Mutation-class dependent signatures outweigh disease-associated processes in cystic fibrosis cells.

BACKGROUND: The phenotypic heterogeneity observed in Cystic Fibrosis (CF) patients suggests the involvement of other genes, besides CFTR. Here, we combined transcriptome and proteome analysis to understand the global gene expression patterns associated with five prototypical CFTR mutations. RESULTS: Evaluation of differentially expressed genes and proteins unveiled common and mutation-specific changes revealing functional signatures that are much more associated with the specific molecular defects associated with each mutation than to the CFTR loss-of-function phenotype. The combination of both datasets revealed that mutation-specific detected translated-transcripts (Dtt) have a high level of consistency. CONCLUSIONS: This is the first combined transcriptomic and proteomic study focusing on prototypical CFTR mutations. Analysis of Dtt provides novel insight into the pathophysiology of CF, and the mechanisms through which each mutation class causes disease and will likely contribute to the identification of new therapeutic targets and/or biomarkers for CF.

Exploring YAP1-centered networks linking dysfunctional CFTR to epithelial-mesenchymal transition.

Mutations in the CFTR anion channel cause cystic fibrosis (CF) and have also been related to higher cancer incidence. Previously we proposed that this is linked to an emerging role of functional CFTR in protecting against epithelial-mesenchymal transition (EMT). However, the pathways bridging dysfunctional CFTR to EMT remain elusive. Here, we applied systems biology to address this question. Our data show that YAP1 is aberrantly active in the presence of mutant CFTR, interacting with F508del, but not with wt-CFTR, and that YAP1 knockdown rescues F508del-CFTR processing and function. Subsequent analysis of YAP1 interactors and roles in cells expressing either wt- or F508del-CFTR reveal that YAP1 is an important mediator of the fibrotic/EMT processes in CF. Alongside, five main pathways emerge here as key in linking mutant CFTR to EMT, namely, (1) the Hippo pathway; (2) the Wnt pathway; (3) the TGFß pathway; (4) the p53 pathway; and (5) MYC signaling. Several potential hub proteins which mediate the crosstalk among these pathways were also identified, appearing as potential therapeutic targets for both CF and cancer.

Knee anteromedial compartment dissection: Final results and anterior oblique ligament description.

The anteromedial region of the knee is little explored in the literature and may play an important role in anteromedial rotatory instability. The purpose of this study is to describe a ligamentous structure in the anteromedial region of the knee identified in a series of anatomical dissections of cadaveric specimens. Twenty-one cadaveric knees were dissected to study the medial compartment. Exclusion criteria were signs of trauma, previous surgery, signs of osteoarthritis, and poor preservation state. The main structures of this region were identified during medial dissection. After releasing the superficial medial collateral ligament of the tibia, the anterior oblique ligament (AOL) was isolated. The morphology of the structure and its relationship with known anatomical parameters were determined. For the statistical analysis, the means and standard deviations were calculated for continuous variables. A 95% confidence interval was defined as significant. Student's t-tests were used for continuous variables. After dissection, a distinct ligamentous structure (AOL) was found in the medial region of the knee. This structure was found in 100% of the cases, was located extracapsularly and originated in the anterior aspect of the medial epicondyle, running obliquely toward the tibia. When crossing the joint, the ligament presented a fan-shaped opening, exhibiting a larger area at the tibial insertion. The AOL had a mean thickness of 6.83 ± 1.51 mm at its femoral origin and 13.39 ± 2.64 at its tibial insertion. It had a significantly (p = 0.0001) longer mean length with the knee at 90° of flexion (35.27 ± 6.59 mm) than with the knee in total extension (27.89 ± 5.46 mm), indicating that the ligament is tensioned in flexion. A new structure was identified in the anteromedial compartment of the knee with a ligamentous appearance. Further studies are necessary to identify its importance on knee stability. This study demonstrates the anatomy of a new medial structure of the knee. As a result, there will be a better understanding of the stability of the knee.

Frequency of respiratory pathogens other than SARS-CoV-2 detected during COVID-19 testing.

