Copper(II) complex-loaded castor oil-based nanostructured lipid carriers used against Mycobacterium tuberculosis : Development, characterisation, in vitro and in vivo biological assays.

A copper(II) complex-loaded castor oil-based nanostructured lipid carrier was evaluated to enhance the poor water solubility of antimicrobial compounds, improving their biological properties and antimicrobial activity against Mycobacterium tuberculosis. Nanostructured lipid carriers were composed of the castor oil, polyoxyethylene 40 stearate and caprylic/capric triglyceride, poloxamer 407, cetyltrimethylammonium bromide and three different copper(II) complexes. The systems were ultrasonicated at an amplitude of 8% for 20 min and an ice bath was used throughout the procedure. The blank nanostructured lipid carrier (F5) and nanostructured lipid carriers loaded with copper(II) complex 1, 2 and 3 (F5.1, F5.2 and F5.3, respectively) for 45 days presented values of mean diameter, poly dispersity index and zeta potential ranging from 186 to 199 nm, 0.14 to 0.2 and 24 to 30 mV, respectively. Atomic force microscopy indicated that the nanostructured lipid carriers were distributed at the nanoscale, corroborating the mean diameter data. Differential scanning calorimetry determined the melting points of the constituents of the nanostructured lipid carriers. The antimicrobial activity of copper(II) complexloaded F5 against M. tuberculosis H37Rv showed better anti-tuberculosis activity than the free complexes. In vivo biological assays of complex-loaded F5 demonstrated reduced toxicity. Our results suggest that nanostructured lipid carriers could be a potential nanotechnological strategy to optimise tuberculosis treatment.

The 4G/4G polymorphism of the plasminogen activator inhibitor-1 (PAI-1) gene as an independent risk factor for placental insufficiency, which triggers fetal hemodynamic centralization.

OBJECTIVE: To describe a case report of 4G/4G polymorphism of the plasminogen activator inhibitor-1 (PAI-1) gene as an independent risk factor for placental insufficiency. DESIGN: Case report. SETTING: Department of Public Health, State University of Ceará (UECE), Fortaleza-CE, Brazil. CASE REPORT: Hereditary hypofibrinolysis, which is mediated by 4G/4G homozygosity for the PAI-1 gene, is an independent risk factor for pregnancy complications, probably acting through thrombotic induction of placental insufficiency. We report a case of a low risk pregnancy, which separately presented placental insufficiency and fetal centralization at the beginning of the third trimester, without any other clinical manifestations during pregnancy. However, immediately after childbirth, the patient had a deep vein thrombosis of a lower limb. The anatomopathological examination of the placenta showed old and recent placental infarcts. Homozygosity for the 4G allele of PAI-1 gene was subsequently diagnosed as the sole probable causal factor.

Genetic variability in the ITS and IGS regions of the ribosomal DNA of Acremonium cavaraeanum exhibiting antimicrobial activity.

Endophytic microorganisms represent promising alternatives for obtaining new drugs of biotechnological importance. In this study, the endophytic species Acremonium cavaraeanum (A1a) isolated from Cocos nucifera was cultivated for the production of secondary metabolites, and its extracts and fractions were evaluated by the dilution method (MIC). The EtOAc extracts and MeOH fractions were tested against Gram-positive and -negative bacteria, and had an MIC of 125 µg/mL when evaluated in the EtOAc extract (EBI). The EtOAc extract (EBII) had an MIC of 62.25 µg/mL for Staphylococcus aureus and an MIC between 125 and 250 µg/mL for Gram-negative bacteria. The methanolic fractions showed activity with MIC between 125 and 250 µg/mL for all bacteria tested. The IGS region of the rDNA repeat unit of genomic DNA was analyzed by PCR/RFLPs, including endonucleases PstI, BamHII, HinfI, and EcoRI. The physical maps showed different restriction sites for the 6 Acremonium sp isolates, and revealed 5 RFLP patterns. The results showed that isolates of the same Acremonium species exhibited variation in this specific region. The sequences of ITS1-5.8S-ITS2 regions were aligned by Clustal W using the neighbor joining method, which grouped the isolates into 5 distinct clusters. This study aimed to evaluate the genetic diversity of A. cavaraeanum crops exhibiting antibacterial activity. The results of this study indicate that different fungal genetic isolates have biotechnological potential for the production of active bio-compounds against several human pathogens.

