Electrophysiological evidence for the presence of NR2C subunits of N-methyl-D-aspartate receptors in rat neurons of the nucleus tractus solitarius

The nucleus tractus solitarius (NTS) plays an important role in the control of autonomic reflex functions. Glutamate, acting on N-methyl-D-aspartate (NMDA) and non-NMDA ionotropic receptors, is the major neurotransmitter in this nucleus, and the relative contribution of each receptor to signal transmission is unclear. We have examined NMDA excitatory postsynaptic currents (NMDA-EPSCs) in the subpostremal NTS using the whole cell patch clamp technique on a transverse brainstem slice preparation. The NMDA-EPSCs were evoked by stimulation of the solitary tract over a range of membrane potentials. The NMDA-EPSCs, isolated pharmacologically, presented the characteristic outward rectification and were completely blocked by 50 µM DL-2-amino-5-phosphonopentanoic acid. The I-V relationship of the NMDA response shows that current, with a mean (± SEM) amplitude of -41.2 ± 5.5 pA, is present even at a holding potential of -60 mV, suggesting that the NMDA receptors are weakly blocked by extracellular Mg2+ at near resting membrane potentials. This weak block can also be inferred from the value of 0.67 ± 0.17 for parameter delta obtained from a fit of the Woodhull equation to the I-V relationship. The maximal inward current measured on the I-V relationship was at -38.7 ± 4.2 mV. The decay phase of the NMDA currents was fitted with one exponential function with a decay time constant of 239 ± 51 and 418 ± 80 ms at a holding potential of -60 and +50 mV, respectively, which became slower with depolarization (e-fold per 145 mV). The biophysical properties of the NMDA receptors observed in the present study suggest that these receptors in the NTS contain NR2C subunits and may contribute to the synaptic signal integration.

The modest but growing Brazilian presence in psychiatric, psychobiological and mental health research: assessment of the 1998-2002 period

The objective of the present survey was to assess the Brazilian scientific production in psychiatry, psychobiology, and mental health during the 1998-2002 period. The universities' graduate programs concentrate the vast majority of the scientific production in Brazil. We assessed the annual reports from the graduate programs to the Brazilian Ministry of Education concerning master's and doctoral theses and the articles published in journals indexed by the Institute of Scientific Information (ISI). There are nine Master's and Doctoral graduate programs dedicated to research in psychiatry, neuropsychiatry, psychobiology, and mental health in the country, seven being located in southern states. During the 5-year period, from 1998 to 2002, 186 students received their doctorate degree (37/year). The programs published 637 articles in journals indexed by ISI, the majority of them in journals with an impact factor higher than 2. The research advisors' productivity varied among graduate programs, ranging from 0.6 to 2.0 articles per year in ISI-indexed journals. Despite the substantial barriers faced by the Brazilian scientific community (mainly financial and writing difficulties), Brazil's scientific mental health production is on the rise. The number of articles published in ISI-indexed journals has doubled without a significant increase in the number of graduate theses, suggesting that there was an improvement in both the quality of the scientific production and the productivity of the graduate programs. Based on these data, it is reasonable to predict a tendency to an increase in production over the next few years.

p16INK4 expression in precursor lesions of squamous cell cervical cancer related to the presence of HPV-DNA

