Dynein-dynactin segregate meiotic chromosomes in C. elegans spermatocytes.

The microtubule motor cytoplasmic dynein 1 (dynein) and its essential activator dynactin have conserved roles in spindle assembly and positioning during female meiosis and mitosis, but their contribution to male meiosis remains poorly understood. Here, we characterize the G33S mutation in the C. elegans dynactin subunit DNC-1, which corresponds to G59S in human p150Glued that causes motor neuron disease. In spermatocytes, dnc-1(G33S) delays spindle assembly and penetrantly inhibits anaphase spindle elongation in meiosis I, which prevents the segregation of homologous chromosomes. By contrast, chromosomes segregate without errors in the early dnc-1(G33S) embryo. Deletion of the DNC-1 N-terminus shows that defective meiosis in dnc-1(G33S) spermatocytes is not due to the inability of DNC-1 to interact with microtubules. Instead, our results suggest that the DNC-1(G33S) protein, which is aggregation prone in vitro, is less stable in spermatocytes than the early embryo, resulting in different phenotypic severity in the two dividing tissues. Thus, the dnc-1(G33S) mutant reveals that dynein-dynactin drive meiotic chromosome segregation in spermatocytes and illustrates that the extent to which protein misfolding leads to loss of function can vary significantly between cell types.

Regulation of human T-cell leukemia virus type 1 antisense promoter by myocyte enhancer factor-2C in the context of adult T-cell leukemia and lymphoma.

Adult T-cell leukemia and lymphoma (ATLL) is an intractable T-cell neoplasia caused by a retrovirus, namely human T-cell leukemia virus type 1 (HTLV-1). Patients suffering from ATLL present a poor prognosis and have a dearth of treatment options. In contrast to the sporadic expression of viral transactivator protein Tax present at the 5' promoter region long terminal repeats (LTR), HTLV-1 bZIP gene (HBZ) is encoded by 3'LTR (the antisense promoter) and maintains its constant expression in ATLL cells and patients. The antisense promoter is associated with selective retroviral gene expression and has been an understudied phenomenon. Herein, we delineate the activity of transcription factor MEF (myocyte enhancer factor)-2 family members, which were found to be enriched at the 3'LTR and play an important role in the pathogenesis of ATLL. Of the four MEF isoforms (A to D), MEF-2A and 2C were highly overexpressed in a wide array of ATLL cell lines and in acute ATLL patients. The activity of MEF-2 isoforms were determined by knockdown experiments that led to decreased cell proliferation and regulated cell cycle progression. High enrichment of MEF-2C was observed at the 3'LTR along with cofactors Menin and JunD resulting in binding of MEF-2C to HBZ at this region. Chemical inhibition of MEF-2 proteins resulted in the cytotoxicity of ATLL cells in vitro and reduction of proviral load in a humanized mouse model. Taken together, this study provides a novel mechanism of 3'LTR regulation and establishes MEF-2 signaling a potential target for therapeutic intervention for ATLL.

Increased cholinergic activity under conditions of low estrogen leads to adverse cardiac remodeling.

Cholinesterase inhibitors are used in postmenopausal women for the treatment of neurodegenerative diseases. Despite their widespread use in the clinical practice, little is known about the impact of augmented cholinergic signaling on cardiac function under reduced estrogen conditions. To address this gap, we subjected a genetically engineered murine model of systemic vesicular acetylcholine transporter overexpression (Chat-ChR2) to ovariectomy and evaluated cardiac parameters. Left-ventricular function was similar between Chat-ChR2 and wild-type (WT) mice. Following ovariectomy, WT mice showed signs of cardiac hypertrophy. Conversely, ovariectomized (OVX) Chat-ChR2 mice evolved to cardiac dilation and failure. Transcript levels for cardiac stress markers atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) were similarly upregulated in WT/OVX and Chat-ChR2/OVX mice. 17ß-Estradiol (E2) treatment normalized cardiac parameters in Chat-ChR2/OVX to the Chat-ChR2/SHAM levels, providing a link between E2 status and the aggravated cardiac response in this model. To investigate the cellular basis underlying the cardiac alterations, ventricular myocytes were isolated and their cellular area and contractility were assessed. Myocytes from WT/OVX mice were wider than WT/SHAM, an indicative of concentric hypertrophy, but their fractional shortening was similar. Conversely, Chat-ChR2/OVX myocytes were elongated and presented contractile dysfunction. E2 treatment again prevented the structural and functional changes in Chat-ChR2/OVX myocytes. We conclude that hypercholinergic mice under reduced estrogen conditions do not develop concentric hypertrophy, a critical compensatory adaptation, evolving toward cardiac dilation and failure. This study emphasizes the importance of understanding the consequences of cholinesterase inhibition, used clinically to treat dementia, for cardiac function in postmenopausal women.