Prevalence of clue cells suggestive for Gardnerella vaginalis in population-based cervical screening in the public versus private health care in Maringá, Paraná, Brazil.

OBJECTIVES: To determine the prevalence of clue cells suggestive for Gardnerella vaginalis in population-based cervical screening in the public (SUS) versus private (RP) health care utilization in Maringá, Paraná, Brazil. METHODS: A retrospective study on the results of 133,966 cytology examinations of patients from the Brazilian Public Health System-SUS (better socio-economic status-SES) and from the Private Network-RP (low SES) done by the Souza Anatomy Laboratory in Maringá, Brazil, from 2004 to 2007. RESULTS: In patients using the SUS, the prevalence of clue cells was 6.75% (n = 3,573), higher (p < 0.05) than for RP users (3.53%, n = 2,813). The clue cells predominated in age range of the 20-49 years from the SUS and RP (p > 0.05). In both the populations, the prevalence and age range did not vary among the 4 years (p > 0.05) and the prevalence declines in age above 45 years (p < 0.05). CONCLUSIONS: The prevalence of clue cells was low and stable in both populations in the 4 years, indicating equilibrium in terms of numbers of bacterial vaginosis (BV) in this localization. The results indicate that SES may influence the epidemiology of BV and that the prevalence declines expressively in older women.

The centrality of workers to sustainability based on values: Exploring ergonomics to introduce new rationalities into decision-making processes.

This article presents the results of an academic research project connecting the discipline of ergonomics (and work-related issues) with the theme of sustainability. Despite the stated aim of creating value for stakeholders, including employees, companies face difficulties in introducing effective sustainability policies. The research question addressed in this article is the following: How can companies improve their decision-making processes to increase workers' wellbeing using policies integrating issues related to corporate sustainability and ergonomics? Currently, corporate sustainability is focused mainly on the triple bottom line (TBL) concept. In this context, the integration of ergonomics is fragmented and arguably separate from strategic human-resource functions (which have largely been the primary promoter of the internal-social component of corporate sustainability). This research argues that corporate sustainability requires a new step, improving the decision-making process, with the inclusion of more types of rationalities and the recognition of the centrality of workers in the process of creating sustainable action. When corporate sustainability policies focus on worker centrality, they open space for the integration of ergonomics as a pillar of an organization's corporate sustainability strategy. Based on a complex view of work systems, ergonomics can introduce values aligned with sustainability and promote cooperation in organizations. Different stakeholders working at companies can make use of different concepts proposed by ergonomics and other work sciences to support changes in their decision-making processes. In view of broader sustainability objectives, integrating different work-related considerations can help improve performance, including productivity, quality and health.

Furoxan derivatives demonstrated in vivo efficacy by reducing Mycobacterium tuberculosis to undetectable levels in a mouse model of infection.

OBJECTIVES: The most recent survey conducted by the World Health Organization described Tuberculosis (TB) as one of the top 10 causes of death and the leading cause of death from a single infectious agent. The increasing number of TB-resistant cases has contributed to this scenario. In light of this, new strategies to control and treat the disease are necessary. Our research group has previously described furoxan derivatives as promising scaffolds to be explored as new antitubercular drugs. RESULTS: Two of these furoxan derivatives, (14b) and (14c), demonstrated a high selectivity against Mycobacterium tuberculosis. The compounds (14b) and (14c) were also active against a latent M. tuberculosis strain, with MIC90 values of 6.67 µM and 9.84 µM, respectively; they were also active against monoresistant strains (MIC90 values ranging from 0.61 to 20.42 µM) and clinical MDR strains (MIC90 values ranging from 3.09 to 42.95 µM). Time-kill experiments with compound (14c) showed early bactericidal effects that were superior to those of the first- and second-line anti-tuberculosis drugs currently used in therapy. The safety of compounds (14b) and (14c) was demonstrated by the Ames test because these molecules were not mutagenic under the tested conditions. Finally, we confirmed the safety, and high efficacy of compounds (14b) and (14c), which reduced M. tuberculosis to undetectable levels in a mouse aerosol model of infection. CONCLUSION: Altogether, we have identified two advanced lead compounds, (14b) and (14c), as novel promising candidates for the treatment of TB infection.