The frequencies of 19 respiratory pathogens other than SARS-CoV-2 were assessed in 6,"?>235 Brazilian individuals tested for COVID-19. Overall, only 83 individuals who tested positive for SARS-CoV-2 had codetection of other pathogens. Individuals infected with Rhinovirus/Enterovirus, Human Coronavirus (HCoV)-HKU1, HCoV-NL63, HPIV-4, Influenza A (-H1N1 and other subtypes), Influenza B, Human Respiratory Syncytial Virus and Human Metapneumovirus were less likely to test positive for SARS-CoV-2. Infection with Streptococcys pyogenes, Chlamydophila pneumoniae, Mycoplasma pneumoniae, and Bordetella pertussis were more frequent in individuals who tested negative for SARS-CoV-2, but without significancy. We found 150 individuals infected with ≥2 pathogens other than SARS-CoV-2, only 3 out of whom tested positive for COVID-19. The codetection frequency was low in individuals diagnosed with COVID-19. Other viral infections may provide a cross-reactive, protective immune response against SARS-CoV-2. Screening for bacterial respiratory infections upon COVID-19 testing is important to drive suitable therapeutic approaches and avoid unnecessary antibiotic prescription.

Prospective and Randomized Clinical Evaluation of Hamstring Versus Patellar Tendon Autograft for Anterior Cruciate Ligament Reconstruction in Soccer Players.

BACKGROUND: Anterior cruciate ligament (ACL) reconstruction is usually performed with autologous bone-patellar tendon-bone (PT) or hamstring tendon (HT) graft. There has been only 1 randomized clinical trial examining ACL reconstruction with these grafts specifically in soccer players, and more studies comparing these graft types within a homogenous cohort such as soccer athletes may better highlight differences in outcomes. PURPOSE: To compare the results of ACL reconstruction with PT versus HT autograft in soccer players and to evaluate objective and subjective outcomes. STUDY DESIGN: Randomized controlled trial; Level of evidence, 1. METHODS: A total of 62 professional or semiprofessional soccer players (mean age, 25.1 years) with ACL injury were randomized to undergo reconstruction with PT or HT autograft by a single orthopaedic surgeon (n = 31 in each group). Outcome measures were recorded preoperatively and at 2 years postoperatively. The primary outcome was the modified Cincinnati Knee Rating System, and secondary outcomes were the objective and subjective International Knee Documentation Committee scores, Lachman test, pivot-shift test, anterior drawer test, and Lysholm score. The following variables were also evaluated postoperatively: return to soccer, level at return, graft rerupture, postoperative complications, anterior knee pain, patellar tendinitis, difficulty sprinting, and loss of kicking power. RESULTS: The PT and HT groups were homogenous in terms of age, sex distribution, injured side, and time from injury to surgery, and there was no difference between them on any preoperative outcome score. At 2 years postoperatively, there were no differences between the groups on any outcome score; however, there were significantly fewer patients with anterior knee pain in the HT group compared with the PT group (7 [22.6%] vs 15 [48.4%], respectively; P = .03). Two patients from each group (2/31; 6.5%) sustained rerupture. CONCLUSION: There were no differences between soccer players who underwent different types of ACL reconstruction with the exception of anterior knee pain, which was more frequent in players who underwent reconstruction with PT graft.Registration: NCT02642692 (ClinicalTrials.gov).

Personalized Medicine Based on Nasal Epithelial Cells: Comparative Studies with Rectal Biopsies and Intestinal Organoids.

As highly effective CFTR modulator therapies (HEMT) emerge, there is an unmet need to find effective drugs for people with CF (PwCF) with ultra-rare mutations who are too few for classical clinical trials and for whom there are no drug discovery programs. Therefore, biomarkers reliably predicting the benefit from CFTR modulator therapies are essential to find effective drugs for PwCF through personalized approaches termed theranostics. Here, we assess CFTR basal function and the individual responses to CFTR modulators in primary human nasal epithelial (pHNE) cells from PwCF carrying rare mutations and compare these measurements with those in native rectal biopsies and intestinal organoids, respectively, in the same individual. The basal function in pHNEs shows good correlation with CFTR basal function in rectal biopsies. In parallel, CFTR rescue in pHNEs by CFTR modulators correlates to that in intestinal organoids. Altogether, results show that pHNEs are a bona fide theranostic model to assess CFTR rescue by CFTR modulator drugs, in particular for PwCF and rare mutations.

Return to Sports after ACL Reconstruction with Resection or Remnant-Preserving Technique.