The purpose of the present study was to identify the expression of p16INK4 in cervical cancer precursor lesions by immunohistochemistry and to correlate it with lesion grade and presence of human papillomavirus (HPV) infection. Cervical specimens from 144 women seen consecutively at the gynecology outpatient clinic of our institution from December 2003 to May 2005 were analyzed by cytopathology, histopathology, polymerase chain reaction for HPV-DNA, and p16INK4 immunostaining. Histologically normal biopsies, HPV-DNA negative by polymerase chain reaction, were used as control. HPV-DNA prevalence, including the control group, was 68.1% and the prevalence of p16INK4 expression was 55.0%. The percentage of cells stained by p16INK4 ranged from 10 to 100%, both in the group consisting of cervical intraepithelial neoplasia (CIN)1/HPV specimens and in the group of CIN2/CIN3 specimens with P value of 0.0001. p16INK4 expression was 48.3% in the CIN1/HPV group, as opposed to 94.3% in the CIN2/CIN3 group (P = 0.001), showing a statistically significant difference between the two groups. The quantitative method used here is simple and less subjective than the different semiquantitative methods described in the literature. In view of the different definitions of a p16INK4-positive case, it is almost impossible to compare the findings reported by different investigators. This study confirms the association between p16INK4 and CIN2 and CIN3 lesions. Moreover, it shows that some low grade lesions expressed high levels of this protein. This may indicate that such low grade lesions may be predisposed to progress to high grade lesions. This means that p16INK4 may be a strong marker for "neoplastic lesions" induced by HPV and not just an infection marker.

Hydrogen peroxide induces a specific DNA base change profile in the presence of the iron chelator 2,2’ dipyridyl in Escherichia coli

Pretreatment of Escherichia coli cultures with the iron chelator 2,2’-dipyridyl (1 mM) protects against the lethal effects of low concentrations of hydrogen peroxide (<15 mM). However, at H2O2 concentrations equal to or greater than 15 mM, dipyridyl pretreatment increases lethality and mutagenesis, which is attributed to the formation of different types of DNA lesions. We show here that pretreatment with dipyridyl (1 mM) prior to challenge with high H2O2 concentrations (≥15 mM) induced mainly G:C→A:T transitions (more than 100X with 15 mM and more than 250X with 20 mM over the spontaneous mutagenesis rate) in E. coli. In contrast, high H2O2 concentrations in the absence of dipyridyl preferentially induced A:T→T:A transversions (more than 1800X and more than 300X over spontaneous mutagenesis for 15 and 20 mM, respectively). We also show that in the fpg nth double mutant, the rpoB gene mutation (RifS-RifR) induced by 20 mM H2O2 alone (20X higher) was increased in 20 mM H2O2 and dipyridyl-treated cultures (110X higher), suggesting additional and/or different lesions in cells treated with H2O2 under iron deprivation. It is suggested that, upon iron deprivation, cytosine may be the main damaged base and the origin of the pre-mutagenic lesions induced by H2O2.

Anti-parasitic action and elimination of intracellular Toxoplasma gondii in the presence of novel thiosemicarbazone and its 4-thiazolidinone derivatives

Toxoplasma, which infects all eukaryotic cells, is considered to be a good system for the study of drug action and of the behavior of infected host cells. In the present study, we asked if thiosemicarbazone derivatives can be effective against tachyzoites and which morphological and ultrastructural features of host cells and parasites are associated with the destruction of Toxoplasma. The compounds were tested in infected Vero cell culture using concentration screens (0.1 to 20 mM). The final concentration of 1 mM was chosen for biological assay. The following results were obtained: 1) These new derivatives decreased T. gondii infection with an in vitro parasite IC50% of 0.2-0.7 mM, without a significant effect on host cells and the more efficient compounds were 2, 3 (thiosemicarbazone derivatives) and 4 (thiazolidinone derivative); 2) The main feature observed during parasite elimination was continuous morphological disorganization of the tachyzoite secretory system, progressive organelle vesiculation, and then complete disruption; 3) Ultrastructural assays also revealed that progressive vesiculation in the cytoplasm of treated parasites did not occur in the host cell; 4) Vesiculation inside the parasite resulted in death, but this feature occurred asynchronously in different intracellular tachyzoites; 5) The death and elimination of T. gondii was associated with features such as apoptosis-like stage, acidification and digestion of parasites into parasitophorous vacuoles. Our results suggest that these new chemical compounds are promising for the elimination of intracellular parasites by mainly affecting tachyzoite development at 1 mM concentration for 24 h of treatment.

