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Bovine mastitis is an important and costly disease to dairy cattle. Diagnostic methods usually performed in Brazil are somatic cell counts (SCC) and milk microbiology. Low bacteria shedding in milk implies no colony growth in microbiological tests and false negative results. Streptococcus agalactiae and Staphylococcus aureus are principal pathogens of mixed mastitis. However, S. agalactiae has a higher bacterial release from the mammary gland than S. aureus , affecting microbiological sensitivity to diagnose S. aureus . This study aimed to estimate the SCC and total bacterial count (TBC) from cows according to pathogen isolated in milk and to evaluate variation in S. aureus diagnosis by a microbiological test during S. agalactiae treatment, which is called blitz therapy. Both S. agalactiae and S. aureus presented high SCC means, although S. agalactiae showed shedding of bacteria 2.3 times greater than S. aureus . Microbiological sensitivity to S. aureus increased for 5 months during S. agalactiae treatment. The prevalence of S. agalactiae fell after 5 months of therapeutic procedures. The prevalence of S. aureus increased to 39.0. The results showed that due to high sensitivity, the polymerase chain reaction (PCR) could be used at the beginning of blitz therapy with the goal of S. agalactiae eradication from the dairy herd.

Staphylococcus aureus is one of the agents of bovine mastitis of hardest control due to a complex pathogenesis comprising a variety of virulence factors, which ensures its persistence in the mammary gland, causing significant health and economic losses. Therefore, understanding the pathogenesis of this agent is imperative. Galleria mellonella has stood out as an invertebrate animal model for the study of infectious diseases that affect several hosts. This work aimed to evaluate G. mellonella larvae as an experimental model for the study of virulence phenotypes in an S. aureus population isolated from bovine mastitis. Thirty genetically divergent S. aureus strains were chosen based on PFGE analysis. After experimental infection, larvae survival rates, bacterial growth in hemolymph, melanization intensity of the dorsal vessel, and histological characteristics of the infected tissues were evaluated. The G. mellonella model showed a clear diversity in the S. aureus pathogenicity pattern, allowing the differentiation of strains with virulence phenotypes ranging from high to low degrees. Histological analysis confirmed that the strains tested were capable of inducing the formation of nodules and melanization spots in the dorsal vessels of the larvae in different magnitudes. The strains 16S-717, 19C-828, and 31S-1443 presented the highest virulence intensity among the bacteria tested and will be used further for the generation of S. aureus mutant populations to prospect genetic targets aimed to develop control strategies of bovine mastitis. Altogether, our results suggest that G. mellonella is an attractive and low-cost animal model for characterizing virulence phenotypes of large S. aureus populations., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Objective: To report the experience of offering the Quality End of Life Care for All (QELCA) Programme, highlighting the distinct methodology for the training of health professionals., Development: The QELCA Programme, intellectual property of St Christopher's Hospice, was offered to seven health professionals working in the hospital palliative care unit at the National Cancer Institute, between June and December 2022, with the support of Premier Institute. The programme, which originates in the UK, has been evaluated there and is currently being evaluated in Hong Kong, and is delivered in two phases: (1) a 5-day immersion programme; (2) monthly sessions of Action Learning for 6 months. Participants realised that communication between members of the multidisciplinary team, as well as between health professionals and patients/loved ones, was one of the key challenges for achieving quality of death in the hospital palliative care unit. This insight empowered them to drive forward significant changes in practice that promise to improve quality of care., Conclusion: The QELCA Programme enabled participants to engage in active problem-solving to promote the relief of suffering of patients and their families in end-of-life care., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2023. No commercial re-use. See rights and permissions. Published by BMJ.)

