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This in-silico investigation evaluated the mechanical impact of Morse tape implant-abutment interface and retention system (with and without screw) and restorative materials (composite block and monolithic zirconia) by means of a three-dimensional finite element analysis (3D-FEA). Four 3D models were designed for the lower first molar. A dental implant (4.5 × 10 mm B&B Dental Implant Company) was digitized (micro CT) and exported to computer-aided design (CAD) software. Non-uniform rational B-spline surfaces were reconstructed, generating a 3D volumetric model. Four different models were generated with the same Morse-type connection, but with a different locking system (with and without active screw) and a different crown material made of composite block and zirconia. The D2 bone type, which contains cortical and trabecular tissues, was designed using data from the database. The implants were juxtaposed inside the model after Boolean subtraction. Implant placement depth was simulated for the implant model precisely at crestal bone level. Each acquired model was then imported into the finite element analysis (FEA) software as STEP files. The Von Mises equivalent strains were calculated for the peri-implant bone and the Von Mises stress for the prosthetic structures. The highest strain values in bone tissue occurred in the peri-implant bone interface and were comparable in the four implant models (8.2918e-004–8.6622e-004 mm/mm). The stress peak in the zirconia crown (64.4 MPa) was higher than in the composite crown (52.2 MPa) regardless of the presence of the prosthetic screw. The abutment showed the lowest stress peaks (99.71–92.28 MPa) when the screw was present (126.63–114.25 MPa). Based on this linear analysis, it is suggested that the absence of prosthetic screw increases the stress inside the abutment and implant, without effect on the crown and around the bone tissue. Stiffer crowns concentrate more stress on its structure, reducing the amount of stress on the abutment.

Human lower-limb kinematic measurements are critical for many applications including gait analysis, enhancing athletic performance, reducing or monitoring injury risk, augmenting warfighter performance, and monitoring elderly fall risk, among others. We present a new method to estimate lower-limb kinematics using an error-state Kalman filter that utilizes an array of body-worn inertial measurement units (IMUs) and four kinematic constraints. We evaluate the method on a simplified 3-body model of the lower limbs (pelvis and two legs) during walking using data from simulation and experiment. Evaluation on this 3-body model permits direct evaluation of the ErKF method without several confounding error sources from human subjects (e.g., soft tissue artefacts and determination of anatomical frames). RMS differences for the three estimated hip joint angles all remain below 0.2 degrees compared to simulation and 1.4 degrees compared to experimental optical motion capture (MOCAP). RMS differences for stride length and step width remain within 1% and 4%, respectively compared to simulation and 7% and 5%, respectively compared to experiment (MOCAP). The results are particularly important because they foretell future success in advancing this approach to more complex models for human movement. In particular, our future work aims to extend this approach to a 7-body model of the human lower limbs composed of the pelvis, thighs, shanks, and feet.

The success of the application of granular fertilizers (GFs) in planting rows depends on the uniformity and performance of product dispensing systems, which are influenced by external factors. The objective of this study was to determine the outflow rates of two GF formulations (GF1 04-14-08 and GF2 04-30-10) using three types of fertilizer spreader—with one spiral roller (A), two spiral rollers (B), or a fluted roller (C)—and three operating speeds (1,11, 1.94, and 2.77 m s-1). The following parameters were determined in GFs: density, angle of repose, water content, and segregation (particle size). In the designed test bench, GFs were transferred from a reservoir to a spreader, and ultimately to a container, where they were weighed, and data were transmitted to the data acquisition system (DAS). A total of 7,560 outflow data points were collected (g s-1) and subjected to descriptive analysis of measures of central tendency, dispersion, asymmetry, and kurtosis, and Shewhart control charts were generated. Particle density and segregation were significantly different between the GFs, whereas the angle of repose and water content were not significantly different. The bench design and the DAS allowed measuring the outflow of GFs in different spreaders and demonstrated that this parameter was influenced by particle segregation. The segregation of GF1 was higher than that of GF2. The outflow variability at the speed of 1.11 m s-1 was lower, and the spreader with a fluted roller had the highest uniformity and was the most suitable for application with variable rates.

