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Neurodegeneration occurring in multiple sclerosis (MS) contributes to the progression of disability. It is therefore important to identify and neutralize the mechanisms that promote neurodegeneration in MS. Here, we report that oxidized phosphatidylcholines (OxPCs) found in MS lesions, previously identified as end-product markers of oxidative stress, are potent drivers of neurodegeneration. Cultured neurons and oligodendrocytes were killed by OxPCs, and this was ameliorated by microglia. After OxPC injection, mouse spinal cords developed focal demyelinating lesions with prominent axonal loss. The depletion of microglia that accumulated in OxPC lesions exacerbated neurodegeneration. Single-cell RNA sequencing of lesioned spinal cords identified unique subsets of TREM2 high mouse microglia responding to OxPC deposition. TREM2 was detected in human MS lesions, and TREM2 -/- mice exhibited worsened OxPC lesions. These results identify OxPCs as potent neurotoxins and suggest that enhancing microglia-mediated OxPC clearance via TREM2 could help prevent neurodegeneration in MS.

Isoflavone is a phytoestrogen found in different types of food that can act as endocrine disrupters leading to testicular dysfunction. Currently, fragmented data on the action of this compound in the testicles make it difficult to assess its effects to define a safe dose. Thus, we systematically reviewed the preclinical evidence of the impact of isoflavone on testicular function. We also determined which form (aglycones or glycosylated) was the most used, which allowed us to understand the main biological processes involved in testicular function after isoflavone exposure. This systematic review was carried out according to the PRISMA guidelines using a structured search on the biomedical databases MEDLINE (PubMed), Scopus, and Web of Science, recovering and analyzing 22 original studies. The bias analysis and the quality of the studies were assessed by the criteria described in the risk of bias tool developed by SYRCLE (Systematic Review Centre for Laboratory Animal Experimentation). The aglycones and glycosylated isoflavones proved to be harmful to the reproductive health, and the glycosylates at doses of 50, 100, 146, 200, 300, 500, and 600 mg/kg, in addition to 190 and 1000 mg/L, appear to be even more harmful. The main testicular pathologies resulting from the use of isoflavones are associated with Leydig cells resulting from changes in molecular functions and cellular components. The most used isoflavone to evaluate testicular changes was the genistein/daidzein conjugate. The consumption of high doses of isoflavones promotes changes in the functioning of Leydig cells, inducing testicular changes and leading to infertility in murine models., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2021 Amanda Alves Lozi et al.)

Objective: To evaluate the compliance of the care process involving insertion of central vascular catheter (CVC) in hemodialysis., Method: Cross-sectional quantitative approach developed at the hemodialysis service of a reference hospital in Sergipe, Brazil. Sample consisting of 1,342 actions evaluated, corresponding to 122 forms for monitoring and control of CVC insertion. Data collection was held from July to December 2016., Results: The adherence rate to the use of the insertion form was 54.9%. The procedure evaluated achieved 93% overall compliance. Of the 11 specific actions observed, seven (64%) presented 100% compliance. The density of the overall incidence of primary bloodstream infections reduced from 10.6 to 3.1 infections per 1,000 patients/day., Conclusion: Although the observed actions reached specific desired conformities, the use of the checklist was lower than expected. Strategies for monitoring, coaching and educational and organizational actions can contribute to safe care.

Objective: To compare the effectiveness of aquatic exercises with patient-education in individuals with knee osteoarthritis., Design: Randomized controlled trial with blinded assessor and intention-to-treat analysis., Setting: Aquatic Physiotherapy Centre and Primary Health Care Unit., Subjects: A total of 60 patients, aged 68.3 (SD = 4.8) with clinical symptoms and radiographic grading (Kellgren-Lawrence 1-4) of knee osteoarthritis were included., Interventions: An eight-week treatment protocol of aquatic exercise ( n = 31) (16 individual sessions, twice a week) and an educational program (group sessions, once a week) ( n = 29)., Main Measures: Before, after eight-week intervention, and a three-month follow-up with results for the following outcome measures: pain, function, quality of life, functional mobility, and depression., Results: At the end of treatment, the WOMAC (Western Ontario and McMaster Universities Osteoarthritis Index) functional capacity values reduced in favour of the aquatic exercise group for both the total score MD (mean difference) = -14.2; 95% CI (confidence interval) (-18; -10.5), P = 0.04 and the pain domain MD = -3.8 points; 95% CI (-8.71; -1), P = 0.021. The total score also reduced in the follow-up: MD = -12.3 points; 95% CI (-24.7; -6.1), P = 0.017. No differences were found for the outcomes functional mobility or depression., Conclusion: Aquatic exercise improved pain and function after eight weeks, and function at the three-month follow-up compared to the patient-education program.