Objectives To analyze the results of anterior cruciate ligament (ACL) reconstruction with remnant-preserving versus remnant-resecting technique, concerning the return to pre-lesion activity level. Methods The present retrospective cohort study has assessed adults > 18 years old who underwent ACL anatomical reconstruction between 2010 and 2014. The main outcomes assessed were: level of physical activity (4-point scale), sports participation rate, ACL rerupture defined as documented lesion requiring revision surgery and the numeric pain scale rate (NPSR). Results A total of 83 individuals were included in the study, with a mean age of 31.8 years old and follow-up mean time of 4.2 years after the surgery. A total of 34 patients underwent ACL reconstruction with remnant-preserving technique, and 49 without remnant preservation. No statistically significant difference was found between groups in all outcomes assessed: level of physical activity before the lesion and after the surgery, ACL rerupture rates and postoperative pain level. Subgroup analysis has shown a statistically significant decrease in the activity level in both groups. The most practiced sport was football; 72% of patients in the remnant group have resumed football activity versus 52.6% of the control group. Conclusion Based in these findings, the comparison between ACL reconstruction with remnant preserving technique and remnant resecting technique has shown no differences concerning the return to prelesion activity level.

Regulation of TMEM16A by CK2 and Its Role in Cellular Proliferation.

Casein kinase 2 (CK2) is a highly ubiquitous and conserved serine/threonine kinase that forms a tetramer consisting of a catalytic subunit (CK2α) and a regulatory subunit (CK2ß). Despite being ubiquitous, CK2 is commonly found at higher expression levels in cancer cells, where it inhibits apoptosis, and supports cell migration and proliferation. The Ca2+-activated chloride channel TMEM16A shows similar effects in cancer cells: TMEM16A increases cell proliferation and migration and is highly expressed in squamous cell carcinoma of the head and neck (HNSCC) as well as other malignant tumors. A microscopy-based high-throughput screening was performed to identify proteins that regulate TMEM16A. Within this screen, CK2 was found to be required for proper membrane expression of TMEM16A. small interfering (si) RNA-knockdown of CK2 reduced plasma membrane expression of TMEM16A and inhibited TMEM16A whole cell currents in (cystic fibrosis bronchial epithelial) CFBE airway epithelial cells and in the head and neck cancer cell lines Cal33 and BHY. Inhibitors of CK2, such as TBB and the preclinical compound CX4549 (silmitasertib), also blocked membrane expression of TMEM16A and Ca2+-activated whole cell currents. siRNA-knockout of CK2 and its pharmacological inhibition, as well as knockdown or inhibition of TMEM16A by either niclosamide or Ani9, attenuated cell proliferation. Simultaneous inhibition of CK2 and TMEM16A strongly potentiated inhibition of cell proliferation. Although membrane expression of TMEM16A is reduced by inhibition of CK2, our data suggest that the antiproliferative effects by inhibition of CK2 are mostly independent of TMEM16A. Simultaneous inhibition of TMEM16A by niclosamide and inhibition of CK2 by silmitasertib was additive with respect to blocking cell proliferation, while cytotoxicity was reduced when compared to solely blockade of CK2. Therefore, parallel blockade TMEM16A by niclosamide may assist with anticancer therapy by silmitasertib.

Assessment of Distinct Electrophysiological Parameters in Rectal Biopsies for the Choice of the Best Diagnosis/Prognosis Biomarkers for Cystic Fibrosis.

Most cases of Cystic Fibrosis (CF) are diagnosed early in life. However, people with atypical CF forms pose diagnosis dilemmas, requiring laboratory support for diagnosis confirmation/exclusion. Ex vivo analysis of fresh rectal biopsies by Ussing chamber has been the best discriminant biomarker for CF diagnosis/prognosis so far. Here we aimed to evaluate different electrophysiological parameters from Ussing chamber analysis of rectal biopsies from people with CF (PwCF) to establish the one with highest correlations with clinical features as the best CF diagnosis/prognosis biomarker. We analyzed measurements of CFTR-mediated Cl- secretion in rectal biopsies from 143 individuals (∼592 biopsies), the largest cohort so far analyzed by this approach. New parameters were analyzed and compared with the previous biomarker, i.e., the IBMX (I)/Forskolin (F)/Carbachol (C)-stimulated short-circuit current (I'sc-I/F/C). Correlations with clinical features showed that the best parameter corresponded to voltage measurements of the I/F + (I/F/CCH) response (VI/F+I/F/C), with higher correlations vs. I'sc-I/F/C for: sweat chloride (59 vs. 52%), fecal elastase (69 vs. 55%) and lung function, measured by FEV1 (27 vs. 20%). Altogether data show that VI/F+I/F/C is the most sensitive, reproducible, and robust predictive biomarker for CF diagnosis/prognosis effectively discriminating classical, atypical CF and non-CF groups.

An Epidemiological Profile of CrossFit Athletes in Brazil.