Adherence to drug therapy in kidney disease

Non-adherence to drug therapy has not been extensively studied in patients with chronic kidney disease (CKD). The objective of the present study was to identify determinants of non-adherence to drug therapy in patients with CKD, not on dialysis. A prospective cohort study involving 149 patients was conducted over a period of 12 months. Adherence to drug therapy was evaluated by the self-report method at baseline and at 12 months. Patients who knew the type of drug(s) and the respective number of prescribed pills in use at the visit preceding the interview were considered to be adherent. Patients with cognitive decline were assessed by interviewing their caregivers. Mean patient age was 51 ± 16.7 years. Male patients predominated (60.4%). Univariate analysis performed at baseline showed that non-adherence was associated with older age, more pills taken per day, worse renal function, presence of coronary artery disease, and reliance on caregivers for the administration of their medications. In multivariate analysis, the factors that were significantly associated with non-adherence were daily use of more than 5 pills and drug administration by a caregiver. Longitudinal evaluation showed an increase in non-adherence over time. Medication non-adherence was lower (17.4%) at the baseline period of the study than after 1 year of the study (26.8%). Compared to the baseline period, the percentage of adherent patients who became non-adherent (22%) was lower than the percentage of non-adherent patients who became adherent (50%). In CKD patients not on dialysis, non-adherence was significantly associated with the number of pills taken per day and drug administration by third parties. Adherence is more frequent than non-adherence over time.

Disentangling the effects of insomnia and night work on cardiovascular diseases: a study in nursing professionals

Cardiovascular diseases (CVDs) are known to be associated with poor sleep quality in general populations, but they have not been consistently associated with specific work schedules. Studies of CVD generally do not simultaneously consider sleep and work schedules, but that approach could help to disentangle their effects. We investigated the association between insomnia and a self-reported physician diagnosis of CVD in day and night workers, considering all sleep episodes during nocturnal and diurnal sleep. A cross-sectional study was conducted in 1307 female nursing professionals from 3 public hospitals, using baseline data from the “Health and Work in Nursing - a Cohort Study.” Participants were divided into two groups: i) day workers with no previous experience in night shifts (n=281) and whose data on insomnia were related to nocturnal sleep and ii) those who worked exclusively at night (n=340) and had data on both nocturnal and diurnal sleep episodes, as they often sleep at daytime. Multiple logistic regression analysis was performed. Among day workers, insomnia complaints increased the odds of CVD 2.79-fold (95% CI=1.01-6.71) compared with workers who had no complaints. Among night workers, reports of insomnia during both nocturnal and diurnal sleep increased the odds of reported CVD 3.07-fold (95% CI=1.30-7.24). Workers with insomnia had similar probabilities of reporting CVD regardless of their work schedule, suggesting a relationship to insomnia and not to night work per se. The results also highlighted the importance of including evaluation of all sleep episodes (diurnal plus nocturnal sleep) for night workers.

Accuracy of dose planning for prostate radiotherapy in the presence of metallic implants evaluated by electron spin resonance dosimetry

Radiotherapy is one of the main approaches to cure prostate cancer, and its success depends on the accuracy of dose planning. A complicating factor is the presence of a metallic prosthesis in the femur and pelvis, which is becoming more common in elderly populations. The goal of this work was to perform dose measurements to check the accuracy of radiotherapy treatment planning under these complicated conditions. To accomplish this, a scale phantom of an adult pelvic region was used with alanine dosimeters inserted in the prostate region. This phantom was irradiated according to the planned treatment under the following three conditions: with two metallic prostheses in the region of the femur head, with only one prosthesis, and without any prostheses. The combined relative standard uncertainty of dose measurement by electron spin resonance (ESR)/alanine was 5.05%, whereas the combined relative standard uncertainty of the applied dose was 3.35%, resulting in a combined relative standard uncertainty of the whole process of 6.06%. The ESR dosimetry indicated that there was no difference (P>0.05, ANOVA) in dosage between the planned dose and treatments. The results are in the range of the planned dose, within the combined relative uncertainty, demonstrating that the treatment-planning system compensates for the effects caused by the presence of femur and hip metal prostheses.

