Anatomical features of the urethra and urinary bladder catheterization in female mice and rats. An essential translational tool

PURPOSE: To present fundamental anatomical aspects and technical skills necessary to urethra and urinary bladder catheterization in female mice and rats. METHODS: Urethral and bladder catheterization has been widely utilized for carcinogenesis and cancer research and still remains very useful in several applications: from toxicological purposes as well as inflammatory and infectious conditions to functional aspects as bladder dynamics and vesicoureteral reflux, among many others. RESULTS: Animal models are in the center of translational research and those involving rodents are the most important nowadays due to several advantages including human reproducibility, easy handling and low cost. CONCLUSIONS: Although technical and anatomical pearls for rodent urethral and bladder access are presented as tackles to the advancement of lower urinary tract preclinical investigation in a broaden sight, restriction to female animals hampers the male microenvironment, demanding future advances.

Electromyographic analysis of masseter and anterior temporalis muscle in sleep bruxers after occlusal splint wearing

Bruxism is widely defined as an anxiety response to environmental stress. Occlusal splints are frequently used in sleep bruxism, to protect teeth from damage resulting from the contraction force of mandibular muscles, or to reduce the orofacial pain by relaxing masticatory muscles. Surface electromyography (EMG) of the right and left masseter and temporalis muscles was performed in 15 women presenting sleep bruxism and temporomandibular disorders related to occupational stress, after nocturnal use of the occlusal splint. The EMG signals were recorded twice per patient: After a work shift (pre-splint) and after a night of sleep with the occlusal splint (post-splint) before a new workday. The parametric t-paired test was used to compare differences of the RMS amplitude between pre and post-splint records, for resting and maximal clenching effort. The level of significance for each comparison was set to p < 0.05. The results of the study supports the premise that the use of occlusal splint reduces EMG activity in the masseter and anterior temporalis muscles, in patients who presented with sleep bruxism related to occupational stress. © 2011 Elsevier Ltd.

Cytogenetic analysis in Thoracocharax stellatus (Kner, 1858) (Characiformes, Gasteropelecidae) from Paraguay river basin, Mato Grosso, Brazil

Thoracocharax stellatus (Characiformes, Gasteropelecidae) is a small Neotropical species of fish, widely distributed in several rivers of South America. Evidence for karyotype heteromorphysm in populations from different geographical regions has been reported for this species. In this way, populations of T. stellatus from the Paraguay River basin were cytogenetically characterized and the results were compared with other studies performed in the same species but from different basins. The results showed a diploid number of 2n = 54 for T. stellatus, with chromosomes arranged in 6 metacentric (m), 6 submetacentric (sm), 2 subtelocentric (st) and 40 acrocentric (a), for both sexes, with a simple Nucleolus Organiser Region (NOR) system reported by the techniques of silver nitrate impregnation and fluorescent in situ hybridisation (FISH) using 18S rDNA sequences as probe. The distribution of constitutive heterochromatin, observed by the C-band technique and Chromomycin A3 staining showed great similarity among the analyzed populations and consists mainly of discrete blocks in the pericentromeric and telomeric regions of most chromosomes. The presence of female heterogamety was alsoobserved indicating a ZZ/ZW system with W chromosome almost totally heterochromatic. The results also show cytogenetic diversity of the group and are useful to understand the mechanisms of karyotype evolution of the family. © Edson Lourenço da Silva et al.

Proposal of a simplified process to correct the phase decoupling using modal analysis

Modal analysis is widely approached in the classic theory of transmission line modeling. This technique is applied to model the three-phase representation of conventional electric systems taking into account their self and mutual electrical parameters. However the methodology has some particularities and inaccuracies for specific applications which are not clearly described in the basic references of this topic. This paper provides a thorough review of modal analysis theory applied to line models followed by an original and simple procedure to overcome the possible errors embedded in the modal decoupling through the three-phase system modeling. © 2012 IEEE.

