Predicting maximal lactate steady state from lactate thresholds determined using methods based on an incremental exercise test in beagle dogs: A study using univariate and multivariate approaches.

The reliability of four lactate threshold (LT) methods to estimate the maximal lactate steady state (MLSS), defined as the highest intensity that can be maintained without plasma lactate ([La-]) accumulation over time, was determined in Beagle dogs. Six male Beagle dogs performed a standardized incremental exercise test on a treadmill when plasma lactate ([La-]) measurements were performed. The LTs for predicting MLSS, were determined by visual inspection (LTV), using a bi-segmented linear regression model (LTBI), or using a polynomial function on the [La-]/velocity ratio (LTP) by considering the vertices of the curve and calculating the point that yields the maximal distance from a curve representing [La-] as a function of velocity to the line formed by the two endpoints of the curve (LTDMAX method). The agreement was assessed using Bland-Altman plots and ordinary least products (OLP) regression among the velocities corresponding to the LTs identified using different methods (VLTv, VLTBI, VLTP, and VLTDMAX) and the velocity corresponding to the MLSS (VMLSS). A principal component (PC) analysis approach was performed to detect the degree of co-relatedness among the variables. The mean ± SD [La-] at MLSS was 1.03 ± 0.24 mM. VMLSS had a lower mean bias with VLTv, followed by VLTBI. The VLTDMAX underestimated MLSS. VLTv and VLTBI had the lowest limits of agreement with the VMLSS. The VLTP and VLTDMAX showed relatively high limits of agreement with MLSS. VLTv, VLTBI, and VMLSS had more collinearity and were dominantly aligned with the second component (PC2). VLTv and VLTBI can be used as simple methods to objectively determine aerobic fitness in Beagle dogs.

Effect of sublethal concentrations of the antiparasitic ivermectin on the polychaeta species Hediste diversicolor: biochemical and behavioral responses.

Pharmaceutical drugs have emerged as major micropollutants in aquatic ecosystems. Their presence has been systematically reported in monitoring surveys, and their wide distribution and constant presence in the wild is a direct consequence of their massive use, in both human and veterinary therapeutics. Drugs used to treat parasitic infections in livestock are major contaminants, given the amounts in which they are administered, and reach the aquatic compartment in high amounts, where they may affect non target species. Some of these drugs are prone to find their final deposit in sediments of estuarine areas, exerting their toxic effects preferentially at these locations. Sediment dwelling organisms of coastal areas, such as polychaetas, are especially prone to have their major physiological functions compromised after being exposed to pharmaceutical drugs. Ivermectin is one of the most used antiparasitic drugs, and its effects are not limited to biochemical traits, but also behavioral features may be compromised considering their neurotoxic actions. Despite these putative effects, little is known about their toxicity on polychaetas. The present study aimed to characterize the toxicity of realistic levels of ivermectin on the polychaeta Hediste diversicolor, in biochemical and behavioral terms. The obtained results showed that low levels of ivermectin are capable of causing significant disturbances in mobility and burrowing activity of exposed worms, as well as alterations of metabolic and anti-oxidant defense efficacy of exposed animals, suggesting that its environmental presence may mean a major environmental concern.

Systemic inflammatory biomarkers and somatic cell count in dairy cows with subclinical mastitis.

Systemic parameters and its relationship with Somatic Cell Count (SCC) in dairy cows were evaluated. Cows that presented subclinical mastitis (SMC; n = 16) and healthy cows (HC; n = 6) were selected and identified by the California Mastitis Test and SCC in milk. SCC results were logarithmically transformed into somatic cell score (SCS). HC presented SCS 〈 4.32 while SMC presented SCS 〉 4.32. Milk and blood samples were collected in three days: D1 (first day of sampling), D2 (48 h after D1) and D3 (7 days after D1), to determine White Blood Cells (WBC), albumin, total protein, total bilirubin and malondialdehyde (MDA). Results were expressed as mean ± standard deviation. It was considered significant at P < 0.05. The data of SCS on D1, D2 and D3 in SMC were: 6.8 ± 1.7, 6.4 ± 1.8, and 6.3 ± 2.0, respectively. In SMC the MDA (nmolL-1) were: D1 - 9.3 ± 2.6, D2 - 8.6 ± 2.4, and D3 - 11.5 ± 3.5. The MDA on D3 in SMC (11.5 ± 3.5) were increase when compared to HC (6.0 ± 1.3) (P < 0.001). No significant difference was found in WBC, TP, ALB, and TB between groups. It was observed a positive correlation between MDA-SCS (ρ = 0.4) and between WBC-SCS (ρ = 0.3) in the SMC group. It was concluded that the systemic repercussion damage in the mammary gland promoted by subclinical mastitis in dairy cows can be assessed using the MDA and WBC biomarkers.

