Molecular identification of cetaceans from the West Atlantic using the E3-I5 region of COI.

Molecular identification is very useful in cases where morphology-based species identification is not possible. Examples for its application in cetaceans include the identification of carcasses of stranded animals in advanced state of decomposition and body parts that are illegally traded. One DNA region that is often used for molecular identification is the Folmer region of the mitochondrial gene cytochrome c oxidase subunit I (COI) (locus 48 to 705 bp). This locus has been used for the identification of several animal species, including whales and dolphins. The goal of the present study was to evaluate the usefulness of another region of COI, the E3-I5 (locus 685 to locus 1179; 495 bp) as a marker for identification of cetaceans from northeastern Canada and northeastern Brazil. The identification markers were successfully obtained for seven cetacean species after performing percent identity and Basic Local Alignment Search Tool analyses. The obtained markers are now publicly available and are useful for the identification of the endangered blue whale (Balaenoptera musculus), common minke whale (B. acutorostrata), vulnerable sperm whale (Physeter macrocephalus), harbor porpoise (Phocoena phocoena), common bottlenose dolphin (Tursiops truncatus), Guiana dolphin (Sotalia guianensis), and melon-headed whale (Peponocephala electra).

Effect of cyclodextrin-loaded cholesterol conjugates on plasma membrane viability of Piau swine breed frozen/thawed spermatozoa.

The objective of this study was to investigate the effect of cyclodextrin-loaded cholesterol conjugates addition to freezing extenders on plasma membrane viability of frozen-thawed spermatozoa of the Piau swine breed. Twenty semen samples were used from five males. The freezing extender was based on lactose-egg yolk extender, added to 2% glycerol, 3% dimethylacetamide. The addition of cyclodextrin-loaded cholesterol conjugates was performed after centrifugation, when semen was diluted with the cooling extender. Four groups were subjected to the following treatment: without addition (group 1); 1.5 mg of cyclodextrin-loaded cholesterol/120 × 10(6) sperm (group 2); 1.5 mg of cyclodextrin-loaded cholestanol/120 × 10(6) sperm (group 3); 1.5 mg of cyclodextrin-loaded desmosterol/120 × 10(6) sperm (group 4). To check post-thawing sperm quality sperm motility and sperm morphology evaluation were used. Additionally, to check sperm viability the hypoosmotic swelling test, supravital staining, and fluorescent assay were used. The mean values recorded for total sperm motility of semen immediately after thawing were 54.5 ± 5.8, 55.5 ± 5.3, 53.7 ± 6.7, and 52.5 ± 6.6% respectively for groups one to four, without difference between themselves (p > 0.05). Regarding fluorescent assay the results were 28.3 ± 13.2, 26.9 ± 12.2, 22.2 ± 11.4, and 32.0 ± 15.3% respectively for groups one to four, also without difference between groups (p > 0,05). Similarly, complementary tests for evaluating the integrity and functionality of the plasma membrane showed no difference between treatments (p > 0.05). In conclusion, use of cyclodextrin-loaded cholesterol conjugates added to the plasma membrane of sperm did not demonstrate any additive effect on increasing and/or maintaining sperm motility.

Reward deficiency and anti-reward in pain chronification.

Converging lines of evidence suggest that the pathophysiology of pain is mediated to a substantial degree via allostatic neuroadaptations in reward- and stress-related brain circuits. Thus, reward deficiency (RD) represents a within-system neuroadaptation to pain-induced protracted activation of the reward circuits that leads to depletion-like hypodopaminergia, clinically manifested anhedonia, and diminished motivation for natural reinforcers. Anti-reward (AR) conversely pertains to a between-systems neuroadaptation involving over-recruitment of key limbic structures (e.g., the central and basolateral amygdala nuclei, the bed nucleus of the stria terminalis, the lateral tegmental noradrenergic nuclei of the brain stem, the hippocampus and the habenula) responsible for massive outpouring of stressogenic neurochemicals (e.g., norepinephrine, corticotropin releasing factor, vasopressin, hypocretin, and substance P) giving rise to such negative affective states as anxiety, fear and depression. We propose here the Combined Reward deficiency and Anti-reward Model (CReAM), in which biopsychosocial variables modulating brain reward, motivation and stress functions can interact in a 'downward spiral' fashion to exacerbate the intensity, chronicity and comorbidities of chronic pain syndromes (i.e., pain chronification).

