Effects of photobiomodulation and swimming on gene expression in rats with the tibialis anterior muscle injury.

The aim of the present study was to evaluate the effects of photobiomodulation (low-level laser therapy (LLLT)) and aquatic exercise on the expression of genes related to muscle regeneration in rats. Wistar rats were divided into five groups: control group (n = 15), non-treated injury group (n = 15), injury+LLLT group (n = 15), injury+aquatic exercise group (n = 15), and injury+LLLT+aquatic exercise group (n = 15). Cryoinjury was performed on the belly of the tibialis anterior (TA) muscle. LLLT was performed daily with an AlGaAs laser (830 nm; beam spot of 0.0324 cm2, output power of 100 mW, energy density of 180 J/cm2, and 58-s exposure time). Animals were euthanized at 7, 14, and 21 days. The TA muscles were removed for gene expression analysis of TGF-ß, Myogenin, and MyoD. The results were statistically analyzed at a significance level of 5%. The cryoinjury increased the expression of genes related to muscle regeneration-MyoD, Myogenin, and TGF-ß-compared to the control group (p < 0.05); the photobiomodulation increased the expression of these genes at day 7 (p < 0.05), decreasing until day 21; and the aquatic exercise increases the expression of the three genes over time. When the two treatments were combined, the expression of the analyzed genes also increased over time. In summary, the results of our study suggest that photobiomodulation (LLLT), when applied alone in cryoinjury, is able to increase the gene expression of MyoD, Myogenin, and TGF-ß at the acute phase, while when combined with aquatic exercises, there is an increase in expression of these genes specially at the long-term treatment.

Quality assessment of systematic reviews regarding the effectiveness of zygomatic implants: an overview of systematic reviews.

BACKGROUND: Oral rehabilitation of atrophic maxillae features high complexity, for which there are several therapeutic modalities reported on scientific literature. Zygomatic implant placement is a viable option that features low morbidity and allows immediate prosthetic loading. The purpose of the present study was to determine the methodological quality of systematic reviews that assessed the effectiveness of zygomatic implants placed in atrophic maxillae. MATERIAL AND METHODS: Searches were conducted on Medline via Pubmed, LILACS, Dare Cochrane, Scopus, and Sigle via Open Grey up to June 2019. RESULTS: Seven systematic reviews were eligible for Overview and comprised a total of 2313 patients, 4812 zygomatic implants, and a 96,72% success rate. Common surgical complications, in decreasing order, were: maxillary sinusitis, peri-implant mucositis, prosthetic fracture, and infections. Methodological quality was assessed using the AMSTAR 2 tool, which revealed that six systematic reviews showed critically low methodological quality and one review was assessed as of low methodological quality. CONCLUSIONS: Zygomatic implants seem to be an adequate option for atrophic maxilla rehabilitation, however, new studies with a higher methodological rigor are needed to provide more reliable results to professionals and patients undergoing this modality of oral rehabilitation.

Dietary Ingestion of Calories and Micronutrients Modulates the DNA Methylation Profile of Leukocytes from Older Individuals.

OBJECTIVE: Several lines of evidence from the last decade support the connection between nutrition and epigenetic mechanisms. In the present study we evaluated the impact of the daily dietary intake of calories and the micronutrients vitamin A, D, B1, B2, B5, C, E, copper, calcium, phosphorus, iron, iodine, selenium, manganese, potassium and sodium on the global DNA methylation profile of blood cells from older individuals. RESEARCH METHODS AND PROCEDURES: The study enrolled 126 physically independent elderly of both sexes (60 men and 66 women). For the molecular analysis, DNA samples were extracted from leukocytes and global DNA methylation was evaluated using a high throughput Elisa-based method. Correlations between global DNA methylation and the daily intake of calorie or micronutrients were evaluated using Prism5 GraphPad Software. RESULTS: A statistically significant correlation was observed between global DNA methylation and the daily caloric value (p=0.019, r=-0.21), and the intake of vitamin A (p=0.03, r=-0.18), Vitamin E (p=0.027, r=-0.20) and copper (p=0.04, r=-0.18). No correlation was observed between global DNA methylation and the daily intake of vitamin D, B1, B2, B5, C, calcium, phosphorus, iron, iodine, selenium, manganese and potassium (p>0.05). CONCLUSION: Our data demonstrate that the daily intake of calories or the micronutrients vitamin A, vitamin E and copper can potentially modulate the global DNA methylation profile of leukocytes in older adults and corroborate the notion of nutritional influences on epigenetic mechanisms.