Caste-specific gene expression underlying the differential adult brain development in the honeybee Apis mellifera.

Apis mellifera adult workers feature more developed key brain regions than queens, which allows them to cope with the broad range of duties they need to perform in a colony. However, at the end of larval development, the brain of queens is largely more developed than that of workers. Major morphogenetic changes take place after metamorphosis that shift caste-specific brain development. Here, we tested the hypothesis that this phenomenon is hormonally governed and involves differential gene expression. Our molecular screening approach revealed a set of differentially expressed genes in Pp (first pharate-adult phase) brains between castes mainly coding for tissue remodelling and energy-converting proteins (e.g. hex 70a and ATPsynß). An in-depth qPCR analysis of the transcriptional behaviour during pupal and pharate-adult developmental stage in both castes and in response to artificially augmented hormone titres of 18 genes/variants revealed that: i. subtle differences in hormone titres between castes might be responsible for the differential expression of the EcR and insulin/insulin-like signalling (IIS) pathway genes; ii. the morphogenetic activity of the IIS in brain development must be mediated by ILP-2, iii. which together with the tum, mnb and caspase system, can constitute the molecular effectors of the caste-specific opposing brain developmental trajectories.

Suplementação com óleos ricos em ácidos graxos poli-insaturados na dieta de leitões na fase de creche: efeitos no desempenho, na resposta inflamatória, no perfil lipídico e no "status" oxidativo / Supplementation with oils rich in polyunsaturated fatty acids in the diet of pigs in the nursery phase: effects on performance, inflammatory response, lipid profile and oxidative "status"

Nos sistemas de produção, os produtos naturais vêm se destacando na substituição a produtos sintéticos, dentre eles podemos ressaltar os óleos vegetais ricos em ácido graxos poli-insaturados (PUFA), que são conhecidos, popularmente, por seus efeitos benéficos. Dessa forma, o objetivo com este trabalho foi avaliar os efeitos da suplementação dos óleos de pequi (Caryocar brasiliense) e girassol (Helianthus annus) sobre parâmetros fisiológicos em leitões na fase de creche. Foram utilizados 180 leitões alojados em granja comercial, distribuídos em três grupos (n=60) conforme suplementação alimentar: óleo de pequi, óleo de girassol e controle negativo. Amostras de sangue e o peso dos animais foram obtidos nos intervalos de quatro trocas de rações da fase de creche para avaliação do desempenho, da resposta inflamatória, do perfil lipídico e do "status" oxidativo. Os parâmetros fisiológicos mensurados demonstraram que os óleos interferiram positivamente na resposta inflamatória sistêmica, por meio dos leucócitos totais e da relação neutrófilo/linfócito (P<0,05); no equilíbrio oxidante-antioxidante, por mensuração de óxido nítrico e do malondialdeído (P<0,05); e no metabolismo lipídico, com a avaliação de colesterol total e triglicérides (P<0,05). Por outro lado, esses óleos vegetais interferiram no ganho de peso e no consumo de ração (P<0,05). Assim, concluí-se que a suplementação com óleos de pequi e girassol melhora a saúde dos animais, mas tem impacto negativo no desempenho zootécnico de leitões na fase de creche.(AU)

In production systems, natural products have been outstanding in replacing synthetic products, among them, we can highlight the vegetable oils rich in polyunsaturated fatty acids (PUFA) that are popularly known for their beneficial effects. Thus, the objective of this study was to evaluate the effects of pequi (Caryocar brasiliense) and sunflower (Helianthus annus) oil supplementation on nursery physiological parameters. We used 180 piglets housed in a commercial farm, distributed in 3 groups (n=60) according to food supplementation: pequi oil, sunflower oil and negative control. Blood samples and animal weight were obtained at intervals of four nursery phase rations to evaluate performance, inflammatory response, lipid profile and oxidative status. The physiological parameters measured showed that the oils positively interfered in the systemic inflammatory response through total leukocytes and neutrophil / lymphocyte ratio (P<0.05), in the oxidant-antioxidant balance by measuring nitric oxide and malondialdehyde (P<0.05). and lipid metabolism with the assessment of total cholesterol and triglycerides (P<0.05). On the other hand, these vegetable oils interfered with weight gain and feed intake (P<0.05). Thus, it can be concluded that supplementation with pequi and sunflower oils improves animal health, but has a negative impact on the piglet's performance in the nursery phase.(AU)

Facial Squamous Cell Carcinoma and Abdominal Peripheral Nerve Sheath Tumour with Rhabdomyoblastic Differentiation in a Rough-toothed Dolphin (Steno bredanensis).