Elongated cells of Listeria monocytogenes in biofilms in the presence of sucrose and bacteriocin-producing Leuconostoc mesenteroides A11

Listeria monocytogenes is a foodborne pathogen which may survive in biofilms and persist in food processing plants. In this study, the ability of Leuconostoc mesenteroides (bac+ and bac-) to inhibit biofilm formation by L. monocytogenes ATCC 19115 was studied with stainless steel coupons immersed in BHI broth and BHI broth plus sucrose in combination with the Lactic Acid Bacteria (LAB). Adhered cells were collected with swabs and enumerated on selective agars (Oxford for listeria and MRS for leuconostoc). Leuconostoc mesenteroides bac+ in co-culture with L. monocytogenes was effective to inhibit biofilm formation by listeria for up to 3 hours of incubation, but at 24 hours, biofilm was present in all conditions tested, as confirmed by observations of stainless steel coupons under Scanning Electron Microscopy (SEM). It was also observed that in the presence of L. mesenteroides bac+ in BHI plus sucrose, a high number of elongated cells of L. monocytogenes was present, which may indicate an adaptation response of the pathogen to stress conditions with important implications for food safety.

Iron availability in the presence of β-carotene in different mixtures

Iron availability in the diet is very important because iron deficiency affects a large population in the world. The matrix where iron is present has an influence in its availability. The presence of β-carotene is a factor that alters the availability of iron. This research aims to estimate the iron availability in the presence of β-carotene in food mixtures: M1 = egg and pumpkin; M2 = spinach and pumpkin; M3 = spinach and cabbage; M4 = egg and cabbage; M5 = spinach and carrot; M6 = egg and carrot; M7 = bean and carrot; M8 = bean and pumpkin and M9 = bean and cabbage. After cooking, the following figures were determined: proximate composition, oxalic acid, phytic acid, tannin, iron, iron availability in vitro and β-carotene. The data were analyzed by Tukey test (5%). There were no significant statistical differences for oxalic acid. Tannin presented greater results in mixtures with spinach (M2 and M5); phytic acid was greater in bean samples (M7, M8 and M9) and with spinach (M2). Mixtures M5 e M7, with carrot, presented more β-carotene than the others. The best result for iron availability appeared in mixture (M6). A positive correlation was verified between protein and iron dialysis, and between lipids and iron dialysis. Dietetic fiber was an inhibitor to iron availability. Mixtures with egg showed greater iron availability

Comparison between the availability of iron in the presence of vitamin a and β-carotene in foods and medications

The objective of this work was to verify the availability of iron in the presence of vitamin A as components of foods and in combinations with medicines. The iron available was measured in the presence of vitamin A in foods - common bean (B), beef liver (Li) and carrot (C) - and medicines - Fer-In-Sol® (Fer) (Mead Johnson), Arovit® (A) (Roche) and Neutrofer® (N) (Sigma Pharma) - as well as in combinations of both. β-carotene, vitamin A, total iron, heme and non heme iron, percentage of dialyzable iron and amount of dialyzable iron was determined. Vitamin A and β-carotene had a positive effect on the percentage of iron dialysis. Carrot and liver had a better percentage of dialyzable iron than their respective medicine at similar concentrations. Therefore, we can conclude that there has been an influence of vitamin A over the dialysis of iron, being the mixtures containing liver the ones which achieved the highest concentrations of dialyzable iron, and also that, according to the amounts needed to obtain the daily recommended intake of iron, they are good for consumption.

