T-type Ca2+ channels and their relationship with pre-neoplastic and neoplastic lesions in the human breast

The expression of T-type voltage-dependent Ca2+ channels (Cav3) has been previously observed in breast cancer, but their expression and subcellular localization were not evaluated in pre-neoplastic lesions. Therefore, this work aimed to evaluate protein expression and subcellular localization of T-type channel isoforms in human breast tissue samples. Protein expressions of CaV3.1, CaV3.2, and CaV3.3 were evaluated by immunohistochemistry in breast without alteration, in proliferative non-neoplastic lesions, and in neoplastic ductal epithelial lesions of the human breast. CaV3.1, CaV3.2, and CaV3.3 nuclear expressions were decreased in advanced stages of neoplastic transformation, whereas CaV3.1 and CaV3.2 cytoplasmic expression increased. Also, the decrease in nuclear expression was correlated with an increase in cytoplasmic expression for CaV3.1 isoform. The change in CaV3 protein expression and subcellular localization are consistent with the neoplastic transformation stages of mammary epithelial cells, evident in early neoplastic lesions, such as ductal carcinomas in situ. These results suggest a possible involvement of CaV3 in the carcinogenic processes and could be considered as a potential pharmacological target in new therapies for breast cancer treatment.

Influência da dificuldade de inseminação, temperamento e cortisol plasmático sobre a taxa de concepção de vacas e novilhas da raça Nelore inseminadas em tempo fixo / Influence of difficulty of insemination, temperament and plasma cortisol on conception rate of Nelore cows and heifers in timed AI

O objetivo do presente experimento foi avaliar efeito do estresse e da dificuldade de inseminação (DifIA) sobre a taxa de concepção (TC) de vacas (n=93) e novilhas (n= 72) Nelore submetidas à IATF. No D9, anotou-se nota de temperamento (NTe) e tempo da saída do brete (TSB) de todos os animais e coletou-se sangue das novilhas. No dia da IATF (D11), anotou-se NTe, TSB, DifIA e tempo de IA. A TC foi 36% para vacas e 46% para novilhas (P>0,05). Não foi observado efeito de Nte sobre TC (P>0,05). Porém, houve tendência para maior (P<0,10) TC nos animais que não apresentaram dificuldade de inseminação (DifIA1; TP=42%) em comparação aos animais com moderada ou alta dificuldade (DifIA2+DifIA3; TP=27%). Foi observado menor (P<0,05) tempo de IA para animais DifIA1 (17:31±06:02s) que animais DifIA2-3 (30:10±15:45s). Novilhas com maiores (P<0,05) níveis de cortisol apresentaram maior NTe (P<0,05). Entretanto, TC (59%) das novilhas menos reativas (cortisol=4,12±1,12ng/mL; NTe=3,2±0,6) não diferiu da TC (41%; P>0,05) das mais agitadas (cortisol=7,76±1,33ng/mL; NTe=3,82±0,79). Concluiu-se que avaliações de temperamento se relacionaram com nível de estresse, embora esses parâmetros não tenham afetado a TC deste trabalho. A maior dificuldade e/ou tempo necessário para se completar a IA demonstrou ser um potencial fator para a redução da fertilidade na IATF.(AU)

The objective was to evaluate the influence of stress and difficulty of insemination (DifIA) on conception rate (CR) of Nellore cows (n= 93) and heifers (n= 72) in Timed-AI. On D9, temperament (NTe) and time for chute exit (TSB) were recorded for all animals, and blood samples were colected from heifers. On the day of Timed-AI (D11), NTe, TSB, DifIA and time for AI were recorded. For cows, CR was 36% and for heifers 46% (P> 0.05). No effect (P> 0.05) of NTe was observed on CR. However, a tendency (P< 0,10) for higher CR was observed in animals with no difficulty for insemination (DifIA1; CR=42%) compared to animals that presented moderate or high difficulty (DifIA2+DifIA3, CR=27%). Time required for AI was lower (P< 0.05) in animals DifIA1 (17:31±06:02sec) than in animals DifIA2-3 (30:10±15:45sec). Heifers with greater (P< 0.05) cortisol levels presented higher Nte (P< 0.05). However, CR (59%) of less reactive heifers (cortisol=4,12±1,12ng/mL; NTe=3,2±0,6) did not differ from CR (41%; P> 0.05) of stressed animals (cortisol=7,76±1,33ng/mL; NTe=3,82±0,79). It was concluded that assessments of temperament were related to stress level, although these parameters did not affect the CR of this study. However, the higher difficulty and/or time to complete AI showed to be a potential factor for reducing fertility after timed-AI.(AU)