We report the pathological features of a facial squamous cell carcinoma (SCC) and an abdominal peripheral nerve sheath tumour (PNST) with rhabdomyoblastic differentiation in an aged free-ranging rough-toothed dolphin (Steno bredanensis). The animal was found stranded dead in poor body condition. On external examination, there was a 25 × 7 × 3 cm extensively ulcerated area on the right maxillary region of the rostrum, involving the oral mucocutaneous junction with prominent nodular edges, severe soft tissue loss and extensive maxillary and premaxillary bone lysis. On abdominal dissection, a 5 × 4 × 3.5 cm pale tan to red, raised mass expanded the inner aspect of the right transverse abdominis muscle. Microscopically, the aggressive facial lesion was an acantholytic SCC with extensive osteolysis; there was no evidence of metastasis in the tissues examined. The abdominal mass had cytohistomorphological features compatible with a localized PNST, including whorling, Antoni A and Antoni B areas and Verocay bodies intermixed with rhabdomyoblastic components, as suggested by phosphotungstic acid haematoxylin stain. This neoplasm was locally infiltrative, yet no metastases were observed in the tissues examined. No immunohistochemical investigations could be performed due to lack of tissue availability. Total DNA from the formalin-fixed and paraffin wax-embedded SCC was extracted and tested by polymerase chain reaction for herpesvirus and papillomavirus genetic material. There was no amplification for either of these genera. Other pathological findings observed in this animal were related to the 'live-stranding stress response'. The severity and extent of the facial SCC likely related to anorexia and poor body condition and might have played a role in the stranding and death of this dolphin. These two tumour subtypes add to the relatively uncommon reports of neoplasia in cetaceans. Specifically, these appear to be the first neoplasia records for rough-toothed dolphins, including the first documentation of a PNST with features compatible with rhabdomyoblastic differentiation in a marine mammal.

Candidemia in a Portuguese tertiary care hospital: Analysis of a 2-year period.

BACKGROUND: Candidemia is a nosocomial infection of increasing importance, associated with high morbidity and mortality. The aim of this study is to describe the species distribution, risk factors, management and outcomes of patients with candidemia. METHODS: We conducted a retrospective study at Centro Hospitalar Universitário de São João, Portugal, between January 2016 and December 2017. RESULTS: A total of 117 candidemia episodes (n=114 patients) were included. Median age was 65 years, with an increased prevalence of older ages. Candida albicans (51.3%) was the most prevalent species, followed by C. glabrata (22.2%), C. parapsilosis (15.4%), C. tropicalis (4.3%) and C. lusitaniae (2.6%). Forty-two patients (35.9%) did not receive antifungal drugs after diagnosis of candidemia. Echinocandins were used as first-line drug therapy in half of the treated patients (50.7%). The median EQUAL Candida Score was 6/17 (IQR 6-9) for patients without central venous catheter (CVC) and 11/20 (IQR 6-14) for patients with CVC. The 30 days-mortality was 31,6% and was not significantly associated with the timing of antifungal therapy and the EQUAL Candida Score. CONCLUSION: The distribution of Candida species has changed in recent years, with an increase in the proportion of C. albicans and C. glabrata. Rapid diagnostic tests, empiric antifungal therapy and source control are essential to improve the prognosis of patients with candidemia. More multicentric prospective studies are needed to evaluate the association of mortality with the timing of antifungal therapy or the EQUAL Candida Score.

Aspergillus species collected from environmental air samples in Portugal-molecular identification, antifungal susceptibility and sequencing of cyp51A gene on A. fumigatus sensu stricto itraconazole resistant.

