Recovery of Mollicutes from the reproductive tract of dairy cattle in the state of Pernambuco, Brazil

Abstract: The aim of the present study was to report the occurrence of members of the Mollicutesclass in the reproductive system of dairy cattle in Brazil. Five farms containing dairy cattle were visited in January of 2012. In total, 100 cows of different ages, breeds and stages of lactation were examined in the present study. The cows were part of intensive or semi-intensive management systems and were submitted to mechanical milking or hand milking. The samples were collected after washing the vulvar region with water and soap, and then drying it with paper towels and disinfecting the area with alcohol (70°GL). Vaginal mucous was collected using a sterile alginate cotton swab, which was rubbed on the vagina, as well as the lateral and internal walls. Vulvovaginal mucous samples were cultured in both liquid and solid modified Hayflick´s medium, for mycoplasmas, and UB medium, for ureaplasmas. The PCR assays for Mollicutesand Ureaplasmaspp. were performed according to the standard protocols described in the current literature. During isolation, the frequency of Mycoplasmaspp. was of 13.0% (13/100) and for Ureaplasmaspp. was of 6.0% (6/100). In the PCR assays the frequency of Mollicuteswas of 26.0% (26/100) and for Ureaplasmaspp. was of 13.0% (13/100) in the dairy cattle studied. This is the first report of these agents in reproductive system of bovine of the Pernambuco state. Further studies are necessary to determine the pathogenic potential and species of these field isolates.

Variable structure position control of an industrial robotic manipulator

The use of a robust position controller for a robotic manipulator moving in free space is presented. The aim is to implement in practice a controller that is robust to uncertainties in the model of the system, as well as being inexpensive from a computational point of view. Variable structure theory provides the technique for the design of such controller. The design steps are presented, first from a theoretical perspective and then applied to the control of a two degree-of-freedom manipulator. Simulation results that backed the implementation are presented, followed by the experiments conducted and the results that were obtained. The conclusion is that variable structure control is readily applicable to industrial robots for the robust control of positions.

Influência do ácido giberélico no desenvolvimento, produção e qualidade do fruto em cultura de pimentão (Capsicum annuum L.)

Foi aplciado, na cultura de pimentão variedade híbrida Cascadura x Agronômico 10 G, ácido giberélico (GA3), 10, 20 e 40 ppm, pulverizado, uma, três e seis vezes, com intervalos de uma semana, iniciando-se a primeira aplicação sete dias após o transplante , com a finalidade de se estudar a influência desse fitorregulador no desenvolvimento, produção e qualidade do fruto, nessa cultura. Conclui-se que a aplicação de 10 ppm, uma semana após o transplante, resultou em maior produção; maior peso médio do fruto e relativa precocidade. A dose de 40 ppm em se is aplicações consecutivas, resultou em pequena produção, mais tardia, com as plantas mostrando maior desenvolvimento vegetativo, folhas maiores e de coloração verde amarelada.

Efeito de regulador de crescimento na cultura de trigo submetido a diferentes doses de nitrogênio e densidades de plantas

Com o objetivo de avaliar os efeitos de um regulador de crescimento no cultivar de trigo IAPAR-53, em diferentes densidades de plantas e submetido a diferentes doses de nitrogênio, instalou-se um experimento na Fazenda-Escola da UEPG em Ponta Grossa-PR, no ano de 1999. O delineamento experimental foi o de blocos ao acaso, em esquema fatorial 2 x 3 x 4, com quatro repetições. Os tratamentos constaram da aplicação de 125 g i.a. ha-1 de trinexapac-ethyl e testemunha sem aplicação, em densidades de 44, 60 e 75 plantas m-1, no espaçamento de 0,17 m entre fileiras e nas doses de 0, 45, 90 e 135 kg ha-1 de nitrogênio em cobertura. A aplicação de trinexapac-ethyl resultou em plantas com menor massa seca, entrenós mais curtos, maior diâmetro do caule e aumento do número de espigas m-1, de grãos por espiga e produtividade. Com o aumento da dose de nitrogênio ocorreu aumento da estatura das plantas, do número de espigas m-1, do peso de mil grãos e da produtividade. Com o aumento da densidade de plantas, o diâmetro do caule, a massa seca das plantas e o número de grãos por espiga diminuíram e o número de espigas m-1 e o peso de mil grãos aumentaram, sem efeitos na produtividade. Não foi observado acamamento em nenhum dos tratamentos.

