Loss of Methylation at H19 DMD Is Associated with Biallelic Expression and Reduced Development in Cattle Derived by Somatic Cell Nuclear Transfer

Although cloning of mammals has been achieved successfully, the percentage of live offspring is very low because of reduced fetal size and fewer implantation sites. Recent studies have attributed such pathological conditions to abnormal reprogramming of the donor cell used for cloning. The inability of the oocyte to fully restore the differentiated status of a somatic cell to its pluripotent and undifferentiated state is normally evidenced by aberrant DNA methylation patterns established throughout the genome during development to blastocyst. These aberrant methylation patterns are associated with abnormal expression of imprinted genes, which among other genes are essential for normal embryo development and gestation. We hypothesized that embryo loss and low implantation rates in cattle derived by somatic cell nuclear transfer (SCNT) are caused by abnormal epigenetic reprogramming of imprinted genes. To verify our hypothesis, we analyzed the parental expression and the differentially methylated domain (DMD) methylation status of the H19 gene. Using a parental-specific analysis, we confirmed for the first time that H19 biallelic expression is tightly associated with a severe demethylation of the paternal H19 DMD in SCNT embryos, suggesting that these epigenetic anomalies to the H19 locus could be directly responsible for the reduced size and low implantation rates of cloned embryos in cattle.

Consequences of Simulated Loss of Open Cerrado Areas to Bird Functional Diversity

Over the past 35 years, more than two thirds of the Cerrado`s original expanse has been taken by agriculture. Even if some attempts have been made to conserve closed cerrado physiognomies, open cerrado physiognomies, richer in species and more fragile, have been systematically ignored. These open physiognomies are used by almost half of the Cerrado bird species, many of which being endemics. Using data from 11 surveys carried out in Cerrado landscapes, we asked what would happen to bird functional diversity if open cerrado species became extinct. Open cerrado birds would be able to keep on average 59% of the functional diversity. If they became extinct, on average 27% of the functional diversity would be lost. In this case, the remaining functional diversity would be lower than what would be expected by chance in five sites. Although many functions were shared by both open cerrado and forest species, there was some degree of complementarity between them, highlighted by the decrease in functional diversity when the former became extinct. Destruction of open cerrado physiognomies would lead to a habitat simplification, decrease in bird functional diversity, and, ultimately, to a considerable impact on community functioning. Thus, open cerrado physiognomies must receive much more conservation attention than they are currently receiving, because they maintain a high bird functional diversity that would otherwise be considerably diminished Were open cerrado species to become extinct.

Emergence and loss of assortative mating in sympatric speciation

We have studied an agent model which presents the emergence of sexual barriers through the onset of assortative mating, a condition that might lead to sympatric speciation. In the model, individuals are characterized by two traits, each determined by a single locus A or B. Heterozygotes on A are penalized by introducing an adaptive difference from homozygotes. Two niches are available. Each A homozygote is adapted to one of the niches. The second trait, called the marker trait has no bearing on the fitness. The model includes mating preferences, which are inherited from the mother and subject to random variations. A parameter controlling recombination probabilities of the two loci is also introduced. We study the phase diagram by means of simulations, in the space of parameters (adaptive difference, carrying capacity, recombination probability). Three phases are found, characterized by (i) assortative mating, (ii) extinction of one of the A alleles and (iii) Hardy-Weinberg like equilibrium. We also make perturbations of these phases to see how robust they are. Assortative mating can be gained or lost with changes that present hysteresis loops, showing the resulting equilibrium to have partial memory of the initial state and that the process of going from a polymorphic panmictic phase to a phase where assortative mating acts as sexual barrier can be described as a first-order transition. (C) 2009 Published by Elsevier Ltd.

Effect of dietary energy and stocking density on the performance and sensible heat loss of broilers reared under tropical winter conditions

The objective of the present study was to evaluate the effects of different dietary energy levels and stocking densities on the thermoregulating parameters, live performance, and carcass traits of broilers reared under tropical winter conditions at different times of the day. In total, 1,312 one-d-old male broilers were used. Birds were allotted to three different stocking densities (10, 14 or 18 birds/m²) and two dietary energy levels (2900 or 3200 kcal ME/kg). The following parameters were evaluated:radiant heat load (RHL), rectal temperature (RT), feed intake (FI), weight gain (WG), feed conversion ratio (FCR), livability (L), production of live weight per area (WA), and carcass yield. Stocking density did not affect sensible heat loss (SRL) or rectal temperature (RT); however, as expected, sensible heat loss (SRL) and RT were influenced by time of the day, with higher values in the morning and in the afternoon, respectively. There was no effect of treatment (p>0.05) on carcass or parts yield. Feed intake was reduced in 3%, whereas weight gain and feed conversion ratio improved in 8 and 10%, respectively, as dietary energy level increased. on the other hand, stocking density did not influence live performance or carcass traits. Based on the present results, it is concluded that sensible heat loss depends on dietary energy levels and particularly on time of the day. Therefore, environmental house management is suggested during tropical winters in order to reduce differences between broiler skin and environmental temperatures in the morning and in the afternoon.

