Minimal Functionalization of Ruthenium Compounds with Enhanced Photoreactivity against Hard-to-Treat Cancer Cells and Resistant Bacteria.

Metallocompounds have emerged as promising new anticancer agents, which can also exhibit properties to be used in photodynamic therapy. Here, we prepared two ruthenium-based compounds with a 2,2'-bipyridine ligand conjugated to an anthracenyl moiety. These compounds coded GRBA and GRPA contain 2,2'-bipyridine or 1,10-phenathroline as auxiliary ligands, respectively, which provide quite a distinct behavior. Notably, compound GRPA exhibited remarkably high photoproduction of singlet oxygen even in water (Ï•Δ = 0.96), almost twice that of GRBA (Ï•Δ = 0.52). On the other hand, this latter produced twice more superoxide and hydroxyl radical species than GRPA, which may be due to the modulation of their excited state. Interestingly, GRPA exhibited a modest binding to DNA (Kb = 4.51 × 104), while GRBA did not show a measurable interaction only noticed by circular dichroism measurements. Studies with bacteria showed a great antimicrobial effect, including a synergistic effect in combination with commercial antibiotics. Besides that, GRBA showed very low or no cytotoxicity against four mammalian cells, including a hard-to-treat MDA-MB-231, triple-negative human breast cancer. Potent activities were measured for GRBA upon blue light irradiation, where IC50 of 43 and 13 nmol L-1 were seen against hard-to-treat triple-negative human breast cancer (MDA-MB-231) and ovarian cancer cells (A2780), respectively. These promising results are an interesting case of a simple modification with expressive enhancement of biological activity that deserves further biological studies.

Equal rates of bone healing and reduced surgical time with iliac crest allograft compared to autograft in medial opening wedge high tibial osteotomy: a randomized controlled clinical trial.

INTRODUCTION: Iliac crest autograft is frequently used to fill in bone defects after osteotomies. Nonetheless, surgery for bone autograft procurement is associated with morbidity and pain at the donor site. Alternatives to it have been explored, but there is no consensus to guide their application as a routine practice in several orthopedic procedures. Thus, this study was designed to compare the efficacy and safety between iliac crest autograft and allograft in medial opening wedge high tibial osteotomy. MATERIALS AND METHODS: Forty-seven patients with a symptomatic unilateral genu varum and an indication for high tibial osteotomy were randomly assigned to receive either autograft or allograft to fill the osteotomy site. Operative time, bone healing, and complication rates (delayed union, nonunion, superficial and deep infection, loss of correction, and hardware failure) were recorded after a one-year follow-up. Data were expressed as Mean ± Standard Deviation and considered statistically significant when p < 0.05. RESULTS: The time to radiologic union was similar between both groups (Allograft: 2.38 ± 0.97 months vs. Autograft: 2.45 ± 0.91 months; p = 0.79). Complication rates were also similar in both groups, with one infection in the allograft group and two in the autograft group, two delayed unions in the allograft group, and three in the autograft group. The operative time differed by 11 min between the groups, being lower in the allograft group (Allograft: 65.4 ± 15.1 min vs. Autograft: 76.3 ± 15.2 min; p = 0.02). CONCLUSION: Iliac crest allografts can be safely and effectively used in medial opening wedge high tibial osteotomy as it promotes the same rates of bone union as those achieved by autologous grafts, with the benefits of a shorter operative time. TRIAL REGISTRATION NUMBER: U1111-1280-0637 1 December 2022, retrospectively registered.

Açaí (Euterpe oleracea Mart.) Seed Oil and Its Nanoemulsion: Chemical Characterisation, Toxicity Evaluation, Antioxidant and Anticancer Activities.

This study explores a nanoemulsion formulated with açaí seed oil, known for its rich fatty acid composition and diverse biological activities. This study aimed to characterise a nanoemulsion formulated with açaí seed oil and explore its cytotoxic effects on HeLa and SiHa cervical cancer cell lines, alongside assessing its antioxidant and toxicity properties both in vitro and in vivo. Extracted from fruits sourced in Brazil, the oil underwent thorough chemical characterization using gas chromatography-mass spectrometry. The resulting nanoemulsion was prepared and evaluated for stability, particle size, and antioxidant properties. The nanoemulsion exhibited translucency, fluidity, and stability post centrifugation and temperature tests, with a droplet size of 238.37, PDI -9.59, pH 7, and turbidity 0.267. In vitro assessments on cervical cancer cell lines revealed antitumour effects, including inhibition of cell proliferation, migration, and colony formation. Toxicity tests conducted in cell cultures and female Swiss mice demonstrated no adverse effects of both açaí seed oil and nanoemulsion. Overall, açaí seed oil, particularly when formulated into a nanoemulsion, presents potential for cancer treatment due to its bioactive properties and safety profile.