Toxicidade do herbicida S-metolachlor em plantas de milho provenientes de sementes com diferentes formatos e dimensões

Este trabalho teve como objetivo avaliar a toxicidade do herbicida S-metolachlor em plantas de milho oriundas de sementes com diferentes características morfológicas. O experimento foi conduzido em casa de vegetação, localizada no município de Capão do Leão, RS. Os tratamentos foram compostos pelas combinações de três grupos de tamanhos de sementes, classificadas em peneiras de crivo oblongo [sementes retidas nas peneiras de largura 14/64" (peneira 14), 18/64" (peneira 18) e 21/64" (peneira 21)], de dois formatos de sementes (chata e redonda) e de cinco doses do herbicida S-metolachlor (0,00; 0,48; 0,96; 1,44; e 1,92 kg ha-1), totalizando-se 30 tratamentos. O delineamento experimental utilizado foi o de blocos casualizados em esquema fatorial (3 x 2 x 5), com quatro repetições. Após a separação e classificação das sementes, procedeu-se à semeadura em vasos preenchidos com solo homogeneizado. Em cada vaso foram semeadas oito sementes de milho, na profundidade de 3,0 cm, realizando-se, 24 horas depois, a aplicação do S-metolachlor em pré-emergência. Foram realizadas as seguintes avaliações: velocidade de emergência das plântulas; número total de plântulas emergidas; toxicidade visual e altura de plantas aos 7, 14 e 21 dias após a emergência (DAE); e biomassa seca das raízes e da parte aérea aos 21 DAE. O formato das sementes se mostrou fator importante na tolerância das plantas aos efeitos tóxicos do herbicida S-metolachlor, aplicado em pré-emergência, quando as sementes de milho são de menor tamanho, sendo observada maior toxicidade quando as plantas eram provenientes de sementes redondas. O aumento das doses aplicadas do S-metolachlor ocasionou reduções na aquisição de biomassa seca tanto da parte aérea como das raízes de plantas de milho, porém maior decréscimo se observou quanto ao acúmulo de biomassa seca das raízes.

Intoxicação de espécies de eucalipto submetidas à deriva do glyphosate

O glyphosate é o herbicida mais utilizado em áreas de reflorestamento de eucalipto. Nessas áreas tem sido freqüente a verificação de sintomas de intoxicação devido à deriva. Entretanto, trabalhos de pesquisa e observações de campo indicam comportamento diferencial entre as espécies e os clones de eucalipto quando em contato com o glyphosate. O presente trabalho teve como objetivo avaliar os efeitos da deriva simulada de glyphosate, por meio de doses reduzidas, no crescimento de cinco espécies de eucalipto. Utilizou-se o esquema fatorial 5x5, sendo cinco espécies (Eucalyptus urophylla, E. grandis, E. pellita, E. resinifera e E. saligna) e cinco doses (0; 43,2; 86,4; 172,8; e 345,6 g ha-1 de glyphosate), no delineamento em blocos casualizados com quatro repetições. A aplicação do herbicida foi feita sobre as plantas, de modo que não atingisse o terço superior, 23 dias após o plantio destas. Os sintomas de intoxicação causados pelo glyphosate foram semelhantes nas diferentes espécies, sendo caracterizados por murcha, clorose e enrolamento foliar, e, no caso de maiores doses, por necroses e senescência foliar. Plantas submetidas a doses acima de 86,4 g ha-1 de glyphosate foram severamente intoxicadas, afetando o seu crescimento, resultando em menor altura, diâmetro do caule e massa seca aos 45 dias após aplicação do herbicida. Entre as espécies estudadas, E. resinifera foi mais tolerante à deriva de glyphosate, apresentando menores valores de intoxicação e maior incremento em altura e diâmetro, mesmo nas plantas submetidas às maiores doses, o que não foi observado nas demais espécies.