Prospective molecular markers for the identification of illegally traded angelsharks (Squatina) and dolphin (Sotalia guianensis).

Endangered angelsharks and a protected dolphin species are illegally traded in Brazil. In this study, we determined prospective molecular markers for detecting these species in the trade of angelshark carcasses and 'dolphin' eyeball amulets. We compiled publicly available as well as new and unpublished cytochrome b (cyt b) DNA sequences for species involved in these trades. These sequences were digested in silico using restriction enzymes. We then described prospective polymerase chain reaction (PCR)-restriction fragment length polymorphism markers for distinguishing between protected species and the species whose trade was legally allowed in these two trade groups. The prospective marker for identifying angelshark carcasses consists of cyt b PCR and digestion by BstXI, BsgI, BspMI, BsrDI, and HaeII restriction enzymes. The prospective marker for identifying eyeball amulets consists of cyt b PCR and digestion by ApoI, BtsI, HindII, BsaAI, BplI, and SspI restriction enzymes. This is the first study to deposit in GenBank cyt b sequences for the angelshark species Squatina argentina, Squatina guggenheim, and Squatina occulta. Moreover, the S. argentina haplotype is the first DNA sequence for this species deposited in GenBank.

Association between amygdala reactivity and a dopamine transporter gene polymorphism.

Essential for detection of relevant external stimuli and for fear processing, the amygdala is under modulatory influence of dopamine (DA). The DA transporter (DAT) is of fundamental importance for the regulation of DA transmission by mediating reuptake inactivation of extracellular DA. This study examined if a common functional variable number tandem repeat polymorphism in the 3' untranslated region of the DAT gene (SLC6A3) influences amygdala function during the processing of aversive emotional stimuli. Amygdala reactivity was examined by comparing regional cerebral blood flow, measured with positron emission tomography and [(15)O]water, during exposure to angry and neutral faces, respectively, in a Swedish sample comprising 32 patients with social anxiety disorder and 17 healthy volunteers. In a separate US sample, comprising 85 healthy volunteers studied with blood oxygen level-dependent functional magnetic resonance imaging, amygdala reactivity was assessed by comparing the activity during exposure to threatening faces and neutral geometric shapes, respectively. In both the Swedish and the US sample, 9-repeat carriers displayed higher amygdala reactivity than 10-repeat homozygotes. The results suggest that this polymorphism contributes to individual variability in amygdala reactivity.

Publication date and author spelling for the hidden angelshark Squatina occulta.

Two dates of publication have been associated with the hidden angelshark Squatina occulta. Additionally, the name of the second author of this species has been cited with different spellings in different publications. Both inconsistencies are addressed in this study. It is suggested that the hidden angelshark be consistently cited as Squatina occulta Vooren & da Silva 1991.

Identification and validation of differentially expressed genes from pig skeletal muscle.

Pig is an important animal for meat production; this is generally associated with characteristics determined prenatally during myogenesis. Expressed sequence tags (EST) can provide direct information on the transcriptome and indirect information on the relation between the genome and phenotype, giving information about differentially expressed genes (DEG). In this work, the identification and annotation of DEG from EST libraries of three pig breeds (Duroc, Large White and Local Breed Piau) were performed followed by real-time PCR analyses during pre- and postnatal stages (21, 40, 70 and 90 days of pregnancy and 107, 121 and 171 days postnatal) from commercial breed animals for analysis of genes expression levels. Therefore, 34 genes differentially expressed were identified, of which 21 grouped in a network related with muscle development. From this, the expression profile of 13 genes was measured, to confirm their relationship with myogenesis like ANKRD2, MYBPC1, NEB and MYL2. These genes showed a prenatal high expression in this study. Besides, novels candidates for muscle development (TP53 and DCTN1) were listed. These findings can contribute to better explaining gene function mechanism and are helpful in uncovering the pathways that mediate pre- and postnatal skeletal muscle development in vertebrates.

Non-carious cervical lesions: influence of morphology and load type on biomechanical behaviour of maxillary incisors.