AIMS: Aspergillus sp. are ubiquitous saprophytic fungi and their conidia easily inhaled. This is particularly important in immunocompromised patients, more susceptible to developing invasive aspergillosis. In addition to A. fumigatus sensu stricto, cryptic species, many resistant to antifungal drugs, have been associated with invasive infections, making it important to assess their presence and diversity in different environments. Therefore, the aim of this study was to assess the presence, diversity and susceptibility to antifungal drugs of airborne fungi. Moreover, in azole-resistant A. fumigatus sensu stricto isolates the presence of underlying molecular mechanisms of resistance was investigated. METHODS AND RESULTS: Eighty-four Aspergillus isolates were collected from the environment air in hospitals and the Water Treatment Plant. The use of molecular tools allowed to detect 12 different cryptic species, showing a prevalence of 21·4%. The majority of isolates (69·0%) belonged to A. fumigatus complex and 65·4% were A. fumigatus sensu stricto. Among these, 21·8% were resistant to itraconazole (ITZ), 38·2% to posaconazole and 87·3% to isavuconazole; none of them were resistant to voriconazole or amphotericin B. Sequencing of the cyp51A gene on the 12 A. fumigatus sensu stricto ITZ-resistant isolates revealed the presence of mutations. CONCLUSION: Our study reports a large number of environmental-resistant Aspergillus species, including A. fumigatus sensu stricto that display an important role in invasive fungal infections. None of the environmental isolates showed mutations on cyp51A gene related to azole resistance. SIGNIFICANT AND IMPACT OF THE STUDY: This study is the first assessment of molecular resistance mechanisms in A. fumigatus sensu stricto environmental isolates, in Portugal. Since TR34/L98H and TR46/Y121F/T289A cyp51A mutations were already reported in the clinical setting in Portugal (Monteiro et al. J Glob Antimicron Resist 13: 190-191, 2018; Pinto et al. Front Microbiol 9: 1656, 2018), and have been linked to environmental route, it is utmost importance to perform surveillance network for azole-resistant A. fumigatus.

Avaliação do estresse e do desempenho de suínos na fase de creche, empregando-se técnicas de enriquecimento ambiental / Evaluation of stress and performance of pigs during the nursery phase, using environmental enrichment techniques

O enriquecimento ambiental é uma ferramenta importante dentro dos sistemas de produção, a fim de promover o bem-estar e favorecer a saúde dos animais. Portanto, o objetivo deste trabalho foi avaliar o efeito do enriquecimento ambiental sobre o estresse de suínos na fase de creche. Foram utilizados 32 leitões, alojados em granja experimental, distribuídos em quatro grupos (n= 8): corda, corrente, garrafa PET e controle negativo. Amostras de sangue foram coletadas no início e no final do experimento para contagem de leucócitos circulantes e determinação de antioxidantes não enzimáticos, óxido nítrico, malondialdeído, e de saliva para avaliação do cortisol. Foi aplicado etograma e fez-a ganho médio de peso diário e a conversão alimentar. Os parâmetros avaliados no primeiro dia do experimento não variaram entre os grupos (P>0,05). No último dia do experimento, os valores de neutrófilos e da relação neutrófilo/linfócito foram mais elevados nos leitões do grupo corrente, assim como os valores de cortisol salivar (P<0,05). O ácido úrico apresentou-se mais elevado nos leitões do grupo corrente e o malondialdeído (MDA) nos do grupo garrafa (P< 0,05). Os enriquecimentos ambientais estimularam comportamentos positivos nos leitões, tendo a corda se destacado como o mais atrativo. Por outro lado, a corrente apresentou efeito negativo sobre a fisiologia dos animais, gerando estresse, assim como a garrafa, que induziu a peroxidação lipídica e um menor ganho de peso nos leitões.(AU)

Environmental enrichment is an important tool within production systems to promote welfare and animal health. The aim of this study was to evaluate the effect of enrichment objects on stress of piglets in nursery phase. 32 piglets housed in experimental farm were distributed in 4 groups (n= 8): rope, chain, pet bottle and negative control. Blood samples were collected at the beginning and end of the experiment to count circulating leukocytes and determine non-enzymatic antioxidants, nitric oxide, malondialdehyde, and saliva to evaluate cortisol. At the same time, an etogram and evaluation of mean daily gain and feed conversion were applied. The evaluated parameters on first day of the experiment did not vary between groups (P> 0.05). On the last day, neutrophil and neutrophil /lymphocyte ratios were higher in chain group piglets, as were salivary cortisol values (P< 0.05). Uric acid was higher in chain group and MDA in bottle group (P< 0.05). Environmental enrichment stimulated positive behaviors in piglets, where the rope stood out as the most attractive. On the other hand, chain had a negative effect on animal physiology, generating stress, as well as the bottle that induced lipid peroxidation and a lower weight gain in piglets.(AU)

Hormonal control and target genes of ftz-f1 expression in the honeybee Apis mellifera: a positive loop linking juvenile hormone, ftz-f1, and vitellogenin.

