Cryopreservation of cell laden natural origin hydrogels for cartilage regeneration strategies

Statement of Purpose: An increase of scientific published literature and clinical experience supports the requirement of providing products like cultured cells and tissues to the market. One of the main prospects of cartilage tissue engineering is the possibility of developing custom-made regenerative medicine solutions on an individual patient basis. The efficient preservation and storage procedure will provide products available as needed which could be adapted to an autologous immediate solution. Thus, the aim of this study was to examine the effects of the cryopreservation on the chondrogenic differentiation characteristics of human mesenchymal stem cells isolated from adipose tissue (hASCs). Furthermore, we also propose to determine hASCs-hydrogel stability and confirm the potential of these bioengineered constructs to be applied in cartilage regeneration. The results obtained show that the hydrogels withstand the cryopreservation process maintaining their structural integrity, with good cell content after cryopreservation. Thus, cell encapsulation systems of natural based hydrogels may be an interesting approach for the long term preservation of cartilage tissue engineered products. Methods: The κ-carrageenan (κCR) hydrogels were produced using an ionotropic gelation method. Then, stem cells, namely human adipose derived stem cells (hASCs), were encapsulated in κCR discs (5 mm dia. x 3 mm height) at a density of 5x106 cell/cm3 and cultured for 21 days in standard basal (BM) or chondrogenic media (ChM). The cell hydrogels were cryopreserved in liquid nitrogen for up to 30 days. The overall morphology of κCR discs with encapsulated hASCs was observed under light microscope. hASCs viability and proliferation rate was determined by double stand DNA quantification. Additionally, chondrogenic differentiation of hASCs encapsulated in the hydrogels is being characterized by histological staining for selective cartilage staining and real time PCR analysis (Sox9, aggrecan, and collagen: type I, type II and type X). DMA analysis allowed determining the mechanical properties of κ-carrageenan hydrogels, namely storage (elastic) and loss (viscous) while immersed in wet state at 24 °C and throughout a physiological relevant range of frequencies. The described characterization assays were performed both before (BC) and after cryopreservation/freezing (AC) time points. Results: The cell morphology, distribution and appearance of the hydrogels are clearly observed from the microscopic light images (Figure 1A). It is possible to observe the smooth and homogeneous surface, the well defined and stable shape before and after the freezing process. Encapsulated hASCs were able to maintain cellular content, despite an expected decrease observed upon cryopreservation (Figure 1B), which is associated to a recovery time after thawing. The microscopic images and biological evaluation of κCR hydrogel revealed that the cryopreservation process did not change the cellular morphology; the surface and integrity of the hydrogel disc and enables maintenance of hASCs after exposure to low temperatures environments. Figure 1. (A) Representative optical micrographs of hASCs encapsulated in κCR hydrogels and cultured in ChM and BM before and after cryopreservation and (B) cell proliferation results, based on the quantification of dsDNA content. Scale bar represent 100 μm. Conclusions: The results obtained so far indicated the feasibility of hASCs-κCR system in cartilage tissue engineering regeneration strategies due to its ability to support hASCs viability before and after cryopreservation. Ongoing studies on the assessment of chondrogenic features of these cryopreserved systems will provide information on the effect of cryopreservation indicative of a stable chondrocyte phenotype. In summary, this study provided information on the potential of ASCs-hydrogel constructs for a long term storage and ready to use bioengineered tissue substitutes for cartilage regeneration strategies. References: (Popa EG. Biomacromolecules 2011;12:3952-3961

Development of new chitosan/carrageenan nanoparticles for drug delivery applications