Association of public versus private health care utilization and prevalence of Trichomonas vaginalis in Maringá, Paraná, Brazil.

OBJECTIVES: To determine the association of public versus private health care utilization and prevalence of Trichomonas vaginalis in Maringá, Paraná, Brazil. METHODS: A retrospective study on the results of 133,966 cytology examinations of patients from the Brazilian Public Health System (better socio-economic status, SES) and from the Private Network (low SES) done by the Souza Anatomy Laboratory in Maringá, Brazil, from 2004 to 2007. RESULTS: In patients using the SUS, the prevalence of trichomoniasis was 1.44% (n = 782), higher (P < 0.0001) than for RP users (0.18%, n = 147). The infection predominated in age ranges of the 30-39 years and 40-49 years old from the SUS (P < 0.0001) and in ages ranges from 40 to 49 years old from the RP (P < 0.0001). The infection prevalence and age range did not vary among the 4 years (P > 0.05). CONCLUSIONS: The prevalence of Trichomonas spp. was low and stable in both populations in the 4 years, indicating an equilibrium in terms of numbers of this agent in Maringá, Paraná, Brazil. The results corroborate the hypothesis that SES may influence the epidemiology of trichomoniasis. The prevalence of the infection increases with age, a phenomenon not observed with other sexually transmitted infections, such as gonorrhoea or Chlamydia trachomatis.

Prevalence of Candida sp. in the cervical-vaginal cytology stained by Harris-Shorr.

OBJECTIVES: To determine the prevalence of Candida sp. by cervical-vaginal cytology in patients from the Brazilian Public Health System (SUS) and from the Private Network (RP). METHODS: A retrospective study on the results of cytology exams from the Souza Anatomy Laboratory of Maringá, Paraná state, Brazil, from 2004 to 2006. RESULTS: A total of 94,203 reports of cytology were evaluated, with a prevalence of Candida sp. of 4.94% (n = 4,655), with no significant variation in the two populations or among the 3 years studied (P > 0.05). Candida sp. predominated in both populations, aged between 20 and 39 years (P < 0.0001). The bacterial flora linked to Candida sp. was of lactobacilli (P < 0.05). CONCLUSIONS: With the results obtained, it was noticed that the socio-economical level does not seem to influence the epidemiology of the Candida sp. and pointed to a balance in terms of numbers of this agent in Maringá, Paraná, Brazil.

Knowledge Among University Students in Rio de Janeiro Regarding Transplant Organ Donation.

INTRODUCTION: Organ transplantation is often the only possible treatment to save the lives of patients with end-stage organ failure. Limiting factors include failure to notify in cases of patients with brain death, the inefficient procurement and distribution of organs, the lack of specific educational policies for health care professionals, lack of knowledge on the organ transplantation process, and family refusal for organ donation. OBJECTIVE: To evaluate the knowledge of students enrolled in different undergraduate university courses in Rio de Janeiro on the regulations and strategies governing transplant organ donation in Brazil. METHODS: This qualitative, observational study used a 10-item questionnaire aimed at obtaining data on respondents' general knowledge regarding organ donation and transplantation. The questionnaire was applied using either a printed or an electronic version developed on Google Forms and was completed anonymously. RESULTS: Overall, 587 questionnaires were completed. The participants were divided into 3 groups according to their field of study: 256 (43.6%) from courses related to arts and humanities, 159 (27.1%) from science and technology-related courses, and 172 (29.3%) from the biomedical field. Most respondents (396; 67.5%) were unaware of the criteria required to be an organ transplant donor. CONCLUSION: There is a significant lack of knowledge among university students on issues related to the organ donation and transplantation process in Brazil.

Fish hemoglobins.