Purpose Clinical pathways in total knee arthroplasty (TKA) consist of general guidelines, including several topics as early rehabilitation and antibiotic systematization, which are used to improve patient’s management, decrease complication rates and enhance clinical outcomes. The primary purpose of this study was to assess whether the use of a clinical pathway for TKA can contribute to reduce LOS and healthcare costs in a private hospital, without an increase in the hospital readmission rate. We also aimed to assess whether care providers adhered to the recommendations mainly antibiotic use and physical therapy. Methods Retrospective cohort study of 485 patients who underwent TKA at private hospital. Patients were analyzed in two groups: Group I (GI), composed by 220 TKA patients, prior to the clinical pathway implementation, and Group 2 (GII), with 265 TKA patients post-clinical pathway. Several outcomes were analyzed: length of hospital stay, time from use of prophylactic antibiotic therapy, readmission within 30 days, physical therapy and costs associated to procedures and hospitalization rates. Results The implementation of the clinical pathway was related with the reduction of the length of hospital stay from 6.3 days to 4.9 days (p = 0.021) without increase in readmissions. The physical therapy on the first postoperative day was most frequent in GII than GI (96.2% vs 78.1%, p < 0.001). Prophylactic ATB 60 minutes prior the surgery was significantly more used in GII than GI (99.2% vs 87.4%, p < 0.001). In addition, ATB suspension within 48 hours was significantly more frequent in GII than GI (84.7% vs. 51.6%, p < 0.001). The cost procedure of TKA showed a reduction of US$1,252.00 in GII when compared with GI (p

Background Parkinson’s disease (PD) is a progressive multifactorial neurodegenerative condition. Epidemiological studies have shown that patients with type 2 diabetes mellitus (T2DM2) are at increased risk for developing PD, indicating a possible insulin-modulating role in this latter condition. We hypothesized that drugs similar to glucagon-like peptide-1 (GLP-1) and gastric inhibitory polypeptide (GIP), used in the treatment of T2DM2, may play a role in PD. Objectives The purpose of this study is to systematically review and meta-analyze data of preclinical and clinical studies evaluating the efficacy and safety of GLP-1 and GIP drugs in the treatment of PD. Methods Two reviewers will independently evaluate the studies available in the Ovid Medline, Ovid Embase, Web of Science, Cochrane Central Register of Controlled Trials, Cinahl, and Lilacs databases. Preclinical rodent or non-human primate studies and randomized controlled human clinical trials will be included, without language or publication period restrictions. Outcomes of interest in preclinical studies will be primarily locomotor improvements and adverse effects in animal models of PD. For clinical trials, we will evaluate clinical improvements rated by the Movement Disorders Society Unified Parkinson’s Disease Rating Scale–parts I, II, III, and IV, and adverse effects. The risk of bias of preclinical studies will be assessed by the SYRCLE tool and CAMARADES checklist and the clinical studies by the Cochrane tool; the certainty of the evidence will be rated by GRADE. Discussion and conclusion There is an urge for new PD treatments that may slow the progression of the disease rather than just restoring dopamine levels. This study will comprehensively review and update the state of the art of what is known about incretin hormones and PD and highlight the strengths and limitations of translating preclinical data to the clinic whenever possible. Systematic review registration PROSPERO registration number CRD42020223435.

The magnitude and nature of the COVID-19 pandemic prevents public health policies from relying on coercive enforcement. Practicing social distancing, wearing masks and staying at home becomes voluntary and conditional on the behavior of others. We present the results of a large-scale survey experiment in nine countries with representative samples of the population. We find that both empirical expectations (what others do) and normative expectations (what others approve of) play a significant role in compliance, beyond the effect of any other individual or group characteristic. In our vignette experiment, respondents evaluate the likelihood of compliance with social distancing and staying at home of someone similar to them in a hypothetical scenario. When empirical and normative expectations of individuals are high, respondents’ evaluation of the vignette’s character’s compliance likelihood goes up by 55% (relative to the low expectations condition). Similar results are obtained when looking at self-reported compliance among those with high expectations. Our results are moderated by individuals’ trust in government and trust in science. Holding expectations high, the effect of trusting science is substantial and significant in our vignette experiment (22% increase in compliance likelihood), and even larger in self-reported compliance (76% and 127% increase before and after the lockdown). By contrast, trusting the government only generates modest effects. At the aggregate level, the country-level trust in science, and not in government, becomes a strong predictor of compliance.