Background and Aims: Ambulatory blood pressure monitoring (ABPM) allows the assessment of cardiovascular risk markers that cannot be obtained by casual measurements; however, the evidence on the association between food consumption and blood pressure (BP) assessed by ABPM is scarce. We aimed to evaluate the association between food consumption by degree of processing and ambulatory BP., Methods and Results: Cross-sectional analysis (2012-2014) of data from a subsample (n = 815) of ELSA-Brasil cohort participants who performed 24-h ABPM was conducted. Systolic (SBP) and diastolic (DBP) BP means and variability during the 24 h and subperiods (sleep and wake), nocturnal dipping, and morning surge were evaluated. Food consumption was classified according to NOVA. Associations were tested by generalized linear models. The consumption of unprocessed, minimally processed foods, and culinary ingredients (U/MPF&CI) was 63.1% of daily caloric intake, 10.8% of processed (PF), and 24.8% of ultraprocessed (UPF). A negative association was found between U/MPF&CI consumption and extreme dipping (T2: odds ratio [OR] = 0.56, 95% confidence interval [CI] = 0.55-0.58; T3: OR = 0.55; 95% CI = 0.54-0.57); and between UPF consumption and nondipping (T2: OR = 0.68, 95% CI = 0.55-0.85) and extreme dipping (T2: OR = 0.63, 95% CI = 0.61-0.65; T3: OR = 0.95, 95% CI = 0.91-0.99). There was a positive association between PF consumption and extreme dipping (T2: OR = 1.22, 95% CI = 1.18-1.27; T3: OR = 1.34, 95% CI = 1.29-1.39) and sleep SBP variability (T3: Coef = 0.56, 95% CI = 0.03-1.10)., Conclusions: The high consumption of PF was associated with greater BP variability and extreme dipping, while the U/MPF&CI and UPF consumption were negatively associated with alterations in nocturnal dipping., Competing Interests: Conflict of interest The authors declare no conflicts of interest., (Copyright © 2023 The Italian Diabetes Society, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.)

We compared the virulence profile and REP-PCR genotypes of Escherichia coli strains isolated from subclinical and clinical mastitis cases and dairy farm environments in Minas Gerais State, Brazil, to determine virulence factors and genotypes potentially associated with subclinical persistence in the udder. The virulence profile was obtained by the search for three virulence genes: lpf A (long polar fimbriae), fli C (flagella), and esc N (type III secretion system). Subclinical isolates exhibited mainly the fliC gene (33.33%) and fliC + escN genes (30.30%). Clinical isolates exhibited mainly fliC + escN genes (50%) and environmental isolates the lpfA + escN genes (58.04%). Strains isolated from subclinical mastitis showed 6.75 times more positivity to fli C than environmental isolates. Thirty-four genotypes were observed in the REP-PCR analysis, and clinical mastitis isolates indicated more genetic proximity to dairy farm environment isolates than subclinical mastitis isolates. In conclusion, the results suggested that flagella may be an important virulence factor for mammary persistent E. coli infection in cattle, however, none of the E. coli REP-PCR genotypes were associated with subclinical infection.

Background: Minas artisanal cheese (MAC) from the Serro region is a Brazilian intangible cultural heritage. Produced from raw milk, it may carry zoonotic pathogens such as Brucella. This study included a randomized survey for the prevalence of Brucella-positive MAC and its associated factors., Methods: MAC samples (n=55), each one from a different rural family-based cheese-processing agroindustry, were analysed for Brucella by direct polymerase chain reaction (PCR) species-specific DNA detection and cultivation-based approaches., Results: Among 55 MACs that were analysed, we found 17 Brucella DNA-positive samples (30.9% [95% confidence interval {CI} 18.7 to 43.1]) by PCR and, for the first time, from one MAC (1.8% [95% CI 0.5 to 9.7]), viable Brucella abortus was recovered by cultivation. Higher values for two variables, the number of lactating cows per herd (p=0.043) and daily milk production per herd (p=0.043), were each associated with Brucella-positive MAC, which concentrated in three high-risk and one low-risk spatial clusters., Conclusions: MAC may be a source of Brucella for humans, since the positive samples were from batches that were sold by cheesemakers. This should be of concern and encourage cooperation between the health and agriculture sectors in order to mitigate this public health risk through One Health integrated approaches., (© The Author(s) 2022. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.)