Objective: To evaluate factors predictive for relapse in a cohort of adult patients with acute promyelocytic leukemia monitored by molecular methods during consolidation and during at least one month of maintenance therapy., Methods: The charts and laboratory data of 65 adult patients with acute promyelocytic leukemia treated according to the International Consortium on Acute Promyelocytic Leukemia 2006 protocol were reviewed. The identification of the promyelocytic leukemia-retinoic acid receptor-alpha gene rearrangement at diagnosis, post-induction, post-consolidation and during maintenance treatment was performed by qualitative and quantitative reverse transcription polymerase chain reaction., Results: Eighty-nine patients were diagnosed with acute promyelocytic leukemia over a seven-year period and of these 65 were eligible for treatment with the protocol. Among the 45 patients who received consolidation and maintenance treatment, six (13%) relapsed, three of whom presented hematologic and three presented molecular relapse. The first relapses occurred at a median of 39 months. Relapsed patients were from all risk groups (low, intermediate and high) and both morphological types (M3 and M3variant) were found. Three of these patients are alive and in molecular remission after salvage treatment. There were no statistically significant differences regarding gender, age, risk group, morphology, promyelocytic leukemia breakpoint cluster region, use of all-trans retinoic acid, development of differentiation syndrome and number of days to complete remission between the patients who relapsed and those who did not., Conclusion: Our results reinforce the importance of prolonged monitoring of acute promyelocytic leukemia patients using molecular methods to detect relapse early., (Copyright © 2016 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier Editora Ltda. All rights reserved.)

Objectives: The liver pretransplant phase requires the interaction of -multidisciplinary teams for optimal patient outcomes, including dental -professionals. In this study, we sought to estimate the prevalence of complications related to dental extraction in liver transplant candidates., Material and Methods: This study is a case series of patients eligible for liver transplants referred for dental treatment between 2012 and 2014. Eligible patients for this study underwent at least one tooth extraction, had a complete blood count at the time of the procedure, and were candidates for liver transplantation., Results: Forty-three patients, 12 women (28%) and 31 men (72%), with an -average age of 50.9 ± 11.4 years, had received a total of 116 dental extractions. The prevalence of postoperative complications was 1.7% including two bleeding episodes in the same patient. Seven patients presented a platelet count below 50 × 10³/μL and received platelet transfusions preoperatively., Conclusions: Postoperative complications following dental extractions in liver transplant candidates are unusual., Clinical Relevance: Low postoperative complications prevalence indicates that dental surgical procedures may be safety performed in patients eligible for liver transplantation after careful -preoperative evaluation., (© 2016 Special Care Dentistry Association and Wiley Periodicals, Inc.)

The Rh blood group system is one of the most complex, polymorphic and immunogenic blood group systems in humans. Some individuals produce a weak or a partial D as a result of RHD and RHCE gene conversion events and RHD point mutations. Because the incidence of RHD variants differs considerably among ethnic groups, the objective of this study was to establish the frequency of blood donors carrying some weak and partial RHD, at the molecular level, in 400 blood donors from the North/Northwest of the state of Parana, Southern Brazil. Another 30 blood donors whose RhD typing results in serology were inconclusive were also included. In this mixed Brazilian population, the most frequent weak D types were 1, 4, 3 and 2 (frequencies of 4.35%, 2.32%, 1.46% and 0.29%, respectively; total of 8.41%) and partial D was found in 2.90% of samples carrying the RHD gene. For samples with inconclusive RhD typing, 53.33% of them presented weak and partial RHD, and 43.75% had concomitantly more than one RHD variant. Our results demonstrate the presence of Caucasian and African D variants. This knowledge can contribute to the safety of transfusion strategies in this ethnic admixture population., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Dengue is responsible for a wide range of clinical manifestations, ranging from asymptomatic infections to severe cases. The alteration of cytokine levels correlated with clinical characteristics can help determine prognostic markers of the disease and the identification of targets for immunotherapy. We measured the viral load, serotype, and cytokine levels of 212 serum samples from patients with acute dengue infection during days 1-4 after the onset of symptoms. The patients were classified as either with hemorrhagic manifestations (HM) or with no hemorrhagic manifestations (NHM). The cytokines interleukin-6 (IL-6), IL-8, and IL-10 were increased (P < 0.05) in the dengue virus+ group, compared with the control group. A higher viral load (P < 0.05) and IL-6 was detected in the HM group compared with the NHM group. Interestingly, the NHM group demonstrated a significant positive correlation between inflammatory (IL-6 and 8) and anti-inflammatory (IL-10) cytokines, whereas the HM group did not. These findings suggest that a disturbance in the balance of inflammatory cytokines IL-6 and IL-8 with the anti-inflammatory cytokine, IL-10, combined with the high levels of IL-6 and viral load, characterize possible mechanisms related to the formation of HM., (© The American Society of Tropical Medicine and Hygiene.)

