PRECOGx: exploring GPCR signaling mechanisms with deep protein representations.

In this study we show that protein language models can encode structural and functional information of GPCR sequences that can be used to predict their signaling and functional repertoire. We used the ESM1b protein embeddings as features and the binding information known from publicly available studies to develop PRECOGx, a machine learning predictor to explore GPCR interactions with G protein and ß-arrestin, which we made available through a new webserver (https://precogx.bioinfolab.sns.it/). PRECOGx outperformed its predecessor (e.g. PRECOG) in predicting GPCR-transducer couplings, being also able to consider all GPCR classes. The webserver also provides new functionalities, such as the projection of input sequences on a low-dimensional space describing essential features of the human GPCRome, which is used as a reference to track GPCR variants. Additionally, it allows inspection of the sequence and structural determinants responsible for coupling via the analysis of the most important attention maps used by the models as well as through predicted intramolecular contacts. We demonstrate applications of PRECOGx by predicting the impact of disease variants (ClinVar) and alternative splice forms from healthy tissues (GTEX) of human GPCRs, revealing the power to dissect system biasing mechanisms in both health and disease.

Baseline and stress-induced changes in plasma bacterial killing ability against gram-negative bacteria are partially mediated by the complement system in Rhinella diptycha toads.

The plasma bacterial killing ability (BKA) is modulated by the stress response in vertebrates, including amphibians. The complement system is an effector mechanism comprised of a set of proteins present in the plasma that once activated can promote bacterial lysis. Herein, we investigated whether changes in plasma BKA as a result of the acute stress response and an immune challenge are mediated by the complement system in Rhinella diptycha toads. Additionally, we investigated whether the observed changes in plasma BKA are associated with changes in plasma corticosterone levels (CORT). We subjected adult male toads to a restraint or an immune challenge (with three concentrations of Aeromonas hydrophila heat inactivated), and then evaluated the plasma BKA against A. hydrophila, in vitro. We determined the complement system activity on plasma BKA, by treating the plasma (baseline, 1 h and 24 h post-restraint, and after the immune challenge) with ethylenediaminetetraacetic acid, heat, or protease. Our results showed increased CORT 1 h and 24 h after restraint and decreased plasma BKA 24 h post-restraint. The inhibitors of the complement system decreased the plasma BKA compared with untreated plasma at all times (baseline, 1 h, and 24 h after restraint), demonstrating that the plasma BKA activity is partially mediated by the complement system. The immune challenge increased CORT, with the highest values being observed in the highest bacterial concentration, compared with control. The plasma BKA was not affected by the immune challenge but was demonstrated to be partially mediated by the complement system. Our results demonstrated that restraint and the immune challenge activated the hypothalamus-pituitary-interrenal axis, by increasing plasma CORT levels in R. diptycha. Also, our results demonstrated the complement system is participative in the plasma BKA for baseline and post-stress situations in these toads.

Selective laser trabeculoplasty versus micropulse laser trabeculoplasty for intraocular pressure control in patients with primary open angle glaucoma: a 12-month retrospective comparative study.

This study was designed to compare the efficacy of selective laser trabeculoplasty (SLT) and micropulse laser trabeculoplasty (MLT) in patients with primary open angle glaucoma (POAG) requiring additional IOP control. In this retrospective, comparative study, we reviewed the charts of POAG patients requiring additional IOP lowering, who underwent either SLT or MLT and were followed for at least one year. We evaluated mean intraocular pressure (IOP), mean IOP reduction from baseline and mean number of glaucoma medications 1, 3, 6 and 12 months after treatment. Success rates (success defined as IOP ≤ 21 mmHg and ≥ 20% reduction from baseline IOP without additional medications, new laser session, or glaucoma surgery) at one year were also compared. A total of 98 POAG patients were included, 52 individuals in the SLT group and 46 in the MLT group. Laser treatment resulted in significant mean IOP reductions from baseline in both groups (SLT = -6.0 ± 3.3 mmHg (24.9%) and MLT = -5.8 ± 2.6 mmHg (23.4%)) (p < 0.001). However, there was no statistically significant difference between the mean IOP reductions in both groups (p = 0.74). At 12 months, the mean number of glaucoma medications was significantly smaller in the SLT group (1.17 + 0.4) when compared to the MLT group (2.21 + 0.2) (p = 0.001). Additionally, after 12 months, success was obtained in 32 (61.5%) SLT cases compared to 27 (58.7%) MLT-treated eyes (p = 1.0). MLT and SLT are both effective in controlling the IOP in POAG patients requiring additional IOP reduction. However, after 12 months, SLT demonstrated a greater efficacy in reducing medication burden when compared to MLT.Trial registration: CEP/CONEP/MS Brazil 40948620.9.0000.5600.

