Inhibition of iNOS induces antidepressant-like effects in mice: Pharmacological and genetic evidence

Recent evidence has suggested that systemic administration of non-selective NOS inhibitors induces antidepressant-like effects in animal models. However, the precise involvement of the different NOS isoforms (neuronal-nNOS and inducible-iNOS) in these effects has not been clearly defined yet. Considering that mediators of the inflammatory response, that are able to induce iNOS expression, can be increased by exposure to stress, the aim of the present study was to investigate iNOS involvement in stress-induced behavioral consequences in the forced swimming test (FST), an animal model sensitive to antidepressant drugs. Therefore, we investigated the effects induced by systemic injection of aminoguanidine (preferential iNOS inhibitor), 1400W (selective iNOS inhibitor) or n-propyl-L-arginine (NPA, selective nNOS inhibitor) in mice submitted to the FST. We also investigated the behavior of mice with genetic deletion of iNOS (knockout) submitted to the FST. Aminoguanidine significantly decreased the immobility time (IT) in the FST. 1400W but not NPA, when administered at equivalent doses considering the magnitude of their Ki values for iNOS and nNOS, respectively, reduced the IT, thus suggesting that aminoguanidine-induced effects would be due to selective iNOS inhibition. Similarly, iNOS KO presented decreased IT in the FST when compared to wild-type mice. These results are the first to show that selective inhibition of iNOS or its knockdown induces antidepressant-like effects, therefore suggesting that iNOS-mediated NO synthesis is involved in the modulation of stress-induced behavioral consequences. Moreover, they further support NO involvement in the neurobiology of depression. This article is part of a Special Issue entitled 'Anxiety and Depression'. (C) 2011 Elsevier Ltd. All rights reserved.

Analytical method for determination of nitric oxide in zebrafish larvae: Toxicological and pharmacological applications

Zebrafish are currently used at various stages of the drug discovery process and can be a useful and cost-effective alternative to some mammalian models. Nitric oxide (NO) plays an important role in physiology of zebrafish. The availability of appropriate analytical techniques to quantify the NO is crucial for studying its role in physiological and pathological conditions. This work aimed at establishing a high-performance liquid chromatography method for determination of NO levels in zebrafish larvae. Attempts were also made to assess the normal levels of NO at the first days postfertilization and the possible changes under pathological conditions. The method validation was quantitatively evaluated in terms of sensitivity, specificity, precision, accuracy, linearity, and recovery. NO levels from zebrafish larvae at the first days postfertilization and larvae challenged to N(G)-nitro-L-arginine methyl ester, sodium nitroprusside, Escherichia coil lipopolysaccharide, and copper sulfate were analyzed. The samples were derivatized with 2,3-diaminonaphthalene, and fluorescence detection was used for the indirect determination of NO. The method showed a good performance for all validation parameters evaluated and was efficient to monitor changes in NO concentration under physiological and pathophysiological conditions. This method might represent a powerful tool to be applied in NO studies with zebrafish larvae. (C) 2011 Elsevier Inc. All rights reserved.

Kinetic Analysis of Gill (Na+,K+)-ATPase Activity in Selected Ontogenetic Stages of the Amazon River Shrimp, (Decapoda, Palaemonidae): Interactions at ATP- and Cation-Binding Sites

We investigated modulation by ATP, Mg2+, Na+, K+ and NH4 (+) and inhibition by ouabain of (Na+,K+)-ATPase activity in microsomal homogenates of whole zoeae I and decapodid III (formerly zoea IX) and whole-body and gill homogenates of juvenile and adult Amazon River shrimps, . (Na+,K+)-ATPase-specific activity was increased twofold in decapodid III compared to zoea I, juveniles and adults, suggesting an important role in this ontogenetic stage. The apparent affinity for ATP ( (M) = 0.09 +/- A 0.01 mmol L-1) of the decapodid III (Na+,K+)-ATPase, about twofold greater than the other stages, further highlights this relevance. Modulation of (Na+,K+)-ATPase activity by K+ also revealed a threefold greater affinity for K+ ( (0.5) = 0.91 +/- A 0.04 mmol L-1) in decapodid III than in other stages; NH4 (+) had no modulatory effect. The affinity for Na+ ( (0.5) = 13.2 +/- A 0.6 mmol L-1) of zoea I (Na+,K+)-ATPase was fourfold less than other stages. Modulation by Na+, Mg2+ and NH4 (+) obeyed cooperative kinetics, while K+ modulation exhibited Michaelis-Menten behavior. Rates of maximal Mg2+ stimulation of ouabain-insensitive ATPase activity differed in each ontogenetic stage, suggesting that Mg2+-stimulated ATPases other than (Na+,K+)-ATPase are present. Ouabain inhibition suggests that, among the various ATPase activities present in the different stages, Na+-ATPase may be involved in the ontogeny of osmoregulation in larval The NH4 (+)-stimulated, ouabain-insensitive ATPase activity seen in zoea I and decapodid III may reflect a stage-specific means of ammonia excretion since functional gills are absent in the early larval stages.