The optimized tourniquet versus no tourniquet in total knee arthroplasty. Analysis of muscle injury, functional recovery, and knee strength.

Background: Tourniquet is widely used in total knee replacement surgery because it reduces intraoperative hemorrhage and provides a comfortable surgical area for the surgeon. It's possible that its use could lead to impaired postoperative functional and motor recovery, as well as local and systemic complications. Our goal was to compare the outcomes of total knee replacement without ischemia using an optimized protocol, consisting of tourniquet inflation before skin incision and deflation after cementing, with a pressure of one hundred millimeters above systolic blood pressure and without postoperative articular suction drains.). We believed that tourniquet effectively would result in no additional muscle damage and no functional or knee strength impairment compared to no tourniquet. Methods: In a prospective and randomized study, 60 patients with osteoarthritis were evaluated for total knee replacement, divided in two groups: 'without tourniquet' and 'optimized tourniquet'. Outcomes were mean creatine phosphokinase levels, Knee Society Score and knee isokinetic strength. Data were considered significant when p < 0.05. Results: Creatine phosphokinase levels and functional score were similar between groups. There were no differences between groups regarding knee extension strength on the operated limbs, although the knee flexors' peak torque in the operated limb in the optimized tourniquet group was significantly higher at 6 months relative to preoperative and 3 months assessments. Conclusions: The optimized tourniquet protocol use in total knee replacement combines the benefits of tourniquet use without compromising functional recovery and without additional muscle damage and strength deficits compared to surgery without its use.

Correlation between inflammatory mediators and biochemical markers in patients with active pulmonary tuberculosis / Correlação entre mediadores inflamatórios e marcadores bioquímicos em pacientes com tuberculose pulmonar ativa

Objective: Correlate inflammatory mediators and biochemical parameters in patients with active pulmonary tuberculosis (TB) treated at a public hospital in São Luís, MA. Methods: This is a case-control study of patients with a positive diagnosis of active pulmonary TB. Serum samples from patients and the control group were collected for the clinical trials, and epidemiological data were collected through medical records and interviews. The control group consisted of healthy volunteers with no previous contact with TB cases, matched by age and sex to the clinical group. To measure inflammatory cytokines, we used the Human IL-6 ELISA Set and Human IFN-γ ELISA Set kits. Oxidative stress was measured by quantification of thiobarbituric acid reactive substances (TBARS) and nitric oxide (NO). In biochemistry, the levels of uric acid, antistreptolysin "O" (AEO), alanine aminotransferase (ALT), amylase, aspartate aminotransferase (AST), calcium, total cholesterol, gamma-glutamyl transferase (Gamma GT), glucose, alkaline phosphatase, high-density lipoprotein (HDL), C-reactive protein (CRP) and triglycerides were measured. Results: The clinical group consisted of 53 patients. There was a substantial decrease in IFN-γ (p<0.0001) and a significant increase in IL-6 (p<0.0001). TBARS production increased significantly (p= 0.0414). There was no significant difference in NO production (p= 0.3194). In biochemistry, there was a significant increase in ALT (p= 0.0072), AST (p= 0.0016), Gamma GT (p= 0.0011), alkaline phosphatase (p<0.0001), CRP (p<0. .0001) and triglycerides (p= 0.0343), and a significant decrease in calcium (p<0.0001). A significant positive correlation was found between IL-6 and IFN-γ (p= 0.0448), as well as AST and ALT (p<0.0001); CRP and gamma GT (p<0.0001); Gamma GT and ALT (p= 0.0016); Gamma GT and AST (p=0.0004); triglycerides and cholesterol (p= 0.0002); alkaline phosphatase and gamma GT (p<0.0001); CRP and alkaline phosphatase (p<0.0001); triglycerides and calcium (p= 0.0121); cholesterol and calcium (p= 0.0261); glucose and cholesterol (p= 0.0373); and triglycerides and glucose (p= 0.0127) in biochemistry, with a significant negative correlation between glucose and uric acid (p= 0.0092); and CRP and HDL (p=0.0037). The correlation between inflammatory mediators and biochemical markers was positive between IL-6 and gamma GT (p= 0.0011); IL-6 and CRP (p<0.0001); IL-6 and alkaline phosphatase (p=0.0076); and NO and triglycerides (p= 0.0016), and significant negative correlation between IFN-γ and cholesterol (p= 0.0171) and TBARS and cholesterol (p= 0.0138). Conclusion: Immunosuppression of IFN-γ activity was observed. A correlation was found between IL-6 and inflammatory biochemical markers, indicating damage and injury caused by M. tuberculosis (AU).