Background Colorectal cancer patients have a median age of incidence >65years although they are largely under-represented in phase-III trials. This large population contains patients unfit for treatment, those suitable for monotherapy or for doublets and the impact of chemotherapy outside clinical trial is unclear. The aim of the study was to retrospectively analyse Overall Survival(OS) of elderly metastatic colorectal cancer(mCRC) patients treated with chemotherapy in daily practice. Methods Kaplan-Meir method was used for OS, the log-rank or Tarone-Ware test for differences between subgroups, Cox’s proportional hazard model to assess the impact of known prognostic factors and treatment. Results 751 patients with mCRC observed between January 2000 and January 2013 were collected. Median age was 79 year(75–93); Male/Female 61/39%, ECOG-PS 0-1/2 85/15%; colon/rectum 74/26%; multiple metastatic sites 34%, only liver metastasis in 41% of patients. KRAS status was studied in 35% of patients: 44% of them showed gene mutation. 20.5% of patients did not received any kind of treatment including surgery. Comorbidities observed: cardiovascular 34%, diabetes 14%, hypertension 50%. Primary tumor was resected in 80.6%; surgery of liver metastasis was done in 19% of patients (2.3% of patients >80years). 78% of patients underwent chemotherapy. Median follow up was 12 months(range 1–124). Median OS was 17 months (CI 95%15–19);median OS in no-treated patients was 5 months (4–6); mOS of patients with at least one treatment was 20 months (18–22). In KRAS mutated group median OS was 19months (15–23) while in KRAS wild type patients median OS was 25 months (20–30). At multivariate analysis sex(Female), age(80 years old or with a poor performance status (respectively p

Purpose The aims of this study are to describe the epidemiological characteristics of anterior cruciate ligament reconstructions in a private hospital in Brazil and to determine trends in medical practice for comparison with previous studies. Methods We retrospectively reviewed the anterior cruciate ligament institutional register to obtain data from all patients who underwent primary anterior cruciate ligament reconstruction from July 2014 to June 2016. Descriptive statistics were used to summarize the sample. Specific statistical tests were used to assess associations between the meniscal lesion and other variables. Results During the study period, 72.6% out of 500 patients were male. The mean age at surgery was 35.1 years. The mean age was higher among females than among males (37.3 ± 12.1 vs 34.3 ± 10.8 years). The median time from injury to surgery was 44 days. The most common femoral and tibial fixations used were suspensory fixation (60.8%) and interference screw (96%), respectively. The most commonly used graft was hamstring tendon (70.2%), followed by bone-patellar tendon-bone (28.8%). A meniscal lesion was noted in 44.8% of cases. Partial meniscectomy was performed in 69.5% of meniscal lesions, and meniscal repair was performed in 14.1% of lesions. The mean length of hospital stay was 1.4 days. The proportion of men in the group of patients with an associated meniscal lesion was higher than that in the group of patients without a meniscal lesion (p = 0.007). Conclusions In this study, we identified that the vast majority of surgeries were performed in male patients in all age groups, and patients older than 30 years and with a short time from injury to surgery predominated. Concerning surgical technique, we noted a low rate of meniscal repair and a higher preference for the use of hamstring graft and suspensory fixation on the femoral side.