Background: The aim of this study was to evaluate the morphological and functional standards and quality of life of people living with HIV/AIDS., Methods: This was a quasi-experimental study with 15 people living with HIV/AIDS between 35 and 51 years old. The intervention consisted in a program of resisted exercises. The following parameters were evaluated: BMI, WHR, % G, and scapular and hand strength and the Quality of Life (QoL) was evaluated using the WHOQOL HIV BREFS., Results: Significant changes were observed in the %G (17.01±2.66 x 15.12±2.25, P=0.031), scapular force (26.16±2.73 x 29.00±2.76, P=0.007), and hand strength (35.45±9.69 x 37.88±10.84, P=0.039). Significant changes were observed in the following areas in the QL domains: environment (12.90±0.52 x 14.10±0.22, P=0.021), spirituality, religion and personal beliefs (15.60±0.88 x 17.06±0.60, P=0.032), and perception of quality of life and overall health (15.33±0.46 x 17.20±0.47, P=0.005)., Conclusions: The results indicate an improvement in body composition, increasing functional capacity, and improving QL through resisted exercise program.

The Rickettsia bacteria include the aetiological agents for the human spotted fever (SF) disease. In the present study, a SF group Rickettsia amblyommii related bacterium was detected in a field collected Amblyomma sculptum (Amblyomma cajennense species complex) tick from a Brazilian SF endemic site in southeastern Brazil, in the municipality of Juiz de Fora, state of Minas Gerais. Genetic analysis based on genes ompA,ompB and htrA showed that the detected strain, named R. amblyommii str. JF, is related to the species R. amblyommii.

Levan productivity of Bacillus subtilis Natto was evaluated in submerged culture varying the pH, temperature and culture time, using factorial design and response surface methodology. The characterization of levan molecular weight was performed by HPSEC and its antitumor activity against HepG2 cells using metabolomic approach was also evaluated. At first, the variables investigated, as well as their interactions, demonstrated significant effect. Further, a second design using the same variables at different levels was developed. Thus, according to the model, an optimized value corresponding to 5.82 g.L⁻¹.h⁻¹ was achieved at pH 8, 39.5°C in 21 hours, the highest value reported so far. After analysis by HPSEC, two molecular weights were obtained corresponding to 72.37 and 4146 kDa. The levan promoted an increase of acetate, alanine, lactate and phosphocreatine in HepG2 cells suggesting an alteration in the bioenergetics pathways and cellular homeostasis by intracellular accumulation of lactate, justifying its antitumor activity.

Background: The aim of this study was to investigate the distribution of full-length and deleted variants of KIR2DS4 in a population of southern Brazil and compare the results with other populations, as well as comparing two techniques, PCR-SSP and PCR-SSO, for typing of variants., Methods: 258 individuals from southern Brazil were analysed by PCR-SSO ("polymerase chain reaction-sequence specific oligonucleotides", One Lambda, Inc., Canoga Park, CA), of which 161 were also analysed by PCR-SSP., Results: The study population showed similarities with other Caucasian populations; 46.5% of individuals had only KIR2DS4 variants, 21.3% had the full-length form and 25.1% had both forms., Conclusion: The frequencies found in both groups (genotyped by PCR-SSP and PCR-SSO) were 100% concordant., (© 2014 Wiley Periodicals, Inc.)