Objetivo: Correlacionar mediadores inflamatórios e parâmetros bioquímicos em pacientes com tuberculose (TB) pulmonar ativa atendidos em um hospital público, em São Luís, MA. Métodos: Trata-se um caso-controle de pacientes com diagnóstico positivo para TB pulmonar ativa. Amostras de soro dos pacientes e grupo controle foram coletadas para os experimentos clínicos e os dados epidemiológicos foram coletados por meio de prontuários e entrevistas. O grupo controle foi formado por voluntários saudáveis sem contato prévio com casos de TB, pareados com idade e sexo ao grupo clínico. Para dosar citocinas inflamatórias, utilizaram-se os kits Human IL-6 ELISA Set e Human IFN-γ ELISA Set. Mediu-se o estresse oxidativo pela quantificação das espécies reativas do ácido tiobarbitúrico (TBARS) e óxido nítrico (ON). Na bioquímica, mediram-se os níveis de ácido úrico, anti-estreptolisina-O (AEO), alanina aminotransferase (ALT), amilase, aspartato aminotransferase (AST), cálcio, colesterol total, gama glutamil transferase (Gama GT), glicose, fosfatase alcalina, lipoproteína de alta densidade (HDL), proteína C reativa (PCR) e triglicerídeos. A análise estatística foi realizada pelo software Graph Pad Prism 8, com p<0,05 significativo. Re -sultados: O grupo clínico foi formado por 53 pacientes. Houve uma diminuição significativa de IFN-γ (p<0,0001), e aumento significativo de IL-6 (p<0,0001). A produção de TBARS aumentou significativamente (p= 0,0414). Não houve diferença significativa na produção de ON (p= 0,3194). Na bioquímica, houve aumento significativo em ALT (p= 0,0072), AST (p= 0,0016), gama GT (p= 0,0011), fosfatase alcalina (p<0,0001), PCR (p<0,0001) e triglice-rídeos (p= 0,0343), e diminuição significativa de cálcio (p<0,0001). Encontrou-se correlação positiva significativa entre IL-6 e IFN-γ (p= 0,0448), assim como AST e ALT (p<0,0001); PCR e gama GT (p<0,0001); gama GT e ALT (p= 0,0016); gama GT e AST (p= 0,0004); triglicerídeos e colesterol (p= 0,0002); fosfatase alcalina e gama GT (p<0,0001); PCR e fosfatase alcalina (p<0,0001); triglicerídeos e cálcio (p= 0,0121); colesterol e cálcio (p= 0,0261); glicose e colesterol (p= 0,0373); e triglicerídeos e glicose (p= 0,0127) na bioquímica, sendo negativa significativa entre glicose e ácido úrico (p= 0,0092); e PCR e HDL (p= 0,0037). A correlação entre marcadores infla-matório e bioquímicos foi positiva entre IL-6 e gama GT (p= 0,0011); IL-6 e PCR (p<0,0001); IL-6 e fosfatase alcalina (p= 0,0076); e ON e triglicerídeos (p= 0,0016), e negativa significativa entre IFN-γ e colesterol (p= 0,0171) e TBARS e colesterol (p= 0,0138). Conclusões: Observou-se imunossupressão da atividade de IFN-γ. Encontrou-se correlação entre IL-6 e marcadores bioquímicos inflamatórios, indicando dano e lesão causados por M. tuberculosis (AU).

COVID-19 vaccine booster doses provide increased protection against COVID-19 hospitalization compared with previously vaccinated individuals: Interim findings from the REFORCO-Brazil real-world effectiveness study during Delta and Omicron.