Common chromosomal imbalances and stemness-related protein expression markers in endometriotic lesions from different anatomical sites: the potential role of stem cells

Endometriosis is a multifactorial gynecological disease characterized by the presence of functional endometrium-like tissue in ectopic sites. Several studies have focused on elucidating the immunological, endocrine, environmental and genetic factors involved in endometriosis. However, its pathogenesis is still unclear. High-resolution comparative genomic hybridization was applied to screen for genomic imbalances in laser microdissected stromal and epithelial cells from 20 endometriotic lesions and three samples of eutopic endometrium derived from eight patients. The expression of seven stemness-related markers (CD9, CD13, CD24, CD34, CD133, CD117/c-Kit and Oct-4) in endometrial tissue samples was evaluated by immunohistochemistry. Samples of eutopic endometrium showed normal genomic profiles. In ectopic tissues, an average of 68 genomic imbalances was detected per sample. DNA losses were more frequently detected and involved mainly 3p, 5q, 7p, 9p, 11q, 16q, 18q and 19q. Many of the genomic imbalances detected were common to endometriotic stroma and epithelia and also among different endometriotic sites from the same patient. These findings suggested a clonal origin of the endometriotic cells and the putative involvement of stem cells. Positive immunostaining for CD9, CD34, c-Kit and Oct-4 markers was detected in isolated epithelial and/or stromal cells in eutopic and ectopic endometrium in the majority of cases. The presence of shared genomic alterations in stromal and epithelial cells from different anatomical sites of the same patient and the expression of stemness-related markers suggested that endometriosis arises as a clonal proliferation with the putative involvement of stem cells.

COMPARING RAINFALL INTENSITY DURATION RELATIONSHIPS FOR SITES OF THE STATE OF SAO PAULO

Rainfall intensity durations relationships are extremely important in the design of systems for mitigating runoff losses. The objective of this work was to compare rainfall depths generated by the PLUVIO 2.1 software, with depths from the standard intensity duration curves developed by MARTINEZ & MAGNI (1999). It was compared rainfall intensities of 10, 20, 30, 60, 120 and 1440 minute durations for 2, 5, 10, 50 and 100 year return periods for 30 sites in the state of Sao Paulo. The results showed that PLUVIO was effective, except in predicting the 24 hours rainfall from 100 year return period events in four locations in the central and eastern regions of the state.

Nuclear abnormalities in erythrocytes and morphometric indexes in the catfish Cathorops spixii (Ariidae) from different sites on the southeastern Brazilian coast

Nuclear abnormalities in erythrocytes (NAE) were taken as biomarkers in the catfish Cathorops spixii (Ariidae) sampled in an estuary little affected by human activity (Cananeia) and in three regions (Santos Channel: SC, Santos Bay: SB and Sao Vicente Channel: SVC) of the Santos-Sao Vicente estuary impacted by various anthropogenic activities. Increases in NAE were observed in fish from SC and SVC sampled in the summer period as compared with specimens from the Cananeia estuary. These results suggest the presence of genotoxic compounds in these regions. However, the absence of significant differences in micronuclei frequency reflects slight mutagenic effects in these individuals. It is possible that the lower NAE frequency in specimens from SB might be associated with the greater remobilization and dilution of chemicals in this region. The low frequency of NAE in C. spixii from the Cananeia estuary is in accordance with the slight anthropogenic influence in this system, and may be suggestive of the absence of genotoxic and mutagenic effects in these organisms.

Karyotype characterization reveals an up and down of 45S and 5S rDNA sites in Crotalaria (Leguminosae-Papilionoideae) species of the section Hedriocarpae subsection Macrostachyae