Molecular detection of Toxoplasma gondii and Neospora caninum in neotropical primates.

BACKGROUND: Toxoplasma gondii and Neospora caninum infections in primates are potentially fatal and directly impact the conservation of these animals and public health. MATERIALS AND METHODS: A total of 38 blood/clot samples collected from free-living and captive neotropical primates undergoing clinical care or found dead by environmental authorities in the Mato Grosso State, Brazil, were analyzed by PCR for DNA detection of T. gondii and N. caninum. Furthermore, eight animals were submitted to immunohistochemistry for the detection of T. gondii. RESULTS: DNA of T. gondii and N. caninum was amplified in 11 (28.95%) 10 (26.32%) of samples analyzed, respectively. Coinfection was observed in three individuals. One animal returned a positive result in the immunohistochemistry for the detection of T. gondii. CONCLUSION: These findings reflect a concern for the conservation of these animals, as the pathogen-host interaction is unpredictable and infections by these protozoa can lead to animal mortality, which has a substantial impact on endangered species.

Fecal microbiota transplantation in the intestinal decolonization of carbapenamase-producing enterobacteriaceae.

BACKGROUND AND AIMS: fecal microbiota transplantation (FMT) is effective for recurrent Clostridium difficile infection (CDI). Intestinal decolonization of carbapenamase-producing enterobacteriaceae (CPE) can prevent transmission and infection by these agents. The aim of this study was to assess CPE decolonization after FMT. METHODS: this was a case-series study that consecutively included all CPE-carriers that underwent FMT between 2014 and 2019. The indications included refractory/recurrent CDI and CPE-decolonization. RESULTS: out of 21 CPE-carriers, eight were excluded due to incomplete post-FMT testing. CPE decolonization was confirmed in 76.9 % (n = 10). The median decolonization time was 16-weeks (IQR-23) and ranged from two to 53 weeks. CONCLUSION: FMT may be used in the clinical practice for CPE-decolonization as an alternative to combined antibiotic regimens.

Fecal microbiota transplantation in refractory or recurrent Clostridium difficile infection: a real-life experience in a non-academic center.

AIM: this study aimed to describe the efficacy and safety of fecal microbiota transplantation (FMT) for the treatment of refractory and recurrent Clostridium difficile infection (CDI). METHODS: this was an observational study of patients with refractory or recurrent CDI treated with FMT between June 2014 and January 2017. Primary and secondary outcomes were the resolution of diarrhea without CDI recurrence within two months after one or more FMT. A descriptive analysis was performed. RESULTS: thirty-four FMT were performed in 28 patients, 88.2% (n = 30) using an upper route with a gastroscopy and 11.8% (n = 4) with colonoscopy; 50% (n = 17) of FMT were due to recurrent CDI and 50% (n = 17) were due to refractory CDI. The overall cure rate of upper FMT was 87.5% (21/24) and 100% (4/4) when colonoscopy was performed. A cure was achieved after one FMT in 88% (22/25) of cases and after two or more FMT in 8% (2/25) of cases, resulting in an overall cure rate of 96% (24/25). No severe adverse events were reported. CONCLUSION: FMT constitutes an effective and safe approach for the management of refractory and recurrent CDI, with an overall cure rate of 96% and no reported severe adverse events.

Predictors of consistent condom use among Portuguese women attending family planning clinics.