Objective Numerous devices have been designed to support the back during lifting tasks. To improve the utility of such devices, this research explores the use of preparatory muscle activity to classify muscle loading and initiate appropriate device activation. The goal of this study was to determine the earliest time window that enabled accurate load classification during a dynamic lifting task. Methods Nine subjects performed thirty symmetrical lifts, split evenly across three weight conditions (no-weight, 10-lbs and 24-lbs), while low-back muscle activity data was collected. Seven descriptive statistics features were extracted from 100 ms windows of data. A multinomial logistic regression (MLR) classifier was trained and tested, employing leave-one subject out cross-validation, to classify lifted load values. Dimensionality reduction was achieved through feature cross-correlation analysis and greedy feedforward selection. The time of full load support by the subject was defined as load-onset. Results Regions of highest average classification accuracy started at 200 ms before until 200 ms after load-onset with average accuracies ranging from 80% (±10%) to 81% (±7%). The average recall for each class ranged from 69–92%. Conclusion These inter-subject classification results indicate that preparatory muscle activity can be leveraged to identify the intent to lift a weight up to 100 ms prior to load-onset. The high accuracies shown indicate the potential to utilize intent classification for assistive device applications. Significance Active assistive devices, e.g. exoskeletons, could prevent back injury by off-loading low-back muscles. Early intent classification allows more time for actuators to respond and integrate seamlessly with the user.

Prophylactic vaccination using live attenuated classical swine fever (CSF) vaccines has been a very effective method to control the disease in endemic regions and during outbreaks in previously disease-free areas. These vaccines confer effective protection against the disease at early times post-vaccination although the mechanisms mediating the protection are poorly characterized. Here we present the events occurring after the administration of our in-house developed live attenuated marker vaccine, FlagT4Gv. We previously reported that FlagT4Gv intramuscular (IM) administered conferred effective protection against intranasal challenge with virulent CSFV (BICv) as early as 7 days post-vaccination. Here we report that FlagT4Gv is able to induce protection against disease as early as three days post-vaccination. Immunohistochemical testing of tissues from FlagT4Gv-inoculated animals showed that tonsils were colonized by the vaccine virus by day 3 post-inoculation. There was a complete absence of BICv in tonsils of FlagT4Gv-inoculated animals which had been intranasal (IN) challenged with BICv 3 days after FlagT4Gv infection, confirming that FlagT4Gv inoculation confers sterile immunity. Analysis of systemic levels of 19 different cytokines in vaccinated animals demonstrated an increase of IFN-α three days after FlagT4Gv inoculation compared with mock infected controls.

Arboviruses (arthropod borne viruses) have life cycles that include both vertebrate and invertebrate hosts with substantial differences in vector and host specificity between different viruses. Most arboviruses utilize RNA for their genetic material and are completely dependent on host tRNAs for their translation, suggesting that virus codon usage could be a target for selection. In the current study we analyzed the relative synonymous codon usage (RSCU) patterns of 26 arboviruses together with 25 vectors and hosts, including 8 vertebrates and 17 invertebrates. We used hierarchical cluster analysis (HCA) and principal component analysis (PCA) to identify trends in codon usage. HCA demonstrated that the RSCU of arboviruses reflects that of their natural hosts, but not that of dead-end hosts. Of the two major components identified by PCA, the first accounted for 62.1% of the total variance, and among the 59 codons analyzed in this study, the leucine codon CTG had the highest correlation with the first principal component, however isoleucine had the highest correlation during amino acid analysis. Nucleotide and dinucleotide composition were the variables that explained most of the total codon usage variance. The results suggest that the main factors driving the evolution of codon usage in arboviruses is based on the nucleotide and dinucleotide composition present in the host. Comparing codon usage of arboviruses and potential vector hosts can help identifying potential vectors for emerging arboviruses.