Vertebrate hemoglobin, contained in erythrocytes, is a globular protein with a quaternary structure composed of 4 globin chains (2 alpha and 2 beta) and a prosthetic group named heme bound to each one. Having myoglobin as an ancestor, hemoglobin acquired the capacity to respond to chemical stimuli that modulate its function according to tissue requirements for oxygen. Fish are generally submitted to spatial and temporal O2 variations and have developed anatomical, physiological and biochemical strategies to adapt to the changing environmental gas availability. Structurally, most fish hemoglobins are tetrameric; however, those from some species such as lamprey and hagfish dissociate, being monomeric when oxygenated and oligomeric when deoxygenated. Fish blood frequently possesses several hemoglobins; the primary origin of this finding lies in the polymorphism that occurs in the globin loci, an aspect that may occasionally confer advantages to its carriers or even be a harmless evolutionary remnant. On the other hand, the functional properties exhibit different behaviors, ranging from a total absence of responses to allosteric regulation to drastic ones, such as the Root effect.

Monitoring of urban and rural basins: water quality of Mourão basin.

The Mourão River basin is located on the central western region of the Paraná State - Brazil, between coordinates 23º 44' - 24º 25 South latitude and 52º 12' - 52º 30' West longitude, between 270 and 820 m above sea level, and 1,648.21 km2 drainage area. Water quality was evaluated by monitoring physical, chemical and microbiological parameters. Monthly samplings were performed for a year at five sites in the basin for analysis of: pH, temperature, dissolved oxygen, biochemical oxygen demand, total nitrogen, ammoniacal nitrogen, nitrite, nitrate, total phosphorus, turbidity, total solids, volatile solids and fecal coliforms. The results of the evaluated parameters showed higher values than the limits set by CONAMA Resolution 357 from 2005 for Class 2 in some samples. The Water Quality Index (WQI) indicated that 72% of samples had average quality and 28% had good quality for the Mourão River basin. Higher values of WQI were observed after rainfall period with median of 75 compared to the dry period with median of 62. The source of the Mourão River is contaminated with fecal coliforms, evidencing the real need to treat sewage in rural areas.

Avaliação da técnica de vitrificação de ovários de camundongos da linhagem B6D2F1, pertencentes ao Instituto de Ciência e Tecnologia em Biomodelos - ICTB/Fiocruz-RJ / Evaluation of ovary vitrification technique from B6D2F1 strain belonging to the Institute of Biomodels Science and Technology ICTB / Fiocruz-RJ

Objetivou-se testar a vitrificação de ovários de camundongos do ICTB/Fiocruz. Inicialmente, fez-se coleta e maturação in vitro dos oócitos de ovários a fresco e vitrificados, bem como avaliação de estruturas no cultivo embrionário, pós-fertilização in vitro. Fêmeas B6D2F1 foram eutanasiadas para remoção dos ovários (n=60) e divididas em três grupos: grupo 1 (n=30 animais) - oócito de ovários vitrificados, maturados e fertilizados in vitro (120 fragmentos); grupo 2 (n=15) (controle 1) - oócitos coletados a fresco, maturados e fertilizados in vitro; e grupo 3 (n=15) (controle 2) - oócitos maturados in vivo e fertilizados in vitro. A técnica foi verificada no desenvolvimento embrionário in vitro, que foi avaliado pelo teste de qui-quadrado (BioStat 5.0). Recuperaram-se 123, 224 e 328 oócitos nos G1, G2 e G3, respectivamente. Observaram-se diferenças significativas nas taxas de clivagem às 24 horas (embriões ≥ 2 células) entre G1 (8%) e G2 (32%) (P<0,1) e G1 e G3 (49%) (P<0,05), mas não entre G2 e G3 (P>0,05). Para blastocistos, às 96 horas, os grupos G1, G2 e G3 apresentaram, respectivamente, 6%, 11% e 46%, diferindo significativamente entre eles (P<0,05). A vitrificação de ovários, a maturação oocitária e a fertilização in vitro são alternativas para a produção de embriões de camundongos in vitro.(AU)