Women account for 30% of all AIDS cases reported to the Health Ministry in Portugal and most infections are acquired through unprotected heterosexual sex with infected partners. This study analyzed socio-demographic and psychosocial predictors of consistent condom use and the role of education as a moderator variable among Portuguese women attending family planning clinics. A cross-sectional study using interviewer-administered fully structured questionnaires was conducted among 767 sexually active women (ages 18-65). Logistic regression analyses were used to explore the association between consistent condom use and the predictor variables. Overall, 78.7% of the women were inconsistent condom users. The results showed that consistent condom use was predicted by marital status (being not married), having greater perceptions of condom negotiation self-efficacy, having preparatory safer sexual behaviors, and not using condoms only when practicing abstinence. Living with a partner and having lack of risk perception significantly predicted inconsistent condom use. Less educated women were less likely to use condoms even when they perceive being at risk. The full model explained 53% of the variance in consistent condom use. This study emphasizes the need for implementing effective prevention interventions in this population showing the importance of taking education into consideration.

Initial experience with fecal microbiota transplantation in Clostridium difficile infection - transplant protocol and preliminary results.

BACKGROUND AND AIMS: Clostridium difficile infection (CDI) constitutes an important cause of antibiotic-associated diarrhea. Recurrence after first-line treatment with antibiotics is high and fecal microbiota transplantation (FMT) may be effective for refractory and recurrent CDI. This series aims to describe the efficacy of FMT in the treatment of refractory and recurrent CDI. METHODS: A prospectively recorded single-centre case series of patients with persistent or recurrent CDI treated with FMT between June 2014 and March 2015 was analyzed. Primary and secondary outcomes were defined as resolution of diarrhea without recurrence of CDI within 2 months after one or more FMT, respectively. A descriptive analysis was performed. RESULTS: 8 FMT were performed in 6 patients, 3 with refractory CDI and 3 with recurrent CDI. The median age of recipients was 71 years and 66.7% were women. One FMT was delivered through colonoscopy and the remaining 87.5% through esophagogastroduodenoscopy. One upper FMT was excluded due to recurrence of CDI after antibiotic exposure for a respiratory infection. The overall cure rate of FMT was total with lower route and 83.3% with upper route. Primary cure rate was achieved in 83.3% of patients and secondary cure rate was achieved in all patients. Median time to resolution of diarrhea after FMT was 1 day and no complications were reported during follow-up. CONCLUSION: FMT appears to constitute a safe and effective approach in the management of refractory and recurrent CDI. Difference between primary and secondary cure rates may result of insufficient restoration of intestinal microbiota with a single FMT.

Expert Insights from a Delphi-driven Neurologists' Panel: Real-world Mexiletine use in Patients with Myotonic Disorders in Italy.

Background: Myotonic disorders, such as non-dystrophic myotonias (NDMs) and myotonic dystrophies (DMs) are characterized by a delay in muscle relaxation after a contraction stimulus. There is general consensus that protocols to treat myotonia need to be implemented. Objective: Mexiletine is the only pharmacological agent approved for the symptomatic treatment of myotonia in adult patients with NDM and is considered to be the first-line treatment for DMs; however, its production in Italy was halted in 2022 making its availability to patients problematic. Methods: A panel of 8 Italian neurologists took part in a two-round Delphi panel between June and October 2022, analyzing the current use of mexiletine in Italian clinical practice. Results: The panelists assist 1126 patients (69% DM type1, 18% NDM and 13% DM type2). Adult NDM patients receive, on average, 400-600 mg of mexiletine hydrochloride (HCl) while adult DM patients receive 100-600 mg, per day in the long-term. The severity of symptoms is considered the main reason to start mexiletine treatment for both NDM and DM patients. Mexiletine is reckoned to have a clinical impact for both NDM and DM patients, but currently drug access is problematic. Conclusions: Mexiletine treatment is recognized to have a role in the reduction of the symptomatic burden for NDM and DM patients. Patient management could be improved by facilitating access to therapy and developing new drug formulations.

The landscape of cancer-rewired GPCR signaling axes.

We explored the dysregulation of G-protein-coupled receptor (GPCR) ligand systems in cancer transcriptomics datasets to uncover new therapeutics opportunities in oncology. We derived an interaction network of receptors with ligands and their biosynthetic enzymes. Multiple GPCRs are differentially regulated together with their upstream partners across cancer subtypes and are associated to specific transcriptional programs and to patient survival patterns. The expression of both receptor-ligand (or enzymes) partners improved patient stratification, suggesting a synergistic role for the activation of GPCR networks in modulating cancer phenotypes. Remarkably, we identified many such axes across several cancer molecular subtypes, including many involving receptor-biosynthetic enzymes for neurotransmitters. We found that GPCRs from these actionable axes, including, e.g., muscarinic, adenosine, 5-hydroxytryptamine, and chemokine receptors, are the targets of multiple drugs displaying anti-growth effects in large-scale, cancer cell drug screens, which we further validated. We have made the results generated in this study freely available through a webapp (gpcrcanceraxes.bioinfolab.sns.it).