Maternal exposure to fluoxetine during gestation and lactation induces long lasting changes in the DNA methylation profile of offspring's brain and affects the social interaction of rat.

Fluoxetine (FLX) is an antidepressant from the selective serotonin reuptake inhibitor class that has largely been used for the treatment of depression in pregnancy. However, increasing evidences have indicated the potential of early maternal exposure to FLX to induce molecular and neuro functional effects on the offspring. In the present study we evaluated possible long lasting impacts of the maternal exposure to FLX during gestation and lactation. Female Wistar rats were gavaged with 5 mg/kg of FLX during the period that comprehends the first day of pregnancy (PD0) and the last day of lactation (LD21) (Group FLX). Control group (CTL) received a proportional volume of water. On the postnatal day 75 (PND75), male rats were euthanized and hippocampus, cortex, hypothalamus, and periaqueductal gray area (PAG) were removed. Global DNA methylation was quantified using a high-throughput ELISA-based method. In order to address neuro functional changes animals (PND75) were evaluated in the elevated plus maze and social interaction tests as well as submitted to repeated restraint stress. An increase in the global DNA methylation profile of hippocampus (p = 0.0399) was associated with the early exposure to FLX, whereas no significant change was observed in the hypothalamus (p = 0.6556), cortex (p = 0.9402) or PAG (p = 0.3822). Furthermore, early exposure to FLX was also associated with a reduction in the social interaction time (p = 0.0084) and to a decreased in the plasma corticosterone level when animals were submitted to the restraint stress (p < 0.0001). No significant change in the elevated plus maze test was associated with the early exposure to FLX. In summary, our data demonstrate that maternal exposure to FLX during gestation and lactation results in a long lasting impact on the DNA methylation of hippocampus, and affects the social behavior and the corticosterone response to stress.

Effects of chronic restraint stress on the global DNA methylation profile of rat lung cells: Modulation by physical exercise.

The potential of behavioral stress to affect epigenetic mechanisms of non-encephalic tissues is still underestimated. In the present study we evaluated the effects of chronic behavioral stress on the DNA methylation profile of rat lung cells. Furthermore, we evaluated the potential of physical exercise to modulate the changes evoked by behavioral stress in lung cells. Male Wistar rats were divided into four experimental groups: (1) animals submitted to chronic restraint stress (CRS) (ST group) during the period of the 67th-80th postnatal day (PND); (2) animals submitted to physical exercise (EX group) during the 53rd-79th PND; (3) animals submitted to swimming during the 53rd-79th PND and to CRS during the 67th-80th PND (EX-ST group); and (4) animals not submitted to stress or swimming protocols (CTL). Global DNA methylation was quantified using an ELISA-based approach and gene expression was evaluated by real time PCR. A decreased global DNA methylation profile was observed in the ST group, however physical exercise demonstrated protection of lung cells from this stress-related hypomethylation. Increased expression of the Dnmt1 gene was evidenced in the ST group, whereas physical exercise was shown to protect lung cells from this stress-related effect in the EX-ST group. Comparative analysis of the ST and EX groups revealed opposite effects on the expression of Dnmt3a and Dnmt3b; however, a stress-related increase in expression of Dnmt3a and Dnmt3b was not seen in the EX-ST group. Our data showed that behavioral stress induced significant changes in the DNA methylation profile of rat lung cells and that this could be modulated by physical exercise.

