Mitochondrial DNA control region diversity in a population from Parana state-increasing the Brazilian forensic database.

The entire mitochondrial DNA (mtDNA) control region (nucleotide position 16024-576) sequences were obtained through Sanger sequencing method for 122 individuals from Parana state, South of Brazil. We observed a total of 108 different haplotypes of which 97 were unique and 11 were shared by more than one individual. The haplogroups were classified according to the updated mtDNA phylogeny, by EMMA (estimating mitochondrial haplogroups using a maximum likelihood approach). Our results revealed the predominance of Amerindian haplogroups with a frequency of 49.2% of the population sample, followed by European lineages with 38.5% and 12.3% of African lineages. Parana population sample set presented a high haplotype diversity (0.9976) and the random match probability was 0.0106. The phylogenetical findings and the diversity indices confirm the high genetic heterogeneity of this population and suggest a high informativeness of mtDNA analyses in forensic cases. The population data will contribute to increase the Brazilian mtDNA database for forensic purposes and it is available through EMPOP (European DNA Profiling Group mitochondrial DNA population database) under the accession number EMP00714.

Spinal cord injury epidemiological profile in the Sarah Network of Rehabilitation Hospitals-a Brazilian population sample.

STUDY DESIGN: Retrospective cross-sectional study. OBJECTIVE: To describe the epidemiological profile of individuals with spinal cord injury (SCI), who visited the Sarah Network in 2014. SETTING: Brazil, Brasília-Federal District. METHODS: A review of the electronic medical records of persons with SCI, aged over 15 years, was carried out. RESULTS: The medical records of 2076 persons were evaluated: 83% were male, the mean age was 31 years, 40% did not complete elementary school, 67.7% had paraplegia, and 74.4% had complete SCI. The primary etiologies were gunshot wounds (28.4%), motorcycle accidents (24.6%), and motor vehicle accidents (19.1%). In the comparative analysis between genders, differences were observed regarding the level of education and completeness of injury. CONCLUSION: Gunshot wounds were the main cause of SCI in our population, comprising mostly male individuals with thoracic injury, resulting in complete SCI. Data show a high incidence of SCI in young adults owing to violence in Brazil.

Predictors of musculoskeletal pain in the upper extremities of individuals with spinal cord injury.

INTRODUCTION: Increased demands on the upper extremities (UE) have been associated with a higher occurrence of musculoskeletal pain in individuals with spinal cord injury (SCI). STUDY DESIGN: This is a cross-sectional retrospective study. OBJECTIVE: The aim of this study was to characterize SCI subjects with musculoskeletal pain in the UE and to determine which variables could predict musculoskeletal pain in these individuals. SETTING: The study was conducted in SARAH Network of Rehabilitation Hospitals, Brasília, Brazil. METHODS: Five hundred sixty-four electronic medical records were investigated through a retrospective, exploratory and descriptive analysis to identify individuals with UE pain. A logistic regression model was applied to estimate (odds ratios) whether gender, age, the level of injury, severity of injury, time of injury, body mass index, type of mobility and locomotion aid could predict the occurrence of musculoskeletal pain. RESULTS: The prevalence of musculoskeletal pain in the UE was 27.7%. The odds of having musculoskeletal pain were two times higher among woman and tetraplegic patients; those over 41 years of age had twice the frequency of pain than did those <24.7 years; <1 year of injury was a predictor of musculoskeletal pain, compared with the other quartiles (1.1-2.8, 2.9-6.8 and 6.9+ years of injury). There were no differences between the wheelchair and ambulatory individuals. CONCLUSION: Female individuals, those with tetraplegia, aged >40 years and <1 year of injury showed an increased risk of musculoskeletal pain.

Functional regulation of the apurinic/apyrimidinic endonuclease 1 by nucleophosmin: impact on tumor biology.

Nucleophosmin 1 (NPM1) is a nucleolar protein involved in ribosome biogenesis, stress responses and maintaining genome stability. One-third of acute myeloid leukemias (AMLs) are associated with aberrant localization of NPM1 to the cytoplasm (NPM1c+). This mutation is critical during leukemogenesis and constitutes a good prognostic factor for chemotherapy. At present, there is no clear molecular basis for the role of NPM1 in DNA repair and the tumorigenic process. We found that the nuclear apurinic/apyrimidinic endonuclease 1 (APE1), a core enzyme in base excision DNA repair (BER) of DNA lesions, specifically interacts with NPM1 within nucleoli and the nucleoplasm. Cytoplasmic accumulation of APE1 is associated with cancers including, as we show, NPM1c+ AML. Here we show that NPM1 stimulates APE1 BER activity in cells. We provide evidence that expression of the NPM1c+ variant causes cytoplasmic accumulation of APE1 in: (i) a heterologous cell system (HeLa cells); (ii) the myeloid cell line OCI/AML3 stably expressing NPM1c+; and (iii) primary lymphoblasts of NPM1c+ AML patients. Consistent with impaired APE1 localization, OCI/AML3 cells and blasts of AML patients have impaired BER activity. Cytoplasmic APE1 in NPM1c+ myeloid cells is truncated due to proteolysis. Thus, the good prognostic response of NPM1c+ AML to chemotherapy may result from the cytoplasmic relocalization of APE1 and the consequent BER deficiency. NPM1 thus has an indirect but significant role in BER in vivo that may also be important for NPM1c+ tumorigenesis.

