Using the mini-VIDAS(®) Easy Salmonella protocol to assess contamination in transitional and coastal waters.

Classical methodologies for Salmonella detection may be too long in time to assure public safety. Presently, one of the fastest assays for Salmonella detection using the mini-VIDAS(®) system is the Easy Salmonella protocol. This assay, developed for food matrixes analysis, was here assessed for the applicability on the detection of these bacteria in transitional and saltwaters. The presence of Salmonella was detected in 4.2 % of the samples studied. In these transitional waters, the proposed protocol presented an efficiency of 79.1 %, due to a high false positive rate (20.8 %), and a false negative rate of 0 %-implying reducing analysis time, the use of enrichment broths, and making it more cost effective. Despite the multitude of samples nature, the method here described revealed to be an efficient and promising tool for transitional waters analysis.

Oxidized-LDL and paraoxonase-1 as biomarkers of coronary artery disease in patients with sleep-disordered breathing.

UNLABELLED: Coronary Artery Disease (CAD) and OBJECTIVES: Sleep Disordered Breathing (SDB) are both oxidative stress disorders. SDB intermittent hypoxia induces oxidative stress, and reduces NO(·) availability, causing endothelial dysfunction. Low-density lipoprotein (LDL) peroxidation is involved in atherosclerosis, and is reported in SDB. Oxidized LDL (ox-LDL) and malondialdehyde (MDA) are lipid peroxidation markers. High-density lipoprotein (HDL) presents antiatherosclerotic properties related to paraxonase-1 (PON-1) activity. PON-1 hydrolyseyses lipid peroxides as ox-LDL. This study compares the relationship of HDL and PON-1, the lipid peroxidation markers ox-LDL and MDA, and 8-OHdG DNA damage marker in the association of SDB and CAD. DESIGN AND METHODS: 29 controls and 27 cases with CAD (defined as > 30% coronary narrowing) patients were included. The apnea-hypopnea index (AHI), and several lipid and oxidative stress parameters were measured in these patients. RESULTS: AHI is increased in CAD patients, and PON-1 activity and HDL levels are decreased. Regression analyseyses showed that lower PON-1 activity and higher ox-LDL levels are important CAD predictors, compared to HDL or MDA levels and present an age-dependent increase. Nitrites and nitrates, indirect NO(·) markers, are positive vs correlated with PON-1 and are negatively correlated to ox-LDL. SDB is not correlated to PON-1 activity decrease or ox-LDL increase. AHI is inversely correlated to HDL levels. CONCLUSIONS: These results indicate that PON-1 and ox-LDL are important predictors of CAD, however they may not be directly related to SDB.

Application of the mammalian glyceraldehyde-3-phosphate dehydrogenase gene for sample quality control in multiplex PCR for diagnosis of leishmaniasis

Leishmaniasis is a neglected disease endemic in five continents. It is a severe disease that may lead to death, and its early detection is important to avoid severe damage to affected individuals. Molecular methods to detect Leishmania are considered alternatives to overcome the limitations presented by conventional methods. The aim of this study was to develop multiplex PCR systems able to detect small amounts of target DNA of Leishmania infantum and Leishmania braziliensis, and the gene coding for glyceraldehyde-3-phosphate dehydrogenase (G3PD) in mammals, enabling quality evaluation of the sample simultaneously with detection of the specific target. The systems created for G3PD recognition were combined with detection systems for L. infantum and L. braziliensis to compose multiplex PCR systems for visceral (mVL) and cutaneous (mACL) leishmaniasis diagnosis. The multiplex PCR systems developed were assessed in blood samples from five different species of mammal reservoirs involved in the disease cycle in Brazil, and 96 and 52 human samples from patients with suspected visceral leishmaniasis (VL) and cutaneous leishmaniasis (ACL), respectively. Three G3PD detection systems were created (G3PD1, G3PD2 and G3PD3) with different product sizes, G3PD2 was chosen for the formation of multiplex PCR systems. The two multiplex PCR systems (mVL and mACL) were reproducible in all species evaluated. Results of test samples (sensitivity, specificity and efficiency) suggest its use in routine diagnosis, research activities in medicine and veterinary medicine. Additionally, the systems designed to detect the G3PD gene are capable of combining with other targets used for molecular diagnosis of infectious diseases. Concerning leishmaniasis, the multiplex PCR systems can be used in epidemiological studies for the detection of new and classic reservoirs, which may contribute to the reliability of results and development of actions to control the disease.(AU)

Headache is not more frequent among patients with moderate to severe hypertension.