The use of polymeric nanoparticles, especially those composed of natural polymers, has become a very interesting approach in drug delivery., mainly because of the advantages offered by their small dimensions. The aim of this work was to develop a novel formulation of nanoparticles comprised of two natural marine-derived polymers, namely chitosan and carrageenan, and to evaluate their potential for the association and controlled release of macromolecules. Nanoparticles were obtained in a hydrophilic environment, under very mild conditions, avoiding the use of organic solvents or other aggressive technologies for their preparation. The developed nanocarriers presented sizes within 350-650 run and positive zeta potentials of 50-60 mV. Polymeric interactions between nanoparticles' components were evaluated by Fourier transform infrared spectroscopy. Using ovalbumin as model protein, nanoparticles evidenced loading capacity varying from 4% to 17%, and demonstrated excellent capacity to provide a controlled release for up to 3 weeks. Furthermore, nanoparticles have demonstrated to exhibit a noncytotoxic behavior in biological in vitro tests performed using L929 fibroblasts, which is critical regarding the biocompatibility of those carriers. In summary, the developed chitosan-carrageenan nanoparticles have shown promising properties to be used as carriers of therapeutic macromolecules, with potential application not only strictly in drug delivery, but also in broader areas, such as tissue engineering and regenerative medicine. (C) 2009 Wiley Periodicals, Inc. J Biomed Mater Res 92A: 1265-1272, 2010Contract grant sponsor: Portuguese Foundation for Science and Technology (FCT) (POCTI/FEDER programmes)Contract grant sponsor: European Union STREP Project HIPPOCRATES; contract grant number: NMP3-CT-2003505758Contract grant sponsor: European NoE EXPERTISSUES; contract grant number: NMP3-CT-2004-50028

Adapted chester step test can have maximal response characteristics for the assessment of exercise capacity in young women

Chester step test (CST) estimates the exercise capacity through a submaximal response, which can limit its application in the prescription of exercise. This study aimed to assess whether an adaptation of the CST (with a progressive profile) can have maximal response characteristics in young women and compare it to the incremental shuttle walk test (ISWT). Another aim was to determine its within-day test–retest reliability. A cross-sectional study was conducted with 25 women (20.3 ± 1.5 years) who performed the field tests twice on two different days (48 h apart).info:eu-repo/semantics/publishedVersio

Avaliação de Desempenho: Percepção dos Auditores Independentes na Aferição Da Remuneração

O objetivo deste estudo é evidenciar a percepção dos auditores em relação aos critérios utilizados para aferição da sua remuneração. Para tanto foi elaborado um questionário estruturado através da plataforma Google docs que foi disponibilizado para auditores de 7 diferentes escritórios de auditoria entre os meses de maio e junho de 2013. O tratamento estatístico remete a análise descritiva dos dados obtidos na pesquisa de campo, com base na utilização de medidas de posição e dispersão. Foram utilizadas ainda tabelas de contingência para analisar comportamentos de indivíduos em variáveis distintas. A amostra foi composta por 17 indivíduos componentes dos diversos cargos hierárquicos da carreira de auditor independente. A análise dos resultados evidenciou que auditores que investem mais tempo em reuniões com pares também desejam ser remunerados pelo resultado da equipe e que as informações contábeis não são utilizadas para avaliar os esforços da maior parte dos auditores. Verificou-se ainda que a existência de fatores incontroláveis impulsiona os respondentes a reconhecerem a necessidade do uso da subjetividade em suas avaliações de desempenho. Conclui-se que não existe consenso entre os respondentes se critérios contábeis deveriam ser utilizados para avaliar desempenho e que o cargo em que o auditor se encontra influencia na sua percepção em relação a critérios de remuneração variável

Evaluation of the functionality of workers with Repetitive Strain Injury (RSI)/ Work-related musculoskeletal disorders (MSDs): the construction of the ICF Core Set for RSI/MSDs