Ftz-f1 is an orphan member of the nuclear hormone receptor superfamily. A 20-hydroxyecdysone pulse allows ftz-f1 gene expression, which then regulates the activity of downstream genes involved in major developmental progression events. In honeybees, the expression of genes like vitellogenin (vg), prophenoloxidase and juvenile hormone-esterase during late pharate-adult development is known to be hormonally controlled in both queens and workers by increasing juvenile hormone (JH) titres in the presence of declining levels of ecdysteroids. Since Ftz-f1 is known for mediating intracellular JH signalling, we hypothesized that ftz-f1 could mediate JH action during the pharate-adult development of honeybees, thus controlling the expression of these genes. Here, we show that ftz-f1 has caste-specific transcription profiles during this developmental period, with a peak coinciding with the increase in JH titre, and that its expression is upregulated by JH and downregulated by ecdysteroids. RNAi-mediated knock down of ftz-f1 showed that the expression of genes essential for adult development (e.g. vg and cuticular genes) depends on ftz-f1 expression. Finally, a double-repressor hypothesis-inspired vg gene knock-down experiment suggests the existence of a positive molecular loop between JH, ftz-f1 and vg.

The venous patterns of the cubital fossa in subjects from Brazil.

Venous punctures are among the most common procedures performed by healthcare professionals. In particular, the cubital fossa is the site where the venous accesses are frequently made due to the number of superficial veins and the numerous anastomoses in this region. The arrangement of these venous connections is of particular interest for clinical application in several areas, thus, the healthcare professional must possess knowledge about these vessels and their anatomical relationships. The present study aims to analyze the venous pattern of the cubital fossa among individuals from Brazil. This study was approved by a Research Ethics Committee. The sample had 100 healthy individuals (50 men and 50 women). The superficial veins of the cubital fossa were analyzed with the aid of a sphygmomanometer. When inflated, the pressure in the forearm increased and the veins became prominent. It was observed that in the selected sample the types with the highest prevalence were the Type I and Type VII, both with 22% in 200 limbs studied. The chi2 test showed a significant statistical difference between the anastomosis pattern and the sex of the studied sample. The anastomotic pattern of the superficial veins of the studies sample is similar to African, European and Asian populations. The study of these variations is necessary to provide scientific basis for the healthcare professional during a venipuncture in order to avoid iatrogenic errors and damages in cutaneous nerves or neighboring arteries.

Biophysical and biological properties of small linear peptides derived from crotamine, a cationic antimicrobial/antitumoral toxin with cell penetrating and cargo delivery abilities.

Crotamine is a natural polypeptide from snake venom which delivers nucleic acid molecules into cells, besides having pronounced affinity for negatively charged membranes and antifungal activity. We previously demonstrated that crotamine derived short linear peptides were not very effective as antifungal, although the non-structured recombinant crotamine was overridingly more potent compared to the native structured crotamine. Aiming to identify the features necessary for the antifungal activity of crotamine, two linear short peptides, each comprising half of the total positively charged amino acid residues of the full-length crotamine were evaluated here to show that these linear peptides keep the ability to interact with lipid membrane model systems with different phospholipid compositions, even after forming complexes with DNA. Interestingly, the presence of cysteine residues in the structure of these linear peptides highly influenced the antifungal activity, which was not associated to the lipid membrane lytic activity. In addition to the importance of the positive charges, the crucial role of cysteine residues was noticed for these linear analogs of crotamine, although the tridimensional structure and lipid membrane lytic activity observed only for native crotamine was not essential for the antifungal activity. As these peptides still keep the ability to form complexes with DNA molecules with no prejudice to their ability to bind to lipid membranes, they may be potentially advantageous as membrane translocation vector, as they do not show lipid membrane lytic activity and may harbor or not antifungal activity, by keeping or not the semi-essential amino acid cysteine in their sequence.

