Genomic and physiological characterization of Kitasatospora sp. nov., an actinobacterium with potential for biotechnological application isolated from Cerrado soil.

An Actinobacteria - Kitasatospora sp. K002 - was isolated from the soil of Cerrado, a savanna-like Brazilian biome. Herein, we conducted a phylogenetic, phenotypic and physiological characterization, revealing its potential for biotechnological applications. Kitasatospora sp. K002 is an aerobic, non-motile, Gram-positive bacteria that forms grayish-white mycelium on solid cultures and submerged spores with vegetative mycelia on liquid cultures. The strain showed antibacterial activity against Bacillus subtilis, Pseudomonas aeruginosa and Escherichia coli. Genomic analysis indicated that Kitasatospora xanthocidica JCM 4862 is the closest strain to K002, with a dDDH of 32.8-37.8% and an ANI of 86.86% and the pangenome investigations identified a high number of rare genes. A total of 60 gene clusters of 22 different types were detected by AntiSMASH, and 22 gene clusters showed low similarity (< 10%) with known compounds, which suggests the potential production of novel bioactive compounds. In addition, phylogenetic analysis and morphophysiological characterization clearly distinguished Kitasatospora sp. K002 from other related species. Therefore, we propose that Kitasatospora sp. K002 should be recognized as a new species of the genus Kitasatospora - Kitasatospora brasiliensis sp. nov. (type strains = K002).

Cytogenetic study in sand spiders (Sicariidae) from the brazilian Caatinga: sex chromosome system diversity in closely related species / Estudio citogenético en arañas de arena (Sicariidae) de la Caatinga brasileña: diversidad del sistema de cromosomas sexuales en especies estrechamente relacionadas

ABSTRACT In this study, we investigated the chromosomes of three species of Sicarius spiders from the Brazilian Caatinga, using classical and molecular cytogenetic techniques. Based on the phylogenetic approach, we also discussed about the variation of diploid number, types of sex chromosome system and changes in the localization of ribosomal genes of Scytodoidea. Sicarius are Synspermiata spiders that together with the genera Loxosceles and Hexophthalma constitute the family Sicariidae. In this group, the available cytogenetic data showed a low diploid number range (2n♂=18 to 2n♂=23) and the presence of only multiple sex chromosome systems (X1X2Y and X1X20). Mitotic metaphase cells exhibited 2n♂=16+X1X2Y for Sicarius cariri and S. ornatus, and 2n♂=18+XY for S. tropicus. In these species, silver impregnation revealed nucleolar organizer region (Ag-NOR) on the terminal region of pair 1. In S. ornatus and S. tropicus, the results obtained with fluorescent in situ hybridization (FISH) using 18S rDNA probe were similar to Ag-NOR, however in S. cariri, the ribosomal sites were localized in the terminal region of the X1 sex chromosome. In this work, we presented the first description of a simple sex chromosome system for Sicariidae, helping to understand how the XY sex chromosome system evolved from the X1X2Y system. Additionally, FISH data incongruous with Ag-NOR indicate that the cytogenetic studies in Sicariidae allow investigating the relation between the karyotype evolution and the distribution and the activity of rDNA genes.

RESUMEN En este estudio, investigamos los cromosomas de tres especies de arañas Sicarius de la Caatinga brasileña, utilizando técnicas de citogenética clásica y molecular. Usando un enfoque filogenético, también discutimos la variación del número diploide, los tipos de sistema cromosómico sexual y los cambios en la localización de los genes ribosómicos en Scytodoidea. Los Sicarius son arañas Synspermiata que, junto con los géneros Loxosceles y Hexophthalma, constituyen a la familia Sicariidae. En este grupo, los datos citogenéticos disponibles mostraron un rango de número diploide bajo (2n♂=18 a 2n♂=23) y únicamente la presencia de sistemas de cromosomas sexuales múltiples (X1X2Y y X1X20). Las células mitóticas en metafase mostraron 2n♂=16+X1X2Y para Sicarius cariri y S. ornatus, y 2n♂=18+XY para S. tropicus. En estas especies, la impregnación de plata reveló la región organizadora nucleolar (Ag-NOR) en la región terminal del par 1. En S. ornatus y S. tropicus, los resultados obtenidos con la hibridación in situ fluorescente (FISH) utilizando la sonda de ADNr 18S fueron similares a los de Ag-NOR, sin embargo, en S. cariri los sitios ribosomales se localizaron en la región terminal del cromosoma sexual X1. En este trabajo, presentamos la primera descripción de un sistema cromosómico sexual simple para Sicariidae, ayudando a entender cómo el sistema cromosómico sexual XY evolucionó a partir del sistema X1X2Y. Además, los datos de FISH incongruentes con Ag-NOR indican que los estudios citogenéticos en Sicariidae permiten investigar la relación entre la evolución del cariotipo y la distribución y la actividad de los genes de ADNr.