The importance of the 'magic square' in immediate implant placement and provisionalization in postextraction sites.

With changing expectations for dental treatment, esthetics have become an essential factor in defining successful rehabilitation with dental implants. The stability of the gingival contours as well as the color and texture of the surrounding tissue are critical for the long-term maintenance of successful implant treatment outcomes. The aim of the present article is to demonstrate the correlation of the 3D implant position and the adjacent tissue management protocol with the long-term maintenance of immediate implant placement results in postextraction sites. A series of 12 cases with a mean follow-up of 21.91 months is presented to introduce the concept of the 'magic square' (MS), denoting the area formed in the cervical implant region when the ideal 3D implant position is achieved. This position is 3-mm coronoapical from the implant platform to the gingival margin, with the maintenance of a 3-mm vestibulopalatine thickness of the buccal bone (ie, hard tissue gap from the buccal implant surface to the outer portion of the buccal bone wall), and a ≥ 3-mm soft tissue gap at the cervical portion of the implant. The achievement of the MS promotes soft tissue growth and the formation of a thicker peri-implant bone ridge, and ensures the stability of treatment outcomes over time.

EXPANSION: a webserver to explore the functional consequences of protein-coding alternative splice variants in cancer genomics.

Summary: EXPANSION (https://expansion.bioinfolab.sns.it/) is an integrated web-server to explore the functional consequences of protein-coding alternative splice variants. We combined information from Differentially Expressed (DE) protein-coding transcripts from cancer genomics, together with domain architecture, protein interaction network, and gene enrichment analysis to provide an easy-to-interpret view of the effects of protein-coding splice variants. We retrieved all the protein-coding Ensembl transcripts and mapped Interpro domains and post-translational modifications on canonical sequences to identify functionally relevant splicing events. We also retrieved isoform-specific protein-protein interactions and binding regions from IntAct to uncover isoform-specific functions via gene-set over-representation analysis. Through EXPANSION, users can analyze precalculated or user-inputted DE transcript datasets, to easily gain functional insights on any protein spliceform of interest. Availability and Implementation: EXPANSION is freely available at http://expansion.bioinfolab.sns.it/. The code of the scripts used for EXPASION is available at: https://github.com/raimondilab/expansion. Datasets associated to this resource are available at the following URL: https://doi.org/10.5281/zenodo.8229120. The web-server was developed using Apache2 (https://https.apache.org/) and Flask (v2.0.2) (http://flask.pocoo.org/) for the web frontend and for the internal pipeline to handle back-end processes. We additionally used the following Python and JavaScript libraries at both back- and front-ends: D3 (v4), jQuery (v3.2.1), DataTables (v2.3.2), biopython (v1.79), gprofiler-officia l(v1.0.0), Mysql-connector-python (v8.0.31). To construct the API, Fast API library (v0.95.1) was used.

Long-Term Outcomes of Sternal-Sparing Versus Sternotomy Approaches for Mitral Valve Repair: Meta-Analysis of Reconstructed Time-to-Event Data.

OBJECTIVE: Since there are concerns about the durability of mitral valve repair (MVRp) with minimally invasive techniques in patients with mitral regurgitation (MR), we aimed to evaluate the long-term outcomes of these sternal-sparing approaches when compared with conventional approaches with sternotomy in patients undergoing MVRp. METHODS: We performed a systematic review according to a preestablished protocol and performed a pooled analysis of Kaplan-Meier-derived reconstructed time-to-event data from studies with longer follow-up comparing sternal-sparing versus sternotomy approaches for MVRp. Our outcomes of interest were survival, freedom from recurrent MR, and freedom from reoperation. RESULTS: Eleven studies met our eligibility criteria comprising 7,596 patients with follow-up (sternal sparing, n = 4,246; sternotomy, n = 3,350). Patients who underwent sternal-sparing MVRp had a significantly lower risk of mortality over time compared with patients who underwent MVRp with sternotomy (hazard ratio [HR] = 0.29, 95% confidence interval [CI]: 0.23 to 0.36, P < 0.001) in the overall analysis. However, we found no statistically significant difference between the groups in the sensitivity analysis with adjusted populations (HR = 0.85, 95% CI: 0.63 to 1.15, P = 0.301). Regarding the outcomes freedom from recurrent MR and freedom from reoperation, we found no statistically significant differences between the groups in the follow-up in both overall and sensitivity analyses. CONCLUSIONS: In comparison with MVRp with sternotomy approaches, sternal-sparing MVRp was not associated with worse outcomes in terms of survival, recurrent MR, and reoperations over time.