Physical exercise affects the epigenetic programming of rat brain and modulates the adaptive response evoked by repeated restraint stress.

Epigenetics has recently been linked to molecular adaptive responses evoked by physical exercise and stress. Herein we evaluated the effects of physical exercise on global DNA methylation and expression of the Dnmt1 gene in the rat brain and also verified its potential to modulate responses evoked by repeated restraint stress (RRS). Wistar rats were classified into the following experimental groups: (1) physically active (EX): animals submitted to swimming during postnatal days 53-78 (PND); (2) stress (ST): animals submitted to RRS during 75-79PND; (3) exercise-stress (EX-ST): animals submitted to swimming during 53-78PND and to RRS during 75-79PND, and (4) control (CTL): animals that were not submitted to intervention. Samples from the hippocampus, cortex and hypothalamus were obtained at 79PND. The global DNA methylation profile was assessed using an ELISA-based method and the expression of Dnmt1 was evaluated by real-time PCR. Significantly increased methylation was observed in the hypothalamus of animals from the EX group in comparison to CTL. Comparative analysis involving the EX-ST and ST groups revealed increased global DNA methylation in the hippocampus, cortex, and hypothalamus of EX-ST, indicating the potential of physical exercise in modulating the responses evoked by RRS. Furthermore, decreased expression of the Dnmt1 gene was observed in the hippocampus and hypothalamus of animals from the EX-ST group. In summary, our data indicate that physical exercise affects DNA methylation of the hypothalamus and might modulate epigenetic responses evoked by RRS in the hippocampus, cortex, and hypothalamus.

Maternal exposure to fluoxetine during gestation and lactation affects the DNA methylation programming of rat's offspring: modulation by folic acid supplementation.

Fluoxetine is an antidepressant that has been largely used for treatment of depression in pregnancy. In the present study we evaluated the effects of the exposure to fluoxetine during gestation and lactation on DNA methylation of rat brain regions. Female Wistar rats were treated with 5mg/kg of fluoxetine during pregnancy and lactation. In order to assess the effects of fluoxetine in the context of maternal folic acid supplementation we performed an additional combined treatment composed by folic acid (8 mg/kg/day) and fluoxetine (5 mg/kg/day). On the postnatal day 22, male rats were euthanized and hippocampus, cortex, hypothalamus, and periaqueductal gray area were removed. Global DNA methylation was quantified using a high-throughput ELISA-based method. Neurofunctional changes were addressed using validated behavioral tests: hot plate, elevated plus maze and open field. A decrease in the global DNA methylation profile of hippocampus was associated to the exposure to fluoxetine, whereas an increase in methylation was observed in cortex. The combined treatment induced an increase in the methylation of hippocampus indicating the potential of folic acid to modulate this epigenetic alteration. Increase in the latency to the thermal nociceptive response was observed in animals exposed to fluoxetine whereas this effect was abolished in animals from the combined treatment. In summary we demonstrated that exposure to fluoxetine during gestation and lactation affect the DNA methylation of brain and the nociceptive response of rats. Furthermore our data reveal the potential of folic acid to modulate epigenetic and functional changes induced by early exposure to fluoxetine.

Relação entre polimorfismo do gene do hormônio do crescimento e características de precocidade em novilhas da raça Nelore / Relationship between polymorphism of growth hormone and precocity traits in Nellore heifers

Avaliaram-se as relações entre o polimorfismo do gene do hormônio do crescimento (GH) e as características de precocidade, em novilhas da raça Nelore. Amostras de sangue periférico foram obtidas de 181 animais de três rebanhos distintos do estado da Bahia, nas quais foi realizada a extração de DNA e a amplificação por PCR, seguidas por digestão com enzima de restrição AluI. Os fragmentos resultantes da digestão enzimática foram analisados em gel de agarose 2 por cento para determinação dos respectivos genótipos. A frequência do alelo Leu nas amostras analisadas foi estimada em 100 por cento. Em decorrência da alta incidência de homozigose para o alelo Leu, sugere-se que o restriction fragment lenght polymorphism AluI do gene GH não possa ser considerado como marcador molecular para precocidade sexual em novilhas Nelore nesses rebanhos.