Potential role of microRNAs as biomarkers in human glioblastoma: a mini systematic review from 2015 to 2020.

Glioblastoma (GBM) is the most common, aggressive and malignant type of glioma, with poor prognosis, despite advances in medical knowledge and technology. It's known that some microRNAs (miRNAs) can be dysregulated and associated with tumors. We aim to investigate miRNAs that may have a role as potential biomarkers in human glioblastoma. A search was performed using PubMed, LILACS and SCIELO databases to find papers from 2015 to 2020, related to human in vitro and ex vivo data. From 99 articles, 10 were eligible and 13 dysregulated miRNAs were found with description of regulation, target(s), pathway(s) and mechanism(s). The miRNAs of interest were found and seem to be involved in development and progression of glioblastoma and used as target therapies. Understanding the mechanisms in which those miRNAs are involved and their role in epigenetic pathways that lead to cancer, as well as their potential in clinical application, may improve GBM clinical outcome (CRD42020182706, 07/10/2020, retrospectively registered).

Parental Monitoring of Computer Use and its Association with Drug Use among Students in Southern Brazil.

BACKGROUND: Parental control is considered important in preventing drug use among adolescents. OBJECTIVE: This study aimed to explore the association between the perception of parental monitoring of computer use and drug use among adolescent students. METHODS: A cross-sectional school-based study was conducted in 2012 among 2,980 students aged 12-17 years from Southern Brazil. The outcomes investigated were the use of tobacco, alcohol, cannabis, and other illicit drugs in the previous year. The main exposure was the self-reported perception of parental monitoring of computer use. Poisson regression with robust variance was used for the multivariable analysis. RESULTS: The prevalence rates of drugs used were 48.3% (95% confidence interval[CI]: 46.5-50.0) for alcohol, 9.7% (95%CI: 8.7-10.7) for tobacco, 5.1% (95%CI: 4.3-5.9) for cannabis, and 8.7% (95%CI: 7.7-9.7) for other illicit drugs. The absence of monitoring of computer use was reported by 44.3% (95%CI: 42.5-46.1) of the students. After adjusting for potential confounders, we found that students aged 12 to 14 years who reported no parental monitoring of computer use had a higher probability of alcohol (Prevalence Ratio[PR] = 1.26, 95%CI:1.07-1.49) and cannabis use (PR = 3.25, 95%CI:1.11-9.53). In addition, a higher consumption of tobacco was observed among students aged 15 to 17 years who reported no parental monitoring of computer use (PR = 1.75, 95%CI:1.17-2.63). In both age groups, psychiatric morbidity was a covariate significantly associated with drug use throughout analysis. CONCLUSIONS: Parental monitoring of computer use was significantly associated with adolescent drug use, suggesting its importance in preventing drug use.

Association between interferon lambda 3 rs12979860 polymorphism and clinical outcome in dengue virus-infected children.