HDAC inhibition decreases XIST expression on female IVP bovine blastocysts

During initial development, both X chromosomes are active in females, and one of them must be silenced at the appropriate time in order to dosage compensate their gene expression levels to male counterparts. Silencing involves epigenetic mechanisms, including histone deacetylation. Major X chromosome inactivation (XCI) in bovine occurs between hatching and implantation, although in vitro culture conditions might disrupt the silencing process, increasing or decreasing X-linked gene expression. In this study, we aimed to address the roles of histone deacetylase inhibition by trichostatin A (TSA) on female preimplantation development.We tested the hypothesis that by enhancing histone acetylation, TSA would increase the percentage of embryos achieving 16-cell stage, reducing percentage of embryos blocked at 8-cell stage, and interfere with XCI in IVF embryos. We noticed that after TSA treatment, acetylation levels in individual blastomeres of 8-16 cell embryos were increased twofold on treated embryos, and the samewas detected for blastocysts. Changes among blastomere levels within the same embryo were diminished on TSA group, as low-acetylated blastomeres were no longer detected. The percentage of embryos that reached the 5th cleavage cycle 118 h after IVF, analyzed by Hoechst staining, remained unaltered after TSA treatment. Then, we assessed XIST and G6PD expression in individual female bovine blastocysts by quantitative real-time PCR. Even though G6PD expression remained unaltered after TSA exposure, XIST expression was eightfold decreased, and we also detected a major decrease in the percentage of blastocysts expressing detectable XIST levels after TSA treatment. Based on these results, we conclude that HDAC is involved on XCI process in bovine embryos, and its inhibition might delay X chromosome silencing and attenuate aberrant XIST expression described for IVF embryos. © 2013 Society for Reproduction and Fertility.

Comparative analysis between three methods of bone estimating age in individuals with down syndrome by mode of the hand and wrist ray

The wrist and hand region has been the most commonly used for estimating age and osseous development due to the great number of ossification centers. The aim was to determine which method, Tanner & Whitehouse's (TW3), Greulich & Pyle's (GP) or Eklof & Ringertz's, more closely relates to the chronological age in subjects with Down syndrome with chronological ages between 61 and 180 months, using wrist and hand radiographs. The sample consisted of 85 radiographs, 52 of males and 33 of females. Eklof & Ringertz's method was computerized (Radiomemory). Greulich & Pyle's atlas was used and compared with the wrist and hand radiographs. For the TW3 method, 13 ossification centers were evaluated; for each one of them, there are seven or eight development stages to which scores are assigned; these scores are then added and the results are transformed into osseous age values. No statistically significant differences were observed between the male and female genders for methods TW3 and GP, contrasting with the observed differences for the Eklof & Ringertz method. Correlation (r2) between osseous and chronological ages was 0.8262 for TW3 and 0.7965 for GP, while for the method of Eklof & Ringertz, it was 0.7656 for females and 0.8353 for males. The author concluded that the osseous age assessment method that better related to the chronological age was the TW3, followed by Greulich & Pyle's and Eklof & Ringertz's.

Analysis of genetic correlations of hip height with selection indices and mature weight in Nelore cattle

Body size is directly related to the productive and reproductive performance of beef cattle raised under free-range conditions. In an attempt to better plan selection criteria, avoiding extremes in body size, this study estimated the heritabilities and genetic correlations of yearling hip height (YH) and mature hip height (MH) with selection indices obtained at weaning (WI) and yearling (YI) and mature weight (MW). Data from 102,373 Nelore animals born between 1984 and 2010, which belong to 263 farms that participate in genetic evaluation programmes of beef cattle conducted in Brazil and Paraguay, were used. The (co)variance components and genetic parameters were estimated by Bayesian inference in multi-trait analysis using an animal model. The mean heritabilities for YH, MH and MW were 0. 56 ± 0. 06, 0. 47 ± 0. 02 and 0. 42 ± 0. 02, respectively. The genetic correlation of YH with WI (0. 13 ± 0. 01) and YI (0. 11 ± 0. 01) was practically zero, whereas a higher correlation was observed with MW (0. 22 ± 0. 03). Positive genetic correlations of medium magnitude were estimated between MH and WI and YI (0. 23 ± 0. 01 and 0. 43 ± 0. 02, respectively). On the other hand, a high genetic correlation (0. 68 ± 0. 03) was observed between the indicator traits of mature body size (MH and MW). Considering the top 20 % of sire (896 sires) in terms of breeding values for the yearling index, the rank sire correlations between breeding values for MH and MW was 0. 62. In general, the results indicate that selection based on WI and YI should not lead to important changes in YH. However, an undesired correlated response in mature cow height is expected, particularly when selection is performed using YI. Therefore, changes in the body structure of Nelore females can be obtained when MH and MW is used as a selection criterion for cows. © 2012 Institute of Plant Genetics, Polish Academy of Sciences, Poznan.