This study introduces a new method to understand how added load affects human performance across a broad range of athletic tasks (ten obstacles) embedded in an outdoor obstacle course. The method employs an array of wearable inertial measurement units (IMUs) to wirelessly record the movements of major body segments to derive obstacle-specific metrics of performance. The effects of load are demonstrated on (N = 22) participants who each complete the obstacle course under four conditions including unloaded (twice) and with loads of 15% and 30% of their body weight (a total of 88 trials across the group of participants). The IMU-derived performance metrics reveal marked degradations in performance with increasing load across eight of the ten obstacles. Overall, this study demonstrates the significant potential in using this wearable technology to evaluate human performance across multiple tasks and, simultaneously, the adverse effects of body-borne loads on performance. The study addresses a major need of military organizations worldwide that frequently employ standardized obstacle courses to understand how added loads influence warfighter performance. Importantly, the findings and conclusions drawn from IMU data would not be possible using traditional timing metrics used to evaluate task performance.

The study investigated the effect of antibiotic combinations against 20 clinical isolates of A. baumannii (seven colistin-resistant and 13 colistin-susceptible) with different resistance mechanisms. Clinical data, treatment, and patient mortality were evaluated. The following methods were used: MIC, PCRs, and outer membrane protein (OMP) analysis. Synergy was investigated using the checkerboard and time-kill methods. Clonality was evaluated by PFGE. Based on clonality, the whole genome sequence of six A. baumannii isolates was analyzed. All isolates were resistant to meropenem, rifampicin, and fosfomycin. OXA-23 and OXA-143 were the most frequent carbapenemases found. Four isolates showed loss of a 43kDa OMP. The colistin-susceptible isolates belonged to different clones and showed the highest synergistic effect with fosfomycin-amikacin. Among colistin-resistant isolates, the highest synergistic effect was observed with the combinations of colistin-rifampicin followed by colistin-vancomycin. All colistin-resistant isolates harbored blaOXA-23-like and belonged to CC113. Clinical and demographic data were available for 18 of 20 patients. Fourteen received treatment and eight patients died during treatment. The most frequent site of infection was the blood in 13 of 14 patients. Seven patients received vancomycin plus an active drug against A. baumannii; however, mortality did not differ in this group. The synergistic effect was similar for colistin-susceptible isolates of distinct clonal origin presenting with the same resistance mechanism. Overall mortality and death during treatment was high, and despite the high synergism in vitro with vancomycin, death did not differ comparing the use or not of vancomycin plus an active drug against A. baumannii.

Ajudes rebudes: Marie Curie Career Integration Grant; Dexeus Foundation for Women's Health Research; i Contratos Ramón y Cajal CD11c is an α integrin classically employed to define myeloid dendritic cells. Although there is little information about CD11c expression on human T cells, mouse models have shown an association of CD11c expression with functionally relevant T cell subsets. In the context of genital tract infection, we have previously observed increased expression of CD11c in circulating T cells from mice and women. Microarray analyses of activated effector T cells expressing CD11c derived from naïve mice demonstrated enrichment for natural killer (NK) associated genes. Here we find that murine CD11c+ T cells analyzed by flow cytometry display markers associated with non-conventional T cell subsets, including γδ T cells and invariant natural killer T (iNKT) cells. However, in women, only γδ T cells and CD8+ T cells were enriched within the CD11c fraction of blood and cervical tissue. These CD11c+ cells were highly activated and had greater interferon (IFN)-γ secretory capacity than CD11c- T cells. Furthermore, circulating CD11c+ T cells were associated with the expression of multiple adhesion molecules in women, suggesting that these cells have high tissue homing potential. These data suggest that CD11c expression distinguishes a population of circulating T cells during bacterial infection with innate capacity and mucosal homing potential.