BACKGROUND: CrossFit is a conditioning and training program that has been gaining recognition and interest among the physically active population. Approximately 440 certified and registered CrossFit fitness centers and gyms exist in Brazil, with approximately 40,000 athletes. To date, there have been no epidemiological studies about the CrossFit athlete in Brazil. PURPOSE: To evaluate the profile, sports history, training routine, and presence of injuries among athletes of CrossFit. STUDY DESIGN: Descriptive epidemiological study. METHODS: This cross-sectional study was based on a questionnaire administered to CrossFit athletes from various specialized fitness centers in Brazil. Data were collected from May 2015 to July 2015 through an electronic questionnaire that included demographic data, level of sedentary lifestyle at work, sports training history prior to starting CrossFit, current sports activities, professional monitoring, and whether the participants experienced any injuries while practicing CrossFit. RESULTS: A total of 622 questionnaires were received, including 566 (243 women [42.9%] and 323 men [57.1%]) that were completely filled out and met the inclusion criteria and 9% that were incompletely filled out. Overall, 176 individuals (31.0%) mentioned having experienced some type of injury while practicing CrossFit. We found no significant difference in injury incidence rates regarding demographic data. There was no significant difference regarding previous sports activities because individuals who did not practice prior physical activity showed very similar injury rates to those who practiced at any level. CONCLUSION: CrossFit injury rates are comparable to those of other recreational or competitive sports, and the injuries show a profile similar to weight lifting, power lifting, weight training, Olympic gymnastics, and running, which have an injury incidence rate nearly half that of soccer.

Exercises with partial vascular occlusion in patients with knee osteoarthritis: a randomized clinical trial.

PURPOSE: The objective of this study was to evaluate whether women with knee osteoarthritis performing a rehabilitation programme consisting of low-load exercises combined with PVO exhibited the same results in changes in quadriceps strength, pain relief, and functional improvement when compared to women receiving a programme consisting of high-load exercises without PVO. METHODS: Thirty-four women (mean age, 61 years) with a diagnosis of knee osteoarthritis were randomly assigned to a conventional or occlusion group. The women in the conventional group (n = 17) performed a 6-week quadriceps strengthening and stretching programme using a load around 70 % of the 1-repetition maximum (RM). The women in the occlusion group (n = 17) performed the same programme, however, only using a load around 30 % of the 1-RM, while PVO was induced. The PVO was achieved using a pressure cuff applied to the upper third of the thigh and inflated to 200 mmHg during the quadriceps exercise. An 11-point Numerical Pain Rating Scale (NPRS), the Lequesne questionnaire, the Timed-Up and Go (TUG) test, and muscle strength measurement using a hand-held dynamometer were used as outcome measures at baseline (pretreatment) and at the end of the 6-week of treatment. Pain, using the NPRS, was also assessed when performing the quadriceps exercises during the exercise sessions. RESULTS: At baseline, demographic, strength, pain, and functional assessment data were similar between groups. Patients from both the conventional and occlusion groups had a higher level of function (Lequesne and TUG test), less pain (NPRS), and higher quadriceps strength at the 6-week evaluation when compared to baseline (all P < 0.05). However, the between-group analysis showed no differences for all outcomes variables at posttreatment (n.s.). Patients in the occlusion group experienced less anterior knee discomfort during the treatment sessions than those in the high-load exercise group (P < 0.05). CONCLUSION: A rehabilitation programme that combined PVO to low-load exercise resulted in similar benefits in pain, function, and quadriceps strength than a programme using high-load conventional exercise in patients with knee osteoarthritis. However, the use of PVO combined with low-load exercise resulted in less anterior knee pain during the training sessions. LEVEL OF EVIDENCE: I.

Proposta para estruturação de um departamento de Vigilância em Saúde Ambiental / Proposal for structuring a Environmental Health Surveillance

A estruturação do Departamento de Vigilância em Saúde Ambiental requer a dedicação de membros do setor público, em específico colaboradores da secretaria de Saúde. Desta forma foi proposta uma metodologia capaz de abranger os principais pontos a serem observados no cumprimento desta tarefa. A começar pela capacitação e treinamento continuado de seus membros, mediante pesquisas ou reuniões, passando pela estruturação de uma equipe multiprofissional capaz de lidar com questões ambientais, epidemiológicas, sanitárias e também relacionadas à saúde humana, à aquisição de instrumentos técnicos e constituição de base bibliográfica para pesquisa.

Open kinetic chain exercises in a restricted range of motion after anterior cruciate ligament reconstruction: a randomized controlled clinical trial.