Introduction: In Brazil, the Unified Health System (SUS) controls and oversees public health care, and the Family Health Strategy (FHS) is its primary access, with 60% of the population registered in it. The surveillance of risk factors for cardiovascular diseases (CVD) is the responsibility of the FHS. In 2010, the American Heart Association (AHA) proposed the evaluation of seven metrics (smoking, Body Mass Index (BMI), physical activity, diet, total cholesterol, blood pressure and blood glucose) with an aim to monitoring cardiovascular health (CVH). However, the results of the FHS regarding the CVH of the Brazilian population are unascertained., Objective: Evaluate the control of CVH among adult patients treated by the FHS in the city of Aracaju, Sergipe, Brazil., Material and Methods: A cross-sectional study was conducted using the seven metrics recommended by the AHA to evaluate CVH among patients treated by the FHS. The city of Aracaju has a population of 571,149 inhabitants, with 394,267 > 20 years of age; therefore, it was admitted that in a simple random sample, sampling error of 5% with 95% CI, 329 individuals would be needed., Results: Among 400 patients, only 32.5% had controlled CVH. In a univariate analysis, the adjusted multivariate analysis found that being female (aOR: 2.07 IC: 1.20 to 3.60 p : 0.006) under 45 years old (aOR: 1.61 IC: 1.15 to 2.28 p : 0.006) and with the habit of following health advice from family members and neighbors (aOR: 1.28 IC: 1.15 to 2.28 p : 0.040) were associated with control of CVH. On the other hand, those ones who had a greater number of children (aOR: 0.91 IC: 0.84 to 0.95 p : 0.020) were associated with less control of CVH., Conclusions: The study showed that only 32.5% of patients have controlled CVH. Being a woman, young and following health advice from family members and neighbors have a positive influence in controlling CVH. More children reduced controlling these metrics., Competing Interests: Authors MA-S, AS, and JB-F were employed by Rede D'Or São Luiz. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Tavares, Ribeiro, Almeida-Santos, Sousa and Barreto-Filho.)

Background: Global publications on Q fever have increased after the 2007 epidemic in the Netherlands. However, the epidemiology of Q fever/coxiellosis in Brazil is still poorly understood. Accordingly, there have been few studies investigating the presence of Coxiella burnetii in dairy products around the world, especially in Brazil, where consumption of fresh cheese made from raw-milk is very high., Objective: This study was a random survey to assess the prevalence of C. burnetii by PCR in traditional Minas artisanal cheese from the Serro microregion, Brazil, which is manufactured from bovine raw-milk., Methods: DNA extracted from 53 cheese samples were analyzed by nested PCR with C. burnetii-specific primers and the products confirmed by DNA sequencing., Results: Out of the 53 cheese samples five (9.43%) were C. burnetii DNA-positive, each coming from one of the respective randomly selected manufacturing agroindustries. Based on our results, it is estimated that 1.62 tons/day of ready-to-eat cheese made from raw-milk from a total of 16.2 tons produced daily in the study region are contaminated with C. burnetii., Conclusion: To our knowledge, this is the first report of highly heat-resistant zoonotic pathogen in raw-milk Brazilian artisanal cheese. This food safety hazard has been completely neglected in ready-to-eat raw-milk Brazilian artisanal cheese and could imply potential threats to consumers, since C. burnetii survives in artisanal cheese submitted to long ripening periods. Thus, this work established random and representative baseline prevalence of C. burnetii in this food product in Brazil. Further epidemiological studies, monitoring trends and setting control targets are warranted. Finally, these results point out the importance of including C. burnetii in animal and public health surveillance programs., (Copyright © 2020 Sociedade Brasileira de Infectologia. Published by Elsevier España, S.L.U. All rights reserved.)