The association between hypertension and headache has been a contentious issue. Most studies have showed that mild hypertension and headache are not associated, but this may not be the case in patients with hypertension classified at more severe stages. We investigated the association between hypertension classified at moderate to severe stages and headache in a cross-sectional study conducted in the hypertension clinic of a tertiary care University hospital. In total, 1763 referred patients with a medical diagnosis of hypertension in most cases (95.7%) were evaluated by an extensive protocol questionnaire, detailed physical examination, laboratory examination, and had their blood pressure classified according to the VI Joint National Committee (JNC-VI) recommendation. Logistic regression models were used to explore the association between severity of hypertension and pulse pressure with the presence of headache, controlling for several potential confounders. The complaint of headache was referred by 903 (51.3% of whole sample), and a total of 378 patients (21.4%) were classified at the moderate to severe stage (stage III of the JNC-VI report). The diagnosis of moderate to severe hypertension was not associated with the complaint of headache (OR 1.02, 95% CI from 0.79 to 1.30). Pulse pressure and headache were inversely associated (OR 0.91, 95% CI from 0.86 to 0.97, for 10 mmHg). We concluded that headache and hypertension classified at moderate to severe stages were not associated in patients attending to a hypertension clinic. The novel finding of an inverse association between pulse pressure and headache should be addressed in further investigations.

Efeitos do amitraz e da xilazina sobre algumas características fisiológicas de cavalos / Effect of amitraz and xylazine on some physiological variables of horses

Avaliaram-se os efeitos das injeçöes intravenosa (iv) de amitraz (0,1mg/kg) e xilazina (1mg/kg), em cavalos, sobre a atividade cardíaca, freqüência respiratória, atividade motora intestinal, temperatura retal, sudorese e freqüência de apreensäo de alimentos. O amitraz causou diminuiçäo significativa da atividade cardíaca, da freqüência respiratória e da movimentaçäo intestinal, mas esses efeitos näo foram täo pronunciados quanto os causados pela xilazina. O amitraz causou, também, relaxamento significativo da musculatura lisa retal, e um aparente aumento da sudorese e da freqüência de cavalos flagrados mastigando feno. A temperatura retal näo foi influenciada pelo amitraz. Os resultados indicam que o amitraz, na dose utilizada, näo causou efeitos colaterais severos em cavalos

Hepatic regeneration after partial hepatectomy in mice infected with Schistosoma mansoni, at the acute and chronic phases of the disease.

Outbred male albino mice normal or infected with 30 cercariae of Schistosoma mansoni (LE strain) were submitted to 65% hepatectomy during the acute (70 days) and chronic phase (160 days) phases of the disease. A group of the infected animals was treated with 400 mg/kg of oxamniquine during the acute phase before hepatectomy. Non-infected, infected and treated but not hepatectomized animals were kept as controls. Hepatic regeneration was evaluated by incorporation of tritiated thymidine, intraperitoneally injected into non-hepatectomized and hepatectomized animals, 24 hours after surgery. The results showed that removal of 65% of the hepatic parenchyma, during the acute phase, led to a statistically significant increase of thymidine incorporation, when compared with the uninfected hepatectomized controls. This phenomenon was not observed at the chronic phase. Treatment with oxamniquine administered during the acute phase led to a decrease in thymidine incorporation rate 160 days after infection (90 days after treatment) and 24 hours after hepatectomy. The data suggest that infection with S. mansoni represents a considerable stimulus for the regenerative capacity of the liver during the acute, but not the chronic phase of disease.

