Polyphosphate Nanoparticles: Balancing Energy Requirements in Tissue Regeneration Processes.

Nanoparticles of a particular, evolutionarily old inorganic polymer found across the biological kingdoms have attracted increasing interest in recent years not only because of their crucial role in metabolism but also their potential medical applicability: it is inorganic polyphosphate (polyP). This ubiquitous linear polymer is composed of 10-1000 phosphate residues linked by high-energy anhydride bonds. PolyP causes induction of gene activity, provides phosphate for bone mineralization, and serves as an energy supplier through enzymatic cleavage of its acid anhydride bonds and subsequent ATP formation. The biomedical breakthrough of polyP came with the development of a successful fabrication process, in depot form, as Ca- or Mg-polyP nanoparticles, or as the directly effective polymer, as soluble Na-polyP, for regenerative repair and healing processes, especially in tissue areas with insufficient blood supply. Physiologically, the platelets are the main vehicles for polyP nanoparticles in the circulating blood. To be biomedically active, these particles undergo coacervation. This review provides an overview of the properties of polyP and polyP nanoparticles for applications in the regeneration and repair of bone, cartilage, and skin. In addition to studies on animal models, the first successful proof-of-concept studies on humans for the healing of chronic wounds are outlined.

The Physiological Inorganic Polymers Biosilica and Polyphosphate as Key Drivers for Biomedical Materials in Regenerative Nanomedicine.

There is a need for novel nanomaterials with properties not yet exploited in regenerative nanomedicine. Based on lessons learned from the oldest metazoan phylum, sponges, it has been recognized that two previously ignored or insufficiently recognized principles play an essential role in tissue regeneration, including biomineral formation/repair and wound healing. Firstly, the dependence on enzymes as a driving force and secondly, the availability of metabolic energy. The discovery of enzymatic synthesis and regenerative activity of amorphous biosilica that builds the mineral skeleton of siliceous sponges formed the basis for the development of successful strategies for the treatment of osteochondral impairments in humans. In addition, the elucidation of the functional significance of a second regeneratively active inorganic material, namely inorganic polyphosphate (polyP) and its amorphous nanoparticles, present from sponges to humans, has pushed forward the development of innovative materials for both soft (skin, cartilage) and hard tissue (bone) repair. This energy-rich molecule exhibits a property not shown by any other biopolymer: the delivery of metabolic energy, even extracellularly, necessary for the ATP-dependent tissue regeneration. This review summarizes the latest developments in nanobiomaterials based on these two evolutionarily old, regeneratively active materials, amorphous silica and amorphous polyP, highlighting their specific, partly unique properties and mode of action, and discussing their possible applications in human therapy. The results of initial proof-of-concept studies on patients demonstrating complete healing of chronic wounds are outlined.

Cell migration, DNA fragmentation and antibacterial properties of novel silver doped calcium polyphosphate nanoparticles.

To combat infections, silver was used extensively in biomedical field but there was a need for a capping agent to eliminate its cytotoxic effects. In this study, polymeric calcium polyphosphate was doped by silver with three concentrations 1, 3 or 5 mol.% and were characterized by TEM, XRD, FTIR, TGA. Moreover, cytotoxicity, antibacterial, cell migration and DNA fragmentation assays were done to assure its safety. The results showed that the increase in silver percentage caused an increase in particle size. XRD showed the silver peaks, which indicated that it is present in its metallic form. The TGA showed that thermal stability was increased by increasing silver content. The antibacterial tests showed that the prepared nanoparticles have an antibacterial activity against tested pathogens. In addition, the cytotoxicity results showed that the samples exhibited non-cytotoxic behavior even with the highest doping concentration (5% Ag-CaPp). The cell migration assay showed that the increase in the silver concentration enhances cell migration up to 3% Ag-CaPp. The DNA fragmentation test revealed that all the prepared nanoparticles caused no fragmentation. From the results we can deduce that 3% Ag-CaPp was the optimum silver doped calcium polyphosphate concentration that could be used safely for medical applications.

Microfragmented Fat and Biphasic Calcium Phosphates for Alveolar Cleft Repair: Protocol for a Prospective, Nonblinded, First-in-Human Clinical Study.