Involvement of mast cells, CD68+ and VEGF+ expressions in response to Himatanthus drasticus commercial latex in mice wound healing model / Envolvimento de mastócitos, expressão de CD68+ e VEGF+ em resposta ao látex comercial de Himatanthus drasticus em modelo de cicatrização em camundongos

This study aimed to evaluate Himatanthus drasticus latex in a mice wound healing experimental model. Animals were divided into four groups (n=7) according to the treatments: GI - saline 0.9% (control), GII - mineral oil (vehicle), GIII - H. drasticus commercial latex (HdCL) and GIV - H. drasticus mixed isolated fraction (MIF, 1 mg/mL). The treatments were applied topically once daily, 50 µL for 14 consecutive days. Macroscopic lesions were evaluated, considering parameters such as swelling, redness, granulation tissue and reepithelialization. VEGF+, CD68+ expressions and mast cells (Toluidin blue stain) were evaluated. HdCL induced higher contraction and exuberant granulation tissue (P > 0.05). HdCL showed a mild inflammatory process while MIF induced intense infiltrate inflammatory predominantly by lymphocytes, vascular congestion, bleeding and did not presented full reepithelialization. Reorganization of collagen fibers (red picrosirius stain) was observed. CD68+ expression and mast cells were presented as moderate, intense and mild in GI, GIII and GIV, respectively. Neovascularization occurred in all groups, while VEGF+ expression was intense in MIF in relation to HdCL. We concluded that HdCL presents wound healing potential, through modulation of mast cells, CD68+ and VEGF+ expressions that can be associated to triterpenes presence according MIF isolated from HdCL.(AU)

Objetivou-se avaliar o látex de Himatanthus drasticus em feridas induzidas experimentalmente em camundongos. Os animais foram divididos em quatro grupos (n=7): GI - salina 0,9% (controle), GII - óleo mineral (veículo), GIII - látex comercial de H. drasticus (HdCL) e GIV - fração isolada mista de H. drasticus (MIF, 1mg/mL). Os tratamentos foram aplicados topicamente uma vez ao dia (50µL), durante 14 dias consecutivos. Lesões macroscópicas, as expressões de VEGF+, CD68+ e a participação dos mastócitos (coloração azul de toluidina) foram avaliadas. HdCL induziu maior contração e tecido de granulação exuberante (P >0,05). HdCL induziu leve processo inflamatório enquanto MIF promoveu intenso infiltrado inflamatório predominantemente linfocítico, congestão vascular, hemorragia e reepitelização parcial. Observou-se reorganização das fibras colágenas (coloração picrosírius). A expressão de CD68+ e os mastócitos apresentaram-se moderados, intensos e leves em GI, GIII e GIV, respectivamente. A neovascularização foi observada em todos os grupos, enquanto a expressão de VEGF+ foi mais intensa em MIF em relação a HdCL. Conclui-se que HdCL apresenta potencial de cicatrização por meio da modulação dos mastócitos e das expressões de CD68+ e VEGF+, o que pode estar associado à presença de triterpenos de acordo com MIF isolada de HdCL.(AU)

De novo transcriptome assembly of sugarcane leaves submitted to prolonged water-deficit stress.

Sugarcane production is strongly influenced by drought, which is a limiting factor for agricultural productivity in the world. In this study, the gene expression profiles obtained by de novo assembly of the leaf transcriptome of two sugarcane cultivars that differ in their physiological response to water deficit were evaluated by the RNA-Seq method: drought-tolerant cultivar (SP81-3250) and drought-sensitive cultivar (RB855453). For this purpose, plants were grown in a greenhouse for 60 days and were then submitted to three treatments: control (-0.01 to -0.015 MPa), moderate water deficit (-0.05 to -0.055 MPa), and severe water deficit (-0.075 to -0.08 MPa). The plants were evaluated 30, 60, and 90 days after the beginning of treatment. Sequencing on an Illumina platform (RNA-Seq) generated more than one billion sequences, resulting in 177,509 and 185,153 transcripts for the tolerant and sensitive cultivar, respectively. These transcripts were aligned with sequences from Saccharum spp, Sorghum bicolor, Miscanthus giganteus, and Arabidopsis thaliana available in public databases. The differentially expressed genes detected during the prolonged period of water deficit permit to increase our understanding of the molecular patterns involved in the physiological response of the two cultivars. The tolerant cultivar differentially expressed a larger number of genes at 90 days, while in the sensitive cultivar the number of differentially expressed genes was higher in 30 days. Both cultivars perceived the lack of water, but the tolerant cultivar responded more slowly than the sensitive cultivar. The latter requires rapid activation of different water-deficit stress response mechanisms for its survival. This rapid activation of metabolic pathways in response to water stress does not appear to be the key mechanism of drought tolerance in sugarcane. There is still much to clarify on the molecular and physiological pattern of plants in response to drought.

Studying cytokinesis in Drosophila epithelial tissues.