BACKGROUND: Recent studies have shown that an early start of open kinetic chain (OKC) exercises for quadriceps strengthening in a full range of motion (ROM) could increase anterior knee laxity after anterior cruciate ligament (ACL) reconstruction with flexor tendons. However, there are no clinical trials that evaluated outcomes of OKC exercises in a restricted ROM for pain, function, muscle strength, and anterior knee laxity at 1 year after surgery. PURPOSE: To determine if an early start of OKC exercises for quadriceps strength in a restricted ROM would promote a clinical improvement without causing increased anterior knee laxity in patients after ACL reconstruction. STUDY DESIGN: Randomized controlled clinical trial; Level of evidence, 1. METHODS: A total of 49 patients between 16 and 50 years of age who underwent ACL reconstruction with semitendinosus and gracilis autografts were randomly assigned to an early start OKC (EOKC) exercise group or a late start OKC (LOKC) exercise group. The EOKC group (n = 25; mean age, 26 years) received a rehabilitation protocol with an early start of OKC (fourth week postoperatively) within a restricted ROM between 45° and 90°. The LOKC group (n = 24; mean age, 24 years) performed the same protocol with a late start of OKC exercises between 0° and 90° (12th week postoperatively). Quadriceps and hamstring muscle strength, 11-point numerical pain rating scale (NPRS), Lysholm knee scoring scale, single-legged and crossover hop tests, and anterior knee laxity were measured to assess outcomes at the 12-week, 19-week, 25-week, and 17-month postoperative follow-up (range, 13-24 months). RESULTS: No difference (P < .05) was noted between groups with respect to demographic data. Both groups (EOKC and LOKC) had a higher level of function and less pain at the 19-week, 25-week, and 17-month assessments when compared with 12 weeks postoperatively (P < .05). The EOKC group had improved quadriceps muscle strength at the 19-week, 25-week, and 17-month follow-up when compared with 12 weeks postoperatively (P < .05); the LOKC group showed improvement only at the 17-month postoperative assessment. However, the analysis between groups showed no difference for all pain and functional assessments, including anterior knee laxity (P > .05). CONCLUSION: An early start of OKC exercises for quadriceps strengthening in a restricted ROM did not differ from a late start in terms of anterior knee laxity. The EOKC group reached the same findings in relation to pain decrease and functional improvement when compared with the LOKC group but showed a faster recovery in quadriceps strength. The nonweightbearing exercises seem appropriate for patients who have undergone ACL reconstruction, when utilized in a specific ROM. The magnitude of difference in quadriceps strength between the 2 rehabilitation protocols was around 5%; however, this difference was not clinically significant, especially because both groups had equal function on the hop tests.

Anterior tibial stress fractures treated with anterior tension band plating in high-performance athletes.

PURPOSE: The non-surgical treatment of anterior tibial cortex stress fractures requires long periods of abstention from sports activities and often results in non-union. Many different surgical techniques have already been previously described to treat these fractures, but there is no consensus on the best treatment. We describe the outcome of treatment using anterior tibial tension band plating in three high-performance athletes (4 legs) with anterior tibial cortex stress fractures. METHODS: Tibial osteosynthesis with a 3.5-mm locking compression plate in the anterolateral aspect of the tibia was performed in all patients diagnosed with anterior tibial stress fracture after September 2010 at Santa Casa Hospital. RESULTS: All of the fractures were consolidated within a period of 3 months after surgery, allowing for an early return to pre-injury levels of competitive sports activity. There were no infection, non-union, malunion or anterior knee pain complications. CONCLUSIONS: Anterior tibial tension band plating leads to prompt fracture consolidation and is a good alternative for the treatment of anterior tibial cortex stress fractures. Bone grafts were shown to be unnecessary.

LATERAL PAIN IN AN ATHLETE'S KNEE: A RARE CASE OF DISLOCATION OF THE FEMORAL BICEPS TENDON.

Dislocation of the femoral biceps tendon is rare and is described clinically in the literature as a lateral pain in the knee. It was initially reported as an anomalous insertion of the long head of the femoral biceps. Subsequently, it was found to be caused by abnormal mobility of the tendon over the prominence of the fibular head at certain angles of knee flexion. The objective of the present report was to describe and discuss a condition of lateral knee pain in a swimmer who started to present subluxation of the femoral biceps during sports practice, which incapacitated him from taking part in trials and competitions. The case is discussed in the light of the literature surveyed; the likelihood that the etiology for the trauma leading to this condition was repetition; and the surgical treatment instituted, which led to excellent results and the patient's return to his habitual sports practice.