Single nucleotide polymorphisms (SNPs) in immune-related genes have been shown to play a role in driving the development of the severe phenotypes of dengue virus (DENV) infection. We assessed the association between IFNL3 gene SNP (rs12979860) and dengue clinical outcomes in children. Patients with dengue-related symptoms (aged 1-15 years) admitted at a public hospital in Northeast Brazil were invited to participate. The association between rs12979860 polymorphism and dengue classification and clinical signs and symptoms were analysed. A total of 206 DENV-infected children were included: 53.4% of the infections were classified as severe dengue. The T allele carriers had higher risk of developing severe dengue when compared to CC genotype carriers (OR: 1.81; 95% CI: 0.98-3.32 p = .054). The T allele carriers also showed longer fever episodes when compared to patients with the CC genotype (OR: 1.90; 95%CI: 1.07-3.38; p = .027). On the other hand, the ones carrying the CT/TT genotype had 70% lower chance of developing thrombocytopenia when compared to those with the CC genotype (OR: 0.30; 95%CI: 0.08-0.88; p = .042). Our findings demonstrated that the T allele carriers of the IFNL3 gene had higher risk of developing severe dengue, suggesting a link between IFN-λ expression and DENV immunopathogenesis.

Reproductive characteristics of bulls from two breed compositions and their correlations with infrared thermography.

The objective was to evaluate reproductive characteristics of crossbred Girolando (Gyr x Holstein) bulls from two breed compositions and correlate these results with infrared thermography data. Evaluations were performed considering sperm motility, vigor and morphology; scrotal circumference; body morphology and temperament. Infrared thermography was performed to determine surface temperatures of ocular and scrotal areas. Thermoregulation capacity was assessed by differences between air and rectal temperatures, air and maximum temperatures in ocular and scrotal areas, and dorsal and ventral lines of the scrotum. Data analysis was performed using a linear mixed model (breed composition as fixed effect and year of evaluation as random effect). Spearman correlation coefficient was used to associate thermography and reproductive data. Girolando 3/4 Holstein bulls had higher (P ≤ 0.001) scrotal circumference and higher average body morphology and temperament, whereas 5/8 Holstein bulls had a higher (P < 0.001) percentage of major-type and total sperm defects. Girolando 3/4 Holstein bulls had scrotal temperatures 0.8 °C higher (P < 0.001) and 5/8 Holstein bulls had 9.8% and 10.6% higher differences on "rectal - scrotal area" and "ocular area - scrotal area" temperatures, respectively. Ocular area temperatures had negative correlations (P ≤ 0.1) with sperm motility and vigor, and positive correlations (P < 0.001) with minor-type and total sperm defects. Ventral line scrotal temperatures had positive correlations (P < 0.001) with minor-type and total sperm defects. Girolando 3/4 Holstein bulls were found to be superior to 5/8 Holstein bulls for reproductive characteristics. Under non-stressing climatic conditions, semen characteristics of Girolando bulls were more influenced by breed composition than by the capacity for scrotal thermoregulation. Correlations between semen quality and scrotal temperatures can aid in the identification of bulls for breeding, particularly when a large number of animals are in the tests.

Healthcare Experience of People with Acute Spinal Cord Injury: A Phenomenological Study.

Living with spinal cord injury (SCI) is a challenge that begins in the acute phase, when the disease, the limitations, and the treatments fill the days at the hospital. This study aims to understand the healthcare experience of the person with SCI in the acute phase, based on the Activities of Living Nursing Model (ALNM). It is a qualitative and phenomenological study based on the Standards for Reporting Qualitative Research. Data were collected via semi-structured interviews. Content analysis was performed using the ATLAS.ti software and Bardin's methodology. The article was written following the COREQ guidelines. The categories were defined using the Roper-Logan-Tierney Model for Nursing. The sample included 16 people with incomplete SCI, different etiology, and neurological levels. Eleven of the twelve ALNM emerged from the interviews. The activities of mobilizing, eliminating, maintaining a safe environment, and communicating were emphasized the most. Controlling body temperature was not relevant. Mobility deficits and pain increased dependence. Feelings of motivation, encouragement, and frustration were highlighted. Professional expertise, rehabilitation resources, and support equipment promoted independence. The results in this sample revealed that people with SCI in the acute phase have complex challenges related to dependence awareness and treatments, but they always keep recovery expectations in mind.