The relationships between polymorphism of growth hormone gene (GH) and precocity traits in Nellore heifers were evaluated. A total of 181 animals from three different farms of Bahia state, Brazil, were blood sampled. The DNA of each animal was extracted, amplified by PCR, and digested by "AluI" restriction enzyme, and the resultant fragments were analyzed in 2 percent agarose gel for genotype identification. The frequency of Leu allele in the analyzed samples was estimated in 100 percent. Due to the high incidence of homozygose for the Leu allele, it is suggested that the restriction fragment lenght polymorphism AluI of GH gene can not be considered as a molecular marker for sexual precocity in Nellore heifers of those herds.

Abnormal methylation at the KvDMR1 imprinting control region in clinically normal children conceived by assisted reproductive technologies.

Genomic imprinting alterations have been shown to be associated with assisted reproductive technologies (ARTs) in animals. At present, data obtained in humans are inconclusive; however, some epidemiological studies have demonstrated an increased incidence of imprinting disorders in children conceived by ARTs. In the present study, we focused on the effect of ARTs [IVF and intracytoplasmic sperm injection (ICSI)] on the epigenetic reprogramming of the maternally methylated imprinting control region KvDMR1 in clinically normal children. Qualitative and quantitative methylation at KvDMR1 were assessed by the methylation-specific PCR approach and by the methylation-sensitive enzymatic digestion associated with real-time PCR method, respectively. DNA was obtained from peripheral blood of 12/18 and umbilical cord blood and placenta of 6/18 children conceived by IVF or ICSI. The methylation patterns observed in this group were compared with the patterns observed in 30 clinically normal naturally conceived children (negative controls) and in 3 naturally conceived Beckwith-Wiedemann syndrome patients (positive controls). Hypomethylation at KvDMR1 was observed in 3/18 clinically normal children conceived by ARTs (2 conceived by IVF and 1 by ICSI). A discordant methylation pattern was observed in the three corresponding dizygotic twins. Our findings corroborate the hypothesis of vulnerability of maternal imprinting to ARTs. Furthermore, the discordant methylation at KvDMR1 observed between dizygotic twins could be consequent to one of the following possibilities: (i) a differential vulnerability of maternal imprints among different embryos; or (ii) epimutations that occurred during gametogenesis resulting in the production of oocytes without the correct primary imprint at KvDMR1.

Association between birth weight, body mass index and IGF2/ApaI polymorphism.

Insulin-like growth factor II (IGF-II) is a polypeptide that plays a key role in mammalian growth, influencing fetal cell division, differentiation, and possibly metabolic regulation. In adult humans, polymorphisms of the IGF2 gene have been associated with predisposition to obesity. In the present study, we tested the association between IGF2/ApaI genotype and Body Mass Index (BMI) in 294 healthy volunteers (95 men and 199 women; 18-30y) and correlated the results with their birth weights (BW) in order to investigate the relationship between this polymorphic site, fetal life and adult BMI. Blood samples were obtained for DNA extraction, PCR and genotyping. The statistical analyses were performed by the Chi-square, Kolmogorov-Smirnov normality, and Tukey post hoc tests. Although the IGF2 genotype was not significantly associated with BMI and/or BW, we observed a statistically significant correlation of 0.33 (p < 0.023) between BW and BMI in GG subjects whose BW was higher than 3.5 kg (n = 47). We hypothesize that high BWs associated with homozygosis for the G allele of IGF2/ApaI is not a null factor and might be associated with predisposition to high BMI in young adults.

Early gestational hemolytic uremic syndrome: case report and review of literature.