Relevance of AND-ASPEN criteria of malnutrition to predict hospital mortality in critically ill patients: A prospective study.

PURPOSE: Malnutrition is prevalent in the intensive care units (ICU), yet, there is a paucity of validated assessment tools. Subsequently, this study evaluates the validity of the malnutrition AND-ASPEN tool as an ICU mortality predictor. METHODS: Patients admitted to a large mixed ICU (72 beds) from 2014 to 2016, were followed during stay and had electronic health records on sex, age, Apache II and baseline nutrition assessment collected at admission. Patients with shortstay (<48h) and missing data were excluded. The main hypothesis, hospital mortality prediction, was assessed with a logistic regression model. RESULTS: Patients eligible were 375 where 13% were excluded by the adopted criteria. In the eligible group, 94.2% had AND-ASPEN assessment in their files, showing a malnutrition prevalence rate of 29.7%. Logistic regression (n=327, p=0.0001, r2=0.304, Roc (AUC)=0.80) suggested that mortality risk was 2.5× higher (95%CI, 1.38-4.46, p=0.001) in malnourished patients vs non-malnourished (controlled by sex, Apache II, hospital stay and clinical admission), malnutrition crude OR was 3.04 (95% CI, 1.86-4.97). For every 1-point increase in Apache II, mortality risk rises 14% (95%CI 1.10-1.18, p=0.001). CONCLUSION: This study showed the applicability of the AND-ASPEN tool in the ICU setting as a predictor of mortality.

Hematimetria manual e automática em jaguatiricas (Leopardus pardalis - Linnaeus, 1758) / Manual and automated blood cells count in ocelots (Leopardus pardalis - Linnaeus, 1758)

Aparelhos de automação, cujo uso está estabelecido na hematologia dos animais domésticos, ainda não são empregados rotineiramente em felídeos selvagens. Esse estudo objetivou avaliar a técnica automatizada na contagem de células sanguíneas de jaguatiricas (Leopardus pardalis), comparando-a com a técnica manual. Foram coletadas amostras sanguíneas de oito jaguatiricas, que se submeteram à contagem de eritrócitos, leucócitos, plaquetas e à determinação do volume globular pelo método manual padrão e por meio do aparelho BC - 2800 VET® com a configuração para gatos domésticos. Os resultados foram avaliados por meio do teste t para dados pareados, e as técnicas submetidas à análise de correlação de Pearson. A técnica automatizada demonstrou resultados estatisticamente semelhantes para eritrócitos e leucócitos. Apenas para volume globular houve diferença significativa entre as técnicas manual e automatizada, mas a correlação foi alta. Apesar de não haver diferença significativa entre as técnicas para plaquetas, a correlação foi baixa. Conclui-se que o aparelho BC - 2800 VET® com a configuração para gatos domésticos é uma técnica confiável na realização do eritrograma e do leucograma para jaguatiricas. Para a determinação do parâmetro volume globular, o aparelho pode ser utilizado, desde que se faça a correção. Para a contagem de plaquetas, a técnica manual é recomendada.(AU)

Automated equipment, whose use is established in hematology of domestic animals, is not yet routinely used in blood cells count of wildlife, due to lack of studies that validate its use. The purpose of this study was to evaluate the automated technique for blood cells count of ocelots (Leopardus pardalis), comparing it with the manual technique. Blood samples were collected from eight ocelots, which were submitted to counting of erythrocytes, leukocytes, platelets and packed cell volume by the standard manual method and by the device VET ® 2800 BC with the configuration for domestic cats. The results were evaluated using the t test for paired data and the techniques submitted to Pearson correlation. The automated technique showed statistically similar results to erythrocytes and leukocytes. Significant difference was found only for packed cell volume between the manual and automated techniques, but the correlation was high. Although there was no significant difference between the techniques for platelets, the correlation was low. We concluded that the 2800 BC VET ® device with the configuration for domestic cats is a reliable technique in performing the erythrocyte and leukocyte counts for ocelots. The device may be used to determine packed cell volume, provided the correction is made. The manual technique is recommended for the platelet count.(AU)

Chromosomal characteristics of a Brazilian whip spider (Amblypygi) and evolutionary relationships with other arachnid orders.