BACKGROUND: The aim of this study was to measure the effect of simulating two different loads on maxillary incisors displaying eight morphological types of non-carious cervical lesions, unrestored and restored with an adhesive restoration, by quantifying the stress distributions generated using finite element analysis. METHODS: Virtual models of maxillary incisors were generated using the CAD software (RhinoCeros). After composing virtual dental and supporting structures with and without non-carious cervical lesions, each model was meshed using a control mesh device (ANSYS Finite Element Analysis Software). All of the virtual models were subjected to two load types, oblique load and vertical load, to simulate occlusal forces of 100 N each. Comparisons were made between simulated teeth with non-carious cervical lesions, with and without composite resin restorations, and a simulated sound tooth. Data summarizing the stress distributions were obtained in MPa using von Mises criteria. RESULTS: Oblique loading on simulated non-carious cervical lesions resulted in greater stress concentration compared with vertical loading, and non-carious cervical lesions with acute angles displayed higher stress concentrations at the depth of the lesion. Restoring the lesions with an adhesive restoration, such as composite resin, appeared to overcome this stress concentration. CONCLUSIONS: Restoring NCCLs with adhesive restorative materials, such as a nanohybrid composite resin, appears to recover the biomechanical behaviour similar to sound teeth.

Haematological response of curimbas Prochilodus lineatus, naturally infected with Neoechinorhynchus curemai.

This study evaluated the haematological response of curimbas Prochilodus lineatus, naturally infected with Neoechinorhynchus curemai (Acanthocephala: Neoechinorhynchidae). Thirty-seven fish were captured in October 2010 from the Mogi Guaçu River, Porto Ferreira, SP, Brazil. Infected fish presented increased mean corpuscular volume of erythrocytes, and lower thrombocyte and higher monocyte counts than uninfected fish.

Reevaluation of RAPD markers involved in a case of stingray misidentification (Dasyatidae: Dasyatis).

We investigated a reported case of stingray Dasyatis americana misidentification not detected in a published study using the random amplified polymorphic DNA (RAPD) technique. If the referred specimen (landed by fisheries in Ceará, northeastern Brazil) was misidentified (as Dasyatis centroura) in the field, why did its RAPD data fail to clarify the mistake? Was it due to limitations of RAPD markers or perhaps to a taxonomic issue? Contrary to our initial expectations, neither of these hindered the detection of the misidentification. After reanalyzing the primary genetic data associated with the misidentified specimen (PCR gel photographs and/or matrices of presence/absence of markers for six RAPD primers), we found that the RAPD markers were sufficient to correctly assign the misidentified specimen to its proper species identity. In the original study, the specimen misidentification was neither noticed by the authors nor apparent in the published article due to how their results were interpreted and presented.

Transcript profiling of expressed sequence tags from semimembranosus muscle of commercial and naturalized pig breeds.

In general, genetic differences across different breeds of pig lead to variation in mature body size and slaughter age. The Commercial breeds Duroc and Large White and the local Brazilian breed Piau are ostensibly distinct in terms of growth and muscularity, commercial breeds are much leaner while local breeds grow much slower and are fat type pigs. However, the genetic factors that underlie such distinctions remain unclear. We used expressed sequence tags (ESTs) to characterize and compare transcript profiles in the semimembranosus muscle of these pig breeds. Our aim was to identify differences in breed-related gene expression that might influence growth performance and meat quality. We constructed three non-normalized cDNA libraries from semimembranosus muscle, using two samples from each one, of these three breeds; 6902 high-quality ESTs were obtained. Cluster analysis was performed and these sequences were clustered into 3670 unique sequences; 24.7% of the sequences were categorized as contigs and 75.3% of the sequences were singletons. Based on homology searches against the SwissProt protein database, we were able to assign a putative protein identity to only 1050 unique sequences. Among these, 58.5% were full-length protein sequences and 17.2% were pig-specific sequences. Muscle structural and cytoskeletal proteins, such as actin, and myosin, were the most abundant transcripts (16.7%) followed by those related to mitochondrial function (12.9%), and ribosomal proteins (12.4%). Furthermore, ESTs generated in this study provide a rich source for identification of novel genes and for the comparative analysis of gene expression patterns in divergent pig breeds.

Gene expression in swine granulosa cells and ovarian tissue during the estrous cycle.