Intoxicação de eucalipto submetido à deriva simulada de diferentes herbicidas

Além do glyphosate, amplamente usado, outros herbicidas, como o triclopyr e o carfentrazone-ethyl, apresentam potencial de uso na eucaliptocultura. Entretanto, a nãoseletividade destes herbicidas ao eucalipto pode levar à intoxicação da cultura, caso haja contato das folhas com a calda aplicada. Objetivou-se avaliar os efeitos da deriva de alguns herbicidas em plantas de Eucalyptus urophylla, simulada por meio de subdoses: 0, 43,2, 86,4 e 172,8 g ha-1 de glyphosate; 14,4, 28,8 e 57,6 g ha-1 de triclopyr; 0,84, 1,68 e 3,36 g ha-1 de carfentrazone-ethyl; e das misturas: 43,2 g ha-1 de glyphosate + 14,4 g ha-1 de triclopyr; 86,4 g ha¹ de glyphosate + 28,6 g ha-1 de triclopyr; 43,2 g ha-1 de glyphosate + 1,68 g ha-1 de carfentrazone-ethyl; 86,4 g ha-1 de glyphosate + 3,36 g ha-1 de carfentrazone-ethyl. As mudas de eucalipto com aproximadamente 50 cm de altura receberam a aplicação dos herbicidas de modo a não atingir o terço superior das plantas, 45 dias após o transplantio. Entre os tratamentos, a deriva de glyphosate mostrou-se mais danosa ao eucalipto, seguida de carfentrazone-ethyl e triclopyr. Plantas de eucalipto expostas à subdose de 172,8 g ha-1 de glyphosate e às misturas glyphosate + carfentrazone-ethyl tiveram maior porcentagem de intoxicação e menor crescimento aos 15, 30 e 45 dias após aplicação (DAA). Houve recuperação das plantas expostas à deriva do triclopyr, que apresentaram 56,25 e 15,10% de intoxicação aos 15 e 30 DAA, respectivamente, indicando menor risco da deriva do triclopyr em comparação aos demais herbicidas e misturas estudados.

Melatonin modulation of presynaptic nicotinic acetylcholine receptors located on short noradrenergic neurons of the rat vas deferens: a pharmacological characterization

Melatonin, the pineal hormone produced during the dark phase of the light-dark cycle, modulates neuronal acetylcholine receptors located presynaptically on nerve terminals of the rat vas deferens. Recently we showed the presence of high affinity nicotine-binding sites during the light phase, and low and high affinity binding sites during the dark phase. The appearance of the low affinity binding sites was due to the nocturnal melatonin surge and could be mimicked by exposure to melatonin in vitro. The aim of the present research was to identify the receptor subtypes responsible for the functional response during the light and the dark phase. The rank order of potency of agonists was dimethylphenylpiperazinium (DMPP) = cytisine > nicotine > carbachol and DMPP = nicotine = cytisine > carbachol, during the light and dark phase, respectively, due to an increase in apparent affinity for nicotine. Mecamylamine similarly blocked the DMPP response during the light and the dark phase, while the response to nicotine was more efficiently blocked during the light phase. In contrast, methyllycaconitine inhibited the nicotine-induced response only at 21:00 h. Since a7 nicotinic acetylcholine receptors (nAChRs) have low affinity for nicotine in binding assays, we suggest that a mixed population composed of a3ß4 - plus a7-bearing nAChR subtypes is present at night. This plasticity in receptor subtypes is probably driven by melatonin since nicotine-induced contraction in organs from animals sacrificed at 15:00 h and incubated with melatonin (100 pg/ml, 4 h) is not totally blocked by mecamylamine. Thus melatonin, by acting directly on the short adrenergic neurons that innervate the rat vas deferens, induces the appearance of the low affinity binding site, probably an a7 nAChR subtype.