The genus Crotalaria is one of the largest within the family Leguminosae-Papilionoideae, with more than 600 species. However, few karyotypes have been described. In the present paper, five species belonging to the section Hedriocarpae were studied (subsection Machrostachyae), in order to better understand chromosomal evolution in Crotalaria. The results reveals that all species presented 2n = 2x = 16 with symmetrical karyotypes, and slight differences in the chromosome morphology. A secondary constriction was identified at short arm of the pair 1. The 45S rDNA was mapped in the secondary constriction and adjacent heterochromatin (NOR-heterochromatin) and a minor site was identified in C. ochroleuca. The 5S rDNA was mapped linked to 45S rDNA at chromosome 1 short arm in all species. Additional sites for 5S rDNA were identified in C. pallida, C. striata and C. mucronata. Heterochromatin blocks around the centromeres are not CMA(+) neither DAPI(+). The karyotypes of the subsection Macrostachyae are characterized by an inversion at chromosome pair one in relation to previous specialized floral species analyzed. Additional sites of 45S and 5S rDNA were assumed to be a result of transposition events by different ways. The results suggest heterochromatin differentiation and the position of ribosomal genes indicates chromosomal rearrangements during evolution. Karyotype characteristics corroborate the morphological infrageneric classification.

TL dating of pottery fragments from four archaeological sites in Taquari Valley, Brazil

Sixty-three pottery fragments from four archaeological sites, numbered RST110, RST101, RST114 and RST114, in the Taquari Valley, vicinity of the city of Lajeado, Rio Grande do Sul state, southern Brazil, have been dated by the thermoluminescence method. Some of them from RST110 and RST101 are as old as 1400-1200 years, whereas those from RST114 and RST107 are younger than 800 years. This result indicates that RST101 and RST110 were peopled earlier than RST114 and RST107. The recent dates found are 302, 295 and 146 years and they are possible, since the first German immigrants who arrived in this region encountered Tupi-Guarani Indians still living there. One interesting result refers to the glow curves of quartz grains RST110, RST101 and RST114 that differ from the glow curves of RST107 quartz grains.

DNA damage defines sites of recurrent chromosomal translocations in B lymphocytes

Recurrent chromosomal translocations underlie both haematopoietic and solid tumours. Their origin has been ascribed to selection of random rearrangements, targeted DNA damage, or frequent nuclear interactions between translocation partners; however, the relative contribution of each of these elements has not been measured directly or on a large scale. Here we examine the role of nuclear architecture and frequency of DNA damage in the genesis of chromosomal translocations by measuring these parameters simultaneously in cultured mouse B lymphocytes. In the absence of recurrent DNA damage, translocations between Igh or Myc and all other genes are directly related to their contact frequency. Conversely, translocations associated with recurrent site-directed DNA damage are proportional to the rate of DNA break formation, as measured by replication protein A accumulation at the site of damage. Thus, non-targeted rearrangements reflect nuclear organization whereas DNA break formation governs the location and frequency of recurrent translocations, including those driving B-cell malignancies.

Performance of surveillance cultures at different body sites to identify asymptomatic Staphylococcus aureus carriers

The objective was to evaluate the performance of surveillance cultures at various body sites for Staphylococcus aureus colonization in pregnant women and newborns (NB) and the factors associated with nasal colonization. For NB, 4 sites were evaluated: nares, oropharynx, perineum, and umbilical stump (birth, third day, and weekly). For pregnant women, 4 sites during labor: anterior nares, anus, perineum, and oropharynx. Nasally colonized patients were compared with colonized only extranasally. Colonization was 53% of 392 pregnant women (methicillin-resistant S. aureus [MRSA]: 4%) and 47% of 382 NB (MRSA: 9%). For newborn patients, the best body site was the umbilical stump (methicillin-susceptible S. aureus [MSSA]: 64%; MRSA: 68%) and the combination of nares + umbilical (MSSA: 86%; MRSA: 91%). Among pregnant women, the best body site was the anterior nares (MSSA: 59%; MRSA: 67%) and the combination of nares + oropharynx (MSSA: 83%; MRSA: 80%). A smaller number of household members were associated with MRSA carriage in pregnant women (2.2 +/- 0.6 versus 3.6 +/- 1.8; P = 0.04). In conclusion, multiple culture sites are needed. Control programs based on surveillance cultures may be compromised. (C) 2012 Elsevier Inc. All rights reserved.

Life History Aspects of Oxyrhopus trigeminus (Serpentes: Dipsadidae) from Two Sites in Southeastern Brazil

We studied the reproduction, sexual dimorphism, and diet of Oxyrhopus trigeminus from two sites in southeastern Brazil. Oxyrhopus trigeminus from Irape Power Plant (IPP) contained vitellogenic follicles and eggs in both rainy and dry seasons and clutch size was not correlated with female snout vent length (SVL). Sexual dimorphism was evident. Females attain larger SVL but males have longer tails. We found three females from Santa Clara Power Plant (SPP) with vitellogenic follicles, all of them collected in the dry season. Mean SVLs of adult females from IPP and SPP were 717.7 mm and 786 mm, respectively. Mean SVL of adult males from IPP was 553.4 mm and the single adult male from SPP was 507 mm. The diet of O. trigeminus from IPP included rodents (46.7%), lizards (33.3%), and birds (20%). The volume of individual prey items was not correlated with snake SVL. The diet of O. trigeminus from SPP included rodents (37.5%), lizards (37.5%), birds (12.5%), and marsupials (12.5%). It seemed that an ontogenetic shift may occur in individuals of this snake species from IPP.