BACKGROUND: Although COVID-19 booster vaccination is widely recommended, there is limited long-term, population-level, real-world evidence on the magnitude of improved protection against severe COVID-19 conferred by boosting with monovalent COVID-19 vaccines developed against ancestral SARS-CoV-2, especially in low- or middle-income countries. We present interim results from the first large-scale assessment of the relative vaccine effectiveness (rVE) of first and second booster doses against severe COVID-19 in a low-/middle-income country. METHODS: REFORCO-Brazil is an ongoing, test-negative case-control study (NCT05697705) utilizing Brazil national severe acute respiratory syndrome (SARS) surveillance and vaccination data. In SARS hospitalizations from August 1, 2021 to July 31, 2022, we matched test-positive (via SARS-CoV-2 antigen/reverse transcription polymerase chain reaction [RT-PCR]) cases and test-negative case-controls (via RT-PCR) based on admission date, preceding vaccinations, and age. We evaluated the rVEs of four monovalent COVID-19 vaccines (AZD1222, Ad26.COV2.S, CoronaVac, and BNT162b2) as second boosters compared with any first boosters received ≥4 months previously, and as first boosters compared with primary-series vaccinations completed ≥4 months previously. RESULTS: The overall rVE of second boosters, from 5668 (2238 test-positive) evaluated hospitalizations, was 24.7 % (95 % confidence interval [CI]: 12.6-35.1); the overall rVE of first boosters, from 30,272 (12,063 test-positive) hospitalizations, was 46.8 % (95 % CI: 43.3-50.0). The rVEs of AZD1222 and BNT162b2 were similar: 29.4 % (95 % CI: 8.6-45.5) and 25.5 % (95 % CI: 4.2-42.2), respectively, for second boosters; and 42.5 % (95 % CI: 28.0-54.0) and 50.8 % (95 % CI: 47.5-54.0), respectively, for first boosters. In general, rVEs were higher in elderly (≥80 years) and immunocompromised/high-risk individuals. CONCLUSIONS: Our results support the use of AZD1222 and other adenoviral/mRNA vaccine boosters to maintain protection against COVID-19 hospitalization from Omicron subvariants, including in elderly and immunocompromised individuals at increased risk of accelerated waning or severe outcomes.

Treatment of Chondral Lesions in the Knee.

Articular cartilage injuries are common and lead to early joint deterioration and osteoarthritis. Articular cartilage repair techniques aim at forming a cartilaginous neo-tissue to support the articular load and prevent progressive degeneration. Several techniques are available for this purpose, such as microfracture and chondrocyte transplantation. However, the procedural outcome is often fibrocartilage, which does not have the same mechanical resistance as cartilaginous tissue. Procedures with autologous osteochondral graft have a morbidity risk, and tissue availability limits their use. As such, larger lesions undergo osteochondral transplantation using fresh or frozen grafts. New techniques using minced or particulate cartilage fragments or mesenchymal stem cells are promising. This paper aims to update the procedures for treating chondral lesions of the knee.

Treatment of Chondral Lesions in the Knee / Tratamento das lesões condrais no joelho

Abstract Articular cartilage injuries are common and lead to early joint deterioration and osteoarthritis. Articular cartilage repair techniques aim at forming a cartilaginous neo-tissue to support the articular load and prevent progressive degeneration. Several techniques are available for this purpose, such as microfracture and chondrocyte transplantation. However, the procedural outcome is often fibrocartilage, which does not have the same mechanical resistance as cartilaginous tissue. Procedures with autologous osteochondral graft have a morbidity risk, and tissue availability limits their use. As such, larger lesions undergo osteochondral transplantation using fresh or frozen grafts. New techniques using minced or particulate cartilage fragments or mesenchymal stem cells are promising. This paper aims to update the procedures for treating chondral lesions of the knee.

Resumo As lesões da cartilagem articular são comuns e levam à deterioração precoce da articulação e ao desenvolvimento da osteoartrite. As técnicas de reparo da cartilagem articular visam a formação de um neo-tecido cartilaginoso capaz de suportar carga articular e evitar a progressão da degeneração. Há várias técnicas disponíveis para esse fim, como a microfratura e o transplante de condrócitos. Entretanto muitas vezes o desfecho do procedimento é a formação de fibrocartilagem, que não possui a mesma resistência mecânica do tecido cartilaginoso. Em outros procedimentos, nos quais é realizado enxerto osteocondral autólogo, há risco de morbidade associada ao procedimento, além da disponibilidade limitada de tecido. Por esse motivo, o transplante osteocondral, utilizando enxertos a fresco ou congelados tem sido utilizado para lesões de maior volume. Por fim, novas técnicas utilizando fragmentos de cartilagem picada ou particulada, assim como o uso de células tronco mesenquimais se apresentam como promissores. O objetivo desse artigo é realizar uma atualização dos procedimentos para tratamento das lesões condrais do joelho.

High Incidence of Respiratory Syncytial Virus in Children with Community-Acquired Pneumonia from a City in the Brazilian Pre-Amazon Region.