Epithelial tissue cohesiveness is ensured through cell-cell junctions that maintain both adhesion and mechanical coupling between neighboring cells. During development, epithelial tissues undergo intensive cell proliferation. Cell division, and particularly cytokinesis, is coupled to the formation of new adhesive contacts, thereby preserving tissue integrity and propagating cell polarity. Remarkably, the geometry of the new interfaces is determined by the combined action of the dividing cell and its neighbors. To further understand the interplay between the dividing cell and its neighbors, as well as the role of cell division for tissue morphogenesis, it is important to analyze cytokinesis in vivo. Here we present methods to perform live imaging of cell division in Drosophila epithelial tissues and discuss some aspects of image processing and analysis.

MicroRNA signatures characterizing caste-independent ovarian activity in queen and worker honeybees (Apis mellifera L.).

Queen and worker honeybees differ profoundly in reproductive capacity. The queen of this complex society, with 200 highly active ovarioles in each ovary, is the fertile caste, whereas the workers have approximately 20 ovarioles as a result of receiving a different diet during larval development. In a regular queenright colony, the workers have inactive ovaries and do not reproduce. However, if the queen is sensed to be absent, some of the workers activate their ovaries, producing viable haploid eggs that develop into males. Here, a deep-sequenced ovary transcriptome library of reproductive workers was used as supporting data to assess the dynamic expression of the regulatory molecules and microRNAs (miRNAs) of reproductive and nonreproductive honeybee females. In this library, most of the differentially expressed miRNAs are related to ovary physiology or oogenesis. When we quantified the dynamic expression of 19 miRNAs in the active and inactive worker ovaries and compared their expression in the ovaries of virgin and mated queens, we noted that some miRNAs (miR-1, miR-31a, miR-13b, miR-125, let-7 RNA, miR-100, miR-276, miR-12, miR-263a, miR-306, miR-317, miR-92a and miR-9a) could be used to identify reproductive and nonreproductive statuses independent of caste. Furthermore, integrative gene networks suggested that some candidate miRNAs function in the process of ovary activation in worker bees.

FSH supplementation to culture medium is beneficial for activation and survival of preantral follicles enclosed in equine ovarian tissue.

This study investigated the effect of adding different concentrations of bovine recombinant follicle-stimulating hormone on the IVC of equine preantral follicles enclosed in ovarian tissue fragments. Randomized ovarian fragments were fixed immediately (fresh noncultured control) or cultured for 1 or 7 days in α-MEM(+) supplemented with 0, 10, 50, and 100 ng/mL FSH and subsequently analyzed by classical histology. Culture media collected on Day 1 or Day 7 and were analyzed for steroids (estradiol and progesterone) and reactive oxygen species (ROS). After Day 1 and Day 7 of culture, 50-ng/mL FSH treatment had a greater (P < 0.05) percentage of morphologically normal follicles when compared to the other groups, except the 10-ng/mL FSH treatment at Day 1 of culture. The percentage of developing follicles (transition, primary, and secondary), and follicular and oocyte diameters were higher (P < 0.05) in the 50-ng/mL FSH treatment compared to the other groups after Day 7 of culture. Furthermore, estradiol secretion and ROS production were maintained (P > 0.05) throughout the culture in the 50-ng/mL FSH treatment. In conclusion, the addition of 50 ng/mL of FSH promoted activation of primordial follicles to developing follicles, improved survival of preantral follicles, and maintained estradiol and ROS production of equine ovarian tissue after 7 days of culture.

O uso dos métodos cirúrgico e rotatório no tratamento da despigmentação gengival ­ relato de dois casos clínicos / The use of surgical and rotary methods to treat gingival depigmentation ­ report of two clinical cases

Este artigo relatou dois tipos diferentes de tratamento para pacientes com despigmentação melânica gengival. No primeiro caso, uma paciente de 21 anos de idade apresentou-se com intensa pigmentação melânica (marrom escuro) na região entre os caninos. O tratamento consistiu na remoção desta faixa de tecido (raspagem gengival) usando-se apenas anestesia local e uma lâmina de bisturi 15C. No segundo caso, um paciente de 22 anos de idade apresentou-se com pigmentação melânica moderada (marrom médio), também na região intercanina superior. O tratamento consistiu na remoção deste tecido utilizando brocas diamantadas 1016HL sob irrigação com soro fisiológico e finalização com broca diamantada 4138. Em ambos os casos, um cimento cirúrgico foi usado para proteção, a dor foi controlada com dipirona, e as áreas foram limpas com solução de clorexidina 0,12%. Após 14 dias, uma coloração rosácea final foi obtida. As duas técnicas apresentadas possuem eficácia imediata satisfatória, agilidade e custo baixo em relação às técnicas mais sofisticadas. Entretanto, os pacientes precisam estar cientes da possibilidade de recorrência da pigmentação melânica.