Hepatic regeneration after partial hepatectomy in mice infected with Schistosoma mansoni, at the acute and chronic phases of the disease

Outbred male albino mice normal or infected with 30 cercariae of Schistosoma mansoni (LE strain) were submitted to 65 percent hepatectomy during the acute (70 days) and chronic phase (160 days) phases of the disease. A group of the infected animals was treated with 400 mg/kg of oxamniquine during the acute phase before hepatectomy. Non-infected, infected and treated but not hepatectomized animals were kept as controls. Hepatic regeneration was evaluated by incorporation of tritiated thymidine, intraperitoneally injected into non-hepatectomized and hepatectomized animals, 24 hours after surgery. The results showed that removal of 65 percent of the hepatic parenchyma, during the acute phase, led to a statistically significant increase of thymidine incorporation, when compared with the uninfected hepatectomized controls. This phenomenon was not observed at the chronic phase. Treatment with oxamniquine administered during the acute phase led to a decrease in thymidine incorporation rate 160 days after infection (90 days after treatment) and 24 hours after hepatectomy. The data suggest that infection with S. mansoni represents a considerable stimulus for the regenerative capacity of the liver during the acute, but not the chronic phase of disease

[Perception on and registration of alcohol abuse and alcohol-related diseases at a university general hospital]. / Percepção e registro de abuso de álcool e de doenças relacionadas num hospital geral universitário.

BACKGROUND: The high frequency of alcohol related problems highlights the importance of its approach. The association of both the alcohol-abuse and alcohol related diseases has been pointed in the literature and are often the main reason for hospitalization. Physicians use different tools to detect abusive drinkers, and one of them is the CAGE-test, validated by Mayfield et al. in 1974. OBJECTIVES: To study the association between the result of the CAGE-test and the perception and registration of alcohol abuse by the medical staff and the nurses in charge of inpatients of Hospital de Clínicas de Porto Alegre, southern Brazil. MATERIAL AND METHOD: The design involved three phases. First, the application of the CAGE-test to the 385 eligible patients. Second, review of all 51 CAGE-positive patients' records as well the controlled evaluation of 51 randomized CAGE-negative patients, to seek alcoholic habits and alcohol-related diseases. Third, interview with the medical staff to find out their perception about these subjects. RESULTS: We found 51 patients CAGE-positive (prevalence = 13%). When evaluated through the registration and through a standardized interview, medical staff and nurses were not aware of more than half of CAGE-positive patients. No significant differences were found between medical staff registration and perception; the prevalence of alcohol related diseases was the same for both: 26 and 27% (p = .861). The prevalence of alcohol related diseases was higher among CAGE-positive patients (p = .0003). CONCLUSIONS: In spite of the high prevalence among our patients, alcohol abuse is not pursued by our health teams as it should be. As for the future, there is reason to envision a continuing alcoholism rapprochement.

Percepçäo e registro de abuso de álcool e de doenças relacionadas num hospital geral universitário / Perception and registration of alcohol abuse and alcohol related diseases by nurses and medical staff of an university general hospital