Determination of presence and quantification of cadmium, lead and copper in Nile tilapia (Oreochromis niloticus) fillets obtained from three cold storage plants in the state of Parana, Brazil

Pisciculture is an economic activity that is steadily growing in the state of Parana, Brazil, and Nile tilapia (Oreochromis niloticus) is one of the widely cultivated species in this state. Tilapia is not only a very nutritious food, but also an important indicator of environmental contamination. This study aimed to verify contamination by cadmium, copper and lead in tilapia fillets, and to compare the found values to international legislations. Were collected 135 samples of tilapia fillets, between July 2006 and May 2007, in three fish stores located in regions west and north of Paraná State. Samples of tilapia fillet were analyzed in relation to the presence of cadmiun, lead and copper, using atomic absorption spectrophotometry. Lead has not been detected in the analyses. Cadmium has been detected in three samples, on concentrations of 0.012 µg.g-1, 0.011 µg.g-1 and 0.014 µg.g-1. Copper has been detected in all fillets, and the average concentration of each cold storage plant was of 0.122 µg.g-1, 0.106 µg.g-1 and 0.153 µg.g-1. The concentrations found in this study are within the limits allowed by both the European and the Australian legislations.

Stigmastadiene and specific extitntion (270 nm) to evaluate the presence of refined oils in virgin olive oil commercialized in Brazil

The increased marketing of olive oil in Brazil has intensified legal requirements to ensure regulation of this product. The measurement of the specific extinction at 270 nm (E 270) and content of stigmastadiene can be used to assess the presence of refined oils in virgin olive oil. During the vegetable oil refining process, compounds with conjugated double bonds are generated from unsaturated fatty acids that absorb at 270 nm and sterols, such as stigmasta-3,5-diene. To compare these parameters, seven samples of extra virgin olive oil and three samples of olive oil (blend of virgin and refined) were analyzed. Among the samples analyzed, four extra virgin samples had levels of stigmastadiene and E 270 higher than expected, among which two were adulterated with seed oil (rich in linoleic acid) and the other two with olive pomace oil. The results demonstrate the higher sensitivity of stigmastadiene to determine the presence of the refined oil in virgin olive oil and good agreement with determining E 270. The latter technique is a simple, quick, and low cost method of determination that can be easily implemented in laboratories to assist in the screening and regulation of olive oils sold in Brazil.

Molecular screening of bovine raw milk for the presence of Shiga toxin-producing Escherichia coli (STEC) on dairy farms

Milkborne transmission of Shiga toxin- producing Escherichia coli (STEC) has raised considerable concern due to recent outbreaks worldwide and poses a threat to public health. The aim of this study was to develop a sensitive and specific multiplex PCR assay to detect the presence of STEC in bovine raw milk. To identify E. coli (ATCC 25922) contamination, the gene uspA was used, and PCR sensitivity and specificity were accessed by testing diluted samples ranging from 2 to 2.0 × 10(6) CFU/mL. To detect STEC, the stx1 and stx2 genes were selected as targets. After reaction standardization, the multiplex assay was tested in raw milk collected from 101 cows on dairy farms. PCR assay for E. coli detection had a specificity of 100% and sensitivity of 79% (P<0.0001), with a lower detection limit of 2 CFU/mL. Multiplex PCR assay had 100% sensitivity for E. coli positive raw milk samples, and 31.1% were contaminated with STEC, 28.3% of stx2, and 1.9% of stx1. The multiplex PCR assay described in the present study can be employed to identify and screen E. coli harboring stx1 and stx2 genes in raw milk on dairy farms and in industries.

Microbiological quality of meals served in nursing homes in the city of Ponta Grossa, Paraná

Abstract The aim of this study was to analyze the microbiological quality of meals served in nursing homes in the city of Ponta Grossa, Paraná, before and after the training of food handlers. The first stage was to perform a checklist conforming with current legislation. The second consisted of the collection of 4 food samples from each location and a microbiological investigation in accordance with the relevant legislation. The third was the training of food handlers in relation to good food handling practices. The fourth was a further microbiological analysis of new samples. The application of a checklist showed that the locations met the requirements of current legislation. Of the 40 samples analyzed, 17.5% (7 samples) were unfit for consumption. Among the unfit samples 15% (6 samples) had coliforms at 45 °C, 2.5% (1 sample) had coagulase-positive staphylococci, 2.5% (1 sample) had Bacillus cereus and 2.5% (1 sample) had Salmonella sp. The results of this study show the importance of controlling the quality of food served to an age group that is prone to health risks.