Objective: This study investigated the occurrence of the p190 and p210 breakpoint cluster region-Abelson (BCR-ABL) rearrangements in adults with acute lymphoblastic leukemia and possible associations with clinical and laboratory characteristics and survival., Methods: Forty-one over 18-year-old patients with acute lymphoblastic leukemia of both genders followed-up between January 2008 and May 2012 were included in this study. Clinical and laboratory data were obtained from the medical charts of the patients. Reverse transcription polymerase chain reaction (RT-PCR) using specific primers was employed to identify molecular rearrangements., Results: At diagnosis, the median age was 33 years, and there was a predominance of males (61%). The most common immunophenotype was B lineage (76%). BCR-ABL rearrangements was detected in 14 (34%) patients with the following distribution: p190 (28%), p210 (50%) and double positive (22%). Overall survival of patients with a mean/median of 331/246 days of follow up was 39%, respectively, negative BCR-ABL (44%) and positive BCR-ABL (28%)., Conclusion: These results confirm the high frequency of BCR-ABL rearrangements and the low survival rate of adult Brazilian patients with acute lymphoblastic leukemia., (Copyright © 2014 Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular. Published by Elsevier Editora Ltda. All rights reserved.)

Dengue is currently a major public-health problem. Dengue virus (DENV) is classified into four distinct serotypes, DENV 1-4. After 28 years of absence, DENV-4 was again detected in Brazil in 2010 in Roraima State, and one year later, the virus was identified in the northern Brazilian states of Amazonas and Pará, followed by Rio de Janeiro and São Paulo. In Minas Gerais, the first confirmed case of DENV-4 occurred in the municipality of Frutal in 2011 and has now been isolated from a growing number of patients. Although DENV-2 is associated with the highest risk of severe forms of the disease and death due to the infection, DENV-4 has also been associated with severe forms of the disease and an increasing risk of hemorrhagic manifestations. Herein, the first fatal case of confirmed DENV-4 in Brazil is reported. The patient was an 11-year-old girl from the municipality of Montes Claros in northern Minas Gerais State, Brazil. She had idiopathic thrombocytopenic purpura as a comorbid condition and presented with a fulminant course of infection, leading to death due to hemorrhagic complications. Diagnosis was confirmed by detection of Dengue-specific antibodies using IgM capture enzyme-linked immunosorbent assay and semi-nested RT-PCR. Primary care physicians and other health-care providers should bear in mind that DENV-4 can also result in severe forms of the disease and lead to hemorrhagic complications and death, mainly when dengue infection is associated with coexisting conditions.

Polymorphisms of Rh, Kell, Duffy, Kidd and Diego blood group systems were studied in 209 unrelated Brazilian Japanese descendants from South of Brazil. The methods used were multiplex-PCR, AS-PCR and RFLP-PCR. The differences in frequencies among the populations were evaluated using chi-square test. The frequencies for Rh, Kell, Kidd and Diego system were similar to those of the Japanese. RHCE(*)CC, RHCE(*)EE genotypes and FY(*)01 allele were lower and FY(*)01N.01 was higher than Japanese. These differences in the frequencies between Brazilian Japanese descendants and Japanese could indicate a gene flow in Brazilian population and reinforce the importance of this knowledge to achieve safe red blood cells., (Copyright © 2013. Published by Elsevier Ltd.)