Objetivo. Avaliar a associaçao entre o resultado do teste CAGE aplicado em pacientes internados num hospital geral universitário (Hospital de Clínicas de Porto ALegre - HCPA) e a percepçao e registro de abuso de álcool e de doenças relacionadas pelas equipes de saúde. Pacientes e Métodos. O delineamento envolveu três fases: a) aplicaçao do teste CAGE aos 385 pacientes adultos e adolescentes internados considerados elegíveis para o estudo; b) revisao dos prontuários dos pacientes CAGE-positivo e de um número igual de pacientes CAGE-negativo, estratificados por área de internaçao e sorteados, para identificar hábitos alcoólicos e a presença de doenças relacionadas ao uso de álcool; c) entevista com os membros das equipes médicas responsáveis para avaliar sua percepçao sobre o hábito alcoólico em seu paciente. As diferenças encontradas foram analisadas através do teste Qui-quadrado e do teste t de Student, com significância estatística em nível de 5 por cento bicaudal. Resultados. Foram encontrados 51 pacientes CAGE-positivo (13 por cento). A equipe médica registrou abuso de álcool no prontuário de 51 por cento dos pacientes CAGE-positivo e 10 por cento dos CAGE-negativo, e a equipe de enfermagem fez este registro no prontuário de 31 por cento dos pacientes CAGE-positivo e 8 por cento dos CAGE-negativo (p=.724). Nao houve diferença entre o registro e a percepçao da equipe médica (p=.184). A prevalência de doenças relacionadas ao abuso de álcool foi 27 por cento quando se examinou os prontuários médicos e 26 por cento quando se entrevistou a equipe médica (p=.861), sendo maior entre os pacientes CAGE-poistivo (43 por cento) do que entre os CAGE-negativo (10 por cento) (p=.0003). Conclusoes. O abuso de bebidas alcoólicas e a presença de doenças associadas ao mesmo sao condiçoes prevalentes nos pacientes admitidos ao HCPA. Entretanto, mais de metade dos casos nao foram detectados pela equipe de saúde, havendo correspondência entre os registros de prontuário e a entrevista.

Characterization of the antinociceptive and sedative effect of amitraz in horses.

Amitraz, an acaricide used to control ectoparasites in animals has a complex pharmacological activity, including alpha2-adrenergic agonist action. The purpose of this research was to investigate the possible antinociceptive and/or sedative effect of amitraz in horses. The sedative effect of the intravenous (i.v.) injection of dimethylformamide (DMF, 5 mL, control) or amitraz (0.05, 0.10, 0.15 mg/kg), was investigated on the head ptosis test. The participation of alpha2-adrenergic receptors in the sedative effect provoked by amitraz was studied by dosing yohimbine (0.12 mg/kg, i.v.). To measure the antinociception, xylazine hydrochloride (1 mg/kg, i.v., positive control) and the same doses of amitraz and DMF were used. A focused radiant light/heat directed onto the fetlock and withers of a horse were used as a noxious stimulus to measure the hoof withdrawal reflex latency (HWRL) and the skin twitch reflex latency (STRL). The three doses of amitraz used (0.05, 0.10 and 0.15 mg/kg) provoked a dose-dependent relaxation of the cervical muscles. The experiments with amitraz and xylazine on the HWRL showed that after i.v. administration of all doses of amitraz there was a significant increase of HWRL up to 150 min after the injections. Additionally, there was a significant difference between control (DMF) and positive control (xylazine) values up to 30 min after drug injection. On the other hand, the experiments on the STRL show that after administration of amitraz at the dose of 0.15 mg/kg, a significant increase in STRL was observed when compared with the control group. This effect lasted up to 120 min after injection. However, no significant antinociceptive effect was observed with the 0.05 and 0.10 mg/kg doses of amitraz or at the 1.0 mg/kg dose of xylazine.

Influence of an antiprogestin (onapristone) on in vivo and in vitro fertilization.