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)

Situação epidemiológica da brucelose bovina no Estado do Tocantins / Epidemiological situation of bovine brucellosis in the State of Tocantins, Brazil

Realizou-se um estudo para caracterizar a situação epidemiológica da brucelose bovina no Estado do Tocantins, entre fevereiro de 2002 e agosto de 2003. O Estado foi dividido em seis áreas com características produtivas homogêneas (circuitos produtores). Para cada área, foi calculada uma amostragem simples aleatória de 300 propriedades, com o objetivo de estimar a prevalência de focos de brucelose além da prevalência de fêmeas bovinas adultas soropositivas. Para isso, foram amostradas de 10 a 15 vacas com idade superior a dois anos em cada propriedade. Um total de 20.908 soros foi obtido de 1.842 propriedades. A prevalência de focos de brucelose foi de 21,2 por cento [19,3-23,1 por cento] e a prevalência de fêmeas bovinas adultas soropositivas de 4,4 por cento [3,6-5,3 por cento] para o Estado. Quando se considerou o circuito produtor, observou-se que os circuitos 1, 2, 3 e 5 tiveram prevalência de focos significativamente maior que os circuitos 4 e 6. Os resultados da prevalência nos circuitos 1, 2, 3 e 5 foram de: 16,0 por cento [12,1-20,6 por cento], 37,6 por cento [32,1-43,4 por cento], 26,4 por cento [21,5-31,7 por cento] e 29,3 por cento [24,3-34,7 por cento], respectivamente. Nos circuitos 4 e 6, foram de 5,8 por cento [3,5-9,1 por cento] e 8,6 por cento [5,7-12,2 por cento], respectivamente. Em cada propriedade, foi aplicado um questionário epidemiológico, com o objetivo de avaliar o grau de associação de possíveis fatores de risco com a doença. Os fatores de risco (odds ratio, OR) associados à condição de foco de brucelose foram: rebanho com mais de 120 vacas (OR= 2,0) e abate de reprodutores na propriedade (OR= 1,52). Vacinação contra brucelose (OR= 0,37), presença de piquete de parição (OR= 0,72) e exploração de leite (OR= 0,63) apresentaram-se como fatores de proteção.

A study was carried out to characterize the epidemiological situation of brucellosis in the State of Tocantins from February 2002 to August 2003. The State was divided into six regions with a homogeneous productive system. For each region, a simple random sample was calculated to estimate the prevalence both in farms and cows older than two-year. To achieve this, from 10 to 15 adult cows (older than two-year) were sampled. A total of 20,908 sera from 1,842 farms were obtained. For the whole State of Tocantins, the prevalence of positive farms (or farms with at least one positive animal) was 21.2 percent [19.3-23.1 percent]. When the production regions were considered, the prevalences for the regions 1, 2, 3, and 5 were: 16.0 percent [12.1-20.6 percent], 37.6 percent [32.1-43.4 percent], 26.4 percent [21.5-31.7 percent], and 29.3 percent [24.3-34.7 percent], respectively. In the regions 4 and 6, the prevalences were 5.8 percent [3.5-9.1 percent] and 8.6 percent [5.7-12.2 percent], respectively. In each visited farm, a questionnaire was applied, in order to evaluate the association between with possible risk factors and the brucellosis. The risk factors (odds ratio, OR) associated with the infected herds were number of cows above 120 (OR= 2.0) and slaughtering of breeding animals in the farm (OR= 1.52). Vaccinating against brucellosis (OR= 0.37), presence of birth pen (OR= 0.72), and dairy farm (OR= 0.63) presented as protective factors.