Peptide fragments of bradykinin show unexpected biological activity not mediated by B1 or B2 receptors.

BACKGROUND AND PURPOSE: Bradykinin (BK-(1-9)) is an endogenous nonapeptide involved in multiple physiological and pathological processes. Peptide fragments of bradykinin are believed to be biologically inactive. We have now tested the two major peptide fragments of bradykinin in human and animals. EXPERIMENTAL APPROACH: BK peptides were quantified by MS in male rats. NO release was quantified from human, mouse and rat cells loaded with DAF-FM. Rat aortic rings were used to measure vascular reactivity. Changes in BP and HR were measured in conscious male rats. To evaluate pro-inflammatory effects both vascular permeability and nociception were measured in adult mice. KEY RESULTS: BK-(1-7) and BK-(1-5) are produced in vivo from BK-(1-9). Both peptides induced NO production in all cell types tested. However, unlike BK-(1-9), NO production elicited by BK-(1-7) or BK-(1-5) was not inhibited by B1 or B2 receptor antagonists. BK-(1-7) and BK-(1-5) induced concentration-dependent vasorelaxation of aortic rings, without involvement of B1 or B2 receptors. Intravenous or intra-arterial administration of BK-(1-7) or BK-(1-5) induced similar hypotensive response in vivo. Nociceptive responses of BK-(1-7) and BK-(1-5) were reduced compared to BK-(1-9), and no increase in vascular permeability was observed for BK-(1-9) fragments. CONCLUSIONS AND IMPLICATIONS: BK-(1-7) and BK-(1-5) are endogenous peptides present in plasma. BK-related peptide fragments show biological activity, not mediated by B1 or B2 receptors. These BK fragments could constitute new, active components of the kallikrein-kinin system.

Spirocamallanus krameri and Contracaecum sp. thirdstage larvae (Nematoda) in Hoplerythrinus unitaeniatus (Erythrinidae) of eastern Amazon (Brazil).

This study aimed to investigate the presence of nematodes Spirocamallanus krameri and Contracaecum sp. in Hoplerythrinus unitaeniatus caught in a lake in the state of Pará, in northern Brazil, eastern Amazon region (Brazil). From 92 hosts, 83.7% were infected, and a total of 401 parasites were covered. There was a high prevalence of S. krameri, while the predominance was of Contracaecum sp. larvae. Both parasite species presented aggregated dispersion, and abundance of these parasites presented positive correlation with weight and length of the hosts. Morphological and morphometric data of third-stage larvae of Contracaecum sp. are described.

Adoption of Precision Technologies by Brazilian Dairy Farms: The Farmer's Perception.

The use of precision farming technologies, such as milking robots, automated calf feeders, wearable sensors, and others, has significantly increased in dairy operations over the last few years. The growing interest in farming technologies to reduce labor, maximize productivity, and increase profitability is becoming noticeable in several countries, including Brazil. Information regarding technology adoption, perception, and effectiveness in dairy farms could shed light on challenges that need to be addressed by scientific research and extension programs. The objective of this study was to characterize Brazilian dairy farms based on technology usage. Factors such as willingness to invest in precision technologies, adoption of sensor systems, farmer profile, farm characteristics, and production indexes were investigated in 378 dairy farms located in Brazil. A survey with 22 questions was developed and distributed via Google Forms from July 2018 to July 2020. The farms were then classified into seven clusters: (1) top yield farms; (2) medium-high yield, medium-tech; (3) medium yield and top high-tech; (4) medium yield and medium-tech; (5) young medium-low yield and low-tech; (6) elderly medium-low yield and low-tech; and (7) low-tech grazing. The most frequent technologies adopted by producers were milk meters systems (31.7%), milking parlor smart gate (14.5%), sensor systems to detect mastitis (8.4%), cow activity meter (7.1%), and body temperature (7.9%). Based on a scale containing numerical values (1-5), producers indicated "available technical support" (mean; σ2) (4.55; 0.80) as the most important decision criterion involved in adopting technology, followed by "return on investment-ROI" (4.48; 0.80), "user-friendliness" (4.39; 0.88), "upfront investment cost" (4.36; 0.81), and "compatibility with farm management software" (4.2; 1.02). The most important factors precluding investment in precision dairy technologies were the need for investment in other sectors of the farm (36%), the uncertainty of ROI (24%), and lack of integration with other farm systems and software (11%). Farmers indicated that the most useful technologies were automatic milk meters systems (mean; σ2) (4.05; 1.66), sensor systems for mastitis detection (4.00; 1.57), automatic feeding systems (3.50; 2.05), cow activity meter (3.45; 1.95), and in-line milk analyzers (3.45; 1.95). Overall, the concerns related to data integration, ROI, and user-friendliness of technologies are similar to those of dairy farms located in other countries. Increasing available technical support for sensing technology can have a positive impact on technology adoption.