Guanylin peptides: cyclic GMP signaling mechanisms

Guanylate cyclases (GC) serve in two different signaling pathways involving cytosolic and membrane enzymes. Membrane GCs are receptors for guanylin and atriopeptin peptides, two families of cGMP-regulating peptides. Three subclasses of guanylin peptides contain one intramolecular disulfide (lymphoguanylin), two disulfides (guanylin and uroguanylin) and three disulfides (E. coli stable toxin, ST). The peptides activate membrane receptor-GCs and regulate intestinal Cl- and HCO3- secretion via cGMP in target enterocytes. Uroguanylin and ST also elicit diuretic and natriuretic responses in the kidney. GC-C is an intestinal receptor-GC for guanylin and uroguanylin, but GC-C may not be involved in renal cGMP pathways. A novel receptor-GC expressed in the opossum kidney (OK-GC) has been identified by molecular cloning. OK-GC cDNAs encode receptor-GCs in renal tubules that are activated by guanylins. Lymphoguanylin is highly expressed in the kidney and heart where it may influence cGMP pathways. Guanylin and uroguanylin are highly expressed in intestinal mucosa to regulate intestinal salt and water transport via paracrine actions on GC-C. Uroguanylin and guanylin are also secreted from intestinal mucosa into plasma where uroguanylin serves as an intestinal natriuretic hormone to influence body Na+ homeostasis by endocrine mechanisms. Thus, guanylin peptides control salt and water transport in the kidney and intestine mediated by cGMP via membrane receptors with intrinsic guanylate cyclase activity.

Estradiol decreases iodide uptake by rat thyroid follicular FRTL-5 cells

Estradiol has well-known indirect effects on the thyroid. A direct effect of estradiol on thyroid follicular cells, increasing cell growth and reducing the expression of the sodium-iodide symporter gene, has been recently reported. The aim of the present investigation was to study the effect of estradiol on iodide uptake by thyroid follicular cells, using FRTL-5 cells as a model. Estradiol decreased basal iodide uptake by FRTL-5 cells from control levels of 2.490 ± 0.370 to 2.085 ± 0.364 pmol I-/µg DNA at 1 ng/ml (P<0.02), to 1.970 ± 0.302 pmol I-/µg DNA at 10 ng/ml (P<0.003), and to 2.038 ± 0.389 pmol I-/µg DNA at 100 ng/ml (P<0.02). In addition, 4 ng/ml estradiol decreased iodide uptake induced by 0.02 mIU/ml thyrotropin from 8.678 ± 0.408 to 7.312 ± 0.506 pmol I-/µg DNA (P<0.02). A decrease in iodide uptake by thyroid cells caused by estradiol has not been described previously and may have a role in goiter pathogenesis.

HHV-8 infection in patients with AIDS-related Kaposi's sarcoma in Brazil

The aims of the present study were to determine the prevalence of human herpesvirus type 8 (HHV-8) in HIV-positive Brazilian patients with (HIV+/KS+) and without Kaposi's sarcoma (HIV+/KS-) using PCR and immunofluorescence assays, to assess its association with KS disease, to evaluate the performance of these tests in detecting HHV-8 infection, and to investigate the association between anti-HHV-8 antibody titers, CD4 counts and staging of KS disease. Blood samples from 66 patients, 39 HIV+/KS+ and 27 HIV+/KS-, were analyzed for HHV-8 viremia in peripheral blood mononuclear cells by PCR and HHV-8 antigenemia for latent and lytic infection by immunofluorescence assay. Positive samples for latent nuclear HHV-8 antigen (LNA) antibodies were titrated out from 1/100 to 1/409,600 dilution. Clinical information was collected from medical records and risk behavior was assessed through an interview. HHV-8 DNA sequences were detected by PCR in 74.3% of KS+ patients and in 3.7% of KS- patients. Serological assays were similar in detecting anti-LNA antibodies and anti-lytic antigens in sera from KS+ patients (79.5%) and KS- patients (18.5%). HHV-8 was associated with KS whatever the method used, i.e., PCR (odds ratio (OR) = 7.4, 95% confidence interval (CI) = 2.16-25.61) or anti-LNA and anti-lytic antibodies (OR = 17.0, 95%CI = 4.91-59.14). Among KS+ patients, HHV-8 titration levels correlated positively with CD4 counts (rho 0.48, P = 0.02), but not with KS staging. HHV-8 is involved in the development of KS in different geographic areas worldwide, as it is in Brazil, where HHV-8 is more frequent among HIV+ patients. KS severity was associated with immunodeficiency, but no correlation was found between HHV-8 antibody titers and KS staging.