INTRODUCTION: Although fewer children have been affected by the severe form of the coronavirus disease 2019 (COVID-19), community-acquired pneumonia (CAP) continues to be the leading global cause of child hospitalizations and deaths. AIM: This study investigated the incidence of respiratory syncytial virus (RSV) as well its subtypes (RSV A and B), adenovirus (ADV), rhinovirus (HRV), metapneumovirus (HMPV), coronavirus (NL63, OC43, 229E and HKU1), parainfluenza virus subtypes (PI1, PI2 and PI3), bocavirus and influenza A and B viruses (FluA and FluB) in children diagnosed with CAP during the COVID-19 pandemic. METHODS: A total of 200 children with clinically confirmed CAP were initially recruited, of whom 107 had negative qPCR results for SARS-CoV-2 and were included in this study. Viral subtypes were identified using a real-time polymerase chain reaction in the nasopharyngeal swab samples. RESULTS: Viruses were identified in 69.2% of the patients. RSV infections were the most frequently identified (65.4%), with type RSV B being the most prevalent (63.5%). In addition, HCoV 229E and HRV were detected in 6.5% and 3.7% of the patients, respectively. RSV type B was associated with severe acute respiratory infection (ARI) and a younger age (less than 24 months). CONCLUSIONS: New strategies for preventing and treating viral respiratory infections, particularly RSV infections, are necessary.

New nitrosyl ruthenium complexes with combined activities for multiple cardiovascular disorders.

Nitrosyl ruthenium complexes are promising platforms for nitric oxide (NO) and nitroxyl (HNO) release, which exert their therapeutic application. In this context, we developed two polypyridinic compounds with the general formula cis-[Ru(NO)(bpy)2(L)]n+, where L is an imidazole derivative. These species were characterized by spectroscopic and electrochemical techniques, including XANES/EXAFS experiments, and further supported by DFT calculations. Interestingly, assays using selective probes evidenced that both complexes can release HNO on reaction with thiols. This finding was biologically validated by HIF-1α detection. The latter protein is related to angiogenesis and inflammation processes under hypoxic conditions, which is selectively destabilized by nitroxyl. These metal complexes also presented vasodilating properties using isolated rat aorta rings and demonstrated antioxidant properties in free radical scavenging experiments. Based on these results, the new nitrosyl ruthenium compounds showed promising characteristics as potential therapeutic agents for the treatment of cardiovascular conditions such as atherosclerosis, deserving further investigation.

Optimized Tourniquet Use in Primary Total Knee Arthroplasty: A Comparative, Prospective, and Randomized Study.

BACKGROUND: The results of recent studies investigating tourniquet (TNQ) use for knee arthroplasty are controversial. Therefore, this study aimed to compare patients undergoing total knee arthroplasty who did not have a TNQ to those in whom an optimized TNQ protocol was applied. METHODS: We prospectively evaluated 127 patients who had knee osteoarthritis who had undergone total knee arthroplasty and randomized them into two groups: "without TNQ" and "optimized TNQ" (TNQ inflation before skin incision, deflation after cementing, with pressure one hundred millimeters of mercury above the systolic blood pressure, and without articular suction drain usage). The means of surgery and TNQ duration, blood loss, number of blood transfusions, degree of pain, edema, range of motion (ROM), functional score over time, and postoperative complications were compared between the groups. Statistical significance was set at P < .05. RESULTS: No significant differences were found in terms of surgical timing, blood loss, thigh and knee pain, edema, ROM, functional scores, and complications between the "without TNQ" and "optimized TNQ" groups. CONCLUSION: The use of an optimized TNQ in primary total knee arthroplasty presents similar clinical results to surgery without a TNQ and did not increase the incidence of postoperative complications. Its use allowed surgery to occur with the benefits of a clean and dry surgical field provided by TNQ without increasing procedure-related comorbidities.

Nitroprusside─Expanding the Potential Use of an Old Drug Using Nanoparticles.

For more than 70 years, sodium nitroprusside (SNP) has been used to treat severe hypertension in hospital emergency settings. During this time, a few other clinical uses have also emerged such as in the treatment of acute heart failure as well as improving mitral incompetence and in the intra- and perioperative management during heart surgery. This drug functions by releasing nitric oxide (NO), which modulates several biological processes with many potential therapeutic applications. However, this small molecule has a short lifetime, and it has been administered through the use of NO donor molecules such as SNP. On the other hand, SNP also has some setbacks such as the release of cyanide ions, high water solubility, and very fast NO release kinetics. Currently, there are many drug delivery strategies that can be applied to overcome many of these limitations, providing novel opportunities for the use of old drugs, including SNP. This Perspective describes some nitroprusside properties and highlights new potential therapeutic uses arising from the use of drug delivery systems, mainly silica-based nanoparticles. There is a series of great opportunities to further explore SNP in many medical issues as reviewed, which deserves a closer look by the scientific community.

HEROHE Challenge: Predicting HER2 Status in Breast Cancer from Hematoxylin-Eosin Whole-Slide Imaging.