Increased Cholinergic Tone Causes Pre-synaptic Neuromuscular Degeneration and is Associated with Impaired Diaphragm Function.

In vertebrates, muscle activity is dependent on acetylcholine (ACh) released from neuromuscular junctions (NMJs), and changes in cholinergic neurotransmission are linked to a variety of neuromuscular diseases, including congenital myasthenic syndromes (CMS). The storage and release of ACh depends on the activity of the Vesicular Acetylcholine Transporter (VAChT), a rate-limiting step for cholinergic neurotransmission whose loss of function mutations was shown to cause human congenital myasthenia. However, we know much less about increased VAChT activity, due to copy number variations, for example. Therefore, here we investigated the impact of increased VAChT expression and consequently ACh levels at the synaptic cleft of the diaphragm NMJs. We analyzed structure and function of nerve and muscles from a mouse model of cholinergic hyperfunction (ChAT-ChR2-EYFP) with increased expression of VAChT. Our results showed a significant increase of ACh released under evoked stimuli. However, we observed deleterious changes in synaptic vesicles cycle (impaired endocytosis and decrease in vesicles number), together with structural alterations of NMJs. Interestingly, ultrastructure analyses showed that synaptic vesicles from ChAT-ChR2-EYFP mice NMJs were larger, which might be related to increased ACh load. We also observed that these larger synaptic vesicles were less rounded in comparison with control. Finally, we showed that ChAT-ChR2-EYFP mice NMJs have compromised safety factor, possible due to the structural alterations we described. These findings reveal that physiological cholinergic activity is important to maintain the structure and function of the neuromuscular system and help to understand some of the neuromuscular adverse effects experienced by chronically increased NMJ neurotransmission, such as individuals treated with cholinesterase inhibitors.

Dense optical flow software to quantify cellular contractility.

Cell membrane deformation is an important feature that occurs during many physiological processes, and its study has been put to good use to investigate cardiomyocyte function. Several methods have been developed to extract information on cardiomyocyte contractility. However, no existing computational framework has provided, in a single platform, a straightforward approach to acquire, process, and quantify this type of cellular dynamics. For this reason, we develop CONTRACTIONWAVE, high-performance software written in Python programming language that allows the user to process large data image files and obtain contractility parameters by analyzing optical flow from images obtained with videomicroscopy. The software was validated by using neonatal, adult-, and human-induced pluripotent stem-cell-derived cardiomyocytes, treated or not with drugs known to affect contractility. Results presented indicate that CONTRACTIONWAVE is an excellent tool for examining changes to cardiac cellular contractility in animal models of disease and for pharmacological and toxicology screening during drug discovery.

[Epidemiological and socio-demographic profile of primary care workers in the South and Northeast of Brazil]. / Perfil sócio-demográfico e epidemiológico dos trabalhadores da atenção básica à saúde nas regiões Sul e Nordeste do Brasil.

