Beneficial effect of fluorocarbon emulsion media on the function of neuromuscular preparations in vitro

The effects of liquid fluorocarbons as bathing media were determined by use of in vitro neuromuscular preparations. Rat hemidiaphragms were bathed in either oxygenated fluorocarbon (FC) emulsion or standard oxygenated Krebs solution. Contractile force in response to simple supramaximal nerve stimuli as well as to high frequency stimulation was greater, while twitch:tetanus ratio was smaller in FC emulsion. With such medium, post-tetanic potentiation of contraction was also more consistently observed. Indirectly stimulated diaphragms survived longer in FC emulsion. After cessation of oxygenation, oxygen tension (ρO(2)) of the medium declined more rapidly with Krebs than with FC emulsion; ρO(2) directly correlated with force of contraction. Similarly, in the chick biventer cervicis preparation, FC emulsion enhanced nerve-stimulated force of contraction; returning the preparation to standard Krebs solution reversed this phenomenon. Dose-resonse curves of muscle contraction in response to acetycholine and KCl administration were shifted upward during FC emulsion superfusion. Frequency of miniature endplate potentials was lower in FC emulsion than that observed in Krebs solution, measured from the same cell of the rat diaphragm. Resting membrane potentials were also greater in muscle cells sampled from FC emulsion-bathed preparations. These data suggest that FC emulsion is superior to standard Krebs solution as a bathing medium for in vitro neuromuscular preparations by virtue of the high solubility of oxygen in it.     

Sugar cane buds as an efficient explant for plantlet regeneration

An efficient and reproducible protocol for regeneration of plantlets at a high frequency was developed by using sugar cane buds. Disinfected buds were firstly submerged in ethanol sodium hypochlorite solution with 0.1 % polyvinylpyrrolidone, 1.5 % ascorbic acid and 1.75 % citric acid as antioxidants and subsequently treated with solution of agrimicin:captan (1:1). The upper stalk segment was better to obtain bud in vitro culture compared to lower segments. The medium for induction of multiple shoots consisted of Murashige and Skoog basal medium (MS) supplemented with 2 mg dm⁻³ thidiazuron and 1 mg dm⁻³ naphthalene acetic acid. An average of 24 shoots per bud was obtained for cv. Mex 68-P23 within four weeks and 29 shoots for cv. MY 55-14 within six weeks. Indole-3-butyric acid induced more roots in both cultivars compared to the untreated plantlets. Plantlets transferred to soil showed normal growth with up to four axilliary buds in each node. It was concluded that the germplasm obtained through the above mentioned technique generated stalks with more buds in each node which would give farmers more vegetative material for plantations in field with 100 % germination.This research was funded by Fundacion Produce Chiapas A.C. (Mexico).     

A new karyotype in Callicebus torquatus (Cebidae, Primates)

We describe a new karyotype of Callicebus torquatus using conventional staining, G-banding with Wright Stain, CBG, Ag-NOR staining and fluorescence in situ hybridization (FISH) with human telomere probes and comparative analysis with the previously reported karyotype of C. torquatus torquatus (2n = 20). We studied a female specimen maintained in captivity at the Centro Nacional de Primatas (Para, Brazil). This titi monkey presented 2n = 22, with four large biarmed and six acrocentric autosome pairs; the X chromosome is a medium submetacentric. C-bands were revealed at the centromeric region of all acrocentrics and X chromosome; punctual C-bands also are visualized at the centromeric region in the large biarmed pairs. The NOR site was located at the long arm of pair 4, at the position of a conspicuous secondary constriction. Hybridization signals were detected exclusively at the terminal region of all chromosomes. The karyotype described here has one acrocentric pair more than that found in the literature and also differs by amount and distribution of constitutive heterochromatin. Our data support the notion that the torquatus group may be composed of distinct species, each with its own karyotype.     

Thesaurus-based disambiguation of gene symbols

Background  

Massive text mining of the biological literature holds great promise of relating disparate information and discovering new knowledge. However, disambiguation of gene symbols is a major bottleneck.     

Functional studies of signaling pathways in peri-implantation development of the mouse embryo by RNAi

Background

Studies of gene function in the mouse have relied mainly on gene targeting via homologous recombination. However, this approach is difficult to apply in specific windows of time, and to simultaneously knock-down multiple genes. Here we report an efficient method for dsRNA-mediated gene silencing in late cleavage-stage mouse embryos that permits examination of phenotypes at post-implantation stages.

Results

We show that introduction of Bmp4 dsRNA into intact blastocysts by electroporation recapitulates the genetic Bmp4 null phenotype at gastrulation. It also reveals a novel role for Bmp4 in the regulation the anterior visceral endoderm specific gene expression and its positioning. We also show that RNAi can be used to simultaneously target several genes. When applied to the three murine isoforms of Dishevelled, it leads to earlier defects than previously observed in double knock-outs. These include severe delays in post-implantation development and defects in the anterior midline and neural folds at headfold stages.

Conclusion

Our results indicate that the BMP4 signalling pathway contributes to the development of the anterior visceral endoderm, and reveal an early functional redundancy between the products of the murine Dishevelled genes. The proposed approach constitutes a powerful tool to screen the functions of genes that govern the development of the mouse embryo.     