Molecular detection and genetic characterization of Ehrlichia canis and Ehrlichia sp. in neotropical primates from Brazil.

The Anaplasmataceae family includes obligate, arthropod-transmitted intracellular bacteria that can be zoonotic and potentially fatal. Studies focusing on the interaction between neotropical primates and the agents of this family are scarce. The present study aimed to identify agents of the Anaplasmataceae family in the whole blood of free-living and captive neotropical primates in the State of Mato Grosso, Central-West Brazil. Thirty-eight samples of six nonhuman primate (NHP) species were collected in seven municipalities and analysed through polymerase chain reaction (PCR), nucleotide sequencing, and phylogenetic analysis of the dsb, groEL, 16S rRNA, and gltA genes. DNA fragments similar to those of Ehrlichia canis were detected in Sapajus apella and Ehrlichia chaffeensis from Mico melanurus. The sequences generated in this study and homologous sequences retrieved from GenBank® were used for phylogenetic analyses to characterize the Ehrlichial agents detected in NHPs. The agents were then grouped into clades corresponding to different isolates from the NHP species. In addition, an Anaplasma sp. closely related to Anaplasma marginale was identified in two S. apella individuals. These findings shed light on the susceptibility of neotropical NHPs to Anaplasmataceae agents. These bacteria are known to be transmitted by ticks, which can also serve as possible sources of infection for other animals, including humans.

Head and neck basal cell carcinoma prevalence in individuals submitted to childhood X-ray epilation for tinea capitis treatment.

BACKGROUND: A higher prevalence for basal cell carcinoma (BCC) has been associated with radiation, namely with tinea capitis epilation treatment. OBJECTIVE: To evaluate the prevalence of head and neck basal cell carcinoma (BCC) and to identify the major risk factors for BCC in individuals irradiated in childhood for tinea capitis treatment. METHODS: We clinically observed 1,308 individuals from an original cohort of 5,356 irradiated between 1950 and 1963, registering previous skin lesions excisions and proposing for surgery all the suspicious lesions detected. In 585 participants, 47 with BCC, the skin pigmentation was measured. RESULTS: The overall prevalence of BCC was 8.0% and of multiple BCC was 2.4%. Both total (14.7%) and multiple BCC (6.6%) were significantly more common in the individuals who had received a higher radiation dose. Multiple BCC was more prevalent (3.7%) in younger irradiated individuals and total BCC (9.4%) in women. Participants with BCC and without BCC presented similar skin pigmentation. CONCLUSION: Younger age at irradiation, higher dose and female gender increased the risk of developing BCC in these irradiated individuals.

Dispositional mindfulness, emotional regulation and perceived stress among nursing students.

OBJECTIVE: To investigate the correlation between dispositional mindfulness, emotional regulation and perceived stress and to verify factors associated with dispositional mindfulness among nursing students. METHOD: A correlational, cross-sectional study with public undergraduate students. The following instruments were used: Mindful Attention Awareness Scale, Emotional Regulation Questionnaire and Perceived Stress Scale. Analyzes were performed using t tests, Pearson's correlation and multiple linear regression. RESULTS: The study included 330 students. There was no correlation between dispositional mindfulness and general emotional regulation score and/or perceived stress and a weak correlation with the emotional regulation dimension emotional suppression. Being in psychological treatment and psychoactive substance use were associated with decreased dispositional mindfulness. Age, sufficient sleep hours and emotional suppression were associated with an increase in this variable. CONCLUSION: There was a relationship between dispositional mindfulness only with emotional suppression, in addition to the connection of this variable with the perception of sufficient sleep hours, age, use of alcohol or psychoactive substances, undergoing psychological/psychiatric follow-up and emotional suppression.

