with at least two samples were selected. Demographic characteristics were evaluated. From a total of 371 patients included, 68.19% (n=253) were seropositive for anti-JCV antibodies (JCV+). There was a significant difference in anti-JCV antibody seropositivity concerning gender (male 76.27% vs. female 64.43%, p=0.023), but not age. To evaluate seroconversion, 85 patients who were initially seronegative (JCV-) were selected. The annual rate of seroconversion in the first two years was stable, but after that there was a significant increase with treatment duration (ρ=0.90, p=0.037): in the first year it was 5.88% (n=5/85); in the second, 5.71% (n=4/70); in the third, 6.82% (n=3/44); in the fourth, 10.34% (n=3/29); and in the fifth, 15.0% (n=3/20). The mean index variability was higher in patients who experienced seroconversion (1.16±0.97), followed by JCV+ patients (0.44±0.48), compared to JCV- patients (0.08±0.05). In conclusion, anti-JCV antibody prevalence in our population is comparable to other reported cohorts. The seroconversion rate increased with treatment duration. We found a high fluctuation in the antibody index in JCV+ patients.

Long-term effectiveness and safety of natalizumab in a Portuguese population.

OBJECTIVES: Natalizumab long-term effectiveness data in real-world relapsing-remitting multiple sclerosis (RRMS) is needed. Our objective is to report the long-term effectiveness and safety of natalizumab in a cohort of RRMS patients. METHODS: This is a retrospective study of natalizumab treatment for two years or longer in RRMS. Annualized relapse rate, Expanded Disability Status Scale (EDSS), brain magnetic resonance imaging T2 lesion volume, JC virus antibody status, previous treatments and adverse events were analysed. RESULTS: Seventy-one patients were included with a mean treatment duration of 44.86±17.39months. Over the treatment duration there was a significant decrease in annualized relapse rate (88.37%) and EDSS (28.57%); no evidence of clinical disease activity in 73.24% and 61.97% after one and two-years respectively; and brain magnetic resonance imaging T2 lesion volume remained stable. Forty patients suspended natalizumab, in 85% due to high risk of developing progressive multifocal leukoencephalopathy (PML). The major complication was PML (n=3). CONCLUSIONS: Natalizumab showed effectiveness in the long-term follow up period of our cohort, with reduction of ARR, EDSS, and MRI lesion load stabilization. PML was the major complication.

Effect of dietary lysine on performance and expression of electron transport chain genes in the pectoralis major muscle of broilers.

The aim of this study was to evaluate the effect of dietary lysine on performance, protein deposition and respiratory chain gene expression in male broilers. A total of 252 Cobb 500 broilers were distributed, in a completely randomized design, into four treatments with seven replicates of nine birds per experimental unit. Experimental treatments consisted of diets based on corn and soybean meal, with four levels of digestible lysine: 1.016%, 1.099%, 1.182% and 1.265%. The increase in the level of digestible lysine in the diet provided higher weight gains, feed efficiency and body protein deposition. Birds fed the lowest level of dietary lysine (1.016%) showed a lower expression of genes such as NADH dehydrogenase subunit I (ND1), cytochrome b (CYTB) and cytochrome c oxidase subunits I (COX I), II (COX II) and III (COX III), displaying the worst performance and body protein deposition. This demonstrates the relationship existing between the expression of the evaluated genes and the performance responses. In conclusion, results indicate that broilers fed diets with higher levels of digestible lysine have increased messenger RNA expression of some genes coded in the mitochondrial electron transport chain (ND1, CYTB, COX I, COX II and COX III). It may be stated that diets with proper levels of digestible lysine, within the 'ideal protein' concept, promote the expression of genes, which increases the mitochondrial energy, thereby fostering body protein deposition and the performance of broilers in the starter phase.

The effectiveness of fingolimod in a Portuguese real-world population.