Hemolytic uremic syndrome (HUS) is a rare condition which most frequently follows gastrointestinal or respiratory infection episodes in young children, but it can also occur in other settings such as the postpartum period and during use of drugs such as oral contraconceptives, immunosuppressors, and antineoplastics. In early pregnancy, however, its frequency is thought to be very low. The authors report a case of a 30-year-old woman who developed HUS early in her first pregnancy. She had persistent aqueous diarrhea from the beginning of the pregnancy. At the 21st week she developed hypertension which in 2 weeks was followed by seizures, oliguria, and acute pulmonary edema despite intensive medical efforts to control her blood pressure. Surgical intervention for fetal delivery was performed. The patient was initially kept on continuous hemodialysis (CVVHD) followed by an alternate-day conventional hemodialysis schedule. A peripheral blood analysis showed a microangiopathic hemolytic anemia with thrombocytopenia; blood coagulation tests were completely normal. A brain CT scan and an abdominal MRI showed no major abnormalities. HUS was confirmed by a percutaneal kidney biopsy, performed at the 21st day of anuria. Techniques for identification of verotoxin-producing E. coli were not available. Renal function did not recover and the patient has been undergoing regular maintenance hemodialysis for a year.

Mal epilético como apresentaçäo de síndrome de Melas: relato de um caso / Status epilepticus as the presentation of Melas syndrome: a case report

É relatado e discutido o caso de uma paciente admitida no CTI devido a estado de mal epiléptico, atribuído a síndrome de Melas (miopatia mitocondrial, encefalopatia, acidose lática e episódios AVC-símile). A paciente era uma mulher jovem, com história pregressa de hopoacusia bilateral, crises episódicas de enxaqueca com vômitos, que apresentou-se em como, com sinais de irritaçäo meníngea e sem sinais de localizaçäo. Causas estruturais, infecciosas e auto-imunes foram afastadas através de exames laboratoriais e de imagem. Ao invés disso, demonstravam acidose lática grave, aumento de enzimas musculares, azotemia, hiperglicemia e leucocitose e presença de calcificaçöes dos gânglios da base e áreas isquêmicas na tomografia de crânio. Foi levantada a hipótese de Melas, a qual foi confirmada através de biópsia muscular que demonstrou fibras "ragged red" fortemente coradas pelo succinato desidrogenase (SDH). Os autores dirigem atençäo ao fato de uma síndrome rara também deve ser incluída no diagnóstico diferencial de um paciente jovem com convulsäo e acidose lática mesmo quando de uma apresentaçäo inicial.

Surgical repair of transposition of great arteries and total anomalous pulmonary venous return to the coronary sinus (TGA with TAPVR).

The association of transposition of the great arteries (TGA) and total anomalous pulmonary venous return (TAPVR) is very rare; we report one case of this malformation with an intracardiac type of TAPVR draining into the coronary sinus. The surgical repair was performed directing the coronary sinus blood flow to the tricuspid valve. The proximity of the anatomical structures allowed an intra-atrial patch repair similar to a modified Mustard procedure. In the postoperative period the child developed low cardiac output for 2 days, requiring inotropic support and mechanical ventilation. Her recovery was otherwise uneventful. The postoperative echocardiogram showed an intact repair with perfect flow through the patch from the pulmonary veins to the right ventricle. Almost 2 years after surgery, the patient underwent cardiac catheterization that showed integrity of the surgical repair with normal pressures in all heart chambers. To our knowledge this is the first report in the medical literature of surgical treatment of this association.

[The physician in the pharmaceutical industry]. / O médico na indústria farmacêutica.

The physician's role in the pharmaceutical industry has changed over the past years. This change is a consequence of several factors, namely the evolution of the industry itself, the legislative and regulatory changes in this particular area, the development of medicine, new techniques and research, and finally the physician's needs. The changes that this activity has undergone not only create new stimuli for the physicians in the industry, but also new challenges and needs for the complete achievement of their career. This is a review article of the main functions and responsibilities of the physician in the pharmaceutical industry as well as the new challenges that this group of physicians faces. Finally, special attention is given to post-graduate courses and their relation with the academic structure.