This study presents the ICF Core Set for Repetitive Strain Injury (RSI)/ Work-related musculoskeletal disorders (MSDs) and its construction process. The ICF Core Set for RSI/MSDs was created through successive consensuses among specialists in the workers' healthcare area throughout six phases, which involved from the assessment and the study of the ICF, followed by its reading and discussion, to the establishment of codes and comparison with the core sets of generalized pain and depression. For the "body functions" component, the chosen codes were related to the following aspects: mental, sensory, pain and neuromusculoskeletal functions, as well as movement functions. For the "body structures" component, the chosen codes were related to the following aspects: nervous system structures and those related to movement. For the "activity and participation" component, the chosen codes were related to: mobility, personal care, domestic life, interpersonal relations and interactions and the main areas of life. For "environmental factors", the codes chosen were related to: support and relationships, attitudes and services, systems and policies. The environment component was the most limited for the description within the work context. This study represents the effort of constructing a core set based on an interdisciplinary approach, viable to be applied during the treatment and rehabilitation process of workers presenting RSI/MSDs and can contribute to bring Brazil into the international discussion that deals with the consequences of human disease from a social-medical model, displacing the scientific debate and the creation of public policies from the context of disability/incapacity into the health context.O presente artigo apresenta o Core set da CIF para LER/DORT e seu processo de construção.  O Core set da CIF para LER/DORT foi elaborado por meio de consensos sucessivos entre especialistas do campo da saúde do trabalhador em seis etapas, que envolveram desde a aproximação e estudo da CIF, leitura, discussão até a eleição de códigos e comparação com os core sets de dor generalizada e de depressão. Para o componente ‘funções do corpo’ foram escolhidos códigos relacionados aos aspectos: funções mentais, sensoriais, de dor e neuromusculoesqueléticas e relacionadas ao movimento. Para ‘estruturas do corpo’ foram escolhidos códigos relacionados aos aspectos: estruturas do sistema nervoso e relacionadas ao movimento. Para ‘atividade e participação’ foram escolhidos códigos relacionados à: mobilidade, cuidado pessoal, vida doméstica, relações e interações interpessoais e áreas principais da vida. Para ‘fatores ambientais’ foram escolhidos códigos relacionados à: apoio e relacionamentos, atitudes e serviços, sistemas e políticas. O componente ambiente foi o mais limitado para a descrição dentro do contexto do trabalho. Este artigo apresenta o esforço de construção de um core set, a partir de uma abordagem interdisciplinar, viável à aplicação no processo de tratamento e reabilitação de trabalhadores com LER/DORT e poderá contribuir para inserir o Brasil na discussão internacional que trata das conseqüências do adoecimento humano a partir do modelo sócio-médico, deslocando o debate científico e a produção de políticas públicas do contexto da deficiência/incapacidade para o contexto da saúde

Efeitos da intervenção fisioterapêutica nas respostas sensoriais e funcionais de diabéticos neuropatas

The purpose of this study was to elaborate and apply a physical therapy treatment for diabetic neuropathic patients, comparing their sensorial, motor and functional responses before and after treatment to those of a healthy control group. Ten healthy subjects (CG) and 10 neuropathic diabetes patients (DG) clinically diagnosed were studied. Motor, functionaland sensorial evaluations were performed pre and post-intervention in both groups. Treatment was individually applied in 45-minute sessions twice a week, during 5 weeks. Therapy program consisted in stretching exercises for hamstrings, triceps surae and tibialis anterior muscles, actively-resisted exercises for intrinsic and extrinsic foot and ankle muscles, practice of daily activities and feet self-care orientations. Data were statistically analysed. When comparing DG post treatment data to DG-pre and CG, thermal sensitivity showed improvement in heel, hallux, lateral and medial forefoot; DG ankle dorsiflexion and eversion ROM reached values similar to those of CG's; DG showed significant improvement in ankle extension and inversion; and DG-post reached the same muscular functions observed in CG in intrinsic feet muscles, tibialis anterior and triceps surae. The physical therapy treatment showed effective in reducing distal numbness, tingling and pricking and it was also effective in preventing muscle function and mobility limitations in diabetic neuropathic patients.O objetivo foi elaborar e aplicar um tratamento fisioterapêutico para diabéticos neuropatas e comparar suas respostas sensoriais, motoras e funcionais, pré e pós-intervenção, com um grupo de sujeitos não-diabéticos assintomáticos. Participaram do estudo 10 sujeitos controle (GC) e 10 diabéticos neuropatas (GD) diagnosticados clinicamente. Realizou-se uma avaliação motora, funcional e sensorial nos dois grupos pré e pós-intervenção. O tratamento foi aplicado individualmente duas vezes por semana, por 45 minutos, durante 5 semanas. Foram feitos alongamentos de cadeia posterior e tibial anterior; exercícios ativos resistidos para musculatura intrínseca do pé e tornozelo; treino de atividades de vida diária e fornecidas orientações de autocuidado com os pés. Os dados foram analisados estatisticamente. Comparando-se os dados do GD pós-tratamento com os do GC, verificou-se melhora na sensibilidade térmica nas regiões de calcanhar, hálux, antepé lateral e medial; a amplitude ativa de dorsiflexão e eversão do tornozelo igualou-se à do GC; houve melhora significativa da extensão e inversão do pé; e o GD alcançou funções musculares (musculatura intrínseca do pé,tríceps sural e tibial anterior) semelhantes às do GC. Em diabéticos neuropatas, o tratamento fisioterapêutico proposto mostrou-se eficaz na atenuação dos sintomas dormência, formigamento e queimação, além de contribuir para a mobilidade e prevenção de limitações de função muscular