INTRODUCTION: Fingolimod is an oral treatment for Relapsing-Remitting Multiple Sclerosis (RRMS) with established efficacy in clinical trials. Post-marketing studies are important to assess its effectiveness in real-world populations. OBJECTIVES: To report the effectiveness and safety of fingolimod in a real-world population. METHODS: A retrospective study of patients with RRMS treated with fingolimod for at least six months. The demographic characteristics, Annualized Relapse Rate (ARR), Expanded Disability Status Score (EDSS), previous treatments and Adverse Events (AE) were analysed. RESULTS: 104 patients were included, with a mean treatment duration of 21.06 months. First-line disease modifying therapy failure patients (n=56) had an ARR decrease of 68.53% (1.43 vs. 0.45, p<0.001), 66.07% of them were relapse-free, EDSS significantly decreased (2.5 vs. 2.0, p<0.001) and 91.07% showed no disability progression. In patients previously treated with natalizumab as a second-line drug mainly switched due to safety concerns (n=41), although the differences were not statistically significant, both the ARR and EDSS increased in 41.46% and 19.51% of patients, respectively. In treatment-naive patients (n=7) the ARR decreased 94.90% (1.57 vs. 0.08, p=0.027) and there was no disability progression. 56.7% of all patients experienced AE not considered serious in any of the cases. CONCLUSION: In this population, fingolimod was an effective treatment after first-line treatment failure, decreasing both the ARR and EDSS, and may be an effective option after natalizumab.

Queen volatiles as a modulator of Tetragonisca angustula drone behavior.

Tetragonisca angustula mating occurs during the virgin queen nuptial flight, usually in the presence of a drone congregation area (DCA). The presence of virgin queen pheromone is considered the trigger for DCA establishment, although this has not been demonstrated experimentally. We established meliponaries, in different habitats, with T. angustula virgin queens during the main drone reproduction period. Eight DCAs were observed in urban areas, and all established outside or near colonies containing at least one virgin queen. The accumulation of drones in the DCAs occurred from 08:00 to 18:00 h and over 3-35 days. The number of drones in DCAs ranged from 60 to 2,000. In field trials, drones were attracted to virgin queens and also, unexpectedly, to physogastric queens. Volatiles collected from both virgin and physogastric queens elicited strong electoantennogram (EAG) responses from drones. Virgin and physogastric queen volatiles were qualitatively similar, but quantitatively different, in chemical composition. The queen's abdomen was the principal source of these compounds. Isopropyl hexanoate (IPH), the most abundant compound in virgin queen volatiles and one of the most abundant in physogastric queen volatiles, was identified as one of the compounds that elicited EAG responses and was demonstrated to attract drones in a field test.

Independent patterns of damage to retinocortical pathways in multiple sclerosis without a previous episode of optic neuritis.

Asymptomatic visual loss is a feature of multiple sclerosis (MS) but its relative impact on distinct retinocortical pathways is still unclear. The goal of this work was to investigate patterns of subclinical visual impairment in patients with MS with and without clinically associated previous optic neuritis (ON). We have used functional methods that assess parvo-, konio- and magnocellular pathways in order to compare pathophysiological mechanisms of damage in a population of 44 subjects with MS (87 eyes), with and without a previous episode of ON. These methods included chromatic contrast sensitivity across multiple chromatic axes (Cambridge Colour Test-parvo/konio pathways), perimetric achromatic contrast sensitivity for the magno pathway [frequency doubling technique (FDT)] and pattern visual evoked potentials (VEP). These measures were correlated with field sensitivity measures obtained using conventional automated static perimetry (ASP) and were also compared with conventional clinical chromatic/achromatic contrast sensitivity chart-based measures. We have found evidence for uncorrelated damage of all retinocortical pathways only in patients with MS without ON. VEP evidence for axonal damage was found in this group supporting the emerging notion of axonal damage even in sub-clinical stages of ON/MS pathophysiology. Only in this group was significant correlation of functional measures with disease stage observed, suggesting that distinct pathophysiological milestones are present before and after ON has occurred.

Phytosterol compounds having antiviral efficacy.