The aim of this study was to estimate the economic impact of mastitis at the herd level and the weight (percent) of the components of this impact in a Holstein dairy herd under tropical conditions. Three estimates of the economic impact of mastitis were performed. In estimates 1 and 2 the real production and economic indices from February 2011 to January 2012 were considered. In the estimate 1, indices for mastitis classified as ideal were considered, whereas in the estimate 2, the mastitis indices used were those recorded at the farm and at Holstein Cattle Association of Minas Gerais State database (real indices). Ideal mastitis indices were bulk milk somatic cell counts less than 250,000 cells/mL, incidence of clinical mastitis less than 25 cases/100 cows/year, number of culls due to udder health problems less than 5% and the percentage of cows with somatic cell counts greater than 200,000 cells/mL less than 20%. Considering the ideal indices of mastitis, the economic impact was US$19,132.35. The three main components of the economic impact were culling cows (39.4%) and the reduction in milk production due to subclinical and clinical mastitis (32.3% and 18.2%, respectively). Estimate 2 using real mastitis indices showed an economic impact of US$61,623.13 and the reduction in milk production due to mastitis (77.7%) and milk disposal (14.0%) were the most relevant components. The real impact of culling cows was approximately 16 times less than the weight that was considered ideal, indicating that this procedure could have been more frequently adopted. The reduction in milk production was 27.2% higher than the reduction in Estimate 1, indicating a need to control and prevent mastitis. The estimate 3 considered the same indices as estimate 2, but for the period from February 2012 to January 2013. Its economic impact was US$91,552.69. During this period, 161 treatments of cows with an intramammary antibiotic were performed to eliminate Streptococcus agalactiae, and eight cows chronically infected with Staphylococcus aureus were culled. The reduction in milk production due to mastitis was the main component of the economic impact (54.9%). The culling of cows with chronic infection was associated with an increase in the economic impact of mastitis and a reduction in the average productivity per cow. At the herd level reduction in milk production was the component that presented the largest weight in the economic impact of the disease., (Copyright © 2017 Elsevier B.V. All rights reserved.)

This Regional Research Communication describes the characterisation of ampicillin, penicillin and tetracycline resistance in Staphylococcus aureus isolated from bovine subclinical mastitis in Minas Gerais State, Brazil. Ninety S. aureus isolates from bovine mastitis exhibiting phenotypic resistance to ampicillin, penicillin and/or tetracycline were selected for this study. The minimum inhibitory concentration (MIC) of each antibiotic was determined using the E-Test® and the production of beta-lactamase was determined by cefinase disks. The resistance genes blaZ, tet(K), tet(L), tet(M), and tet(O) were investigated by PCR in all of the isolates. The MIC results classified 77, 83 and 71% of the isolates as resistant to ampicillin, penicillin and tetracycline, respectively. The MIC50 and MIC90 were, respectively, 1 and 2 µg/ml for ampicillin, 0·5 and 1 µg/ml for penicillin and 32 and 64 µg/ml for tetracycline. Eighty-six per cent of beta-lactamase producing isolates were detected. Of the 90 isolates investigated, 97% amplified blaZ, 84% amplified tet(K), 9% amplified tet(L), 2% amplified tet(M) and 1% amplified tet(O). Seventy-nine isolates (88%) showed blaZ together with at least one tet gene. S. aureus isolates showed high MIC50 and MIC90 values for the three antimicrobials. The blaZ and tet(K) genes were widespread in the herds studied, and most of the isolates harboured blaZ and tet(K) concomitantly.

Staphylococcus aureus is a pathogen that frequently causes mastitis in bovine herds worldwide. This pathogen produces several virulence factors, including cell-associated adhesins, toxic and cytolytic exoproteins, and capsular polysaccharides. The aim of the present study was to test for the presence of genes involved in capsular polysaccharide production and biofilm formation in S. aureus isolated from bovine mastitis samples collected from 119 dairy herds located in three different Brazilian regions, as well as to assay the production of capsular polysaccharides and biofilm, in vitro. The detection of the cap, icaAD, and bap genes was performed using PCR. The detection and quantification of capsular polysaccharide production was performed using ELISA assays. The ability of the isolates to form a biofilm was examined using the polystyrene surface of microtiter plates. All 159 S. aureus isolates investigated harboured the cap gene: 80 % carried the cap5 gene and 20 % carried the cap8 gene. Sixty-nine percent of the isolates expressed capsular polysaccharide (CP) in vitro, 58 % expressed CP5 and 11 % expressed CP8. All of the isolates harboured the icaA and icaD genes, and 95.6 % of the isolates carried the bap gene. Of the 159 isolates analysed, 97.5 % were biofilm producers. A significant association between the capsular genotype and phenotype and the amount of biofilm formation was detected: cap5/CP5 isolates tended to form more biofilm and to produce a thinner CP layer than cap8/CP8 isolates. The results indicate a high potential for pathogenicity among S. aureus isolated from bovine milk collected from three different regions in Brazil.