Analysis of HCV quasispecies dynamic under selective pressure of combined therapy

Background: The quasispecies composition of Hepatitis C virus (HCV) could have important implications with regard to viral persistence and response to interferon-based therapy. The complete NS5A was analyzed to evaluate whether the composition of NS5A quasispecies of HCV 1a/1b is related to responsiveness to combined interferon pegylated (PEG-IFN) and ribavirin therapy.Methods: Viral RNA was isolated from serum samples collected before, during and after treatment from virological sustained responder (SVR), non-responder (NR) and the end-of-treatment responder patients (ETR). NS5A region was amplified, cloned and sequenced. Six hundred and ninety full-length NS5A sequences were analyzed.Results: This study provides evidence that lower nucleotide diversity of the NS5A region pre-therapy is associated with viral clearance. Analysis of samples of NRs and the ETRs time points showed that genetic diversity of populations tend to decrease over time. Post-therapy population of ETRs presented higher genetic distance from baseline probably due to the bottleneck phenomenon observed for those patients in the end of treatment. The viral effective population of those patients also showed a strong decrease after therapy. Otherwise, NRs demonstrated a continuous variation or stability of effective populations and genetic diversity over time that did not seem to be related to therapy. Phylogenetic relationships concerning complete NS5A sequences obtained from patients did not demonstrate clustering associated with specific response patterns. However, distinctive clustering of pre/post-therapy sequences was observed. In addition, the evolution of quasispecies over time was subjected to purifying or relaxed purifying selection. Codons 157 (P03), 182 and 440 (P42), 62 and 404 (P44) were found to be under positive selective pressure but it failed to be related to the therapy.Conclusion: These results confirm the hypothesis that a relationship exists between NS5A heterogeneity and response to therapy in patients infected with chronic hepatitis C. © 2013 Jardim et al.; licensee BioMed Central Ltd.

Determination of disease biomarkers in Eucalyptus by comprehensive two-dimensional gas chromatography and multivariate data analysis

In this paper is reported the use of the chromatographic profiles of volatiles to determine disease markers in plants - in this case, leaves of Eucalyptus globulus contaminated by the necrotroph fungus Teratosphaeria nubilosa. The volatile fraction was isolated by headspace solid phase microextraction (HS-SPME) and analyzed by comprehensive two-dimensional gas chromatography-fast quadrupole mass spectrometry (GC. ×. GC-qMS). For the correlation between the metabolic profile described by the chromatograms and the presence of the infection, unfolded-partial least squares discriminant analysis (U-PLS-DA) with orthogonal signal correction (OSC) were employed. The proposed method was checked to be independent of factors such as the age of the harvested plants. The manipulation of the mathematical model obtained also resulted in graphic representations similar to real chromatograms, which allowed the tentative identification of more than 40 compounds potentially useful as disease biomarkers for this plant/pathogen pair. The proposed methodology can be considered as highly reliable, since the diagnosis is based on the whole chromatographic profile rather than in the detection of a single analyte. © 2013 Elsevier B.V..

Electroglottographic analysis of actresses and nonactresses' voices in different levels of intensity