BACKGROUND: Biphasic calcium phosphates (BCP) may serve as off-the-shelf alternatives for iliac crest-derived autologous bone in alveolar cleft reconstructions. To add osteoinductivity to the osteoconductive BCPs to achieve similar regenerative capacity as autologous bone, a locally harvested buccal fat pad will be mechanically fractionated to generate microfragmented fat (MFAT), which has been shown to have high regenerative capacity due to high pericyte and mesenchymal stem cell content and a preserved perivascular niche. OBJECTIVE: Our primary objectives will be to assess the feasibility and safety of the BCP-MFAT combination. The secondary objective will be efficacy, which will be evaluated using radiographic imaging and histological and histomorphometric evaluation of biopsies taken 6 months postoperatively, concomitant with dental implant placement. METHODS: Eight patients with alveolar cleft (≥15 years) will be included in this prospective, nonblinded, first-in-human clinical study. MFAT will be prepared intraoperatively from the patient's own buccal fat pad. Regular blood tests and physical examinations will be conducted, and any adverse events (AEs) or serious EAs (SAEs) will be meticulously recorded. Radiographic imaging will be performed prior to surgery and at regular intervals after reconstruction of the alveolar cleft with the BCP-MFAT combination. Biopsies obtained after 6 months with a trephine drill used to prepare the implantation site will be assessed with histological and histomorphometric analyses after methylmethacrylate embedding and sectioning. RESULTS: The primary outcome parameter will be safety after 6 months' follow-up, as monitored closely using possible occurrences of SAEs based on radiographic imaging, blood tests, and physical examinations. For efficacy, radiographic imaging will be used for clinical grading of the bone construct using the Bergland scale. In addition, bone parameters such as bone volume, osteoid volume, graft volume, and number of osteoclasts will be histomorphometrically quantified. Recruitment started in November 2019, and the trial is currently in the follow-up stage. This protocol's current version is 1.0, dated September 15, 2019. CONCLUSIONS: In this first-in-human study, not only safety but also the histologically and radiographically assessed regenerative potential of the BCP-MFAT combination will be evaluated in an alveolar cleft model. When an SAE occurs, it will be concluded that the BCP-MFAT combination is not yet safe in the current setting. Regarding AEs, if they do not occur at a higher frequency than that in patients treated with standard care (autologous bone) or can be resolved by noninvasive conventional methods (eg, with analgesics or antibiotics), the BCP-MFAT combination will be considered safe. In all other cases, the BCP-MFAT combination will not yet be considered safe. TRIAL REGISTRATION: Indonesia Clinical Trial Registry INA-EW74C1N; https://tinyurl.com/28tnrr64. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/42371.

Evaluation of a novel triplex immunochromatographic test for rapid simultaneous detection of norovirus, rotavirus, and adenovirus on a single strip test.

BACKGROUND: Acute gastroenteritis is one of the major causes of morbidity and mortality in young children worldwide. Among these, rotavirus, norovirus, and adenovirus have been reported as the primary viral pathogens associated with the disease. Rapid diagnosis of viral pathogens is crucial when diarrhea outbreaks occur to ensure the timely administration of appropriate treatment and control measures. METHODS: We evaluated three immunochromatographic test kits designed for the detection of norovirus, rotavirus, and adenovirus in 71 stool specimens collected from children with diarrhea who visited clinics in Japan. The first kit is a triplex immunochromatographic test kit designed for simultaneous detections of norovirus, rotavirus, and adenovirus on a single strip (this kit was referred to as IC-A). The other two immunochromatographic test kits are a dual detection kit for rotavirus and adenovirus, and a single detection kit for norovirus (IC-B). The RT-PCR/PCR was used as the gold standard method. RESULTS: The results revealed that both IC-A and IC-B kits exhibited the same level of sensitivity of detection for rotavirus (72.7%) and adenovirus (22.7%), although the detection rate was lower than that of the RT-PCR/PCR method. However, there was a slight difference in the sensitivity of detection for norovirus between IC-A and IC-B, at 86.7% and 93.3%, respectively. The sensitivity of detection for adenovirus of both kits was relatively lower than those of RT-PCR method. This could be due to low viral load of adenovirus in clinical specimens below the detection limit of IC-A and IC-B kits. However, both immunochromatographic test kits (IC-A and IC-B) exhibited 100% specificity for norovirus, rotavirus, and adenovirus. CONCLUSIONS: The triplex immunochromatographic test kit (IC-A) designed for simultaneous detection of norovirus, rotavirus, and adenovirus has been proved to be more practical and convenient than the use of single or dual detection kits with more or less the same sensitivity and specificity of detections.

Is rest-activity rhythm prospectively associated with all-cause mortality in older people regardless of sleep and physical activity level? The 'Como Vai?' Cohort study.