This work aimed test ovarian vitrification of hybrid mouse from ICTB/Fiocruz. Protocol collection and oocyte in vitro maturation from fresh and vitrified ovaries was established and embryos were evaluated after fertilization. B6D2F1 females were euthanized for ovarian removal (n= 60) and divided into 3 groups: G1 (n= 30) - ovaries fragmented (n= 120), vitrified, matured and fertilized; G2 (n= 15) - in vitro fertilization of oocytes matured in vitro from fresh ovaries; G3 (n= 15) - ampulla region oocytes in vitro fertilizated. Viability was verified by thawing, oocyte in vitro maturation and fertilization. In vitro embryo development of each group was evaluated by Chi-square test (BioStat 5.0). 123, 224 and 328 oocytes were recovered from G1, G2 and G3, respectively. Significant differences were observed in cleavage rates at 24 hours (embryos with 2 cells or more) between G1 (8%) and G2 (32%) (P< 0.1) and G1 and G3 (49%) (P< 0.05) but not between G2 and G3 (P> 0.05). Blastocysts at 96 hours presented 6%, 11% and 46%, respectively for G1, G2 and G3, differing significantly (P< 0.05). Ovary vitrification, oocyte in vitro maturation and in vitro fertilization were available for the production of in vitro mouse embryos.(AU)

Optimization of dental CBCT exposures through mAs reduction.

OBJECTIVES: To investigate the effect of tube current-exposure time (mAs) reduction on clinical and technical image quality for different CBCT scanners, and to determine preliminary minimally acceptable values for the mAs and contrast-to-noise ratio (CNR) in CBCT. METHODS: A polymethyl methacrylate (PMMA) phantom and an anthropomorphic skull phantom, containing a human skeleton embedded in polyurethane, were scanned using four CBCT devices, including seven exposure protocols. For all protocols, the mAs was varied within the selectable range. Using the PMMA phantom, the CNRAIR was measured and corrected for voxel size. Eight axial slices and one coronal slice showing various anatomical landmarks were selected for each CBCT scan of the skull phantom. The slices were presented to six dentomaxillofacial radiologists, providing scores for various anatomical and diagnostic parameters. RESULTS: A hyperbolic relationship was seen between CNRAIR and mAs. Similarly, a gradual reduction in clinical image quality was seen at lower mAs values; however, for several protocols, image quality remained acceptable for a moderate or large mAs reduction compared with the standard exposure setting, depending on the clinical application. The relationship between mAs, CNRAIR and observer scores was different for each CBCT device. Minimally acceptable values for mAs were between 9 and 70, depending on the criterion and clinical application. CONCLUSIONS: Although noise increased at a lower mAs, clinical image quality often remained acceptable at exposure levels below the manufacturer's recommended setting, for certain patient groups. Currently, it is not possible to determine minimally acceptable values for image quality that are applicable to multiple CBCT models.

Characterisation of lymphocyte response and cytokine patterns in patients with dengue fever.

It is believed that the pathogenesis of dengue is generated by a deregulation of the immunological response. Dengue virus-infected monocytes/macrophages are likely to secrete monokines, which play a role in clinical features observed in patients with dengue haemorrhagic fever or dengue shock syndrome. This is a report on a study on 45 individuals presenting clinical and laboratory characteristics of dengue virus infection. During the acute phase of infection, immunophenotyping of peripheral mononuclear leukocytes was carried out in 19 patients and demonstrated a reduced frequency of CD2+ lymphocytes and their CD4+ and CD8+ subsets. Normal ratios were recovered during convalescence. Also, during the acute phase, mononuclear cells proliferated poorly in response to mitogens and dengue antigens as detected by incorporation of radiolabeled thymidine. During convalescence the lymphoproliferative response was re-established. In addition, the presence of circulating cytokines was investigated in the plasma of the same 45 patients. Concentrations of tumour necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), interleukin-10 (IL-10) and soluble tumor necrosis factor receptor (sTNF-Rp75) were found to be significantly elevated in patients when compared to normal controls. The increase in TNF-alpha was correlated with haemorrhagic manifestations and the increase in IL-10 with platelet decay. The data demonstrate that during the acute phase of dengue infection subsets of T lymphocytes are depressed in terms of both rate and function and provide evidence that circulating pro-inflammatory cytokines, such as TNF-alpha, are important in the pathogenesis and severity of dengue. IL-10 may be downregulating lymphocyte and platelet function.

Coexpression of cytokeratin and vimentin in mice trophoblastic giant cells.