Aim: Biological disease-modifying antirheumatic drugs (bDMARDs) are used to treat rheumatoid arthritis (RA) with adalimumab and etanercept the most used bDMARDs in Brazil. This open prospective cohort study evaluated their effectiveness and safety among RA patients in the Brazilian Public Health System given their costs., Methods: The Clinical Disease Activity Index was primarily used to assess their effectiveness after 6 and 12 months of follow-up. The Health Assessment Questionnaire and EuroQol-5D were also used., Results: A total of 266 RA patients started treatment with adalimumab or etanercept. Adalimumab was the most widely used bDMARD (70%). In total, 46% achieved remission or low-disease activity at 12 months with no difference in effectiveness between them (p = 0.306). bDMARDs were more effective in patients who had better functionality at treatment onset and had spent longer in education., Conclusion: This real-world study demonstrated that adalimumab and etanercept are equal alternatives for RA treatment and both were well tolerated., Competing Interests: Financial & competing interests disclosure AM Kakehasi has received educational grants from Abbvie, AstraZeneca, BMS, Roche, Eli-Lilly, Pfizer, Roche, Janssen, Novartis and Sanofi. This work was supported by National Council for Scientific and Technological Development, Brazil (grant number 471819/2013-1) and Fapemig, the Minas Gerais State Research Foundation, Brazil (grant number PPM-0015-15). The write-up was in part supported by a Newton Advanced Fellowship awarded to AA Guerra Jr by the Academy of Medical Sciences, through the UK Government's Newton Fund program. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.

Bacteria of the genus Staphylococcus are one of the major pathogens causing bovine mastitis. In recent decades, resistance of this genus to oxacillin (methicillin) has been a matter of concern due to the possibility of reducing the effectiveness of mastitis treatments and the transfer of resistance determinants to other bacteria. Oxacillin resistance was studied in 170 staphylococci from bovine milk samples, including 79 Staphylococcus aureus and 91 coagulase-negative staphylococci (CNS). The susceptibility profile of 10 antimicrobial agents used in veterinary practice was determined by the Etest method. In addition to the Etest, the phenotypic characterization of oxacillin resistance was tested using the cefoxitin disk diffusion test. All isolates were screened by PCR to detect the mecA gene in 2 different regions of the gene. The isolates with an oxacillin minimum inhibitory concentration ≥0.5 µg/mL or resistant to cefoxitin were identified by sequencing a 536-bp fragment of the 16S rRNA gene. This group of isolates was also evaluated for the presence of blaZ and mecC genes. Molecular analysis of the mecA gene was carried out by typing of the staphylococcal cassette chromosome mec (SCCmec). The relatedness of the mecA-positive isolates was evaluated by macrorestriction of chromosomal DNA followed by pulsed-field gel electrophoresis. With the exception of penicillin and oxacillin, 86% of the isolates showed susceptibility to cephalothin, gentamicin, erythromycin, sulfonamide, trimethoprim-sulfamethoxazole, and tetracycline. All S. aureus isolates were susceptible to oxacillin, whereas 47% (n=43) of the CNS isolates were resistant. The CNS isolates showed a higher resistance to cephalothin, erythromycin, tetracycline, and gentamicin in comparison with S. aureus. The mecA gene was only detected in 10 CNS isolates, identified as Staphylococcus epidermidis, and classified into 3 pulsotypes (A, B, and C) and 4 subtypes (A1, B1, B2, and B3). Among the isolates with an oxacillin resistance phenotype, 12 were positive for the blaZ gene, and 9 of them were mecA-positive. Two of the oxacillin-resistant isolates amplified the mecA homolog gene of Staphylococcus sciuri and none amplified mecC. Three SCCmec types, I, IV, and V, were found. Our results suggest that Staphylococcus epidermidis can be a reservoir for mecA for other Staphylococcus species. Studies investigating the molecular and phenotypic profile of antimicrobial resistance in staphylococcal species should be performed for controlling the spread of resistance and the selection of appropriate therapeutic measures., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background: Through rhythmic variations, the diaphragm influence lower esophageal sphincter (LES) pressure acting as an external sphincter. LES pressure recording is characterized by increased pressure in inspiration due to contraction of the diaphragmatic crura that involves the sphincter., Aim: To describe a method of measuring LES pressure during standardized inspiratory maneuvers with increasing loads., Methods: The study population comprised of eight healthy female volunteers (average age of 31.5 years). An esophageal high-resolution manometry and impedance system was used for measuring the LES pressure during 3-second inspiratory efforts under 12, 24 and 48 cm H2O loads (Threshold maneuvers)., Results: There was a significant difference between the average maximum LES pressure and the average maximum basal LES pressure during the first (76.19±17.92 difference, p=0.0008), second (86.92±19.01 difference, p=0.0004), and third seconds of the maneuver (90.86±17.93 difference, p=0.0002), with 12, 24 and 48 cmH2O loads., Conclusion: This maneuver is a standardization of the inspiratory LES pressure and may better differentiate patients with reflux disease from healthy individuals, and may also be useful for monitoring the treatment of these patients through inspiratory muscle training.