This article reports on two different treatments for gingival pigmentation. In the fi rst case, a 21 years-old female patient presented with intense pigmentation (dark-brown) at the maxillary canine region. The treatment consisted on removal of this tissue band (gingival peeling) by applying local anesthesia and using a 15C scalpel blade. In the second case, a 22 years-old male patient demonstrated moderate gingival pigmentation (middle-brown) also in the maxillary canine region. The treatment was made by rotary removal with diamond bur 1016HL, application of saline solution, and finishing with a 4138 tapered diamond bur. In both cases, a surgical dressing was applied to protect the operated area. Pain control was provided with dipyrone, and a 0.12% digluconate chlorhexidine solution recommended in the postoperative period. After 14 days, a pink, gingival aspect was generated. Both techniques have immediate efficacy, are fast, and low cost compared to more sophisticated treatments. However, the patients need to be aware of recurrence of gingival pigmentation.

Recovery of indium from LCD screens of discarded cell phones.

Advances in technological development have resulted in high consumption of electrical and electronic equipment (EEE), amongst which are cell phones, which have LCD (liquid crystal display) screens as one of their main components. These multilayer screens are composed of different materials, some with high added value, as in the case of the indium present in the form of indium tin oxide (ITO, or tin-doped indium oxide). Indium is a precious metal with relatively limited natural reserves (Dodbida et al., 2012), so it can be profitable to recover it from discarded LCD screens. The objective of this study was to develop a complete process for recovering indium from LCD screens. Firstly, the screens were manually removed from cell phones. In the next step, a pretreatment was developed for removal of the polarizing film from the glass of the LCD panels, because the adherence of this film to the glass complicated the comminution process. The choice of mill was based on tests using different equipment (knife mill, hammer mill, and ball mill) to disintegrate the LCD screens, either before or after removal of the polarizing film. In the leaching process, it was possible to extract 96.4 wt.% of the indium under the following conditions: 1.0M H2SO4, 1:50 solid/liquid ratio, 90°C, 1h, and stirring at 500 rpm. The results showed that the best experimental conditions enabled extraction of 613 mg of indium/kg of LCD powder. Finally, precipitation of the indium with NH4OH was tested at different pH values, and 99.8 wt.% precipitation was achieved at pH 7.4.

Prevalência da doença periodontal em diabéticos controlados / Prevalence of periodontal disease in controlled diabetics

Objetivo: o presente estudo teve como objetivo pesquisar, utilizando os parâmetros clínicos periodontais, a prevalência da doença periodontal em indivíduos portadores de diabetes mellitus tipo 2 metabolicamente controlados. Material e métodos: o controle metabólico foi avaliado utilizando como parâmetro o ensaio da hemoglobina glicosilada, cuja média apresentada pela amostra foi de 5,79%. Esta foi constituída por 105 indivíduos com média de idade de 55,8 anos, apresentando diabetes tipo 2, assistidos pelo programa Hiperdia, da Prefeitura de Camaçari, no período entre 2007 e 2009. A periodontite foi caracterizada por, no mínimo, uma bolsa de 4 mm associada à perda de inserção maior ou igual a 4 mm e sangramento gengival. Já para determinação da gengivite, foi considerada a presença de vermelhidão gengival e o sangramento a sondagem em 25% dos sítios. Resultados: a porcentagem de faces coradas foi de 61,15% e a de sangramento a sondagem foi de 62%. Em relação à medida de profundidade de sondagem dos sítios sondados, 89,92% foram enquadradas no subgrupo associado à saúde; 9,19% à profundidade de sondagem moderada; e 0,89% à severa. Sobre a perda de inserção, 76,65% não apresentaram perda de inserção ou perda leve; 14,28% apresentaram perda de inserção moderada e 9,02% apresentaram perda grave; 84 pacientes (80%) apresentaram periodontite crônica; 12 pacientes (11,4%) foram enquadrados nos critérios de diagnóstico para gengivite e nove (8,6%) registraram saúde periodontal. Conclusão: através dos parâmetros clínicos periodontais utilizados neste estudo, foi possível observar uma prevalência de 80% de periodontite e 11,4% de gengivite entre os indivíduos diabéticos controlados metabolicamente.