OBJECTIVE: This study aims to test the association of rest-activity rhythm (intradaily variability and interdaily stability) with all-cause mortality in an older adult cohort in Brazil. It also assesses whether the amount of time spent at each intensity level (i.e., physical activity and nocturnal sleep) interferes with this association. METHODS: This cohort study started in 2014 with older adults (≥60 years). We investigated deaths from all causes that occurred until April 2017. Rest-activity rhythm variables were obtained using accelerometry at baseline. Intradaily variability indicates higher rhythm fragmentation, while interdaily stability indicates higher rhythm stability. Cox proportional-hazard models were used to test the associations controlling for confounders. RESULTS: Among the 1451 older adults interviewed in 2014, 965 presented valid accelerometry data. During the follow-up period, 80 individuals died. After adjusting the analysis for sociodemographic, smoking, morbidity score, and number of medicines, an increase of one standard deviation in interdaily stability decreased 26% the risk of death. The adjustment for total sleep time and inactivity did not change this association. On the other hand, the association was no longer significant after adjusting for overall physical activity and moderate to vigorous physical activity. CONCLUSION: Rest-activity rhythm pattern was not associated with mortality when physical activity was considered, possibly because this pattern could be driven by regular exercise. Promoting physical activity remains a relevant strategy to improve population health.

IFNγ-IL12 axis regulates intercellular crosstalk in metabolic dysfunction-associated steatotic liver disease.

Obesity is a major cause of metabolic dysfunction-associated steatohepatitis (MASH) and is characterized by inflammation and insulin resistance. Interferon-γ (IFNγ) is a pro-inflammatory cytokine elevated in obesity and modulating macrophage functions. Here, we show that male mice with loss of IFNγ signaling in myeloid cells (Lyz-IFNγR2-/-) are protected from diet-induced insulin resistance despite fatty liver. Obesity-mediated liver inflammation is also attenuated with reduced interleukin (IL)-12, a cytokine primarily released by macrophages, and IL-12 treatment in vivo causes insulin resistance by impairing hepatic insulin signaling. Following MASH diets, Lyz-IFNγR2-/- mice are rescued from developing liver fibrosis, which is associated with reduced fibroblast growth factor (FGF) 21 levels. These results indicate critical roles for IFNγ signaling in macrophages and their release of IL-12 in modulating obesity-mediated insulin resistance and fatty liver progression to MASH. In this work, we identify the IFNγ-IL12 axis in regulating intercellular crosstalk in the liver and as potential therapeutic targets to treat MASH.

Caryophyllene-type sesquiterpenes from the endophytic fungus Pestalotiopsis lespedezae through an OSMAC approach.

Two new caryophyllene-type sesquiterpenes pestalotiopsins U and V (1 and 2) and three known compounds pestalotiopsin B (7), pestaloporinate B (8), and pestalotiopsin C (9) were isolated by the cultivation of the endophytic fungus Pestalotiopsis lespedezae on solid rice medium, while four additional new caryophyllene pestalotiopsins W-Z (3-6) were obtained when 3.5% NaI was added to the fungal culture medium. The structures of the new compounds were determined by HRESIMS and 1D/2D nuclear magnetic resonance data. Compounds 1-9 were tested for cytotoxicity against the mouse lymphoma cell line L5178Y, but only 6 displayed significant activity with an IC50 value of 2.4 µM.

Características das instalações e equipamentos para a prática de atividade física em escolas da zona rural de Pelotas, Rio Grande do Sul / Facilities and equipments for physical activity practice in rural schools of Pelotas, Rio Grande do Sul

O objetivo do estudo foi descrever as instalações e equipamentos para atividade física nas escolas da zona rural de Pelotas, Rio Grande do Sul. Trata-se de um estudo transversal, de base escolar, realizado no ano de 2014, em todas as 26 escolas localizadas na zona rural da cidade de Pelotas. Foi utilizado um instrumento com questões relacionadas às políticas de Educação Física e avaliação das estruturas e materiais para a prática de atividade física, elaborado a partir do Physical Activity Resource Assessment (PARA), adaptado para o ambiente escolar. Observou-se que o professor e a disciplina de Educação Física estiveram presentes em todas as escolas. Mesmo com a maior frequência de estruturas como quadras esportivas (34,6% a 65,4%), pracinha/parquinhos (76,9%) e materiais como bolas (50,0% a 84,6%), cones (80,8%), cordas (88,5%), colchonetes (80,8%) e arcos (76,9%), as condições dos atributos foram regulares (7,7% a 46,2%) e ruins (11,5% a 50,0%). Poucas instituições ofereceram instalações e equipamentos em condições adequadas para a prática de atividade física. A ausência de ginásios, quadras cobertas e materiais em melhores condições chama a atenção para a necessidade de investimentos e intervenções nesses espaços