Altres ajuts: Marie Curie Career Integration Grant i una beca Fundació Dexeus Salut de la Dona Efforts to develop vaccines that can elicit mucosal immune responses in the female genital tract against sexually transmitted infections have been hampered by an inability to measure immune responses in these tissues. The differential expression of adhesion molecules is known to confer site-dependent homing of circulating effector T cells to mucosal tissues. Specific homing molecules have been defined that can be measured in blood as surrogate markers of local immunity (e.g. α4β7 for gut). Here we analyzed the expression pattern of adhesion molecules by circulating effector T cells following mucosal infection of the female genital tract in mice and during a symptomatic episode of vaginosis in women. While CCR2, CCR5, CXCR6 and CD11c were preferentially expressed in a mouse model of Chlamydia infection, only CCR5 and CD11c were clearly expressed by effector T cells during bacterial vaginosis in women. Other homing molecules previously suggested as required for homing to the genital mucosa such as α4β1 and α4β7 were also differentially expressed in these patients. However, CD11c expression, an integrin chain rarely analyzed in the context of T cell immunity, was the most consistently elevated in all activated effector CD8+ T cell subsets analyzed. This molecule was also induced after systemic infection in mice, suggesting that CD11c is not exclusive of genital tract infection. Still, its increase in response to genital tract disorders may represent a novel surrogate marker of mucosal immunity in women, and warrants further exploration for diagnostic and therapeutic purposes.

BACKGROUND: High-density electroencephalography (hd-EEG) combined with transcranial magnetic stimulation (TMS) provides a direct and non-invasive measure of cortical excitability and connectivity in humans and may be employed to track over time pathological alterations, plastic changes and therapy-induced modifications in cortical circuits. However, the diagnostic/monitoring applications of this technique would be limited to the extent that TMS-evoked potentials are either stereotypical (non-sensitive) or random (non-repeatable) responses. Here, we used controlled changes in the stimulation parameters (site, intensity, and angle of stimulation) and repeated longitudinal measurements (same day and one week apart) to evaluate the sensitivity and repeatability of TMS/hd-EEG potentials. METHODOLOGY/PRINCIPAL FINDINGS: In 10 volunteers, we performed 92 single-subject comparisons to evaluate the similarities/differences between pairs of TMS-evoked potentials recorded in the same/different stimulation conditions. For each pairwise comparison, we used non-parametric statistics to calculate a Divergence Index (DI), i.e., the percentage of samples that differed significantly, considering all scalp locations and the entire post-stimulus period. A receiver operating characteristic analysis showed that it was possible to find an optimal DI threshold of 1.67%, yielding 96.7% overall accuracy of TMS/hd-EEG in detecting whether a change in the perturbation parameters occurred or not. CONCLUSIONS/SIGNIFICANCE: These results demonstrate that the EEG responses to TMS essentially reflect deterministic properties of the stimulated neuronal circuits as opposed to stereotypical responses or uncontrolled variability. To the extent that TMS-evoked potentials are sensitive to changes and repeatable over time, they may be employed to detect longitudinal changes in the state of cortical circuits.

Metabolic disorders have been established as major risk factors for ocular complications and poor vision. However, little is known about the inverse possibility that ocular disease may cause metabolic dysfunction. To test this hypothesis, we assessed the metabolic consequences of a robust dietary challenge in several mouse models suffering from retinal mutations. To this end, mice null for melanopsin (Opn4-/-), the photopigment of intrinsically photosensitive retinal ganglion cells (ipRGCs), were subjected to five weeks of a ketogenic diet. These mice lost significantly more weight than wild-type controls or mice lacking rod and cone photoreceptors (Pde6brd1/rd1). Although ipRGCs are critical for proper circadian entrainment, and circadian misalignment has been implicated in metabolic pathology, we observed no differences in entrainment between Opn4-/- and control mice. Additionally, we observed no differences in any tested metabolic parameter between these mouse strains. Further studies are required to establish the mechanism giving rise to this dramatic phenotype observed in melanopsin-null mice. We conclude that the causality between ocular disease and metabolic disorders merits further investigation due to the popularity of diets that rely on the induction of a ketogenic state. Our study is a first step toward understanding retinal pathology as a potential cause of metabolic dysfunction.