The effects of a progesterone antagonist (onapristone) on heat synchronization, luteinizing hormone (LH) surge, ovulation, oocyte maturation and fertilization of superovulated ewes were studied. Its effects on in vitro bovine oocyte maturation and fertilization were also studied. Estrus synchronization and superovulation treatments were applied to 39 adult ewes using an intravaginal sponge with fluorgestone acetate for 9 days with injections of prostaglandin F2 alpha and pregnant mare's serum gonadotrophin given 24 h before sponge withdrawal. The animals were randomly assigned to four different groups; T1 receiving only the synchrony treatment (n = 11); T2 ewes received two injections of onapristone (1 mg kg-1, i.v.) 12 h apart from 3 h after sponge withdrawal (n = 10); T3 ewes received two injections of progesterone 12 h apart from sponge withdrawal (n = 10); and, T4 ewes received both onapristone and progesterone as described (n = 8). Ewes were mated by a fertile male during estrus. Progesterone and LH were measured during the superovulation period in plasma samples taken every 4 h. Uterine flushings for ova recovery were performed at 5 days (n = 25), 48 h (n = 5) and 24 h (n = 5). Non-fertilized oocytes collected at 24 and 48 h were checked for meiosis resumption. The effects of two doses of onapristone (D1 and D2) on in vitro bovine oocyte maturation (control = 100, D1 = 100 and D2 = 100) and fertilization (control = 107, D1 = 40 and D2 = 75) were also studied. The percentage of animals showing heat signs was significantly lower in group T3 (50% vs. 100%). The onset of oestrus (27.6, 24.8, 68.8 and 25.5 h, respectively for T1, T2, T3 and T4) and an LH surge (32.3, 28.8, 76.5 and 30.5 h, respectively for T1, T2, T3 and T4) after sponge withdrawal were significantly delayed in group T3. There were no significant differences in the intervals between estrus and LH surge among groups (4.61 +/- 0.75 h). The response and ovulation rates until 40 h after sponge withdrawal (group T3 excluded) were similar among groups, but the fertilization rates were significantly lower in groups T2 and T4 when compared with T1 (2% and 3% vs. 41%, respectively; P < 0.001) due to sperm arrest in the cervix. Ova recovery rate decreased significantly from 24-48 h to 5 days and was not affected by treatments (76.9% vs. 37.1% respectively). Onapristone did not affect the resumption of meiosis. Fertilization of bovine oocytes in vitro decreased significantly only in group D2 when compared to control (48% vs. 62.6%, respectively). In conclusion, onapristone treatment during the preovulatory period did not interfere with normal synchronization of estrus, ovulation and oocyte maturation but severely compromised fertilization by arresting spermatozoa in the cervix.

Atherosclerosis and acute arterial thrombosis in rabbits: a model using balloon desendothelization without dietary intervention.

Acute thrombosis can be induced in rabbits by a triggering protocol using Russell's viper venom and histamine given after 8 months of a 1% cholesterol diet and balloon desendothelization. In the present study, we tested the hypothesis that aortic desendothelization performed 4 months before the triggering protocol without a high cholesterol diet is a highly effective and less expensive way of producing arterial atherosclerosis and thrombosis. Nineteen male New Zealand white rabbits on a normal diet were studied. The control group (N = 9) received no intervention during the 4-month observation period, while the other group (N = 10) was submitted to aortic balloon desendothelization using a 4F Fogarty catheter. At the end of this period, all animals were killed 48 h after receiving the first dose of the triggering treatment. Eight of 10 rabbits (80%) in the balloon-trauma group presented platelet-rich arterial thrombosis while none of the animals in the control group had thrombus formation (P < 0.01). Thus, this model, using balloon desendothelization without dietary manipulation, induces arterial atherosclerosis and thrombosis and may provide possibilities to test new therapeutic approaches.

Atherosclerosis and acute arterial thrombosis in rabbits: a model using ballon desendothelization without dietary intervention

Acute thrombosis can be induced in rabbits by a triggering protocol using Russell's viper venom and histamine given after 8 months of a 1 per cent cholesterol diet and balloon desendothelization. In the present study, we tested the hypothesis that aortic desendothelization performed 4 months before the triggering protocol without a high cholesterol diet is a highly effective and less expensive way of producing arterial atherosclerosis and thrombosis. Nineteen male New Zealand white rabbits on a normal diet were studied. The control group (N = 9) received no intervention during the 4-month observation period, while the other group (N = 10) was submitted to aortic balloon desendothelization using a 4F Fogarty catheter. At the end of this period, all animals were killed 48 h after receiving the first dose of the triggering treatment. Eight of 10 rabbits (80 per cent) in the balloon-trauma group presented platelet-rich arterial thrombosis while none of the animals in the control group had thrombus formation (P<0.01). Thus, this model, using balloon desendothelization without dietary manipulation, induces arterial atherosclerosis and thrombosis and may provide possibilities to test new therapeutic approaches.