Comparison of INAA elemental composition data between Lago Grande and Osvaldo archaeological sites in the central Amazon: a first perspective

In this work, 50 ceramic fragments from the Lago Grande and 30 from the Osvaldo archaeological site were compared to assess elemental similarities. The aim is to perform a preliminary comparison between the sites, which are located in the central Amazon, Brazil. The analytical technique employed to obtain the ceramics elemental composition was instrumental neutron activation analysis (INAA). The data set obtained was explored by the multivariate statistical techniques of cluster, principal component and discriminant analysis. The analyzed elements were: Na, Lu, U, Yb, La, Th, Cr, Cs, Sc, Fe, Eu, Ce and Hf. The results showed the existence of at least two compositional groups for Lago Grande and Osvaldo. Each compositional group of Osvaldo archaeological site matches with one group of Lago Grande. Correlated with the archaeological background, the results suggest commercial or cultural exchange in the region, which is an indicative of socio-cultural interactions between those sites.

A Combined Study Using Ligand-Based Design, Synthesis, and Pharmacological Evaluation of Analogues of the Acetaminophen Ortho-Regioisomer with Potent Analgesic Activity

A ligand-based drug design study was performed to acetaminophen regioisomers as analgesic candidates employing quantum chemical calculations at the DFT/B3LYP level of theory and the 6-31G* basis set. To do so, many molecular descriptors were used such as highest occupied molecular orbital, ionization potential, HO bond dissociation energies, and spin densities, which might be related to quench reactivity of the tyrosyl radical to give N-acetyl-p-benzosemiquinone-imine through an initial electron withdrawing or hydrogen atom abstraction. Based on this in silico work, the most promising molecule, orthobenzamol, was synthesized and tested. The results expected from the theoretical prediction were confirmed in vivo using mouse models of nociception such as writhing, paw licking, and hot plate tests. All biological results suggested an antinociceptive activity mediated by opioid receptors. Furthermore, at 90 and 120 min, this new compound had an effect that was comparable to morphine, the standard drug for this test. Finally, the pharmacophore model is discussed according to the electronic properties derived from quantum chemistry calculations.

Dyslipidemia: evidence of efficacy of the pharmacological and non-pharmacological treatment in the elderly

The clinical decision to control risk factors for cardiovascular disease (CVD) in the elderly takes the followings into consideration: (1) the elderly life expectancy; (2) the elderly biological age and functional capacity; (3) the role of cardiovascular disease in the elderly group; (4) the prevalence of risk factors in the elderly; and (5) The effectiveness of treatment of risk factors in the elderly. A large number of studies showed the efficacy of secondary and primary prevention of dyslipidemia in the elderly. However, the only trial that included patients over 80 years was the Heart Protection Study (HPS). Statins are considered the first line therapy for lowering low-density lipoprotein cholesterol (LDL-C). Because lifestyle changes are very difficult to achieve, doctors in general tend to prescribe many drugs to control cardiovascular risk factors. However, healthy food consumption remains a cornerstone in primary and secondary cardiovascular prevention and should be implemented by everyone.

Haplotype frequencies based on eight polymorphic sites at the 3' untranslated region of the HLA-G gene in individuals from two different geographical regions of Brazil

The Brazilian population represents an admixture of native Amerindians, Portuguese settlers and Africans who were brought as slaves during the colonization period that began in the 16th century and was followed by waves of immigrations of Europeans and Asians in the 20th century. The contribution of these different ethnic groups to the constitution of Brazilian populations from different geographic regions is variable and, in addition to environmental factors, might act by determining different allele profiles among Brazilian populations from different regions. We studied polymorphic sites at the 3' untranslated region of the HLA-G gene in individuals from a Northeastern Brazilian region and compared them to our previously published data about a Southeastern Brazilian region, located at a distance of 2589 km. Our results showed that most polymorphic sites present a similar distribution in both populations, except for the lower frequency of the +3003C allele in the Northeastern population compared to the Southeastern population. Although differences in genotypic distribution were only significant for the +3003 locus (P = 0.0201), the diversity of haplotypes was distinct for each population. These results are important for casecontrol studies on the association of human leucocyte antigen-G polymorphism with disease and also in terms of the genetic structure of two distinct Brazilian populations.