Trophoblastic giant cells reach their maximum size and exhibit a conspicuous synthetic and invasive activity during mouse placentation. The cytoskeleton, given the complex functions of the cells, shows a well-developed network of intermediate filament proteins. Immunohistochemistry combined with confocal and conventional immunofluorescence studies of intermediate filaments proteins cytokeratin and vimentin were performed in mice trophoblastic giant cells on days 9-11 of pregnancy. Specimens were fixed in phosphate-buffered formaldehyde and tissues were processed for routine paraffin embedding. Trophoblastic giant cells from antimesometrial, lateral or mesometrial uterine regions, through days 9-11 of pregnancy, expressed the same staining with both immunoperoxidase and immunofluorescent techniques. Cytokeratin filamentous structures were intensely immunoreactive and were detected throughout the cells cytoplasm; a few cells exhibited strongest fluorescence in the peripheral cytoplasm. Vimentin-positive staining was often distributed throughout the cells cytoplasm, most frequently and more intensely in the peripheral region; in some cells, it was present only in the peripheral regions. It is probable that expression of vimentin in midpregnancy trophoblastic giant cells may be associated with the rapid and conspicuous increase in size and synthetic activity of the cells and also with phagocytosis of degraded materials and invasion of decidual tissue.

The intracarotid amobarbital procedure (Wada test) with two protocols combined, Montreal and Seattle.

The intracarotid amobarbital procedure was carried out in 8 male and 7 female candidates to temporal lobectomy, and a female candidate to frontal lesionectomy, aged 18-50 (mean 32.5) years. Language and memory were tested after injection in each hemisphere. Both were measured by the Montreal procedure. In 9 patients language and memory were evaluated with the Seattle procedure too. In 12 patients the left hemisphere was dominant for language; three had bilateral dominance. In 1 patient the Seattle procedure demonstrated the dominant hemisphere by relatively slowness of speech during the drug effect in the left hemisphere. Memory was defined to be in the left hemisphere in 12 patients, in the right in 2, bilateral in 1 and in another lateralization was not possible. In 1 patient memory dominance was determined by the Montreal protocol alone because of lack of cooperation. These early results indicate that the methods may be complementary for determination of language and memory dominance in epilepsy surgery candidates.

Identification of a new hormone-binding site on the surface of thyroid hormone receptor.

Thyroid hormone receptors (TRs) are members of the nuclear receptor superfamily of ligand-activated transcription factors involved in cell differentiation, growth, and homeostasis. Although X-ray structures of many nuclear receptor ligand-binding domains (LBDs) reveal that the ligand binds within the hydrophobic core of the ligand-binding pocket, a few studies suggest the possibility of ligands binding to other sites. Here, we report a new x-ray crystallographic structure of TR-LBD that shows a second binding site for T3 and T4 located between H9, H10, and H11 of the TRα LBD surface. Statistical multiple sequence analysis, site-directed mutagenesis, and cell transactivation assays indicate that residues of the second binding site could be important for the TR function. We also conducted molecular dynamics simulations to investigate ligand mobility and ligand-protein interaction for T3 and T4 bound to this new TR surface-binding site. Extensive molecular dynamics simulations designed to compute ligand-protein dissociation constant indicate that the binding affinities to this surface site are of the order of the plasma and intracellular concentrations of the thyroid hormones, suggesting that ligands may bind to this new binding site under physiological conditions. Therefore, the second binding site could be useful as a new target site for drug design and could modulate selectively TR functions.

Bivariate and multivariate analyses of the correlations between stability of the erythrocyte membrane, serum lipids and hematological variables.

The observation that the fluidity must remain within a critical interval, outside which the stability and functionality of the cell tends to decrease, shows that stability, fluidity and function are related and that the measure of erythrocyte stability allows inferences about the fluidity or functionality of these cells. This study determined the biochemical and hematological variables that are directly or indirectly related to erythrocyte stability in a population of 71 volunteers. Data were evaluated by bivariate and multivariate analysis. The erythrocyte stability showed a greater association with hematological variables than the biochemical variables. The RDW stands out for its strong correlation with the stability of erythrocyte membrane, without being heavily influenced by other factors. Regarding the biochemical variables, the erythrocyte stability was more sensitive to LDL-C. Erythrocyte stability was significantly associated with RDW and LDL-C. Thus, the level of LDL-C is a consistent link between stability and functionality, suggesting that a measure of stability could be more one indirect parameter for assessing the risk of degenerative processes associated with high levels of LDL-C.

Analysis of agonist and antagonist effects on thyroid hormone receptor conformation by hydrogen/deuterium exchange.