The monoterpenoid 1,8-cineole is obtained from the leaves of Eucalyptus globulus and it has important biological activities. It is a cheap natural substrate because it is a by-product of the Eucalyptus cultivation for wood and pulp production. In this study, it was evaluated the potential of three filamentous fungi in the biotransformation of 1,8-cineole. The study was divided in two steps: first, reactions were carried out with 1,8-cineole at 1 g/L for 24 h; afterwards, reactions were carried out with substrate at 5 g/L for 5 days. The substrate was hydroxylated into 2-exo-hydroxy-1,8-cineole and 3-exo-hydroxy-1,8-cineole by fungi Mucor ramannianus and Aspergillus niger with high stereoselectivity. Trichoderma harzianum was also tested but no transformation was detected. M. ramannianus led to higher than 99% of conversion within 24 h with a starting high substrate concentration (1 g/L). When substrate was added at 5 g/L, only M. ramannianus was able to catalyze the reaction, but the conversion level was 21.7% after 5 days. Both products have defined stereochemistry and could be used as chiral synthons. Furthermore, biological activity has been described for 3-exo-hydroxy-1,8-cineol. To the best of our knowledge, this is the first report on the use of M. ramannianus in this reaction.

The β-ketoester benzyl acetoacetate was enantioselectively reduced to benzyl (S)-3-hydroxybutanoate by seven microorganism species. The best result using free cells was obtained with the yeast Hansenula sp., which furnished 97% ee and 85% of conversion within 24 h. After immobilization in calcium alginate spheres, K.marxianus showed to be more stable after 2 cycles of reaction.

Introduction: The treatment of leishmaniasis ischallenging, given the difficulties in drug administration and resistance. Therefore, we chose to test the efficacy of miltefosine combined with pentoxifylline., Methods: Twenty-seven isogenic C57Bl/6 mice were infected with Leishmania (Leishmania) amazonensis, and equally divided into three groups: miltefosine (200mg/kg/day), miltefosine (200mg/kg/day) with pentoxifylline (8mg/kg/day), and untreated. Response to treatment was evaluated using paw diameter and parasitological criteria., Results: The number of viable Leishmania reduced significantly within the miltefosine-pentoxifylline group (p < 0.05)., Conclusions: There is hope that a viable treatment exists for Leishmania infection.

Objective: To assess whether the month of birth in different latitudes of South America might influence the presence or severity of multiple sclerosis (MS) later in life., Methods: Neurologists in four South American countries working at MS units collected data on their patients' month of birth, gender, age, and disease progression., Results: Analysis of data from 1207 MS patients and 1207 control subjects did not show any significant variation in the month of birth regarding the prevalence of MS in four latitude bands (0-10; 11-20; 21-30; and 31-40 degrees). There was no relationship between the month of birth and the severity of disease in each latitude band., Conclusion: The results from this study show that MS patients born to mothers who were pregnant at different Southern latitudes do not follow the seasonal pattern observed at high Northern latitudes.