We analyzed mitotic and meiotic cells of a Brazilian amblypygid, Heterophrynus longicornis, using conventional and molecular cytogenetic techniques (Giemsa staining, C-banding, Ag-NOR, and FISH with rDNA probe). This is the first study that focuses solely on amblypygid chromosomes; it was undertaken to add data on cytogenetic knowledge of this group and contribute to the understanding of chromosome evolution in the Arachnida. We found 2n = 66 for male and female individuals, monocentric chromosomes, and absence of morphologically differentiated sex chromosomes. C-banding showed heterochromatin in the pericentromeric region of most chromosomes. Mitotic and meiotic nuclei submitted to silver impregnation and FISH revealed, respectively, Ag-NORs and ribosomal genes in the terminal region of two chromosome pairs. Most chromosome features that we observed in H. longicornis are shared with species of other arachnid orders; however, the absence of morphologically differentiated sex chromosomes in amblypygid contrasts with the remarkable variety of sex chromosome systems recorded for the Araneae. Consequently, we conclude that analysis of species of the Tetrapulmonata clade is useful for understanding the trends of sex chromosome evolution in this arachnid group.

May genetic factors in fibromyalgia help to identify patients with differentially altered frequencies of immune cells?

There is common agreement that fibromyalgia (FM) is an extremely heterogeneous entity. Patients differ in their clinical symptoms, endocrine and immune parameters. In this study we evaluated endocrine and immunological features of distinct subsets of FM patients. In contrast to previous attempts to identify subsets of FM patients, based solely on their psychological and cognitive features, herein we propose to separate FM patients by genetic features. Allelic expression of the polymorphic promoter region of the serotonin transporter (5-HTTLPR) was analysed as a relevant genetic factor for FM. Seventy-five patients meeting the American College of Rheumatology criteria and 27 healthy age-matched controls participated in this study. All controls and FM patients were submitted to genotyping of 5-HTTLPR. Twenty-seven FM patients, who were able to discontinue hypnotic, sedative or psychotropic prescription medications for at least 2 weeks, were then subdivided into L (homozygote LL) or S groups (genotypes LS and SS). They were evaluated for salivary cortisol levels, absolute number of leucocyte subpopulations, including natural killer (NK) cells and activated T and B lymphocytes. Both groups presented decreased cortisol levels, more intense in the L group, increased all B lymphocytes subsets and reduced CD4+CD25high T lymphocytes. The L group had increased CD4+CD25low activated T lymphocytes, while the S group displayed elevated CD4+ human leucocyte antigen D-related (HLA-DR)+ activated T lymphocytes and decreased NK cells. We demonstrate that genetic factors may help to identify FM individuals with differentially altered frequencies of immune cells.

Identification and quantitation of carotenoids and their metabolites in the tissues of the human eye.

There is increasing evidence that the macular pigment carotenoids, lutein and zeaxanthin, may play an important role in the prevention of age-related macular degeneration, cataract, and other blinding disorders. Although it is well known that the retina and lens are enriched in these carotenoids, relatively little is known about carotenoid levels in the uveal tract and in other ocular tissues. Also, the oxidative metabolism and physiological functions of the ocular carotenoids are not fully understood. Thus, we have set out to identify and quantify the complete spectrum of dietary carotenoids and their oxidative metabolites in a systematic manner in all tissues of the human eye in order to gain better insight into their ocular physiology. Human donor eyes were dissected, and carotenoid extracts from ocular tissues [retinal pigment epithelium/choroid (RPE/choroid), macula, peripheral retina, ciliary body, iris, lens, vitreous, cornea, and sclera] were analysed by high-performance liquid chromatography (HPLC). Carotenoids were identified and quantified by comparing their chromatographic and spectral profiles with those of authentic standards. Nearly all ocular structures examined with the exception of vitreous, cornea, and sclera had quantifiable levels of dietary (3R,3'R,6'R)-lutein, zeaxanthin, their geometrical (E / Z) isomers, as well as their metabolites, (3R,3'S,6'R)-lutein (3'-epilutein) and 3-hydroxy-beta,epsilon-caroten-3'-one. In addition, human ciliary body revealed the presence of monohydroxycarotenoids and hydrocarbon carotenoids, while only the latter group was detected in human RPE/choroid. Uveal structures (iris, ciliary body, and RPE/choroid) account for approximately 50% of the eye's total carotenoids and approximately 30% of the lutein and zeaxanthin. In the iris, these pigments are likely to play a role in filtering out phototoxic short-wavelength visible light, while they are more likely to act as antioxidants in the ciliary body. Both mechanisms, light screening and antioxidant, may be operative in the RPE/choroid in addition to a possible function of this tissue in the transport of dihydroxycarotenoids from the circulating blood to the retina. This report lends further support for the critical role of lutein, zeaxanthin, and other ocular carotenoids in protecting the eye from light-induced oxidative damage and aging.