Presurgical ketoprofen, but not morphine, dipyrone, diclofenac or tenoxicam, preempts post-incisional mechanical allodynia in rats

The treatment of pain before it initiates may prevent the persistent pain-induced changes in the central nervous system that amplify pain long after the initial stimulus. The effects of pre- or postoperative intraperitoneal administration of morphine (2 to 8 mg/kg), dipyrone (40 and 80 mg/kg), diclofenac (2 to 8 mg/kg), ketoprofen (10 and 20 mg/kg), and tenoxicam (10 and 20 mg/kg) were studied in a rat model of post-incisional pain. Groups of 5 to 8 male Wistar rats (140-160 g) were used to test each drug dose. An incision was made on the plantar surface of a hind paw and the changes in the withdrawal threshold to mechanical stimulation were evaluated with Von Frey filaments at 1, 2, 6 and 24 h after the surgery. Tenoxicam was given 12 or 6 h preoperatively, whereas the remaining drugs were given 2 h or 30 min preoperatively. Postoperative drugs were all given 5 min after surgery. No drug abolished allodynia when injected before or after surgery, but thresholds were significantly higher than in control during up to 2 h following ketoprofen, 6 h following diclofenac, and 24 h following morphine, dipyrone or tenoxicam when drugs were injected postoperatively. Significant differences between pre- and postoperative treatments were obtained only with ketoprofen administered 30 min before surgery. Preoperative (2 h) intraplantar, but not intrathecal, ketoprofen reduced the post-incisional pain for up to 24 h after surgery. It is concluded that stimuli generated in the inflamed tissue, rather than changes in the central nervous system are relevant for the persistence of pain in the model of post-incisional pain.

Chronic excitotoxic lesion of the dorsal raphe nucleus induces sodium appetite

We determined if the dorsal raphe nucleus (DRN) exerts tonic control of basal and stimulated sodium and water intake. Male Wistar rats weighing 300-350 g were microinjected with phosphate buffer (PB-DRN, N = 11) or 1 µg/0.2 µl, in a single dose, ibotenic acid (IBO-DRN, N = 9 to 10) through a guide cannula into the DRN and were observed for 21 days in order to measure basal sodium appetite and water intake and in the following situations: furosemide-induced sodium depletion (20 mg/kg, sc, 24 h before the experiment) and a low dose of dietary captopril (1 mg/g chow). From the 6th day after ibotenic acid injection IBO-DRN rats showed an increase in sodium appetite (12.0 ± 2.3 to 22.3 ± 4.6 ml 0.3 M NaCl intake) whereas PB-DRN did not exceed 2 ml (P < 0.001). Water intake was comparable in both groups. In addition to a higher dipsogenic response, sodium-depleted IBO-DRN animals displayed an increase of 0.3 M NaCl intake compared to PB-DRN (37.4 ± 3.8 vs 21.6 ± 3.9 ml 300 min after fluid offer, P < 0.001). Captopril added to chow caused an increase of 0.3 M NaCl intake during the first 2 days (IBO-DRN, 33.8 ± 4.3 and 32.5 ± 3.4 ml on day 1 and day 2, respectively, vs 20.2 ± 2.8 ml on day 0, P < 0.001). These data support the view that DRN, probably via ascending serotonergic system, tonically modulates sodium appetite under basal and sodium depletion conditions and/or after an increase in peripheral or brain angiotensin II.