Inactivation of the pgg2 gene, a polygalacturonase-encoding gene from Penicillium griseoroseum, reduced the total activity of polygalacturonase (PG) by 90 % in wild-type P. griseoroseum, which indicates that the pgg2 gene is the major gene responsible for PG production in this species. To increase PG production, the coding region of the pgg2 gene was cloned under the control of the glyceraldehyde 3-phosphate dehydrogenase (gpd) promoter and the terminator region of the tryptophan synthase (trpC) gene from Aspergillus nidulans (pAN52pgg2 vector). This vector was then used to transform P. griseoroseum. The transformed strains were characterized according to PG production using glucose, sucrose, or sugar cane juice as the carbon sources. The recombinant P. griseoroseum T146 strain contained an additional copy of the pgg2 gene, which resulted in a 12-fold increase in PG activity when compared with that detected in the supernatant of the control PG63 strain. The proteins secreted by the recombinant strain T146 showed a strong band at 38 kDa, which corresponds to the molecular weight of PG of the P. griseoroseum. The results demonstrate the significant biotechnological potential of recombinant P. griseoroseum T146 for use in PG production.

Objective: The aim of this exploratory, cross-sectional study was to evaluate pain in sickle cell disease patients and aspects related to primary healthcare., Methods: Data were obtained through home interviews. The assessment instruments (body diagram, Numerical Pain Scale, McGill Pain Questionnaire) collected information on the underlying disease and on pain. Data were analyzed using the Statistical Package for Social Sciences program for Windows. Associations between the subgroups of sickle cell disease patients (hemoglobin SS, hemoglobin SC, sickle β-thalassemia and others) and pain were analyzed using contingency tables and non-parametric tests of association (classic chi-square, Fisher's and Kruskal-Wallis) with a level of 5% (p-value < 0.05) being set for the rejection of the null hypothesis., Results: Forty-seven over 18-year-old patients with sickle cell disease were evaluated. Most were black (78.7%) and female (59.6%) and the mean age was 30.1 years. The average number of bouts of pain annually was 7.02; pain was predominantly reported by individuals with sickle cell anemia (hemoglobin SS). The intensity of pain (Numeric Pain Scale) was 5.5 and the quantitative index (McGill) was 35.9. This study also shows that patients presented a high frequency of moderately painful crises in their own homes., Conclusion: According to these facts, it is essential that pain related to sickle cell disease is properly identified, quantified, characterized and treated at the three levels of healthcare. In primary healthcare, accurate measurement of pain combined with better care may decrease acute painful episodes and consequently minimize tissue damage, thus improving the patient's overall health.

Background: Sickle cell disease, the most common hereditary blood disease in the world, is the result of an atypical hemoglobin called S (Hb S) which, when homozygous (Hb SS) is the cause of sickle cell anemia. Changes of puberty, correlated with a delayed growth spurt, begin late in both male and female sickle cell anemia individuals with repercussions on sexuality and reproduction. The objectives of this exploratory and descriptive study were to characterize the development of sexuality in adults with sickle cell anemia by investigating the patient's perception of their sex life, as well as the information they had and needed on this subject., Methods: Twenty male and female sickle cell anemia patients treated at the Hemocentro Regional de Uberaba (UFTM) with ages between 19 and 47 years old were enrolled. A socioeconomic questionnaire and a semi-structured interview on sexuality, reproduction and genetic counseling were applied., Results: This study shows that the sickle cell anemia patients lacked information on sexuality especially about the risks of pregnancy and the possible inheritance of the disease by their children. Moreover, the sexual life of the patients was impaired due to pain as well as discrimination and negative feelings experienced in close relationships., Conclusion: The health care of sickle cell anemia patients should take into account not only the clinical aspects of the disease, but also psychosocial aspects by providing counseling on sexuality, reproduction and genetics, in order to give this population the possibility of a better quality of life.

Background: Liver transplantation is the only effective treatment for chronic liver diseases and terminal survival rate has increased in recent decades. However, biliary complications have high incidence and remain as the "Achilles heel" for liver transplantation., Aim: To evaluate retrospectively endoscopic treatment outcomes of biliary complications in post-liver transplantations., Methods: The sample consisted of post-liver transplantation patients for endoscopic retrograde cholangiopancreatography due to suspected biliary complications., Results: Fifteen patients were included (10 male, mean age of 49.57 years) and 36 endoscopic retrograde cholangiopancreatographies were undertaken (2.4/patient). Biliary stricture was diagnosed in 13 patients and endoscopic treatment was successful in 56% (38,46% still in treatment). Biliary leaks were found in one patient and dysfunction of the hepatobilliary ampulla with choledocholithiasis was diagnosed in one patient, both cured by endoscopic treatment., Conclusions: Post-liver transplantation biliary complications are relatively common and endoscopic treatment may result in satisfactory outcome. Stenosis was the more frequent complication in this series.