The components of the insulin-like growth factor (IGF) system appear to be involved in regulation of ovarian follicular growth and atresia in the pig. We investigated the expression pattern of mRNAs for IGF1 (IGF1), its binding proteins (IGFBP1, IGFBP2, IGFBP3, and IGFBP5), and epidermal growth factor in swine follicle cells and ovarian tissue throughout the estrous cycle using the real-time quantitative PCR technique. The results of gene expression were analyzed using linear regression with gene expression as a dependent variable and days of estrous cycle as an independent variable. Additionally, an analysis was made of the correlation of expression levels with plasma concentration of follicle-stimulating hormone, luteinizing hormone, estradiol-17ß, progesterone, and prolactin. Expression of mRNA of all of these genes was detected in granulosa cells and ovarian tissue. IGFBP3 mRNA showed a quadratic expression pattern (P ≤ 0.001) and was significantly and positively correlated with progesterone (r = 0.81; P ≤ 0.01) but negatively correlated with prolactin (r = -0.596; P ≤ 0.05). Expression of the other genes was unaffected by the stage of the estrous cycle. Real-time quantitative PCR effectively detected all transcripts, including the very low levels of IGFBP1 transcripts, and could be used for studies of follicle dynamics.

Ground roasted peanuts leads to a lower post-prandial glycemic response than raw peanuts.

INTRODUCTION: Few studies have evaluated the effect of nuts processing on the glycemic response and satiety. OBJECTIVE: To evaluate the effect of peanut processing on glycemic response, and energy and nutrients intake. METHOD: Thirteen healthy subjects (4 men and 9 women), with a mean age of 28.5 ± 10 years, BMI 22.7 ± 2.5 kg/m², and body fat 23.7 ± 5.7% participated in this randomized crossover clinical trial. After 10-12 h of fasting, one of the following types of test meals were consumed: raw peanuts with skin (RPS), roasted peanuts without skin, ground-roasted peanuts without skin (GRPWS) or control meal. The test meals had the same nutrient composition, and were consumed with 200 ml of water in 15 minutes. Glycemic response was evaluated 2 hours after each meal. Energy and nutrients intake were assessed through diet records reflecting the habitual food intake and food consumption 24 hours after the ingestion of test meal. RESULT: The area under the glycemic response curve after GRPWS was lower (p = 0.02) the one obtained for RPS. There was no treatment effect on energy intake, macronutrients and fiber consumption after the test meal. CONCLUSION: The consumption of ground-roasted peanuts may favor the control and prevention of diabetes due to its reduction on postprandial glucose response. However, more prospective studies are needed to confirm this hypothesis.

T lymphocyte subsets and cytokine production by graft-infiltrating cells in FSGS recurrence post-transplantation.

BACKGROUND: Focal segmental glomerulosclerosis (FSGS) aetiology remains undefined although a derangement of lymphocytes and monocytes macrophages, at least, has been strongly suspected. We report the graft-infiltrating phenotypes and their cytokine production in a case of FSGS recurrence post-transplantation. METHODS: The kidney transplant recipient suffered immediate FSGS recurrence. Aspiration biopsies were done at the first and second week post-surgery and were analysed by flow cytometry. The cytokine analysis was done on aspiration sample culture supernatants and serum by enzyme-linked immunosorbent assay. RESULTS: High expression of CD3CD69, CD3CD71 and CD4CD29 was found on infiltrating lymphocytes. Biopsy cultures pointed to a Th0/Th1 pattern of cytokine production as well as significant synthesis of transforming growth factor-beta1. Interestingly, monocyte chemokines were absent. CONCLUSION: We report evidence of intragraft lymphocyte activation in the early days of FSGS recurrence. Aspiration biopsy cultures showed failure of cyclosporin A to inhibit interleukin-2 (IL-2) production by infiltrating lymphocytes. If our findings are confirmed in similar patients, a trial with anti-IL-2-receptor antibody could be warranted.

Government policy and fertility regulation: unintended consequences and perverse effects.

PIP: Since some argue that the recent and marked fertility decline experienced in Brazil is related to institutional changes resulting from public policies promoted by the federal government since 1964, the author attempts to shed light upon the role played by such policies upon fertility regulation. Fertility in Brazil and the main explanatory theories are first discussed. The paper then considers the role played by the growth of the consumer society, social security coverage, mass media, and the medicalization of society upon changing patterns of fertility regulation in Brazil. The discussion of government policies and fertility regulation includes consideration of consumer credit policy, social security benefit policy, telecommunications policy, and health care policy. One salient conclusion of the analysis is that the lack of a policy to provide fertility regulation mechanisms other than through the marketplace hurts relatively poor populations.^ieng