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.

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.

High pathological response rate in locally advanced esophageal cancer after neoadjuvant combined modality therapy: dose finding of a weekly chemotherapy schedule with protracted venous infusion of 5-fluorouracil and dose escalation of cisplatin, docetaxel and concurrent radiotherapy.

BACKGROUND: This phase I study was aimed at defining the toxicity profile and pathological response rate of a neoadjuvant schedule including weekly docetaxel and cisplatin, protracted venous infusion (PVI) of 5-FU and concomitant radiotherapy (RT) in locally advanced esophageal cancer. PATIENTS AND METHODS: The schedule consisted of a first phase of chemotherapy alone and a second phase of concurrent chemoradiation. Initial doses were: docetaxel and cisplatin 20 mg/m2 on days 1, 8, 15, 29, 36 and 43 plus 5-FU 150 mg/m2 PVI on days 1-21 and 29-49; RT (40 Gy) started on day 29. In the following steps the doses were escalated up to docetaxel 35 mg/m2 and cisplatin 25 mg/m2 on days 1, 8, 15, 29, 36, 43, 50 and 57 plus 5-FU 180 mg/m2 PVI on days 1-21 and 150 mg/m2 PVI on days 29-63 concurrently with RT 50 Gy. RESULTS: Forty-seven patients were enrolled and 46 completed the planned treatment. During the concomitant phase, grade 3-4 hematological toxicities occurred in three patients (6.5%) (or 3/174 cycles) and non-hematological toxicities in six patients (13%) (or 7/179 cycles). A pathological downstaging was obtained in 59.6% of the cases (28/47): complete remission (pCR) in 14 patients, near pCR (residual microfoci on the primary pN0) in eight patients, pT2 pN0 in three patients and partial response on the primary with positive lymph nodes in three patients. Six (13%) and 13 (28%) patients were considered stable and non-responders, respectively. In the last dose level, eight pCR and four near-pCR were obtained out of 15 patients. The maximum tolerable dose was not formally defined because dose escalation was stopped at the last dose level. CONCLUSION: This schedule represents a feasible treatment and the high pathological response rate is extremely encouraging; the doses found in the last dose-level are the basis for an ongoing phase II study at our institution.

A TEOAE screening protocol based on linear click stimuli: performance and scoring criteria.

In order to improve the quality of current TEOAE recording methodologies, we have conducted a comparison of TEOAE neonatal recordings acquired with linear protocols using click stimuli of 68 dB SPL and non-linear protocols using the ILO default stimulus values. From a theoretical standpoint it was expected that the linear recordings would generate responses characterized by higher S/N ratios due to the fact that the stimulus sequence contains four clicks of the same intensity and polarity. The project included recordings from 1,416 neonatal ears (age 48 h). The TEOAE data were compared in terms of correlation, response amplitude, noise, corrected response and S/N ratio in the 1.0-, 2.0-, 3.0-, 4.0- and 5.0-kHz bands, using a paired t-test criterion. We found that windowed (4-14 ms) responses evoked by a linear TEOAE protocol generated superior S/N estimates in the 2.0-, 3.0-, 4.0- and 5.0-kHz TEOAE bands, in addition to superior correlation estimates, and demonstrated lower levels of noise. Clear-cut scoring criteria were established for the S/N ratios at 2.0, 3.0 and 4.0 kHz, by constructing one-sided distribution-free tolerance boundaries.

[Evaluation of a linear TEOAE protocol in hearing screening of neonates: feasibility study]. / Valutazione di un protocollo TEOAE lineare nello screening audiologico neonatale: studio di fattibilità.

The audiological screening of newborns based on recording TEOAEs--the expression of anatomical-functional integrity of the cochlea--has unanimously gained importance. The prevalence of severe of genetic or congenital hearing loss in the healthy infant population and in the population at risk for audiological disorders, as well as the consequent psycholinguist and socialization problems such children have to deal with, have led the authors to set up a preliminary protocol for the audiological screening of neonates. The purpose of this protocol is to improve the feasibility of applying such a program. To this purpose, the preliminary protocol called for the recording of the TEOAE in both non linear (NL) and linear (L) modes. Examination was performed in 347 newborns (30% of all the children born during 1997) the second day of life and during spontaneous sleep. The ILO 92 was used for the screening. The results obtained with the two methods were statistically compared using the 9 parameters considered response indicators. Using the linear method of a function-window and improving the signal-to-noise ratio, the study showed a significant difference in the quality of the TEOAE using the linear method as compared with the non linear method, although this did not modify response reliability. This lead to the definition, through free distribution statistical analysis, of broader than normal criteria by which to evaluate the responses evoked with the L method. All this is aimed at increasing system specificity and reducing the number of false positives which feeds family anxiety.