Background and Objectives: This is an exploratory, descriptive and transversal study aiming to determine the prevalence, characterization, location, and measurement and discuss pharmacological analgesic measures for acute pain management in five inpatient wards of a university hospital., Method: We enrolled 856 subjects in the study, of whom 272 were in pain at the time. Information related to pain was obtained using a bedside structured interview. Numeric pain scale and body diagram were used., Results: Analgesia was assessed through medical records. The overall prevalence of pain was 31.8%, with severe pain in 44.2% and mean of 6.6 on numeric pain scale. The main reason was trauma and the most common site the abdomen. The most widely used analgesic was dipyrone (76.1%) with/without combination. Strong opioid was prescribed to 4.4%. For 27.5% there was no improvement., Conclusion: We conclude that pain is highly prevalent, poorly evaluated, undertreated, with inappropriate use of analgesics., (Copyright © 2012 Elsevier Editora Ltda. All rights reserved.)

Unlabelled: Multiple sclerosis (MS) is an immune-mediated disease that affects the central nervous system. Clinical presentation and prevalence vary widely around the world., Objective: To describe the clinical and epidemiological aspects of patients with MS in Uberaba (MG)., Method: We conducted a transversal descriptive study, with data analysis of 35 patients with MS., Results: Prevalence of MS was 12.5 cases/100,000 inhabitants, with a predominance in females (71.4%) and Caucasoid (85.7%). The current average age was (43.8 ys). The most common initial symptom was sensory (40%), followed by optical neuritis (25.7%). Expanded Disability Status Scale average score was 2.4. The relapsing-remitting form was predominant (88.6%), most (74.3%) were on immunomodulatory treatment and (40%) had college education., Conclusion: Prevalence of MS in Uberaba (MG) is considered average in accordance to Kurtzke and Page and clinical features are consistent with most Brazilian studies.

Previous reports have described pgg2, a polygalacturonase-encoding gene of Penicillium griseoroseum, as an attractive model for transcriptional regulation studies, due to its high expression throughout several in vitro growth conditions, even in the presence of non-inducing sugars such as sucrose. A search for regulatory motifs in the 5' upstream regulatory sequence of pgg2 identified a putative CCAAT box that could justify this expression profile. This element, located 270 bp upstream of the translational start codon, was tested as binding target for regulatory proteins. Analysis of a 170 bp promoter fragment by electrophoretic mobility shift assay (EMSA) with nuclear extracts prepared from mycelia grown in pectin-containing culture medium revealed a high mobility complex that was subsequently confirmed by analyzing it with a double-stranded oligonucleotide spanning the CCAAT motif. A substitution in the core sequence for GTAGG partially abolished the formation of specific complexes, showing the involvement of the CCAAT box in the regulation of the polygalacturonase gene studied.

The pectin lyase (PL) is an industrially important enzyme since it is used for maceration and clarification in the process of fruit juice production in food industries. In order to increase the yields of pectin lyase we cloned the plg1 (pectin lyase 1) from Penicillium griseoroseum gene under the control of the strong constitutive promoter of the glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) and the terminator region of the tryptophan synthetase (trpC) gene from Aspergillus nidulans (plasmid pAN52-Plg1) and transformed this construct into the P. griseoroseum strain PG63. One of the pAN52-Plg1 multi-copy transformants (strain 105) grown in culture medium containing glucose or sugar cane juice showed PL activities of 4,804 or 5,202 U ml(-1) respectively, which represented 57- and 132-fold increases. In addition, the apparent specific activity of PL produced by this strain was much higher than the one observed for a commercial pectinase preparation. Evaluation of the extracellular proteins in the culture supernatant of strain 105 by SDS-PAGE showed the presence of a clear and strong band of approximately 40 kDa that probably corresponds to PL. The enzyme yields reported here demonstrate that the system we developed is able to express pectin lyase at levels comparable to, or exceeding, previously reported data.