Thyroid hormone receptors (TRs) are ligand-gated transcription factors with critical roles in development and metabolism. Although x-ray structures of TR ligand-binding domains (LBDs) with agonists are available, comparable structures without ligand (apo-TR) or with antagonists are not. It remains important to understand apo-LBD conformation and the way that it rearranges with ligands to develop better TR pharmaceuticals. In this study, we conducted hydrogen/deuterium exchange on TR LBDs with or without agonist (T(3)) or antagonist (NH3). Both ligands reduce deuterium incorporation into LBD amide hydrogens, implying tighter overall folding of the domain. As predicted, mass spectroscopic analysis of individual proteolytic peptides after hydrogen/deuterium exchange reveals that ligand increases the degree of solvent protection of regions close to the buried ligand-binding pocket. However, there is also extensive ligand protection of other regions, including the dimer surface at H10-H11, providing evidence for allosteric communication between the ligand-binding pocket and distant interaction surfaces. Surprisingly, C-terminal activation helix H12, which is known to alter position with ligand, remains relatively protected from solvent in all conditions suggesting that it is packed against the LBD irrespective of the presence or type of ligand. T(3), but not NH3, increases accessibility of the upper part of H3-H5 to solvent, and we propose that TR H12 interacts with this region in apo-TR and that this interaction is blocked by T(3) but not NH3. We present data from site-directed mutagenesis experiments and molecular dynamics simulations that lend support to this structural model of apo-TR and its ligand-dependent conformational changes.

Monitoring of urban and rural basins: water quality of Mourão basin / Monitoramento de bacias urbana e rural: análise integrada de qualidade de água da bacia hidrográfica do Rio Mourão

The Mourão River basin is located on the central western region of the Paraná State – Brazil, between coordinates 23º 44’ - 24º 25 South latitude and 52º 12’ - 52º 30’ West longitude, between 270 and 820 m above sea level, and 1,648.21 km2 drainage area. Water quality was evaluated by monitoring physical, chemical and microbiological parameters. Monthly samplings were performed for a year at five sites in the basin for analysis of: pH, temperature, dissolved oxygen, biochemical oxygen demand, total nitrogen, ammoniacal nitrogen, nitrite, nitrate, total phosphorus, turbidity, total solids, volatile solids and fecal coliforms. The results of the evaluated parameters showed higher values than the limits set by CONAMA Resolution 357 from 2005 for Class 2 in some samples. The Water Quality Index (WQI) indicated that 72% of samples had average quality and 28% had good quality for the Mourão River basin. Higher values of WQI were observed after rainfall period with median of 75 compared to the dry period with median of 62. The source of the Mourão River is contaminated with fecal coliforms, evidencing the real need to treat sewage in rural areas.

Resumo A Bacia Hidrográfica do Rio Mourão, localizada na porção centro ocidental do Estado do Paraná – Brasil, entre as coordenadas 23º 44’ - 24º 25’ Latitude Sul e 52º 12’ - 52º 30’ Longitude Oeste, entre 270 e 820 m acima do nível mar, possui área de drenagem de 1.648,21 km2. Nesta bacia foi avaliada a qualidade da água por meio de monitoramento de parâmetros físico-químicos e microbiológicos. Foram realizadas coletas mensais durante um ano em 5 pontos selecionados da bacia, sendo realizadas análises de: potencial Hidrogeniônico, temperatura, oxigênio dissolvido, demanda bioquímica de oxigênio, nitrogênio total, nitrogênio amoniacal, nitrito, nitrato, fósforo total, turbidez, sólidos totais, sólidos voláteis e coliformes fecais. Os resultados dos parâmetros avaliados indicaram valores superiores aos limites preconizados na Resolução CONAMA 357 de 2005 para rios de classe 2 em algumas amostras. Com a análise do Índice de Qualidade das Águas (IQA), obteve-se 72% das amostras com qualidade média e 28% com qualidade boa para a bacia do Rio Mourão. Os maiores valores do IQA foram observados na época de chuvas com mediana de 75 comparados ao período de seca com mediana de 62. Constatou-se que a nascente do rio Mourão está contaminada com coliformes fecais, mostrando a real necessidade de se tratar os esgotos sanitários no meio rural.