Information on reproductive characteristics of wild rodents is scarce in the literature. This study aimed to assess the testis morphometry and stereology of Hylaeamys megacephalus. We used five animals in the study, captured in forest fragments in southwestern Goias State, between April and August 2009. The testes were fixed in Karnovsky solution, dehydrated, and embedded in methacrylate. Two-micrometer-thick sections from each sample were stained with toluidine blue/sodium borate 1%. Images of the testicular parenchyma were obtained from photomicroscope and morphometric and stereological analyses were carried out using the Image Pro-Plus software. The average body weight observed in the specimens of H. megacephalus in the study was 47.84 g, of which, 0.40% is allocated to the gonads (GSI) and 0.36% to the seminiferous tubules (TSI). These parameters suggest promiscuous reproductive behavior, of the polyandrous type, favoring males with higher sperm production and consequently, larger testes. The volume density of the seminiferous tubules was 94.46%, which represented a volume of 0.18 mL. The volume density and volume of the interstitium were 5.54% and 0.011 mL, respectively. The diameter of the seminiferous tubules was 206.5 μm and the height of seminiferous epithelium was 71.27 μm. H. megacephalus presents 5.06 m of seminiferous tubules and an average of 27.96 m of seminiferous tubules per gram of testis. The mitotic and meiotic indexes showed losses of 85 and 42%, respectively and an overall loss of 90% over the full spermatogenic process. The number of Sertoli cells per testis and per gram of testis was 7.8×10(6) and 95.28×10(6), respectively. Most of the morphometric parameters evaluated in H. megacephalus in this study are within the range of values described for most mammals., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Levan is an exopolysaccharide of fructose primarily linked by β-(2→6) glycosidic bonds with some β-(2→1) branched chains. Due to its chemical properties, levan has possible applications in both the food and pharmaceutical industries. Bacillus subtilis is a promising industrial levan producer, as it ferments sucrose and has a high levan-formation capacity. A new strain of B. subtilis was recently isolated from Japanese food natto, and it has produced levan in large quantities. For future pharmaceutical applications, this study aimed to investigate the effects of levan produced by B. subtilis Natto, mainly as potential hypoglycemic agent, (previously optimized with a molecular weight equal to 72.37 and 4,146 kDa) in Wistar male rats with diabetes induced by streptozotocin and non-diabetic rats and to monitor their plasma cholesterol and triacylglycerol levels. After 15 days of experimentation, the animals were sacrificed, and their blood samples were analyzed. The results, compared using analysis of variance, demonstrated that for this type of levan, a hypoglycemic effect was not observed, as there was no improvement of diabetes symptoms during the experiment. However, levan did not affect any studied parameters in normal rats, indicating that the exopolysaccharide can be used for other purposes.

Leprosy is a chronic infectious disease caused by Mycobacterium leprae, which mainly affects the skin and nervous system. The disease has several clinical forms. This study investigated the MICA and HLA-B genes in 223 samples from leprosy patients and 201 samples from healthy individuals matched for age, gender and ethnical background. Of the patients, 153 had multibacillary, 45 paucibacillary and 25 indeterminate leprosy. The aim of this case-control study was to assess whether the MICA alleles influence susceptibility for leprosy or affect the subtype of the disease in a population of southern Brazil. There were significant differences in frequencies of the MICA*027 allele (4.7% vs 1.8%, P-value = 0.01, OR = 0.37; 95% CI = 0.16-0.85) between leprosy patients and controls, and of the MICA*010 (4.5% vs 1.6%, P-value = 0.05, OR = 0.35, 95% CI = 0.13-0.97) and MICA*027 alleles (4.7% vs 1.3%, P-value = 0.01; OR = 0.27; 95% CI = 0.09-0.79) between multibacillary leprosy patients and the control group. There were no significant differences in the frequency of MICA alleles between paucibacillary leprosy patients and controls. Thus, the MICA*027 allele is associated with a protective effect for leprosy per se, while the MICA*010 and MICA*027 alleles are associated with protection against multibacillary leprosy, the most severe clinical subtype., (© 2011 Blackwell Publishing Ltd.)

Congenital haemophilia A is a chromosome-linked recessive disorder caused by the deficiency or reduction of factor VIII (FVIII) pro-coagulant activity. During treatment, some patients develop alloantibodies (FVIII inhibitors) that neutralize the action of exogenously administered FVIII. Currently, the presence of these inhibitors is the most serious adverse event found in replacement therapy. Some studies have suggested that genetic factors influence the development of the FVIII coagulation inhibitors. To identify the class I and II alleles that may be influencing the formation of inhibitors in severe haemophilic patients. Genotyping of the class I (HLA-A, -B and -C) and class II (HLA-DRB1, -DQA1 and -DQB1) alleles of 122 patients with severe haemophilia A, including 36 who had developed antibodies to factor VIII, was performed. After the comparison of the group without inhibitors and the group with inhibitors, HLA-C*16 [Odds ratio (OR) = 7.73; P = 0.0092] and HLA-DRB1*14 (OR = 4.52; P = 0.0174) were found to be positively associated with the formation of the inhibitors. These results confirm that HLA alleles are involved in inhibitor production and could be used as a tool for recognition of groups at high risk of possible inhibitor development in Southern Brazilian haemophilic patients., (© 2011 Blackwell Publishing Ltd.)