The present study is focused on the antiviral action patterns obtained in vitro with synthetic sterolester comprising compositions on virus-bearing host cell-lines. Appropriate cell-lines were infected with HIV-1, human Cytomegalovirus (HCMV) and Herpes simplex virus (HSV). There appears to exist a clear anti-infective efficacy for a selected number of such ester compounds, provided they are formulated into spontaneously dispersible concentrates, which in aqueous dilution engender ultramicro-emulsions having micelles in the lowest nanosize region. A significant protection against HIV-induced cytopathogenic effect was demonstrated employing a methyltetrazolium salt reduction assay on HIV-infected MT4 cells when they were incubated with such concentrates. A similar effect was evidenced with the same concentrates, when preincubating concentrated virus, but not the target cells. Antiviral activity appeared to be remarkable also on HCMV infections in vitro, where a blocking effect on immediate-early antigen expression in fibroblast monolayers could be observed. Similarly, HSV-associated glycoprotein antigen in VERO cells also suggests that virus-cell interaction and/or virus multiplication could have been blocked at a very early point of time. This would be quite different from antiviral action-patterns studied so far and imputed into the current models of explanation. Proper solubilization of the employed phytosterol compounds is essential for achieving the described activity modes. The often recommended liposome formulations would not be well suited for such compounds and such purpose, since after dilution they produce aqueous macro-emulsions, only. Furthermore, liposome formulations tend to coalesce and exhibit Marangoni effects.

Sneddon's syndrome: a vascular systemic disease with kidney involvement?

Sneddon's syndrome is a systemic disease characterized by livedo reticularis and cerebrovascular disease. Other organs may be involved as well. Typical vascular lesions in the skin biopsy and/or digital arteries biopsy can be found. Arterial hypertension, cardiac pathology (ischemic disease, myocardial infarction, valvulopathy), venous thrombosis and even fetal death are also found sometimes. We present a case of Sneddon's syndrome in which typical vascular lesions in the kidney were demonstrated for the first time unequivocally confirming the systemic nature of this syndrome.

[Frontal lobe-type dementia associated with motor neuron disease]. / Demencia do tipo frontal associada a doenca do neuronio motor.

A case of presenile dementia with dominant frontal disfunction, progressive aphasia and Motor Neuron Disease with prominent bulbar signs is reported. Considering the clinical examination, the measurements of the regional cerebral flow (SPECT) and the histological appearances, we suggest the diagnosis of Dementia of Frontal Lobe Type and Motor Neuron Disease. We reviewed other disorders labelled Primary Frontal or Fronto-temporal Dementias and we discuss this new dementia and the difficulty in its classification.

Effects of heparin and its desulfated derivatives on leukocyte-endothelial adhesion.

The effect of heparin and partially desulfated heparin derivatives on thrombin and PAF-induced adhesion of PMNs to the endothelium was studied either by a fluorescence image analysis or by 111In-labeled PMNs. The results observed with these two techniques consistently indicated that heparin and O-des-heparin inhibited PMN adhesion in a dose-dependent manner. Moreover, N-des-Hep and N-O-des-Hep, even if less effective, also inhibited the adhesion of PMNs when used at high concentrations. The effect of heparin and heparin derivatives was not directed to endothelial cells but rather to PMNs, as shown by the absence of inhibitory effects, when heparins were preincubated with endothelium.

Abnormal proteins in the cerebrospinal fluid of a patient with Creutzfeldt-Jakob disease following administration of human pituitary growth hormone.

A Brazilian case of Creutzfeldt-Jakob disease in a hypopituitary patient who had received cadaver-derived human pituitary growth hormone between 1968 and 1977 is reported. The clinical diagnosis was confirmed during his lifetime by the demonstration of two abnormal 30-kDa proteins in the cerebrospinal fluid by two-dimensional gel electrophoresis. These proteins, characteristic of Creutzfeldt-Jakob disease, present isoelectric points of 5.1 and 5.2. Furthermore, both proteins migrate as doublets, each one displaying a molecular weight variant of about 29-kDa. This is one of 16 cases of the disease associated to therapy with cadaver-derived human growth hormone and one of the few examples among such cases of confirmation of the clinical diagnosis by biochemical characterization of abnormal proteins in the cerebrospinal fluid.