Aplicação do lúdico Termotrilha em turmas inclusivas do PIBID

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Activities and mechanisms of oregano, marjoram and rosemary essential oils against Malassezia pachydermatitis isolates from canine and feline otitis

Malassezia pachydermatis is an opportunistic yeast found in the ear canal of small animals; however, the current azole-based therapy applied to it has failed to achieve clinical success due to the antifungal resistance. This issue has encouraged the studies in natural products, such as Origanum vulgare (oregano), Origanum majorana (marjoram) and Rosmarinus officinalis (rosemary) essential oils, although their mechanism of action remains unclear. Malassezia pachydermatis specimens deriving from otitis cases in dogs (n = 22) and cats (n = 2) were subjected to CLSI M27-A3. Sorbitol protection and ergosterol effect were analyzed to investigate their mechanism of action. Fungistatic (MIC) and fungicidal (MFC) activities were observed for oregano (MIC90/MFC90: 0.625 mg/mL); marjoram (MIC90/MFC90: 2.5 mg/mL) and rosemary MIC90/MFC90 > 2.5 mg/mL). Oregano showed superior antifungal effect even at lower MIC and MFC values. All three oils acted on cell wall and at complexation to fungal ergosterol. By gas chromatography (GC-FID), carvacrol was the major compound found in oregano (73.9%); 1,8-cineole was for marjoram and rosemary (20.9% and 49.4%, respectively). These findings support the potential use of these essential oils to treat canine and feline otitis caused by Malassezia pachydermatis

Uso indiscriminado de morfina no cuidado paliativo de pacientes com câncer: uma revisão integrativa