Microbial inactivation kinetics during high-pressure carbon dioxide treatment: nonlinear model for the combined effect of temperature and pressure in apple juice.

Isobaric and isothermal semi-logarithmic survival curves of natural microflora in apple juice treated with high-pressure carbon dioxide at 7, 13, and 16 MPa pressures and 35, 50, and 60 degrees C temperatures were fitted with a nonlinear equation to find the values of the coefficient b(P), b(T), n(P), and n(T). Profiles of the model parameters were obtained as a function of pressure and temperature. The model fitted with good agreement(R(2) > 0.945), the survival curves. An empirical equation was proposed to describe the combined effects of pressure and temperature. The equation, derived from a power law model, was written in the form: log(10) S(t) = -log(e) [C(0)+C(1) x exp (K(T) x (T-T(C))-C(2) x exp (K(P) x (P-P(C))) x t (C(3)xT(2)+C(4)xT+C(5)). The proposed model fitted the experimental data well. At 7 MPa and 50 and 60 degrees C, 13 MPa and 35 and 60 degrees C, 16 MPa and 35 degrees C, the model provided (log)10 reduction residual values (observed value-fitted value) lower than 0.284 showing a good agreement between the experimental and the predicted survival levels.

Gene transcription in Lactarius quietus-Quercus petraea ectomycorrhizas from a forest soil.

Extracting fungal mRNA from ectomycorrhizas (ECMs) and forest soil samples for monitoring in situ metabolic activities is a significant challenge when studying the role of ECMs in biogeochemical cycles. A robust, simple, rapid, and effective method was developed for extracting RNA from rhizospheric soil and ECMs by adapting previous grinding and lysis methods. The quality and yield of the extracted RNA were sufficient to be used for reverse transcription. RNA extracted from ECMs of Lactarius quietus in a 100-year-old oak stand was used to construct a cDNA library and sequence expressed sequence tags. The transcripts of many genes involved in primary metabolism and in the degradation of organic matter were found. The transcription levels of four targeted fungal genes (glutamine synthase, a general amino acid transporter, a tyrosinase, and N-acetylhexosaminidase) were measured by quantitative reverse transcription-PCR in ECMs and in the ectomycorrhizospheric soil (the soil surrounding the ECMs containing the extraradical mycelium) in forest samples. On average, levels of gene expression for the L. quietus ECM root tips were similar to those for the extraradical mycelium, although gene expression varied up to 10-fold among the samples. This study demonstrates that gene expression from ECMs and soil can be analyzed. These results provide new perspectives for investigating the role of ectomycorrhizal fungi in the functioning of forest ecosystems.

Mud bath therapy influences nitric oxide, myeloperoxidase and glutathione peroxidase serum levels in arthritic patients.

Nitric oxide (NO) has recently been proposed as an important mediator in inflammatory phases and in loss of cartilage. In inflammatory arthritis NO levels are correlated with disease activity and articular cartilage is able to produce large amounts of NO with the appropriate inducing factors such as cytokines and/or endotoxin. Neutrophils also play an important role in inflammatory reactions and the level of myeloperoxidase, a constituent of neutrophil granules, is related to the intensity of the inflammation. Because there is evidence that suggests that mud packs influence the main cytokines involved in cartilage damage, we tried to determine whether NO and myeloperoxidase are involved in the mechanisms of action of mud bath treatment. We enrolled 37 subjects and randomly assigned them to two groups: 19 patients underwent mud bath treatment (group A) while 18 patients underwent bath treatment alone. Blood samples were obtained before and after the treatment cycles to assay serum levels of NO, myeloperoxidase (MPO) and glutathione (GSH)-peroxidase. The results showed a statistically significant decrease in NO and myeloperoxidase serum values in groups A and B, while GSH-peroxidase was not significantly increase in either of the groups; no correlation was found between NO, myeloperoxidase and GSH-peroxidase serum values. Mud bath treatment can exert beneficial effects on cartilage homeostasis and inflammatory reactions, influencing NO and decreasing myeloperoxidase serum values. The increase in GSH-peroxidase was not correlated with the reduction of other biochemical markers, suggesting that mud bath treatment has different mechanisms of action.

In vitro effect of inhibitors on the activity of glucosyltransferase, isolated from human dental plaque.

The enzyme glucosyltransferase plays an important role in plaque formation and growth. Therefore, chemical inhibition of glucosyltransferase may become an effective method for plaque control. In this investigation we have evaluated the effects of some antiplaque substances (chlorhexidine, cetylpiridinium chloride, iodine, sodium fluoride and sodium dodecyl sulfate) on glucosyltransferase activity. Our results revealed that iodine was the most effective inhibitor. Based on in vitro glucosyltransferase inhibition we may suggest that topical iodine could be an auxiliary method for plaque control.

A simplified method for the partial purification and enzyme assay of glucosyltransferase from human dental plaque.

A simplified method for the partial purification and enzyme assay of glucosyltransferase from human dental plaque is described. The enzyme obtained has a reasonable specific activity for the assay of soluble and insoluble polysaccharides synthesis. This method may be useful for nonacquainted people with enzymological methods and of great utility in the in vitro evaluation of antiplaque substances.