The development of a Brazilian National Surveillance System in 1975 led to a compulsory reporting of selected infectious diseases aiming to reduce the burden of these events in the country. However, shifts in the epidemiology of these diseases associated with modern life style, demand constant revision of surveillance activities. In this manuscript we present the epidemiology, trends and differential diagnosis of the following compulsory notifiable diseases in Brazil: Aids, dengue fever, hanseniasis, American tegumentary leishmaniasis, measles, rubella and congenital rubella syndrome and syphilis. Additionally, the current challenges for control and prevention of each disease are presented.

This study evaluates the relation between the use of tobacco, alcoholic beverages and illicit drugs and family protective factors. Data from the National School Health Survey (PeNSE) were analyzed in a sample of 60,973 students at the freshman year of high school, from public and private schools of Brazilian state capitals. Most adolescents lived with both their parents and about a third lived in households only with their mothers. Half the parents or responsible parties are aware of what adolescents do in their free time. Living with both parents is a protective factor for smoking, drinking, and drug use. Family supervision is also important for the prevention of such behavior. Sharing a meal with parents or responsible parties most days of the week and the fact that the parents know what the adolescents have done in their free time in the past 30 days are also protective factors. Students that miss classes without telling their parents have higher chances of using tobacco, alcohol, and drugs. The family plays an essential role to prevent tobacco, alcohol, and drug use, and to promote health among teenagers.

Background: Red blood group genes are highly polymorphic and the distribution of alleles varies among different populations and ethnic groups., Aim: To evaluate allele polymorphisms of the Rh, Kell, Duffy and Kidd blood group systems in a population of the State of Paraná, Methods: Rh, Kell, Duffy and Kidd blood group polymorphisms were evaluated in 400 unrelated blood or bone marrow donors from the northwestern region of Paraná State between September 2008 and October 2009. The following techniques were used: multiplex-polymerase chain reaction genotyping for the identification of the RHD gene and RHCE*C/c genotype; allele-specific polymerase chain reaction for the RHDψ and restriction fragment length polymorphism polymerase chain reaction for the RHCE*E/e, KEL, FY-GATA and JK alleles., Results: These techniques enabled the evaluation of the frequencies of Rh, Kell, Duffy and Kidd polymorphisms in the population studied, which were compared to frequencies in two populations from the eastern region of São Paulo State., Conclusion: The RHCE*c/c, FY*A/FY*B, GATA-33 T/T, JK*B/JK*B genotypes were more prevalent in the population from Paraná, while RHCE*C/c, FY*B/FY*B, GATA-33 C/C, JK*A/JK*B genotypes were more common in the populations from São Paulo.

Introduction: To detect dengue virus, eggs of Aedes sp were collected in the city of Belo Horizonte, Brazil, in 2007., Methods: Egg samples were subsequently hatched and the larvae were tested for the presence of dengue virus RNA by RT-PCR., Results: Among the Aedes aegypti larvae samples, 163 (37.4%) out of 435 were positive, including 32 (10.9%) of 293 individual larvae samples concomitantly positive for two serotypes., Conclusions: Virological surveillance detecting coinfected vectors in the field could represent an important strategy for understanding the numerous factors involved in the transmission and clinical presentation of dengue.

Traditional medicine provides strong guidance for scientific experiments involving plant products used by the Brazilian people. The species "cipó-cravo" (Tynnanthus fasciculatus) is a plant commonly used either to combat indigestion and stomach aches, or as a general stimulant and aphrodisiac. In this study, the effects of "cipó-cravo" infusion were investigated within the testicular parenchyma of adult Wistar rats. Rats were divided into 3 groups: a control (distilled water) and two treated groups, which received the plant infusion (100 and 200mg/animal/day). The 200mg dose promoted a significant increase of the testicular parenchyma weight and of the volume and total length of the seminiferous tubules, as well as in total daily sperm production and sperm production per gram of testis.

We evaluated the usefulness of blood group genotyping as a supplement to hemagglutination to determine the red blood cell (RBC) antigen profile of polytransfused patients with hematological diseases and renal failure. Seventy-nine patients were selected. They all received more than three units of blood and eight (10%) had already clinical significant alloantibodies occurring alone or in combination against Rh, K, Fya, and Di antigens. DNA was prepared from blood samples and RHCE*E/e, KEL*01/KEL*02, FY*01/FY*02 and JK*01/JK*02 alleles were determined by using PCR-RFLP. RHD*/RHD*Ψ and RHCE*C/c were tested using multiplex PCR. Discrepancies for Rh, Kell, Duffy, and Kidd systems were found between the phenotype and genotype-derived phenotype in 16 of the 38 chronically transfused patients. The genotypes of these patients were confirmed by DNA array analysis (HEA Beadchip(™); Bioarray Solutions, Warren, NJ). Genotyping was very important for the determination of the true blood groups of the polytransfused patients, helped in the identification of suspected alloantibodies and in the selection of antigen-negative RBCs for transfusion., (J. Clin. Lab. Anal. 24:311-316, 2010. © 2010 Wiley-Liss, Inc.)