Breast cancer is the most common malignancy in women worldwide, and is responsible for more than half a million deaths each year. The appropriate therapy depends on the evaluation of the expression of various biomarkers, such as the human epidermal growth factor receptor 2 (HER2) transmembrane protein, through specialized techniques, such as immunohistochemistry or in situ hybridization. In this work, we present the HER2 on hematoxylin and eosin (HEROHE) challenge, a parallel event of the 16th European Congress on Digital Pathology, which aimed to predict the HER2 status in breast cancer based only on hematoxylin-eosin-stained tissue samples, thus avoiding specialized techniques. The challenge consisted of a large, annotated, whole-slide images dataset (509), specifically collected for the challenge. Models for predicting HER2 status were presented by 21 teams worldwide. The best-performing models are presented by detailing the network architectures and key parameters. Methods are compared and approaches, core methodologies, and software choices contrasted. Different evaluation metrics are discussed, as well as the performance of the presented models for each of these metrics. Potential differences in ranking that would result from different choices of evaluation metrics highlight the need for careful consideration at the time of their selection, as the results show that some metrics may misrepresent the true potential of a model to solve the problem for which it was developed. The HEROHE dataset remains publicly available to promote advances in the field of computational pathology.

Percepção dos profissionais do Programa Mais Médicos quanto à adesão aos instrumentos de comunicação entre as Redes de Atenção à Saúde e a Atenção Primária / he professionals' perception of the "More Doctors" program regarding communication instruments' adherence between healthcare networks and primary care

O artigo objetiva analisar a percepção dos profissionais do Programa Mais Médicos quanto à adesão aos instrumentos de comunicação entre os pontos de atenção das Redes de Atenção à Saúde. Sua importância diz respeito à necessidade de se analisar o grau de adesão aos instrumentos e qualificar a ocorrência da comunicação entre os entes da rede, na medida em que se reconhece a necessidade de manter informado o profissional detentor da coordenação do cuidado e devolver a ele a responsabilidade pela condução dos indivíduos usuários da rede. Os resultados demonstraram a baixa percepção do retorno dos registros dos usuários da rede aos pontos de atenção, independentemente da macrorregião (P = 0,094). Esse resultado demonstra a necessidade de trabalhar a qualificaçãodas atividades da rede de atenção a fim de valorizar a necessidade das informações oriundas do processo de contrarreferência.

The article's purpose is to analyze professionals' perceptions of the "More Doctors" program regarding communication instruments' adherence among the different points inside the healthcare network. It is important to analyze the instruments' adherence degree and qualify the occurrence of communication inside the network entities, recognizing the need to keep care coordinators well informed and to make them responsible for the care of individuals who are users of the network. The results showed a low perception of users' network records arriving at the proper care points, independent of the macroregion (P = 0,094). This result demonstrates the need to work on qualifying the activities of the care network to value the need for information coming from the counter-reference processes.

Ethyl Acetate Fraction of Bixa orellana and Its Component Ellagic Acid Exert Antibacterial and Anti-Inflammatory Properties against Mycobacterium abscessus subsp. massiliense.

Mycobacterium abscessus subsp. massiliense (Mabs) causes chronic infections, which has led to the need for new antimycobacterial agents. In this study, we investigated the antimycobacterial and anti-inflammatory activities of the ethyl acetate fraction of Bixa orellana leaves (BoEA) and ellagic acid (ElAc). In silico analysis predicted that ElAc had low toxicity, was not mutagenic or carcinogenic, and had antimicrobial and anti-inflammatory activities. Apparently, ElAc can interact with COX2 and Dihydrofolate reductase (DHFR) enzymes, which could explain both activities. In vitro analysis showed that BoEA and ElAc exerted antimicrobial activity against Mabs (minimum inhibitory concentration of 1.56, 1.56 mg/mL and bactericidal concentration of 6.25, 3.12 mg/mL, respectively. Clarithromycin showed MIC and MBC of 1 and 6 µg/mL). Treatment with BoEA or ElAc increased survival of Tenebrio molitor larvae after lethal infection with Mabs and reduced carrageenan-induced paw edema in mice, around 40% of edema volume after the fourth hour, similarly to diclofenac. In conclusion, BoEA and ElAc exert antimicrobial effects against Mabs and have anti-inflammatory effects, making them potential sources of antimycobacterial drugs. The biological activities of ElAc may be due to its high binding affinities predicted for COX2 and DHFR enzymes.

Restricted Activation of the NF-κB Pathway in Individuals with Latent Tuberculosis Infection after HIF-1α Blockade.