Background: Previous studies with long-term average spectrum (LTAS) showed the importance of the glottal source for understanding the projected voices of actresses. In this study, electroglottographic (EGG) analysis was used to investigate the contribution of the glottal source to the projected voice, comparing actresses and nonactresses' voices, in different levels of intensity. Method: Thirty actresses and 30 nonactresses sustained vowels in habitual, moderate, and loud intensity levels. The EGG variables were contact quotient (CQ), closing quotient (QCQ), and opening quotient (QOQ). Other variables were sound pressure level (SPL) and fundamental frequency (F0). A KayPENTAX EGG was used. Variables were inputted in a general linear model. Results/Discussion: Actresses showed significantly higher values for SPL, in all levels, and both groups increased SPL significantly while changing from habitual to moderate and further to loud. There were no significant differences between groups for EGG quotients. There were significant differences between the levels only for F0 and CQ for both groups. Conclusion: SPL was significantly higher among actresses in all intensity levels, but in the EGG analysis, no differences were found. This apparently weak contribution of the glottal source in the supposedly projected voices of actresses, contrary to previous LTAS studies, might be because of a higher subglottal pressure or perhaps greater vocal tract contribution in SPL. Results from the present study suggest that trained subjects did not produce a significant higher SPL than untrained individuals by increasing the cost in terms of higher vocal fold collision and hence more impact stress. Future researches should explore the difference between trained and nontrained voices by aerodynamic measurements to evaluate the relationship between physiologic findings and the acoustic and EGG data. Moreover, further studies should consider both types of vocal tasks, sustained vowel and running speech, for both EGG and LTAS analysis. © 2013 The Voice Foundation.

Female germ cell renewal during the annual reproductive cycle in Ostariophysians fish

The objective was to characterize female germ cell renewal during the annual reproductive cycle in two species of ostariophysian fish with distinct reproductive strategies: a siluriform, Pimelodus maculatus, in which oocyte development is group synchronous and the annual reproductive period is short; and a characiform, Serrasalmus maculatus, with asynchronous oocyte development and a prolonged reproductive period. These reproductive strategies result in fish determinate and indeterminate fecundity, respectively. Annual reproductive phases were determined by biometric and histologic analysis of gonads and interpreted according to new proposals for phase classification and stages of oocyte development (with special attention to germinal epithelium activity). Histologically, there were two types of oogonia in the germinal epithelium: single oogonia and those in mitotic proliferation. Oogonial proliferation and their entry into meiosis resulted in formation of cell nests (clusters of cells in the ovarian lamellae). Morphometric analysis was used to estimate germ cell renewal. Based on numbers of single oogonia in the lamellar epithelium, and nests with proliferating oogonia or early prophase oocytes throughout the annual reproductive cycle, oogonial proliferation and entrance into meiosis were more intense during the regenerating phase and developing phase, but decreased sharply (P < 0.05) during the spawning-capable phase. Oogonial proliferation gradually recovered during the regressing phase. We concluded that, independent of species or features of the reproductive cycle, germ cell renewal occurred during the regenerating phase, ensuring availability of eggs for the spawning event. © 2013 Elsevier Inc.

Analysis of the risk factors related to the immune humoral anti-Anaplasma marginale in dairy cattle

Anaplasma marginale is the most prevalent pathogen of cattle transmitted by ticks in the world. This study aimed to evaluate the risk factors for anaplasmosis in dairy cattle. Fifty dairy cattle from the herd of Empresa de Pesquisa Agropecuaria do Estado do Rio de Janeiro were selected by proportional stratified sampling. The risk factors evaluated were: physiological state, race pattern, number of lactations, milk production, infestation by Rhipicephalus microplus and animal density. Antibody activity against A. marginale was determined using the indirect enzyme-linked immunosorbent assay. The percentual values of seroprevalence for A. marginale were submitted to X2 test, and the level of minimum significance, to keep a factor in the model of logistic regression, was fixated in 5%. It was observed that pregnancy and lactation influenced significantly (p<0.05) in the seropositivity of the animals. Bos indicus animals had 5.21 times more chances of being seropositive than B. taurus animals. Primiparous female had 88% more chances of being seropositive than pluriparous female. Animals with high milk production were 63% more positive than low production animals. When infested by ticks the animals had 39% more chance of being seropositive to A. marginale. Bos indicus animals presented 5.21 times more chance of being seropositive than B. taurus animals. Primiparous females presented 88% more chance of being seropositive than the pluriparous ones. High milk production animals were 63% more positive than the low production ones. When infested by ticks the animals had 39% more chance of being seropositive to A. marginale. High density grazing provided for the animals 3.2 times more chances of being seropositive than low density ones. The herd was classified as erratic to A. marginale, even being placed in a steady enzootic area.