In order to describe the profile of primary health care teams in 41 municipalities with more than 100 thousand inhabitants each, a total of 4,749 health workers in two States from the South (1,730) and five from the Northeast (3,019) of Brazil were included from a sample of traditional primary care units and the Family Health Program (FHP). After providing informed consent, the health workers answered a self-applied questionnaire with demographic, work-related, and their own health-related data. The principal differences between the two models involved the structuring of the teams, with the FHP including more community health agents, more women, more young workers, fewer hired on the basis of formal admissions exams, more with a single job, more precarious employment arrangements, less employment satisfaction, less time on the job, larger workloads, greater specialization in the area, and better pay. The FHP also showed worse self-perceived health and more medical appointments. Management efforts are needed to support these workers, who form the basis of the health system and are key protagonists in the development and consolidation of primary care.

[Evaluation of the effectiveness of Primary Health Care in South and Northeast Brazil: methodological contributions]. / Avaliação de efetividade da Atenção Básica à Saúde em municípios das regiões Sul e Nordeste do Brasil: contribuições metodológicas.

The article presents the methodology used in the Baseline Study that evaluated the effectiveness of the Family Health Strategy as compared to traditional primary health care units. The study included 41 municipalities with more than 100 thousand inhabitants each, 21 from the South and 20 from the Northeast of Brazil. As the principal dependent variable and underlying premise for sampling in the Baseline Study, the effectiveness of program activities in the primary care units was analyzed in the population within the respective coverage areas, using an epidemiological survey. The health care model in the primary care units was the principal independent variable. Its effect on program activities was controlled according to geopolitical region, metropolitan area, and population size in the municipalities. Coverage of the activities was characterized according to socioeconomic, demographic, and health-related factors. The use of comparison groups, multiple-stage samples, standardized measures, adjustment for geographic and socio-demographic characteristics, and well-defined criteria for judging the findings are contributions by the methodology employed here for designing future studies to evaluate primary health care.

Results of stapes surgery for otosclerosis with two kinds of prosthesis in residency training.

UNLABELLED: Stapes surgery is one of the approaches indicated to treat conductive hearing loss secondary to otosclerosis. The procedures requires skill and experience from the surgeon and is part of medical residency training. AIMS: To assess which type of prosthesis (Teflon or metal/steel) presents the best results in surgeries performed by residents and the incidence of complications. MATERIALS AND METHODS: we retrospectively assessed 189 interventions that counted on the active participation of resident physicians, and we compared the two types of prosthesis used. Audiometric results were analyzed following the guidelines from the Committee on Hearing and Equilibrium and also according to the Amsterdam Hearing Evaluation Plots. RESULTS: Bone-air gap reduced in an average value of 21.90 dB (p<0.05) after the surgery in the group that received the Teflon prosthesis and 21.37 dB (p<0.05) in the group that received the mixed prosthesis, and gain in SRI was of 22.33 and 26.10 dB (p<0.05), and the air-bone gap was below 20 dB in 80.6% and 85.04%, respectively. CONCLUSIONS: We did not see differences in the audiometry and in the incidence of complications when we compared the type of prosthesis used. We believe it is valid to continue teaching this procedure in medical residency training programs, regardless of the type of prosthesis.

Inferences on specificity recognition at the Malus×domestica gametophytic self-incompatibility system.

In Malus × domestica (Rosaceae) the product of each SFBB gene (the pollen component of the gametophytic self-incompatibility (GSI) system) of a S-haplotype (the combination of pistil and pollen genes that are linked) interacts with a sub-set of non-self S-RNases (the pistil component), but not with the self S-RNase. To understand how the Malus GSI system works, we identified 24 SFBB genes expressed in anthers, and determined their gene sequence in nine M. domestica cultivars. Expression of these SFBBs was not detected in the petal, sepal, filament, receptacle, style, stigma, ovary or young leaf. For all SFBBs (except SFBB15), identical sequences were obtained only in cultivars having the same S-RNase. Linkage with a particular S-RNase was further established using the progeny of three crosses. Such data is needed to understand how other genes not involved in GSI are affected by the S-locus region. To classify SFBBs specificity, the amino acids under positive selection obtained when performing intra-haplotypic analyses were used. Using this information and the previously identified S-RNase positively selected amino acid sites, inferences are made on the S-RNase amino acid properties (hydrophobicity, aromatic, aliphatic, polarity, and size), at these positions, that are critical features for GSI specificity determination.