Degradabilidade in situ da matéria seca de forrageiras tropicais obtidas em diferentes épocas de corte / In situ dry matter degradation of tropical forages harvested at different ages

Estimou-se a degradabilidade in situ de 10 forrageiras tropicais em três idades ao corte (21, 42 e 63 dias) e em três tempos de incubação (6, 24 e 96h). As forrageiras utilizadas foram: Andropogon gayanus cv. Planaltina, Brachiaria brizantha (acessos 3401, 3413 e 3451 CPAC), Cenchrus ciliaris (acessos 79119, 79146, 79148, 80199 e 81302 CPATSA) e Panicum maximum (3616 CPAC). Os acessos de B. brizantha, P. maximum e A. gayanus apresentaram os maiores valores de degradabilidade da matéria seca e os de C. ciliaris os valores mais baixos. Todas as forrageiras apresentaram queda no desaparecimento da matéria seca com o avanço da idade. Os acessos de Brachiaria brizantha 3413 e 3451 apresentaram as maiores degradabilidades efetivas.

CaV 3.1 and CaV 3.3 account for T-type Ca2+ current in GH3 cells

T-type Ca2+ channels are important for cell signaling by a variety of cells. We report here the electrophysiological and molecular characteristics of the whole-cell Ca2+ current in GH3 clonal pituitary cells. The current inactivation at 0 mV was described by a single exponential function with a time constant of 18.32 ñ 1.87 ms (N = 16). The I-V relationship measured with Ca2+ as a charge carrier was shifted to the left when we applied a conditioning pre-pulse of up to -120 mV, indicating that a low voltage-activated current may be present in GH3 cells. Transient currents were first activated at -50 mV and peaked around -20 mV. The half-maximal voltage activation and the slope factors for the two conditions are -35.02 ñ 2.4 and 6.7 ñ 0.3 mV (pre-pulse of -120 mV, N = 15), and -27.0 ñ 0.97 and 7.5 ñ 0.7 mV (pre-pulse of -40 mV, N = 9). The 8-mV shift in the activation mid-point was statistically significant (P < 0.05). The tail currents decayed bi-exponentially suggesting two different T-type Ca2+ channel populations. RT-PCR revealed the presence of a1G (CaV3.1) and a1I (CaV3.3) T-type Ca2+ channel mRNA transcripts.

Effects of lead and/or zinc exposure during the second stage of rapid postnatal brain growth on delta-aminolevulinate dehydratase and negative geotaxis of suckling rats

Lead has been shown to produce cognitive and motor deficits in young rats that could be mediated, at least in part, by inhibition of the zinc-containing heme biosynthetic enzyme delta-aminolevulinate dehydratase (ALA-D). In the present study we investigated the effects of lead and/or zinc treatment during the second stage of rapid postnatal brain development on brain, kidney and blood ALA-D specific activity, as well as the negative geotaxis behavior of rats. Eight-day-old Wistar rats were injected intraperitoneally with saline, lead acetate (8 mg/kg) and/or zinc chloride (2 mg/kg) daily for five consecutive days. Twenty-four hours after treatment, ALA-D activity was determined in the absence and presence of DL-dithiothreitol (DTT). The negative geotaxis behavior was assessed in 9- to 13-day-old rats. Treatment with lead and/or zinc did not affect body, brain or kidney weights or brain- or kidney-to-body weight ratios of the animals. In spite of the absence of effect of any treatment on ALA-D specific activity in brain, kidney and blood, the reactivation index with DTT was higher in the groups treated with lead or lead + zinc than in the control group, in brain, kidney and blood (mean + or - SEM; brain: 33.33 + or - 4.34, 38.90 + or - 8.24, 13.67 + or - 3.41; kidney: 33.50 + or - 2.97, 37.60 + or - 2.67, 15.80 + or - 2.66; blood: 63.95 + or - 3.73, 56.43 + or - 5.93, 31.07 + or - 4.61, respectively, N = 9-11). The negative geotaxis response behavior was not affected by lead and/or zinc treatment. The results indicate that lead and/or zinc treatment during the second stage of rapid postnatal brain growth affected ALA-D, but zinc was not sufficient to protect the enzyme from the effects of lead in brain, kidney and blood.