An Integrative Genomic and Transcriptomic Analysis Reveals Potential Targets Associated with Cell Proliferation in Uterine Leiomyomas

Background: Uterine Leiomyomas (ULs) are the most common benign tumours affecting women of reproductive age. ULs represent a major problem in public health, as they are the main indication for hysterectomy. Approximately 40-50% of ULs have non-random cytogenetic abnormalities, and half of ULs may have copy number alterations (CNAs). Gene expression microarrays studies have demonstrated that cell proliferation genes act in response to growth factors and steroids. However, only a few genes mapping to CNAs regions were found to be associated with ULs. Methodology: We applied an integrative analysis using genomic and transcriptomic data to identify the pathways and molecular markers associated with ULs. Fifty-one fresh frozen specimens were evaluated by array CGH (JISTIC) and gene expression microarrays (SAM). The CONEXIC algorithm was applied to integrate the data. Principal Findings: The integrated analysis identified the top 30 significant genes (P<0.01), which comprised genes associated with cancer, whereas the protein-protein interaction analysis indicated a strong association between FANCA and BRCA1. Functional in silico analysis revealed target molecules for drugs involved in cell proliferation, including FGFR1 and IGFBP5. Transcriptional and protein analyses showed that FGFR1 (P = 0.006 and P<0.01, respectively) and IGFBP5 (P = 0.0002 and P = 0.006, respectively) were up-regulated in the tumours when compared with the adjacent normal myometrium. Conclusions: The integrative genomic and transcriptomic approach indicated that FGFR1 and IGFBP5 amplification, as well as the consequent up-regulation of the protein products, plays an important role in the aetiology of ULs and thus provides data for potential drug therapies development to target genes associated with cellular proliferation in ULs. © 2013 Cirilo et al.

Outbreak of fungemia caused by Candida parapsilosis in a neonatal intensive care unit: Molecular investigation through microsatellite analysis

Background: Opportunistic infections are an increasingly common problem in hospitals, and the yeast Candida parapsilosis has emerged as an important nosocomial pathogen, especially in neonatal intensive care units (NICUs) where it has been responsible for outbreak cases. Risk factors for C. parapsilosis infection in neonates include prematurity, very low birth weight, prolonged hospitalization, indwelling central venous catheters, hyperalimentation, intravenous fatty emulsions and broad spectrum antibiotic therapy. Molecular methods are widely used to elucidate these hospital outbreaks, establishing genetic variations among strains of yeast. Aims: The aim of this study was to detect an outbreak of C. parapsilosis in an NICU at the Hospital das Clinicas , Faculty of Medicine of Botucatu, a tertiary hospital located in São Paulo, Brazil, using the molecular genotyping by the microsatellite markers analysis. Methods: A total of 11 cases of fungemia caused by C. parapsilosis were identified during a period of 43 days in the NICU. To confirm the outbreak all strains were molecularly typed using the technique of microsatellites. Results: Out of the 11 yeast samples studied, nine showed the same genotypic profile using the technique of microsatellites. Conclusions: Our study shows that the technique of microsatellites can be useful for these purposes. In conclusion, we detected the presence of an outbreak of C. parapsilosis in the NICU of the hospital analyzed, emphasizing the importance of using molecular tools, for the early detection of hospital outbreaks, and for the introduction of effective preventive measures, especially in NICUs. © 2012 Revista Iberoamericana de Micología.

Qualitative analysis of preputial and vaginal bacterial microbiota in owl monkeys (Aotus azarai infulatus) raised in captivity

Background: The aim of this study was to identify the aerobic bacteria of the preputial and vaginal microbiota in owl monkeys that have been raised in captivity and to evaluate the antimicrobial susceptibility of these bacteria by gender and social organization. Methods: Thirty clinically healthy Aotus azarai infulatus were used. A total of 134 samples were collected, 60 from the preputial mucosa and 74 from the vaginal mucosa. An automated system of bacterial identification was used. Results and Conclusions: Staphylococcus intermedius and Proteus mirabilis were the microorganisms that were most frequently identified according to gender and social organization. The antimicrobial susceptibility of the isolated gram-positive bacteria was similar in both sexes. However, the gram-negative strains had some differences. The aerobic bacterial population of the vaginal and preputial microbiota is similar in owl monkeys, and there are no differences in the number and bacterial species according to sex and social organization. © 2012 John Wiley & Sons A/S. Published by Blackwell Publishing Ltd.