Objective: To evaluate how the indiscriminate use of morphine affects end-stage cancer patients. Methods: Integrative review study, with a descriptive and qualitative approach, about the effects of indiscriminate use of morphine among patients with end-stage cancer, between 2013 and 2023. Results: Initially, applying criteria of language, year of publication and availability in full, 1,879 articles were considered, of which 1,521 were excluded because they were reviews, documents and books, resulting in 358 articles read in full, in which 14 met all the inclusion criteria. In summary, two studies bring positive aspects of prolonged morphine regarding the analgesic effects and minimal adverse events, however, 12 studies bring different approaches regarding the indiscriminate use of morphine, in order to demonstrate divergences regarding aspects of pain control, however, a better description as to the occurrence of adverse events (especially nausea, constipation and sleepiness), drug addiction/tolerance and risk of death associated with stroke, AEs, and sepsis. Conclusion: There is still literature divergence as to the contribution of morphine in pain reduction, although some authors point out its contribution, however, it is well reported that the indiscriminate use may reflect in problems of AEs level, tolerance and risks to death, therefore, it is necessary to discuss its prescription in order to promulgate an effective medical monitoring and control in administration.   Keywords: Morphine; Cancer; Abuse; Adverse Events.Objetivo: Avaliar como o uso indiscriminado de morfina afeta os pacientes em estágio terminal de câncer. Métodos: Estudo de revisão integrativa, de abordagem descritiva e qualitativa, sobre os efeitos no uso indiscriminado de morfina entre pacientes com câncer em estágio terminal, entre 2013 e 2023. Resultados: Inicialmente, aplicando critério de idioma, ano de publicação e disponibilidade na íntegra, foram considerando 1.879 artigos onde, 1.521 foram excluídos por se tratar de revisão, documentos e livros, resultando em 358 artigos lidos na íntegra, no qual 14 contemplaram todos os critérios de inclusão. Em suma, dois estudos trazem aspectos positivos da morfina prolongada quanto aos efeitos analgésicos e mínimos eventos adversos, contudo, 12 estudos trazem abordagens diferenciadas quanto ao uso indiscriminado da morfina, de forma a demonstrar divergências quanto aspectos de controle da dor, entretanto, uma melhor descrição quanto a ocorrência de eventos adversos (sobretudo náuseas, constipação e sonolência), vicio/tolerância medicamentosa e riscos de óbitos associados a Acidente Vascular Cerebral, Fibrilação Arterial e sepse. Conclusão: Ainda há divergência literária quanto a contribuição da morfina prolongada na redução da dor, embora alguns autores pontuem seu aporte, contudo, é bem relatado que o uso indiscriminado pode refletir em problemas de nível de EAs, tolerância e riscos ao óbito, logo, é necessário discutir a sua prescrição de forma a promulgar um acompanhamento médico efetivo e controle na administração.   Palavras-chave: Morfina; Câncer; Uso abusivo; Eventos Adversos

Telehealth effectiveness for pre-exposure prophylaxis delivery in Brazilian public services : the Combine! Study

Introduction Pre-exposure prophylaxis (PrEP) delivery based on user needs can enhance PrEP access and impact. We examined whether telehealth for daily oral PrEP delivery could change the indicators of care related to prophylactic use in five Brazilian public HIV clinics (testing centres, outpatient clinics and infectious disease hospitals). Methods Between July 2019 and December 2020, clients on PrEP for at least 6 months could transition to telehealth or stay with in-person follow-up. Clients were clinically monitored until June 2021. A desktop or mobile application was developed, comprising three asynchronous consultations and one annual in-person consultation visit. Predictors influencing telehealth preference and care outcomes were examined. The analysis encompassed intent-to-treat (first choice) and adjustments for sexual practices, schooling, age, duration of PrEP use and PrEP status during the choice period. Results Of 470 users, 52% chose telehealth, with the adjusted odds ratio (aOR) increasing over time for PrEP use (aOR for 25–months of use: 4.90; 95% CI: 1.32–18.25), having discontinued PrEP at the time of the choice (aOR: 2.91; 95% CI: 1.40–6.06) and having health insurance (aOR: 1.91; 95% CI: 1.24–2.94) and decreasing for those who reported higher-risk behaviour (aOR for unprotected anal sex: 0.51; 95% CI: 0.29–0.88). After an average follow-up period of 1.6 years (95% CI: 1.5–1.7), the risk of discontinuing PrEP (not having the medication for more than 90 days) was 34% lower with telehealth (adjusted hazard ratio: 0.66; 95% CI: 0.45–0.97). When adjusted by mixed linear regression, no differences in adherence (measured by mean medication possession rate) were found between in-person and telehealth (p = 0.486) or at pre- and post-telehealth follow-ups (p = 0.245). Sexually transmitted infections increased between the pre-follow-up and post-follow-up choices and were not associated with in-person or telehealth (p = 0.528). No HIV infections were observed. Conclusions Our findings indicate that telehealth for PrEP delivery can enhance service rationalization and reinforce the prevention cascade. This approach reduces prophylaxis interruptions and is mainly preferred by individuals with lower demands for healthcare services