Conventional Versus Minimally Invasive Aortic Valve Replacement Surgery: A Systematic Review, Meta-Analysis, and Meta-Regression.

Objective: To assess the potential benefits of minimally invasive aortic valve replacement (MIAVR) compared with conventional AVR (CAVR) by examining short-term outcomes. Methods: A systematic search identified randomized trials comparing MIAVR with CAVR. To assess study limitations and quality of evidence, we used the Cochrane Risk of Bias tool and GRADE and performed random-effects meta-analysis. We used meta-regression and sensitivity analysis to explore reasons for diversity. Results: Thirteen studies (1,303 patients) were included. For the comparison of MIAVR and CAVR, the risk of bias was judged low or unclear and the quality of evidence ranged from very low to moderate. No significant difference was observed in mortality, stroke, acute kidney failure, infectious outcomes, cardiac events, intubation time, intensive care unit stay, reoperation for bleeding, and blood transfusions. Blood loss (mean difference [MD] = -130.58 mL, 95% confidence interval [CI] = -216.34 to -44.82, I2 = 89%) and hospital stay (MD = -0.93 days, 95% CI = -1.62 to -0.23, I2 = 81%) were lower with MIAVR. There were shorter aortic cross-clamp (MD = 5.99 min, 95% CI = 0.99 to 10.98, I2 = 93%) and cardiopulmonary bypass (CPB) times (MD = 7.75 min, 95% CI = 0.27 to 15.24, I2 = 94%) in the CAVR group. In meta-regression analysis, we found that age was the variable with the greatest influence on heterogeneity. Conclusions: MIAVR seems to be an excellent alternative to CAVR, reducing hospital stay and incidence of hemorrhagic events. Despite significantly greater aortic cross-clamp and CPB times with MIAVR, this did not translate into adverse effects, with no changes in the results found with CAVR.

In-Hospital Outcomes of Right Minithoracotomy vs. Periareolar Access for Minimally Invasive Video-Assisted Mitral Valve Repair.

INTRODUCTION: In minimally invasive mitral valve repair, right minithoracotomy is the most widely performed method, providing a good view of the mitral valve. But regarding other techniques and although it offers limited visualization, the periareolar access is a less traumatic alternative. This study's purpose is to compare in-hospital outcomes in patients who underwent video-assisted minimally invasive mitral valve repair via right minithoracotomy and periareolar access. METHODS: This is a retrospective observational study including 37 patients (> 18 years old), without previous right thoracic surgery, who underwent their primary mitral valve repair, with indication for minimally invasive video-assisted approach (via right minithoracotomy or periareolar access), between January 2018 and August 2019. Patients' medical records were consulted to collect demographics data, operative details, and in-hospital outcomes. RESULTS: Twenty-one patients underwent right minithoracotomy, and 16 were operated via periareolar access. The mean patients' age was 62±12 years in the right minithoracotomy group and 61±9 years in the periareolar access group (P=0.2). There are no significant differences in incision length, cardiopulmonary bypass time, aortic cross-clamping time, hematocrit, amount of chest tube drainage, and intensive care unit and in-hospital length of stay. Time to extubation presented significant differences between the right minithoracotomy and the periareolar access group (4.85 hours vs. 5.62 hours, respectively) (P=0.04). CONCLUSION: In this study, we found similar results in the two applied surgical techniques, except for the time to extubation.

Clinical, Tomographic, and Histologic Evaluation of an Autogenous Bone Graft Harvested from the Maxillary Tuberosity for Guided Bone Regeneration: Case Report with a 4-Year Follow-up.

This report presents a case in which autogenous bone grafts were harvested from the maxillary tuberosity for guided bone regeneration and dental implant placement, with long-term follow-ups and assessment at the clinical, tomographic, and histologic levels. Particulate and block autogenous bone grafts were covered with a resorbable collagen membrane. Advanced bone remodeling and good bone quality, enabling dental implant placement, were observed after a short healing time (3 months). The differences in buccal bone plate thickness in the grafted area between the period immediately after implant placement and 4 years thereafter ranged from +0.879 mm to -0.001 mm. The implants osseointegrated uneventfully, and alveolar bone regeneration remained stable with a satisfactory result after 4 years.