The aim of the study was to describe the facilities and equipment for physical activity practice in rural area schools of Pelotas, Rio Grande do Sul. The design was cross-sectional, school-based, carried out in all the 26 schools located in rural areas of Pelotas Southern, Brazil. We used a survey tool to assess the policies for Physical Education and evaluate the facilities and equipment/materials for physical activity. The instrument was based on the Physical Activity Resource Assessment (PARA), adapted for the school environment. We observed that curricular Physical Education and specific teachers were present in all schools. Despite the high number of facilities, such as courts and fields (34.6% to 65.4%), playgrounds (76.9%) and materials such as balls (50.0% to 84.6%), hoops (88.5%), exercise cones (80.8%) and mats (76.9%), the condition of the facilities and apparatus/equipment were fair (7.7% to 46.2%) or poor (11.5% to 50.0%). Few institutions have provided suitable facilities and equipment for physical activity. The absence of gymnasiums, covered courts and materials in better conditions draws attention to the need for investments and interventions in these spaces

Espaços públicos de lazer: distribuição, qualidade e adequação à prática de atividade física / Public open spaces in Pelotas (Brazil): distribution, quality and suitability for physical activity practice

Os espaços públicos de lazer representam uma alternativa de ampliação e democratização do acesso a locais propícios para prática de atividade física. O objetivo do estudo foi descrever os espaços públicos de lazer em termos quanti e qualitativos, assim como sua adequação para a prática de atividades físicas e distribuição na cidade. Foi realizado um estudo observacional descritivo, realizado na zona urbana do município de Pelotas/RS. Aspectos de conforto, estética e adequação à prática de atividades físicas foram avaliados por meio do instrumento Physical Activity Resource Assessment (PARA) em praças, parques e canteiros habitáveis. Informações sobre renda e densidade populacional dos setores censitários foram baseadas no censo demográfico de 2010. Foram identificadas 110 praças e oito parques. Mais deles (57,6%) apresentam algum atributo para prática de atividade física, enquanto nos canteiros este percentual foi de 44,8%. Espaços para prática de futebol e parquinhos foram as estruturas mais comuns em praças/parques (47,5% e 43,2%, respectivamente), enquanto pistas/trilhas de caminhada foram as mais comuns nos canteiros (43,3%). Em geral, 45,7% das estruturas apresentaram boa qualidade. Quanto maior a média de renda domiciliar dos setores censitários, maior o número de espaços de lazer (r = 0,31) e uma relação inversa foi encontrada com densidade populacional dos setores censitários (r = -0,32). Assim, evidenciou-se iniquidades socioeconômicas e demográficas na distribuição dos espaços públicos de lazer, bem como uma pequena diversidade das estruturas para a prática de atividade física e a necessidade de (re)qualificação dos espaços públicos já existentes.

Public open spaces might be an alternative to increase and democratize the access to places for physical activity practice. The study aim was to describe quantitative and qualitatively public open spaces in the city of Pelotas, Brazil, as well as their distribution in the city and their suitability for physical activity practice. An observational descriptive study was performed in the urban area of Pelotas (Brazil). Amenities, incivilities, aesthetic and suitability for physical activity in the public open spaces were evaluated through the Physical Activity Resource Assessment. Mean income and population density of the census tracts were based on the Brazilian demographic census (2010). Most parks (57.6%) presented some attribute for physical activity practice; while among boulevards the percentage was lower (44.8%). Football pitches and playgrounds were the most common attributes found in parks (47.5% and 43.2%, respectively), and walk path/trail was the most frequent in the boulevards (43,3%). In general, 45.7% of the attributes for physical activity practice were considered as presenting high quality. There was a positive correlation between census tracts' mean income and the number of public open spaces (r = 0.31). Otherwise, an inverse correlation was found with population density (r = -0.32). Socioeconomic and demographic iniquities were observed in the distribution of public open spaces. Further, this study highlighted needs of improvements on diversity of attributes for physical activity practice and (re)qualification of the existing public open spaces.