Cytotoxic activity of the dichloromethane fraction from Vernonia scorpioides (Lam.) Pers. (Asteraceae) against Ehrlich's tumor cells in mice

Vernonia scorpioides has been widely used in Brazil to treat skin problems and chronic wounds, such as ulcers of the lower limbs and diabetic lesions. In the present study, we investigated the effect of a dichloromethane (DCM) fraction of V. scorpioides leaf extract on Ehrlich ascitic and solid tumor-bearing mice. The animals were treated once a day with the DCM fraction at a concentration of 5 mg/kg, administered ip during and after the development of the tumor. The lifespan, weight, number and type of leukocytes, number of tumor cells, volume of solid and ascitic tumors were measured. The development of the tumor with pre-treated tumor cells in vitro with the DCM fraction (5 mg/kg) was analyzed and the animals were sacrificed after 7 days. The DCM fraction (5 mg/kg) totally inhibited tumor development when in direct contact with tumor cells, and also ascitic tumor development with in vitro treatment or when administered ip, in loco (after 7 days). Animals treated with the DCM fraction increased their lifespan ca. 2 weeks and maintained their body weight for 30 days. When applied immediately after the inoculation of the tumor cells in vivo, it totally abolished tumor development, with tumor development only decreasing when treatment was started 3 days after the tumor challenge. These data suggest an antineoplastic activity of the fraction. Oral or ip administration of DCM fraction (5 mg/kg) for 7 days did not reduce the solid tumor volume. The cytotoxic activity described here differs from the conventional immune suppressing profile of standard chemotherapy because it increases neutrophil influx to the peritoneal cavity. These results show that, besides exhibiting a tumoricidal activity, the DCM fraction also exhibits inflammatory activity.

Adenosine A1 receptor-mediated inhibition of in vitro prolactin secretion from the rat anterior pituitary

In previous studies, we demonstrated biphasic purinergic effects on prolactin (PRL) secretion stimulated by an adenosine A2 agonist. In the present study, we investigated the role of the activation of adenosine A1 receptors by (R)-N6-(2-phenylisopropyl)adenosine (R-PIA) at the pituitary level in in vitro PRL secretion. Hemipituitaries (one per cuvette in five replicates) from adult male rats were incubated. Administration of R-PIA (0.001, 0.01, 0.1, 1, and 10 µM) induced a reduction of PRL secretion into the medium in a U-shaped dose-response curve. The maximal reduction was obtained with 0.1 µM R-PIA (mean ± SEM, 36.01 ± 5.53 ng/mg tissue weight (t.w.)) treatment compared to control (264.56 ± 15.46 ng/mg t.w.). R-PIA inhibition (0.01 µM = 141.97 ± 15.79 vs control = 244.77 ± 13.79 ng/mg t.w.) of PRL release was blocked by 1 µM cyclopentyltheophylline, a specific A1 receptor antagonist (1 µM = 212.360 ± 26.560 ng/mg t.w.), whereas cyclopentyltheophylline alone (0.01, 0.1, 1 µM) had no effect. R-PIA (0.001, 0.01, 0.1, 1 µM) produced inhibition of PRL secretion stimulated by both phospholipase C (0.5 IU/mL; 977.44 ± 76.17 ng/mg t.w.) and dibutyryl cAMP (1 mM; 415.93 ± 37.66 ng/mg t.w.) with nadir established at the dose of 0.1 µM (225.55 ± 71.42 and 201.9 ± 19.08 ng/mg t.w., respectively). Similarly, R-PIA (0.01 µM) decreased (242.00 ± 24.00 ng/mg t.w.) the PRL secretion stimulated by cholera toxin (0.5 mg/mL; 1050.00 ± 70.00 ng/mg t.w.). In contrast, R-PIA had no effect (468.00 ± 34.00 ng/mg t.w.) on PRL secretion stimulation by pertussis toxin (0.5 mg/mL; 430.00 ± 26.00 ng/mg t.w.). These results suggest that inhibition of PRL secretion after A1 receptor activation by R-PIA is mediated by a Gi protein-dependent mechanism.