Latent heat loss of Holstein cows in a tropical environment: a prediction model

Nove vacas Holandesas lactantes com 526 ± 5 kg de peso corporal (cinco predominantemente pretas e quatro predominantemente brancas), criadas em região tropical e manejadas em pastagens, foram observadas com os objetivos de determinar simultaneamente as taxas de evaporação cutânea e respiratória em ambiente tropical e desenvolver modelos de predição. Para a medição da perda de calor latente pela superfície corporal, utilizou-se uma cápsula ventilada e, para a perda por respiração, utilizou-se uma máscara facial. Os resultados mostraram que as vacas que tinham maior peso corporal (classe 2 e 3) apresentaram maiores taxas evaporativas. Quando a temperatura do ar aumentou de 10 para 36ºC e a umidade relativa do ar caiu de 90 para 30%, a eliminação de calor por evaporação respiratória aumentou de aproximadamente 5 para 57 W m-2 e a evaporação na superfície corporal passou de 30 para 350 W m-2. Esses resultados confirmam que a eliminação de calor latente é o principal mecanismo de perda de energia térmica sob altas temperaturas (>30ºC); a evaporação cutânea é a maior via e corresponde a aproximadamente 85% da perda total de calor, enquanto o restante é eliminado pelo sistema respiratório. O modelo para predizer o fluxo de perda de calor latente baseado em variáveis fisiológicas e ambientais pode ser utilizado para estimar a contribuição da evaporação na termorregulação, enquanto o modelo baseado somente na temperatura do ar deve ser usado apenas para a simples caracterização do processo evaporativo.

Latent heat loss of dairy cows in an equatorial semi-arid environment

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Loss of nuclear poly(A)-binding protein 1 causes defects in myogenesis and mRNA biogenesis

The nuclear poly(A)-binding protein 1 (PABPN1) is a ubiquitously expressed protein that plays a critical role in polyadenylation. Short expansions of the polyalanine tract in the N-terminus of PABPN1 lead to oculopharyngeal muscular dystrophy (OPMD), which is an adult onset disease characterized by eyelid drooping, difficulty in swallowing and weakness in the proximal limb muscles. Although significant data from in vitro biochemical assays define the function of PABPN1 in control of poly(A) tail length, little is known about the role of PABPN1 in mammalian cells. To assess the function of PABPN1 in mammalian cells and specifically in cells affected in OPMD, we examined the effects of PABPN1 depletion using siRNA in primary mouse myoblasts from extraocular, pharyngeal and limb muscles. PABPN1 knockdown significantly decreased cell proliferation and myoblast differentiation during myogenesis in vitro. At the molecular level, PABPN1 depletion in myoblasts led to a shortening of mRNA poly(A) tails, demonstrating the cellular function of PABPN1 in polyadenylation control in a mammalian cell. In addition, PABPN1 depletion caused nuclear accumulation of poly(A) RNA, revealing that PABPN1 is required for proper poly(A) RNA export from the nucleus. Together, these experiments demonstrate that PABPN1 plays an essential role in myoblast proliferation and differentiation, suggesting that it is required for muscle regeneration and maintenance in vivo.

Loss of Y-chromosome does not correlate with age at onset of head and neck carcinoma: a case-control study

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Loss of vision secondary to obsessive-compulsive disorder: a case report

Objective: To report a rare case of self-inflicted eye injury secondary to obsessive-compulsive disorder (OCD).Method: Case report.Results: A 41 year-old patient had significant bilateral visual loss and blunt trauma through touching his ocular cavities due to recurrent doubts regarding their shape and urges to check the format of bones and cartilages. Differential diagnosis with tic disorders is discussed, and the patient's treatment approach is described.Conclusion: Although rare, severe clinical complications can occur in OCD. (C) 2009 Elsevier B.V. All rights reserved.