The purpose of this study was to investigate the effects of the ovarian hormones and the use of oral contraceptive pills (OCP) on cardiac vagal withdrawal at the onset of dynamic exercise. Thirty physically active women aged 19–32 years were divided into two groups: OCP users (n = 17) and non-OCP users (n = 13). Participants were studied randomly at three different phases of the menstrual cycle: early follicular (day 3.6 ± 1.2; range 1–5), ovulatory (day 14.3 ± 0.8; range 13–16) and midluteal (day 21.3 ± 0.8; range 20–24), according to endogenous (in non-OCP users) or exogenous (in OCP users) estradiol and progesterone variations. The cardiac vagal withdrawal was represented by the cardiac vagal index (CVI), which was obtained by the 4-s exercise test. Additionally, resting heart rate, systolic (SBP) and diastolic blood pressure (DBP) were obtained. The CVI was not significantly different between the three phases of the menstrual cycle in either the non-OCP users (early follicular: 1.58 ± 0.1; ovulatory: 1.56 ± 0.1; midluteal: 1.58 ± 0.1, P > 0.05) or OCP users (early follicular: 1.47 ± 0.1; ovulatory: 1.49 ± 0.1; midluteal: 1.47 ± 0.1, P > 0.05) (mean ± SEM). Resting cardiovascular responses were not affected by hormonal phase or OCP use, except that the SBP was higher in the OCP users than non-OCP users in all phases of the cycle (P < 0.05). In summary, our results demonstrate that cardiac vagal withdrawal at the onset of dynamic exercise was not impacted by the menstrual cycle or OCP use in physically active women.

The ability of mammals to feel noxious stimuli lies in a heterogeneous group of primary somatosensory neurons termed nociceptors, which express specific membrane receptors, such as the Transient Receptor Potential (TRP) family. Here, we show that one of the most important nociceptive-like pathways is conserved in the freshwater coelenterate Hydra vulgaris, the most primitive organism possessing a nervous system. In particular, we found that H. vulgaris expresses TRPM3, a nociceptor calcium channel involved in the detection of noxious heat in mammals. Furthermore, we detected that both heat shock and TRPM3 specific agonist (i.e., pregnenolone sulfate) induce the modulation of the heat shock protein 70 (HSP70) and the nitric oxide synthase (NOS), two genes activated by TRP-mediated heat painful stimuli in mammals. As expected, these effects are inhibited by a TRPM3 antagonist (i.e., mefenamic acid). Interestingly, the TRPM3 agonist and heat shock also induce the expression of nuclear transcription erythroid 2-related factor (Nrf2) and superoxide dismutase (SOD), known markers of oxidative stress; noteworthy gene expression was also inhibited by the TRPM3 antagonist. As a whole, our results demonstrate the presence of conserved molecular oxidative/nociceptive-like pathways at the primordial level of the animal kingdom.

Melanopsin has been implicated in the mammalian photoentrainment by blue light. This photopigment, which maximally absorbs light at wavelengths between 470 and 480 nm depending on the species, is found in the retina of all classes of vertebrates so far studied. In mammals, melanopsin activation triggers a signaling pathway which resets the circadian clock in the suprachiasmatic nucleus (SCN). Unlike mammals, Drosophila melanogaster and Danio rerio do not rely only on their eyes to perceive light, in fact their whole body may be capable of detecting light and entraining their circadian clock. Melanopsin, teleost multiple tissue (tmt) opsin and others such as neuropsin and va-opsin, are found in the peripheral tissues of Danio rerio, however, there are limited data concerning the photopigment/s or the signaling pathway/s directly involved in light detection. Here, we demonstrate that melanopsin is a strong candidate to mediate synchronization of zebrafish cells. The deduced amino acid sequence of melanopsin, although being a vertebrate opsin, is more similar to invertebrate than vertebrate photopigments, and melanopsin photostimulation triggers the phosphoinositide pathway through activation of a G(q/11)-type G protein. We stimulated cultured ZEM-2S cells with blue light at wavelengths consistent with melanopsin maximal absorption, and evaluated the time course expression of per1b, cry1b, per2 and cry1a. Using quantitative PCR, we showed that blue light is capable of slightly modulating per1b and cry1b genes, and drastically increasing per2 and cry1a expression. Pharmacological assays indicated that per2 and cry1a responses to blue light are evoked through the activation of the phosphoinositide pathway, which crosstalks with nitric oxide (NO) and mitogen activated protein MAP kinase (MAPK) to activate the clock genes. Our results suggest that melanopsin may be important in mediating the photoresponse in Danio rerio ZEM-2S cells, and provide new insights about the modulation of clock genes in peripheral clocks.