Restriction enzyme and fluorochrome banding analysis of the constitutive heterochromatin of Saguinus species (Callitrichidae, Primates)

Metaphases of Saguinas fuscicollis fuscicollis and Saguinas mystax were subjected to restriction enzyme banding (Alu I, Hae III, Hin fI, Rsa I, Dde I, Mbo I and Msp I) and sequenced C-banding, together with fluorochrome staining (CMA3 and DAPI). Both species showed large C-bands in the pericentromeric regions. S. f. fuscicollis also manifested distal C-bands in both arms of pair 5 and in the short arms of pairs 8-15. In each species the heterochromatin revealed different reactions to the restriction enzymes and fluorochromes. This was related to its location in the genome (centromeric, pericentromeric, distal), making possible the identification of distinct categories of constitutive heterochromatin. In S. f. fuscicollis there were at least five types, namely centromeric in bi-armed chromosomes, centromeric in acrocentrics, pericentromeric, distal, and cryptic bands, detected only with the Alu I. There were three types in S. mystax, viz centromeric in bi-armed chromosomes, centromeric in acrocentric, and pericentromeric chromosomes. Several aspects of their constitution and origin are discussed.     

Behavioural and physiological consequences of acute social defeat in growing gilts: effects of the social environment

Endocrine, behavioural and immunologic processes, together with body growth, were evaluated in gilts that were defeated at 10 weeks of age in resident-intruder tests. Immediately after defeat, gilts were either separated from or reunited with a familiar conspecific (litter-mate; always a barrow). Gilts were assigned to one of four treatments: (a) DI: defeat, followed by isolation (separation from original litter-mate; n=8); (b) I: no defeat, isolation (control group; n=9); (c) DP; defeat, followed by pair-housing (reunion with original litter-mate; n=8); and (d) P: no defeat, pair-housing (control group; n=8). The following general conclusions were derived: (1) social defeat caused pronounced short-term elevations in hypothalamic-pituitary-adrenal (HPA) and sympathetic-adrenal medullary activities, and of prolactin levels. Moreover, as soon as 1h after defeat, percentages of blood lymphocytes and neutrophilic granulocytes were, respectively, decreased and increased; (2) social defeat had some long-lasting influence on behaviour and physiology, but isolation predominantly determined responses in the longer term. Defeat, as well as isolation, resulted in increased cardiovascular activities compared to P controls, as observed in a novel object test (NOT: +7 days) and an aversion test (AVT: +14 days). Moreover, defeated as well as isolated gilts did not habituate to a repeated novel environment test (NET: -7, +2 and +7 days) in terms of frequencies of vocalising, whereas P controls did. Isolation, through the separation from any other pig, was responsible for the other observed long-term characteristics, which developed progressively. Isolated gilts showed high mobilities and high cortisol responses in the repeated NET (+7 days), not being habituated. This contrasted the reactions of pair-housed gilts, which were much reduced. In addition to their high cardiovascular activities in the NOT and the AVT, isolated gilts also displayed higher heart rates in the repeated NET and during human presence following the NOT, compared to pair-housed gilts. Finally, isolated gilts were more inhibited to approach a novel object (in the NOT) than pair-housed pigs; and (3) stress responses of defeated gilts were modulated by the subsequent social environment. Stimulation of the HPA-axis (plasma- and salivary cortisol) was prolonged in those defeated gilts which were isolated (observed in the first hour). Changes in leucocyte subsets were still observed after 3 days in DI, but were 'normalised' within 1 day in DP gilts. Two days after defeat, habituation to the repeated NET in terms of mobility and salivary cortisol responses occurred in control and DP gilts, but not in DI gilts. We argue that these effects of the social environment shortly after defeat were related to a stress-reducing effect of a stable social relationship, i.e. social support.     

Social Mobility and Mental Disorders at 30 Years of Age in Participants of the 1982 Cohort,Pelotas, Rio Grande Do Sul – RS

This study aimed to evaluate the relationship between mental disorders at 30 years of age and social mobility by formally testing three hypotheses: Risk Accumulation; Critical Period; and Social Mobility. The study was performed using data from the 30-year follow-up of the Pelotas Birth Cohort Study, conducted in 1982, and data from previous follow-ups. The tool used to evaluate mental health was the Self Report Questionnaire (SRQ-20). For the statistical analysis, the chi-square test with the Yates correction was used to estimate the prevalence of mental disorder, and the Poisson regression with robust variance was used to formally test the hypotheses according to the Risk Accumulation, Critical Period and Social Mobility Models. The analyses were stratified by gender. The prevalence of Common Mental Disorders (CMDs) was 24.3% (95% CI 22.9–25.7) when the whole sample was considered. The highest prevalence, 27.1% (95% CI 25.1–29.2), was found in women, and the difference between genders was significant (p < 0.001). CMDs were more frequent in participants who remained “poor” in the three follow-ups. In both men and women, the best fit was obtained with the Risk Accumulation Model, with p = 0.6348 and p = 0.2105, respectively. The results indicate the need to rethink public income maintenance policies. Finally, we suggest further studies to investigate the role of different public policies in decreasing the prevalence of mental disorders and thus contribute proposals of new policies that may contribute to the prevention of these disorders.