Loss of samples in the tissue microarray technique: comparison between slides using adhesive tape and silanized slides

The tissue microarray (TMA) technique allows multiple tissue samples in a single block. Commercial adhesive tape is used to avoid the loss of tissue samples during the immunostaining process. Few reports exist in the literature comparing the use of these adhesive tapes to other adhesive techniques. The objective of this study was to compare loss of sections adhered to slides using commercial adhesive tapes versus using silanized only slides. TMA was constructed with varying tissues using a fixed-base device (Beecher Instruments), placing 108 cylinders of 1 mm diameter in duplicate, spaced 1.2 mm apart. Section of 4 mu m were cut from the TMA block and adhered to 30 silanized slides and 30 commercial glass slides using adhesive tape, according to manufacturer's recommendations. Vimentin immunoexpression was evaluated by immunohistochemistry. Antigenic recovery was realized in citrate buffer using a microwave oven. Cylinder loss in the immunohistochemical process was quantified and expressed as: total (>80%), almost complete (75-79%), or partial (50-74%). The commercial adhesive tape group presented lesser total loss (1.1 versus 6.4%), almost complete loss (2.2 versus 3.5%), and partial loss (2.1 versus 3.8%) than the silanized slide group (ANOVA, P < 0.05). The sum of total and almost complete losses in the silanized slide group was 9.9%, greater than the losses in slides using commercial adhesive tapes (3.3%) and less than reported and considered acceptable in the literature (10-30%). In conclusion, the use of silanized only slides presents very satisfactory results, requires less training, and reduces costs significantly, thus justifying their use in research.

STORAGE RESULTS IN LOSS of THE ANTIGENOTOXIC PROPERTIES of LENTINULA EDODES (SHIITAKE MUSHROOM) and DEVELOPMENT of IN VIVO GENOTOXICITY

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Independent clonal origin of multiple uterine leiomyomas that was determined by X chromosome inactivation and microsatellite analysis

Objective: In an attempt to clarify the clonality and genetic relationships that are involved in the tumorigenesis of uterine leiomyomas, we used a total of 43 multiple leiomyomas from 14 patients and analyzed the allelic status with 15 microsatellite markers and X chromosome inactivation analysis.Study design: We have used a set of 15 microsatellite polymorphism markers mapped on 3q, 7p, 11, and 15q by automated analysis. The X chromosome inactivation was evaluated by the methylation status of the X-linked androgen receptor gene.Results: Loss of heterozygosity analysis showed a different pattern in 7 of the 8 cases with allelic loss for at least 1 of 15 microsatellite markers that were analyzed. A similar loss of heterozygosity findings at 7p22-15 was detected in 3 samples from the same patient. X chromosome inactivation analysis demonstrated the same inactivated allele in all tumors of the 9 of 12 informative patients;. different inactivation patterns were observed in 3 cases.Conclusion: Our data support the concept that uterine leiomyomas are derived from a single cell but are generated independently in the uterus. Loss of heterozygosity findings at 7p22-15 are consistent with previous data that suggested the relevance of chromosomal aberrations at 7p that were involved in individual uterine leiomyomas. (C) 2005 Mosby, Inc. All rights reserved.

Polymorphisms of CYP17A1, CYP19, and androgen in Brazilian women with uterine leiomyomas

Background: Uterine leiomyomas are common, benign, smooth muscle tumors representing a significant public health problem. The aim of this study was to investigate CYP17A1, CYP19, and androgen (AR) polymorphisms, their relative risks for uterine leiomyomas and possible associations with clinical parameters.Methods: Uterine leiomyoma tissues and blood samples were obtained from 87 patients, as were peripheral blood samples from 68 control women. Clinical data were recorded in both groups. The CYP17A1 (rs743572) polymorphism was analyzed by PCR-RFLP, and the CYP19 [TTTA](n) repeat and AR [CAG](n) repeat were analyzed using PCR-based GeneScan analysis. AR loss of heterozygosity (LOH) and microsatellite instability were also evaluated, while samples exhibiting LOH were analyzed for X inactivation.Results: Clinical parameters related to disease development did not differ between cases and controls. CYP17A1 *A2/*A2 genotype was prevalent in non-white women. CYP17A1, CYP19, and AR genotypes and alleles did not differ between groups. However, alleles presenting [TTTA](7) repeats in intron 4 of CYP19 were more frequent in the control group (p=0.0550). Shorter and longer [CAG]n repeat alleles of AR were exclusive to the leiomyoma group. The LOH assay showed allele losses at AR locus in four informative tumors and X chromosome inactivation analysis revealed that these tumors retained the active allele.Conclusions: The overall lack of association between uterine leiomyomas with polymorphisms involved in steroidogenesis or steroid metabolism is consistent with the hypothesis that these polymorphisms do not substantially contribute to the development of these tumors.