Barreiras à prática de atividades físicas de adolescentes escolares da zona rural do sul do Rio Grande do Sul / Barriers to physical activity in adolescent students from rural area of southern Rio Grande do Sul

O objetivo do presente estudo foi identificar a prevalência de barreiras à prática de atividades físicas e fatores associados em alunos do ensino médio de escolas da zona rural da região sul do Rio Grande do Sul. Foram convidados a participar do estudo todos os alunos matriculados no ensino médio, das 10 escolas compreendidas pela 5ª Coordenadoria Regional de Educação (5ª CRE-RS). O estudo, de delineamento transversal, investigou 510 alunos adolescentes (34% do total da população) que responderam questionário sobre barreiras à prática de atividades físicas e fatores associados. Dos entrevistados, 59,0% eram do sexo feminino, 80,0% tinham entre 15 e 17 anos, 92,3% eram de cor de pele branca e 37,7% estavam cursando o primeiro ano do ensino médio. Mais da metade (53,3%) dos estudantes era suficientemente ativo no lazer e 82,9% encontravam-se na categoria normal de IMC. As barreiras que apresentaram maior prevalência foram: dias de chuva (79,1%), falta de tempo (75,3%) e falta de local adequado para a prática (52,3%). Entre as meninas que relataram oito e nove barreiras, as prevalências de insuficientemente ativas foi, respectivamente de 55,6% e 66,7%, enquanto que, para os meninos que relataram esse mesmo número de barreiras, não houve indivíduos insuficientemente ativos. A única barreira que permaneceu associada a inatividade física no lazer na análise multivariável foi a falta de dinheiro (p=0,03). Conclui-se que mais estudos com adolescentes residentes no meio rural são necessários para se conhecer as necessidades e assim estruturar políticas públicas de incentivo a atividade física.

The aim of the study was to identify the prevalence of barriers to physical activity and associated factors in high school students from rural area of southern Rio Grande do Sul. All students enrolled in high school were invited to participate in the study, ranging from 10 schools for the 5th CRE-RS. Cross-sectional study interviewed 510 adolescent students (34% of the total of the population) who answered a questionnaire about barriers to physical activity and associated factors. Among them, 59.0% were female, 80.0% were between 15 and 17 years, 92.3% were white color and 37.7% were attending the first year of high school. 53.3% of students were active during leisure time and 82.9% were with normal BMI. The more prevalent barriers were: rainy days (79.1%), lack of time (75.3%) and lack of a proper place to practice (52.3%). Among girls who reported eight and nine barriers, the prevalence of insufficient physical activity was respectively 55.6% and 66.7%, whereas for boys who reported the same number of barriers, there were no insufficient active individuals. The only barrier that remained associated with physical inactivity in the multivariate analysis was the lack of money (p = 0.03). We conclude that further studies with adolescents living in rural areas are required to meet the needs and improve public policies to encourage physical activity.

Isolation and cultivation of fungal strains from in vitro cell cultures of two marine sponges (Porifera: Halichondrida and Haplosclerida)

Despite the large number of reports describing sponge-microbe associations, limited knowledge is available about associated fungi and their relationships with the hosts. In this work, specific fungal strains were obtained directly from in vitro sponge cell cultures (primmorphs) and single sponge cells (cytospins) and compared with those obtained from whole tissue preparations. A total of 27 fungal strains were isolated from the marine sponges Hymeniacidon heliophila and Haliclona melana. Fifteen strains, nine from H. heliophila and six from H. melana, were obtained from whole tissue and were considered as possible mesohyl associated or transient fungi. Twelve strains were isolated from in vitro sponge cell cultures (primmorphs) and were, therefore, considered as cell associated. From these, five different strains were obtained from H. heliophila isolated cells, while five were identified from cytospins and two from primmorphs of H. melana. The fungal strains obtained from cell cultures from both sponge species were different, and none of them were detected in the whole tissue preparations of the same species. Nine H. heliophila and seven H. melana strains shows low similarity with the sequences available in public databases and belong to potentially new species. This is the first report of fungi isolated directly from sponge cells, which allowed the observation and selection of specific strains that probably would not be obtained by usual culture dependent techniques.

Macrodactilia e síndrome de Klippel-Trenaunay-Weber / Macrodactyly and Klippel-Trenaunay-Weber syndrome

Embora a macrodactilia seja uma condiçäo patológica infreqüente, pouco observada pelo reumatologista, ela merece ser revisitada. Apresentamos este caso (com macrodactilia, hemangiomatose cutânea, varizes exuberantes e assimetria dos membros inferiores, ao lado de síndrome do túnel carpiano bilateral), para alertar quanto ao diagnóstico diferencial entre a síndrome de Klippel-Trenaunay-Weber, a neurofibromatose, a acromegalia e a macrodistrofia lipomatosa. É possível que um diagnóstico correto mais precoce possa propiciar melhor controle da enfermidade, assim como prevenir atitudes cirúrgicas inadequadas