Made available in DSpace on 2014-12-03T13:07:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-01-22Bitstream added on 2014-12-03T13:24:34Z : No. of bitstreams: 1 WOS000330283100118.pdf: 187648 bytes, checksum: cb3f43580359a8cc363610bfd86beeaa (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Bladder cancer is one of the most common genitourinary neoplasms in industrialized countries. Multifocality and high recurrence rates are prominent clinical features of this disease and contribute to its high morbidity. Therefore, more sensitive and less invasive techniques could help identify individuals with asymptomatic disease. In this context, we used the micronucleus assay to evaluate whether cytogenetic alterations could be used as biomarkers for monitoring patients with a history of urothelial cell carcinoma (UCC). We determined the frequency of micronucleated urothelial cells (MNC) in exfoliated bladder cells from 105 patients with (n = 52) or without (n = 53) a history of UCC, all of whom tested negative for neoplasia by cytopathological and histopathological analyses. MNC frequencies were increased in patients with a history of UCC (non-smoker and smoker/ex-smoker patients vs non-smoker and smoker/ex-smoker controls; p

Photobacterium profundum is a cosmopolitan marine bacterium capable of growth at low temperature and high hydrostatic pressure. Multiple strains of P. profundum have been isolated from different depths of the ocean and display remarkable differences in their physiological responses to pressure. The genome sequence of the deep-sea piezopsychrophilic strain Photobacterium profundum SS9 has provided some clues regarding the genetic features required for growth in the deep sea. The sequenced genome of Photobacterium profundum strain 3TCK, a non-piezophilic strain isolated from a shallow-water environment, is now available and its analysis expands the identification of unique genomic features that correlate to environmental differences and define the Hutchinsonian niche of each strain. These differences range from variations in gene content to specific gene sequences under positive selection. Genome plasticity between Photobacterium bathytypes was investigated when strain 3TCK-specific genes involved in photorepair were introduced to SS9, demonstrating that horizontal gene transfer can provide a mechanism for rapid colonisation of new environments. Published version

Scedosporium apiospermum is an emerging fungal pathogen that causes both localized and disseminated infections in immunocompromised patients. Glucosylceramides (CMH, GlcCer) are the main neutral glycosphingolipids expressed in fungal cells. In this study, glucosylceramides (GlcCer) were extracted and purified in several chromatographic steps. Using high-performance thin layer chromatography (HPTLC) and electrospray ionization mass spectrometry (ESI-MS), N-2'-hydroxyhexadecanoyl-1-β-D-glucopyranosyl-9-methyl-4,8-sphingadienine was identified as the main GlcCer in S. apiospermum. A monoclonal antibody (Mab) against this molecule was used for indirect immunofluorescence experiments, which revealed that this CMH is present on the surface of the mycelial and conidial forms of S. apiospermum. Treatment of S. apiospermum conidia with the Mab significantly reduced fungal growth. In addition, the Mab also enhanced the phagocytosis and killing of S. apiospermum by murine cells. In vitro assays were performed to evaluate the CMHs for their cytotoxic activities against the mammalian cell lines L.929 and RAW, and an inhibitory effect on cell proliferation was observed. Synergistic in vitro interactions were observed between the Mab against GlcCer and both amphotericin B (AmB) and itraconazole. Because Scedosporium species develop drug resistance, the number of available antifungal drugs is limited; our data indicate that combining immunotherapy with the available drugs might be a viable treatment option. These results suggest that in S. apiospermum, GlcCer are most likely cell wall components that are targeted by antifungal antibodies, which directly inhibit fungal development and enhance macrophage function; furthermore, these results suggest the combined use of monoclonal antibodies against GlcCer and antifungal drugs for antifungal immunotherapy.