Antidepressant-like effect of lead in adult mice

It has been reported that lead can cause behavioral impairment by inhibiting the N-methyl-D-aspartate (NMDA) receptor complex. MK-801, a noncompetitive NMDA receptor antagonist, exhibits an antidepressant-like action in the forced swimming test. The purpose of the present study was to determine whether subacute lead exposure in adult male Swiss mice weighing 30-35 g causes an antidepressant-like action in a forced swimming test. Mice were injected intraperitoneally (ip) with 10 mg/kg lead acetate or saline daily for 7 consecutive days. Twenty-four hours after the last treatment, the saline and lead-treated mice received an injection of MK-801 (0.01 mg/kg, ip) or saline and were tested in forced swimming and in open-field tests. Immobility time was similarly reduced in the saline-MK-801, Pb-saline and Pb-MK-801 groups compared to the saline-saline group (mean + or - SEM; 197.3 + or - 18.5, 193.5 + or - 15.8, 191.3 + or - 12.3 and 264.0 + or - 14.4 s, respectively; N = 9). These data indicate that lead may exert its effect on the forced swimming test by directly or indirectly inhibiting the NMDA receptor complex. Lead treatment caused no deficit in memory of habituation and did not affect locomotor activity in an open-field (N = 14). However, mice that received MK-801 after lead exhibited a deficit in habituation (22 per cent reduction in rearing responses between session 3 and 1; N = 14) as compared to control (41 per cent reduction in rearing responses; N = 15), further suggesting that lead may have affected the NMDA receptor activity. Forced-swim immobility in a basin in two daily consecutive sessions was also significantly decreased by lead exposure (mean + or - SEM; day 1 = 10.6 + or - 3.2, day 2 = 19.6 + or - 3.6; N = 16) as compared to control (day 1 = 18.4 + or - 3.8, day 2 = 34.0 + or - 3.7; N = 17), whereas the number of crossings was not affected by lead treatment, further indicating a specific antidepressant-like action of lead.

Effect of lead acetate on neurobehavioral development of rats

We investigated the effects of lead exposure during the pre- and postnatal period on the neurobehavioral development of female Wistar rats (70-75 days of age, 120-150 g) using a protocol of lead intoxication that does not affect weight gain. Wistar rats were submitted to lead acetate intoxication by giving their dams 1.0 mM lead acetate. Control dams received deionized water. Growth and neuromotor development were assessed by monitoring daily the following parameters in 20 litters: body weight, ear unfolding, incisor eruption, eye opening, righting, palmar grasp, negative geotaxis, cliff avoidance and startle reflex. Spontaneous alternation was assessed on potnatal day 17 using a T maze. The animals'ability to equilibrate on a beaker rim was measured on postnatal day 19. Lead intoxication was confirmed by measuring renal, hepatic and cerebral lead concentration in dams and litters. Lead treatment hastened the day of appearance of the following parameters: eye opening (control: 13.5 + 0.6, N= 88; lead: 12.9 + 0.6, N=72; P<0.05), startle reflex (control: 13.0 + 0.8, N= 88; lead: 12.0 + 0.7, N=72; P<0.05) and negative geotaxis. On the other hand, spontaneous alternation performance was hindered in lead-exposed animals (control: 37.6 + 19.7; lead: 57.5 + 28.3 percent of alternating animals; P<0.05). These results suggest that lead exposure without concomitant undernutrition alters rat development, affecting specific subsets of motor skills.

Salpingites. Consideracoes etiopatogenicas. / Salpingitis. Etiopathogenic considerations

Os autores em revisao sobre as salpingites, acentuam sua atual importancia, comentando alem dos conhecimentos dos agentes etiologicos, as novas cepas de microorganismos, de real significado no contexto patologico. Apontam os inumeros fatores de risco, focalizando depois o quadro clinico, os aspectos patogeneticos e os principais comentarios anatomo-patologicos