[Evoked otoacoustic emissions using clicks in detection of deafness in young children]. / Les oto-émissions acoustiques provoquées (OEAP) par des transitoires dans le dépistage de la surdité du jeune enfant.

Click-evoked otoacoustic emissions (EOAEs) are sounds emitted by the cochlea due to active outer hair cell contraction following stimulation of the stereocilia. The purpose of this study was to evaluate the use of testing for EOAEs as a valid auditory screening method in newborns, and 1074 healthy fullterm babies (2148 ears) with an Apgar score higher than 8 at birth were examined. EOAEs were elicited 4 days after birth and a correlation index higher than 70% represented normal response criteria. If there was no response, EOAEs and brainstem response potentials were tested at the age of one month. On the fourth day after birth. EOAEs specificity and efficiency were found to be 92.3% and 92.4% respectively, while sensitivity was 100%. Specificity and efficiency at one month were rated at 100%. The use of EOAEs offers numerous advantages over traditional auditory screening methods, thus suggesting that this technique is suitable for the mass-screening of infants.

Comparison of the antianginal efficacy of four calcium antagonists and propranolol in stable angina pectoris.

The antianginal effects of propranolol 160 mg/day, diltiazem 240 mg/day, nicardipine 80 mg/day, nifedipine 40 to 80 mg/day and verapamil 320 mg/day were compared in 12 patients with chronic stable angina pectoris using a symptom-limited exercise test. Compared to placebo propranolol and calcium antagonists similarly reduced (p less than 0.001) the frequency of antianginal attacks and nitroglycerin consumption, and increased exercise tolerance and time to greater than or equal to 1 mm S-T segment depression. After propranolol the pressure-rate product at submaximal and maximal exercise was significantly decreased. The calcium antagonists produced a significant reduction in the submaximal pressure-rate product, but no significant change in the peak pressure-rate product. Maximum ST depression was significantly lower after propranolol and was unchanged after the calcium antagonists. None of the drugs caused significant adverse effects. The results indicate that in patients with stable effort angina pectoris, diltiazem, nicardipine, nifedipine and verapamil were as effective as propranolol in improving exercise tolerance and time to ischaemia, and they did not alter the peak pressure-rate product. Different antianginal mechanisms may be operative for the various calcium antagonists.

Effect of dietary fats on hydroperoxide-induced chemiluminescence emission and eicosanoid release in the rat heart.

The effect of diets supplemented with three different fats (olive oil, sunflower oil, pork fat) on the susceptibility of the rat heart to oxidative stress and on the rate of eicosanoid release were studied. Our results show that when fatty-acid unsaturation of heart lipids is increased or vitamin E is decreased, even to a low degree, a marked enhancement of the susceptibility to hydroperoxide-induced oxidative stress (measured by chemiluminescence emission) occurs, which is associated with an increase of eicosanoid release from the heart.

The combined use of hyperbaric oxygen, antibiotics and surgery in the treatment of gas gangrene.

The clinical course and treatment of 21 critically ill patients suffering from gas gangrene are reported. Analysis of the results has underlined the importance of intensive care and hyperbaric management to prevent the evolution of disease and to improve patients' clinical conditions for surgical procedures.

Acute renal failure: biochemical evaluation of total parenteral nutrition with essential L-amino acids.

Plasma amino acid patterns were studied in three groups: one of 10 healthy subjects used as control, one of five patients in acute renal failure treated by haemodialysis plus a standard high-calorie non-protein diet, and one of five patients with the same clinical condition and treated in the same way except that the total parenteral nutrition consisted of a mixture of carbohydrate and essential L-amino acids. Our results indicate that haemodialysis seems to be responsible for some part of the depletion of amino acids and that their administration reduces this effect both for the essential and for the non-essential amino acids.