Abnormal proteins in the cerebrospinal fluid of a patient with Creutzfeldt-Jakob disease following administration of human pituitary growth hormone

A Brazilian case of Creutzfeldt-Jakob disease in a hypopituitary patient who had received cadaver-derived human pituitary growth hormone between 1968 and 1977 is reported. The clinical diagnosis was confirmed during his lifetime by the demonstration of two abnormal 30-kDa proteins in the cerebrospinal fluid by two-dimensional gel electrophoresis. These proteins, characteristic of Creutzfeldt-Jakob disease, present isoelectric points of 5.1 and 5.2. Furthermore, both proteins migrate as doublets, each one displaying a molecular weight variant of about 29-kDa. This is one of 16 cases of the disease associated to therapy with cadaver-derived human growth hormone and one of the few examples among such cases of confirmation of the clinical diagnosis by biochemical characterization of abnormal proteins in the cerebrospinal fluid

[Fast intravenous rehydration with 90 mmol/L of sodium in dehydrated children with diarrhea]. / Rehidratación endovenosa rápida con 90 mmol/L de sodio en niños deshidratados por diarrea.

Twenty-two infants with moderate dehydration due to diarrhea in whom oral rehydration therapy (ORT) was contraindicated or who failed to respond to this method of therapy were treated with rapid intravenous rehydration (RIR). Clinical signs of dehydration without shock allowed us to estimate 5% to 10% of weight loss. Patients were 11 days to 19 months old, and 9 of them were undernourished. A standard solution containing 90 mmol/L sodium, 80 mmol/L chloride, 30 mmol/L bicarbonate, 20 mmol/L potassium and 111 mmol/L glucose was used for all patients. The IV infusion was maintained until the rehydration was completed at a rate of 15 to 20 mL/kg/hour. Complete rehydration was successfully achieved in all patients. A total of 89.5 +/- 25.0 mL/kg (mean +/- SD) was needed and the duration of the IV infusion was 5.1 +/- 1.6 hours. Weight gain achieved was 6.5 +/- 1.6%. None of the patients developed hypernatremia following treatment. The initial base deficit, -9.5 +/- 6.6, was reduced to -3.5 +/- 2.9. All of the patients tolerated refeeding immediately after completion of the IV infusion. Our study suggests that this modality of rehydration is well tolerated, safe and effective and enhances the possibility of an early hospital discharge.

Prognostic significance of the expression of immunohistochemically detectable differentiation markers in laryngeal carcinomas.

Expression of immunocytochemically detectable markers in 100 cases of laryngeal carcinomas, homogeneous for staging and treatment, was correlated with clinical evolution of the disease. Follow-up for a minimum of 5 years was obtained in all cases. Paraffin sections were re-cut and stained in immunoperoxidase with monoclonal KL1, detecting medium-to-low molecular weight keratins, and with monoclonal HMFG2, revealing a surface glycoprotein. Expression of KL1-related antigen did not correlate with prognosis, whereas cases extensively positive for monoclonal HMFG2 (more than 50% cells stained) had a significantly better recurrence-free rate. In a group of tumors classified as Grade 3 (histologically poorly differentiated) and expressing a low degree of HMFG2-detectable surface glycoprotein (less than 50% cells stained), a high rate of recurrences (93%) was observed. This study indicates that the combined use of morphologic and biologic (immunohistochemical) criteria may constitute an independent parameter of primary importance in predicting the evolution of laryngeal carcinomas.

Monoclonal antibodies against laryngeal carcinomas. Preliminary report.

Following types of monoclonal antibodies (MoAb) (HMFG 1, 2 and AI 16.9) were used in twenty patients with laryngeal carcinomas (10 pharyngolaryngeal carcinomas, T4 N+ MO, and 10 vestibulo-glottic carcinomas, T1 T2, N+ N-, MO). The demonstration of MoAb's binding was performed with immunoperoxidase technique. The results demonstrate that the HMFG 1 and 2 antibodies fixed to both T and N, mostly good or very good, while the AI 16.9 antibody fixed in a various way. None of these three MoAb's was specific for the tissue of metastasis-free satellite lymph nodes.