Interaction of the Bezold-Jarisch reflex with the urinary bladder function.

OBJECTIVES: The Bezold-Jarisch reflex involves a marked increase in vagal (parasympathetic) efferent discharge to the heart. During micturition, there is a marked increase in parasympathetic activity to the bladder (pelvic nerve activity) and this coincides with periods of phasic bladder contractions. In recent studies, we have shown that regions of the CNS that modulate pelvic nerve activity also affect cardiovascular variables. Recent anatomical studies have shown neuronal connections between the areas of the CNS that modulate both reflexes. Accordingly, in the present study we investigate if during the Bezold-Jarisch reflex there are simultaneous changes in pelvic nerve activity. METHODS: Experiments were performed in Wistar female rats, anaesthetised with alpha-chloradose i.p. (80 micrograms/Kg) supplemented i.v. as required. Arterial blood pressure, ECG heart rate, urinary bladder pressure and pelvic nerve activity were recorded. RESULTS: Stimulation of cardiac chemosensitive nerve endings with veratridine evoked a decrease in arterial blood pressure from 88.6 +/- 1.72 to 66.1 +/- 2.50 mmHg (n = 30; p < 0.0001) and of heart rate from 420.3 +/- 5.56 to 248.8 +/- 17.58 bpm (n = 13; p < 0.0001) There were simultaneous changes of pelvic nerve activity that preceded an increase in urinary bladder pressure from 10.2 +/- 0.28 to 26.3 +/- 1.52 cmH2O (n = 30; p < 0.0001) and bladder contractions. We believe that the reflex changes resulted from activation of cardiac chemosensory nerve endings, as equivalent injections of warm saline into the left ventricle produced no changes in either cardiovascular variables or pelvic nerve activity. CONCLUSIONS: These observations suggest a significant role for cardiac afferents in modulating bladder contractions. The integration and functional significance of these inputs remain to be elucidated. The clinical importance of this physiological result may contribute to a better understanding of the mechanism of some semiologic findings, notably the association of urgency to micturate with myocardial ischemia and myocardial infarction.

Normal relationships between cardiovascular variables in active orthostatism and clinostatism in postural change.

OBJECTIVE: Autonomic modulation of hemodynamics, essential for the preservation of homeostasis, is well tested by the abrupt postural change from clinostatism to active orthostatism. The aim of this work was to study normal relationships between the cardiovascular variables in active orthostatism and those in clinostatism. METHODS: Hemodynamic parameters in clinostatism and orthostatism were easily measured in 20 healthy subjects of both sexes, aged between 33 and 78 years, without treatment, using the non-invasive thoracic electric bioimpedance method. RESULTS: Cardiovascular variables values in orthostatism are linearly related with their values in clinostatism. CONCLUSIONS: Results show that cardiovascular variables in active orthostatism are linearly related with their values in clinostatism, each variable being specially regulated. A clinostatism and orthostatism intraindividual correlation was obtained, which provides an easily accessible method of detection and interpretation of autonomic dysfunctions, without deleterious consequences for the subjects, which can be very useful for research on physiopathologic mechanisms.

[Heart insufficiency. Physiology and physiopathology of muscle contraction of the heart]. / Insuficiência Cardíaca. Fisiologia e fisiopatologia da contracção do músculo cardíaco.

To understand physiopathology of contractibility in heart failure is needed the previous knowledge of normal muscle contractibility, calcium regulation, methods of assessment of isolated cardiac muscle and in situ heart contraction. Models of calcium movements during rest and contraction are presented and concepts of preload, afterload, Vmax and Po introduced. The importance of assessment of cardiac diastolic function is referred. Biochemical alterations in the failing myocardium are presented.