One-step real-time PCR assay for detection and quantification of RNA HCV to monitor patients under treatment in Brazil

The Brazilian Public Health Service provides freely αPEG-IFN to treat patients infected with HCV. The primary goal of HCV therapy is the long-term elimination of HCV from the blood to reduce the risk of HCV associated complications and death. Patient viremia affects the treatment duration and response, thus influencing clinical decisions. We developed a high-throughput method to perform the quantification of RNA hepatitis C virus (HCV) virus load in plasma samples to monitor patients under treatment. The method is based on a duplex detection, in a one-step real-time RT-PCR assay and it has been validated according to the rules established by the official Brazilian regulatory agency (ANVISA). This new method was compared to a commercial kit (Cobas/Taqman HCV Test v2.0 - Roche), showing virus load results with significant correlation between them (p?=?0,012) using commercial and clinical panels. In addition, 611 samples from patients treated with peguilated alfa-interferon (αPEG-IFN) from different regions of Brazil were analyzed. Our one-step real-time RT-PCR assay demonstrated good performance in viral load measurement and in treatment course monitoring, with acceptable sensitivity and specificity values     

Correlation between static balance and functional autonomy in elderly women

The purpose of the present study was to verify the correlation between static balance and functional autonomy in elderly women. The sample was a random selection of 32 sedentary elderly women (mean age = 67.47 ± 7.37 years, body mass index = BMI = 27.30 ± 5.07 kg/m²), who live in the city of Teresina in the state of Piauí, Brazil. Static balance was analyzed by stabilometric assessment using an electronic baropodometer which measured the average of the amplitude of postural oscillations in the right (RLD) and left (LLD) lateral displacements, anterior (AD) and posterior (PD) displacements, and in the elliptical area (EA) formed by the body's center of gravity. Functional autonomy was evaluated by a battery of tests from the LADEG protocol which is composed of: a 10 m walk (10 mW), getting up from a seated position (GSP), getting up from the prone position (GPP), getting up from a chair and movement around the house (GCMH), and putting on and taking off a shirt (PTS). The Spearman's correlation coefficient (r) indicated a positive and significant correlation between GPP and LLD (r = 0.382; p = 0.031), GPP and PD (r = 0.398; p = 0.024) and GPP and EA (r = 0.368; p = 0.038). These results show that sedentary elderly women who spent the greatest amount of time performing the GPP test achieved the largest mean amplitude of displacement leading to greater levels of instability.     

The influence of red laser irradiation timeline on burn healing in rats

Low-level laser therapy (LLLT) promotes biomodulation of wound healing and literature reports that light delivery during the inflammation could play a different role compared with latter phases of the healing process. The objective of this study was to investigate whether single dose of a red laser (λ?=?660 nm) is different from fractionated delivery protocol in full thickness burns. Two lesions were inflicted on the back of 36 rats. In the fractionated dose group (FG), the lesions were irradiated with 1 J/cm² on days 1, 3, 8, and 10 post-wounding. In the single dose group (SG), the lesions were irradiated with 4 J/cm² on day 1, immediately after injury. Control lesions (CG) received no light and were left to heal spontaneously. Blood flow was measured on days 1, 3, 8, 10, 15, and 21 using laser Doppler flowmetry. Animals were killed on days 3, 8, 10, 15, and 21. Skin specimens were obtained and routinely processed for hematoxylin and eosin. The specimens were evaluated according to differential leukocyte counting and angiogenesis. Statistical analysis was performed, and significance was accepted at p?<?0.05. Irradiated groups showed a peak of new vessels on day 15 while, for CG, the peak was on day 21. On day 21, FG exhibited a significantly greater number of cumulative neutrophils while SG showed a higher number of mononuclear cells. Our results confirm that both protocols used accelerate angiogenesis and stimulate leukocyte chemotaxis on burn treatment. In addition, this work suggests that a single-dose LLLT accelerates the inflammatory phase of skin repair.     

Diaphragmatic Ultrasound Correlates with Inspiratory Muscle Strength and Pulmonary Function in Healthy Subjects

Diaphragm ultrasound (DUS) has been used to identify diaphragm dysfunction. However, its correlations with respiratory strength and lung function are unclear, even in healthy patients. A total of 64 healthy patients (30 males) had lung function and inspiratory strength (maximal inspiratory pressure and sniff nasal inspiratory pressure) measured. Gastric and oesophageal pressures were measured in a subgroup (n?=?40). DUS was characterized by mobility (quiet breathing [QB] and deep breathing [DB]) and thickness (at functional residual capacity [Th_FRC] and total lung capacity [Th_TLC]). We calculated the thickening fraction (TF). During QB, DUS was similar between sexes. However, during DB, females had lower mobility, thickness and TF than males. Mobility at DB, Th_TLC and TF significantly correlated with lung function and inspiratory strength. These correlations were affected by sex. DUS correlated with inspiratory gastric pressure. In healthy patients, DUS correlated with lung function and inspiratory strength during DB. Significant differences between genders were noticeable when DUS was performed during DB.     

Genetic imbalances detected by multiplex ligation-dependent probe amplification in a cohort of patients with oral squamous cell carcinoma—the first step towards clinical personalized medicine

Oral tumors are a growing health problem worldwide; thus, it is mandatory to establish genetic markers in order to improve diagnosis and early detection of tumors, control relapses and, ultimately, delineate individualized therapies. This study was the first to evaluate and discuss the clinical applicability of a multiplex ligation-dependent probe amplification (MLPA) probe panel directed to head and neck cancer. Thirty primary oral squamous cell tumors were analyzed using the P428 MLPA probe panel. We detected genetic imbalances in 26 patients and observed a consistent pattern of distribution of genetic alterations in terms of losses and gains for some chromosomes, particularly for chromosomes 3, 8, and 11. Regarding the latter, some specific genes were highlighted due to frequent losses of genetic material—RARB, FHIT, CSMD1, GATA4, and MTUS1—and others due to gains—MCCC1, MYC, WISP1, PTK2, CCND1, FGF4, FADD, and CTTN. We also verified that the gains of MYC and WISP1 genes seem to suggest higher propensity of tumors localized in the floor of the mouth. This study proved the value of this MLPA probe panel for a first-tier analysis of oral tumors. The probemix was developed to include target regions that have been already shown to be of diagnostic/prognostic relevance for oral tumors. Furthermore, this study emphasized several of those specific genetic targets, suggesting its importance to oral tumor development, to predict patients’ outcomes, and also to guide the development of novel molecular therapies.