The objective of this study was to standardize a method for extracting high-quality DNA from samples of coagulated blood. Forty-eight samples of human coagulated blood were used for DNA extraction by means of the EZ-DNA commercial kit (Biological Industries, Beit Haemek, Israel), the Neoscience column kit (One Lambda Inc., San Diego, CA, USA) and a modified salting-out method. Only the salting-out method was able to extract high concentrations of DNA (mean, 180 ng/(1/4)microl), which were measured using the Qubit fluorescence detector (Invitrogen, USA). This method enabled amplification of HLA (human leukocyte antigen) genes using the Luminex PCR-SSO (polymerase chain reaction - sequence-specific oligonucleotide) technology, which demands good quality DNA, and amplification of KIR (killer-cell immunoglobulin-like receptor) genes using an in-house PCR-SSP (polymerase chain reaction - sequence-specific primer) technique, which demands a specific concentration of DNA (10 ng/(1/4)microl). We concluded that the modified salting-out technique was very efficient, simple and fast for DNA extraction from human coagulated blood samples, with the aim of genotyping the HLA and KIR genes.

Multiple sclerosis (MS) is a chronic autoimmune demyelinating disease of the central nervous system that causes neurological disorders in young adults. Previous studies in various populations highlighted an association between the HLA-DRB1*15 allele and MS. This study investigated the association between HLA-DRB1*15 and other HLA-DRB1 alleles and MS in a Brazilian Caucasian population sample from Londrina, Southern Brazil. HLA-DRB1 alleles were analyzed by polymerase chain reaction with specific sequence oligonucleotide primers in 119 MS patients and in 305 healthy blood donors as a control. Among the MS patients, 89 (75.0%) presented with relapsing remitting MS, 24 (20.0%) with secondary progressive MS and 6 (5.0%) with primary progressive MS. The frequency of the HLA-DRB1*15 allele observed in the MS Brazilian patients was similar to findings reported in previous studies carried out in populations worldwide. However, the results showed a higher frequency of the HLA-DRB1*15 allele in the MS patients compared to the controls, with a relative frequency of 0.1050 (10.50%) and 0.0443 (4.4%), respectively (OR=2.53; 95% CI 1.43-4.46; p=0.0009). A protector allele was also detected. The frequency of the HLA-DRB1*11 allele was reduced in the MS patients compared to the controls, with a relative frequency of 0.1345 (13.4%) and 0.1869 (18.7%), respectively (OR=0.67; 95% CI 0.44-1.03; p=0.0692). The results demonstrated that the HLA-DRB1*15 allele in heterozygosity is positively associated with MS (p=0.0079), and may be considered a genetic marker of susceptibility to the disease. A negative association between the HLA-DRB1*11 allele in homozygosity and MS was also verified (p=0.0418); this allele may be considered a genetic marker of resistance to MS in the Brazilian population.

Most cases of a predisposition to venous thrombosis are caused by resistance to activated protein C, associated in 95% of cases with the Factor V Leiden allele (FVL or R506Q). Several recent studies report a further increased risk of thrombosis by an association between the AB alleles of the ABO blood group and Factor V Leiden. The present study investigated this association with deep vein thrombosis (DVT) in individuals treated at the Hemocentro de Pernambuco in northeastern Brazil. A case-control comparison showed a significant risk of thrombosis in the presence of Factor V Leiden (OR = 10.1), which was approximately doubled when the AB alleles of the ABO blood group were present as well (OR = 22.3). These results confirm that the increased risk of deep vein thrombosis in the combined presence of AB alleles and Factor V Leiden is also applicable to the Brazilian population suggesting that ABO blood group typing should be routinely added to FVL in studies involving thrombosis.