Tuberculous granuloma formation is mediated by hypoxia-inducible factor 1 alpha (HIF-1α), and is essential for establishing latent tuberculosis infection (LTBI) and its progression to active tuberculosis (TB). Here, we investigated whether HIF-1α expression and adjacent mechanisms were associated with latent or active TB infection. Patients with active TB, individuals with LTBI, and healthy controls were recruited, and the expression of cytokine genes IL15, IL18, TNFA, IL6, HIF1A, and A20 in peripheral blood mononuclear cells (PBMCs) and serum vitamin D (25(OH)D3) levels were evaluated. Additionally, nuclear factor kappa B (NF-κB) and tumor necrosis factor-alpha (TNF-α) levels were analyzed in PBMC lysates and culture supernatants, respectively, after HIF-1α blockade with 2-methoxyestradiol. We observed that IL-15 expression was higher in individuals with LTBI than in patients with active TB, while IL-18 and TNF-α expression was similar between LTBI and TB groups. Additionally, serum 25(OH)D3 levels and expression of IL-6, HIF1A, and A20 were higher in patients with active TB than in individuals with LTBI. Moreover, PBMCs from individuals with LTBI showed decreased NF-κB phosphorylation and increased TNF-α production after HIF-1α blockade. Together, these results suggest that under hypoxic conditions, TNF-α production and NF-κB pathway downregulation are associated with the LTBI phenotype.

BMP-4, TGF-β and Smad3 as Modulators of Synovial Fluid Cells Viability / BMP-4, TGF-β e Smad3 como moduladores da viabilidade das células do líquido sinovial

Abstract Objective Our goal was to evaluate the modulation of the synovial fluid cells (SFC) from patients with and without osteoarthritis (OA) by bone morphogenetic protein 4 (BMP-4), Smad-3 and transforming growth factor beta (TGF-β). Methods Synovial fluid was collected from patients submitted to knee arthroscopy or replacement and were centrifuged to isolate cells from the fluid. Cells were cultured for 21 days and characterized as mesenchymal stem cells (MSCs) according to the criteria of the International Society of Cell Therapy. Then, we performed an [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay (MTT) assay after exposing cells with and without OA to TGF-β, Smad3 and BMP-4 pathway inhibitors and to different concentrations of BMP4. Results Exposure to the TGF-β, Smad3 and BMP-4 inhibitors modifies the mitochondrial activity of the SFCs. The activity of the SFCs is modified by influences of increasing concentrations of BMP4, but there is no difference in cellular activity between patients with and without OA. Conclusion TGF-β, Smad3 and BMP-4 modulate the activity of SFCs from patients with and without knee OA.

Resumo Objetivo Nosso objetivo foi avaliar a modulação das células do líquido sinovial (SFCs, na sigla em inglês) de pacientes com e sem osteoartrite (OA) por proteína morfogenética óssea 4 (BMP-4), Smad3 e transformador do fator de crescimento β (TGF-β). Métodos O do líquido sinovial foi coletado de pacientes submetidos a artroscopia ou artroplastia do joelho, e centrifugados para isolar as células do liquido sinovial. As células foram cultivadas por 21 dias e caracterizadas como células-tronco mesenquimais (MSCs, na sigla em inglês) de acordo com os critérios da International Society of Cell Therapy. Em seguida, realizamos um ensaio de brometo de 3-4,5-dimetil-tiazol-2-il-2,5difeniltetrazólio (MTT) depois de expor células com e sem OA para TGF-β, inibidores de via Smad3 e BMP-4 e para diferentes concentrações de BMP-4. Resultados A exposição aos inibidores TGF-β, Smad3 e BMP-4 modifica a atividade mitocondrial das SFCs. A atividade das SFCs é modificada por influências sobre o aumento das concentrações de BMP-4, mas não há diferença na atividade celular entre pacientes com e sem OA. Conclusão TGF-β, Smad3 e BMP-4 modulam a atividade das SFCs de pacientes com e sem OA do joelho.

MCL1 alternative polyadenylation is essential for cell survival and mitochondria morphology.