School health promotion and use of drugs among students in Southern Brazil.

OBJECTIVE: To analyze the relationship between the health promotion conditions in schools and the consumption of alcohol and other drugs by students. METHODS: This is a cross-sectional study with a probabilistic sample of 3,464 students aged 12 to 17 from all schools of the cities of Lajeado and Sapiranga, state of Rio Grande do Sul, Brazil, and 53 managers from the same schools; the data was collected in 2012. Reports of the use of tobacco, alcohol, and illicit drugs in 2012 were used as outcomes, and the health promotion score in the school environment was used as the exposure of interest. We submitted the data to multilevel analysis. RESULTS: The prevalence of the annual use of tobacco was 9.8% (95%CI 8.8-10.8), alcohol was 46.2% (95%CI 44.5-47.8), and other drugs was 10.9% (95%CI 9.9-12.0). In the crude analysis, only the use of tobacco was associated with less health promoting schools (OR = 1.89, 95%CI 1.16-3.09) when compared to those with better conditions. This association lost statistical significance in the adjusted analysis (OR = 1.27, 95%CI 0.74-2.19). CONCLUSIONS: The effects of the school environment on the use of drugs, especially tobacco and alcohol, are manifested mainly by the individual and family conditions of the adolescents.

IL17A Polymorphism Is Not Associated with Human T-Lymphotropic Virus 1-Associated Myelopathy/Tropical Spastic Paraparesis.

The human T-lymphotropic virus 1 (HTLV-1) is the causative agent of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The present study investigated the association between the rs2275913 polymorphism in the IL17A gene and the development of HAM/TSP. Peripheral blood samples were collected from 116 patients (29 symptomatic patients with HAM/TSP and 87 asymptomatic) with a positive diagnosis of HTLV-1. The single nucleotide polymorphism genotyping was carried out by real time PCR using TaqMan probes. In addition, serum levels of IL-2, IFN-γ, TNF-α, IL-4, IL-6, IL-10, and IL-17 were measured in 64 infected individuals from the study (47 asymptomatic and 17 HAM/TSP), using cytometric bead array technique. No significant differences were found in genotypic and allelic frequencies between the groups. Analysis of cytokine levels showed highest concentrations of IFN-γ and TNF-α in HAM/TSP patients. The results of the present study, therefore, suggest a lack of association between the rs2275913 polymorphism in the IL17A gene and the presence of HAM/TSP.

Impact of genetic polymorphisms in key enzymes of homocysteine metabolism on the pathophysiology of sickle cell anemia.

This work aimed at studying a possible influence of methylenetetrahydrofolate reductase (MTHFR; c. 677C>T) and cystathionine ß-synthase (CBS; 844ins68) polymorphisms on overall oxidative status of sickle cell anemia (SCA) patients and on routine markers, correlating them with hydroxycarbamide (HC) treatment. We evaluated 95 unrelated and diagnosed SCA patients. All patients received a prophylactic treatment with folic acid of 5mg/day, while 41 (43.2%) of them were under hydroxycarbamide (HC) treatment (average dose: 22mg/kg/day). MTHFR and CBS polymorphisms were identified by Polymerase Chain Reaction. Biochemical parameters were measured using spectrophotometric and chromatographic methods. Routine markers were developed by specialized laboratory. We did not find any effect of 677T and "I" allele combination on the biomarkers evaluated. On the other hand, MTHFR 677T mutation was related to a depletion of antioxidant capacity, according to the decreased catalase activity and a reduction about 30% of glutathione levels. Moreover, the presence of the insertion was related to about 23% less biomolecule oxidation levels and lower monocytes count, but about 14% higher lactate dehydrogenase activity. These findings may contribute to highlight that the MTHFR and CBS polymorphisms involvement in SCA pathophysiology is likely to be far more complex than it was explored to date.