The crystal and local atomic structure of monoclinic ReO2 (alpha-ReO2) under hydrostatic pressure up to 1.2 GPa was investigated for the first time using both X-ray absorption spectroscopy and high-resolution synchrotron X-ray powder diffraction and a home-built B4C anvil pressure cell developed for this purpose. Extended X-ray absorption fine-structure (EXAFS) data analysis at pressures from ambient up to 1.2 GPa indicates that there are two distinct Re-Re distances and a distorted ReO6 octahedron in the alpha-ReO2 structure. X-ray diffraction analysis at ambient pressure revealed an unambiguous solution for the crystal structure of the alpha-phase, demonstrating a modulation of the Re-Re distances. The relatively small portion of the diffraction pattern accessed in the pressure-dependent measurements does not allow for a detailed study of the crystal structure of alpha-ReO2 under pressure. Nonetheless, a shift and reduction in the (011) Bragg peak intensity between 0.4 and 1.2 GPa is observed, with correlation to a decrease in Re-Re distance modulation, as confirmed by EXAFS analysis in the same pressure range. This behavior reveals that alpha-ReO2 is a possible inner pressure gauge for future experiments up to 1.2 GPa.

The aim of this study was to investigate the role of killer cell immunoglobulin-like receptor (KIR) genes in leprosy immunopathogenesis. Genotyping of KIR and human leukocyte antigen (HLA) genes was performed by polymerase chain reaction with sequence-specific oligonucleotide probes in 165 leprosy patients. Both activating KIR2DS2 and KIR2DS3 frequencies were higher in tuberculoid leprosy (TT) patients than in lepromatous leprosy (LL) patients, and the inhibitory KIR with its ligand, KIR2DL1-C2/C2, was elevated in TT patients in comparison to all other leprosy subgroups and controls. However, a negative association between KIR2DL3-C1 and KIR2DL3-C1/C1 and the TT group was identified. Borderline patients exhibited a higher frequency of KIR3DL2-A3/11 than the controls and LL patients, and a lower frequency of KIR2DL1-C2 than the controls and TT subgroup. Some KIR-HLA genotypes could be associated to the development of clinical forms of leprosy and should be investigated further.

Introduction: Prostate cancer is a significant cause of morbidity and mortality. In Portugal alone, according to a study published in 2003, the rate of new prostate cancer cases were 53 per 100,000 men (in 2000), with an age-standardized mortality rate of about 28 per 100,000 (in 1995). Multiple bone metastases are one of the major complications of advanced prostate cancer. Samarium-EDTMP showed to be a safe and effective alternative for palliative treatment of bone metastases. The goal of this economic study is to assess the cost-effectiveness of Samarium-153-EDTMP for the treatment of pain due to multiple bone metastases in hormone-refractory prostate cancer versus conventional pain therapy, in Portugal., Methodology: Cost-effectiveness study that compares the expected direct costs to the National Health System of managing patients with painful multiple bone metastases with Samarium-153-EDTMP versus conventional pain therapy, in Portugal, in a 4-moths period., Results: The total direct 4 months cost was 2,311.91 euro for a patient treated with Samarium-153-EDTMP versus 2,450.74 euro for a patient under standard treatment. According to the model a patient treated with Samarium-153-EDTMP represents a 138.83 euros saving., Conclusion: Samarium-153-EDTMP was not only a very effective therapeutic option but also an option with less cost than the conventional pain therapy, in patients with pain due to multiple bone metastases, in Portugal.

Acute promyelocytic leukemia (APL) is characterized by the presence of rearrangements involving the retinoic acid receptor alpha (RARalpha) gene and a variable incidence in different populations. The hybrid gene PML-RARalpha, present in 98% of cases, encodes a fusion protein essential to the pathogenesis of the disease. Depending of the PML's gene breakpoint in chromosome 15, the transcript subtypes bcr1, bcr2 and bcr3 may be formed. The correlation between these transcript subtypes and clinical parameters is still controversial. The objective of this study was to determine the frequencies of the PML-RARalpha transcripts and subtypes in a series of 32 APL patients from Northeast Brazil and to evaluate the association of these subtypes to different parameters. The method used was RT-PCR. The frequency of our APL cases is approximately 28% of the acute leukemias. The results showed the presence of PML-RARalpha isoform in all patients and a higher frequency of the bcr1/2 subtype. No significant statistical association was found between molecular subtypes and age, sex, French-American-British (FAB) classification, leukocyte and platelet count, hemoglobin level or coagulation tests. In conclusion, these data suggest similar molecular and biological features for our APL patients at diagnosis in comparison with those reported in current scientific literature.