Alternative polyadenylation in the 3' UTR (3' UTR-APA) is a mode of gene expression regulation, fundamental for mRNA stability, translation and localization. In the immune system, it was shown that upon T cell activation, there is an increase in the relative expression of mRNA isoforms with short 3' UTRs resulting from 3' UTR-APA. However, the functional significance of 3' UTR-APA remains largely unknown. Here, we studied the physiological function of 3' UTR-APA in the regulation of Myeloid Cell Leukemia 1 (MCL1), an anti-apoptotic member of the Bcl-2 family essential for T cell survival. We found that T cells produce two MCL1 mRNA isoforms (pA1 and pA2) by 3' UTR-APA. We show that upon T cell activation, there is an increase in both the shorter pA1 mRNA isoform and MCL1 protein levels. Moreover, the less efficiently translated pA2 isoform is downregulated by miR-17, which is also more expressed upon T cell activation. Therefore, by increasing the expression of the more efficiently translated pA1 mRNA isoform, which escapes regulation by miR-17, 3' UTR-APA fine tunes MCL1 protein levels, critical for activated T cells' survival. Furthermore, using CRISPR/Cas9-edited cells, we show that depletion of either pA1 or pA2 mRNA isoforms causes severe defects in mitochondria morphology, increases apoptosis and impacts cell proliferation. Collectively, our results show that MCL1 alternative polyadenylation has a key role in the regulation of MCL1 protein levels upon T cell activation and reveal an essential function for MCL1 3' UTR-APA in cell viability and mitochondria dynamics.

Modified Pulvertaft on Weave Technique Restores Full Active Knee Extension in Patients With Large Chronic Quadriceps Tendon Rupture: A Case Series.

PURPOSE: We aimed to investigate the clinical and functional outcomes, including maximal and explosive strength, after chronic quadriceps tendon rupture repair with Modified Pulvertaft on Weave (MPW) technique METHODS: Knee joint range of motion (ROM), patella height, thigh circumference, and Lysholm and International Knee Documentation Committee (IKDC) scores were assessed preoperatively and postoperatively. The knee extensors maximal (isokinetic peak torque and isometric maximal voluntary contraction (MVC) torque) and explosive strength-rate of torque development (RTD) early [RTD50 and RTD100] and late [RTD250]-were performed. We assessed the thigh circumference and vastus lateralis muscle thickness (MT) as indicators of quadriceps muscle mass, and the voluntary quadriceps activation using surface electromyography (EMG50). RESULTS: Nine patients (mean age: 53 ± 11 years) took part in the study. We observed a significant increase in the knee active ROM and a decreased extension deficit (both, P < .001), but not for pain (P = .07), IKDC (P = .07), and Lysholm (P = .21) after the surgery. We did not observe a difference between involved (n = 8) and uninvolved (n = 10) limbs for ROM, thigh circumference, and MT. We observed differences for extensors peak torque, MVC torque, and late RTD (all, P < .05). However, we did not observe differences for early RTD and EMG50. Significant positive correlations were observed for RTD50 (ρ = .80) and RTD100 (ρ = .81) vs EMG50. Both the IKDC and Lysholm were better correlated with the early than with later RTD. CONCLUSIONS: The MPW reestablished the active knee extension. The same level of quadriceps muscle mass was observed in both limbs, suggesting a lack of hypotrophy due to the injury. Although the involved limb had demonstrated lower knee extensors maximal strength, they demonstrate an equivalent early RTD when compared to the uninvolved limb. The early RTD seems to be better correlated with the patient's functionality than the later RTD and maximal strength. LEVEL OF EVIDENCE: IV, case series.

Influence of type I collagen polymorphisms and risk of anterior cruciate ligament rupture in athletes: a case-control study.

BACKGROUND: Anterior cruciate ligament (ACL) rupture is a common and severe knee injury in sports and occurs mostly due to noncontact injuries. There is an increasing amount of evidence associating ACL rupture to single nucleotide polymorphisms (SNPs), and SNPs in the collagen type I genes can change its expression and tissue mechanical features. This study aimed to investigate the association between SNPs in COL1A1 and COL1A2 with sports-related ACL tears. METHODS: A total of 338 athletes from multiple sports modalities were analyzed: 146 were diagnosed with ACL rupture or underwent an ACL reconstruction surgery and 192 have no musculoskeletal injuries. SNPs were genotyped using validated TaqMan assays. The association of the polymorphisms with ACL rupture was evaluated by a multivariable logistic regression model, using odds ratios (OR) and 95% confidence intervals (CI). RESULTS: The age, sport modality, and training location were associated with an increased risk of a non-contact ACL tear. COL1A2 SNPs (rs42524 CC and rs2621215 GG) were associated with an increased risk of non-contact ACL injury (6 and 4-fold, respectively). However, no significant differences were detected in the distribution of COL1A1 rs1107946 and COL1A2 rs412777 SNPs between cases and controls. There was a protective association with ACL rupture (OR = 0.25; 95% CI = 0.07-0.96) between COL1A1 rs1107946 (GT or TT) and the wildtype genotypes of the three COL1A2 (rs412777, rs42524, rs2621215). COL1A2 rs42524 and rs2621215 SNPs were associated with non-contact ACL risk. CONCLUSION: The combined analysis of COL1A1-COL1A2 genotypes suggests a gene-gene interaction in ACL rupture susceptibility.