Biomimetic approach for an articular cartilage patch: Combination of decellularized cartilage matrix and silk-elastin-like-protein (SELP) hydrogel.

Articular cartilage degradation due to injury, disease and aging is a common clinical issue as current regenerative therapies are unable to fully replicate the complex microenvironment of the native tissue which, being avascular, is featured by very low ability to self-regenerate. The extracellular matrix (ECM), constituting almost 90% of the entire tissue, plays a critical role in its function and resistance to compressive forces. In this context, the current tissue engineering strategies are only partially effective in restoring the biology and function of the native tissue. A main issue in tissue regeneration is treatment failure due to scarce integration of the engineered construct, often following a gradual detachment of the graft. In this scenario, we aimed to create an adhesive patch able to adequately support cartilage regeneration as a promising tool for the treatment of cartilage injuries and diseases. For this, we produced an engineered construct composed of decellularized ECM (dECM) obtained from horse joint cartilage, to support tissue regeneration, coupled with a Silk-Elastin-Like Proteins (SELP) hydrogel, which acts as a biological glue, to guarantee an adequate adherence to the host tissue. Following the production of the two biomaterials we characterized them by assessing: 1) dECM morphological, chemical, and ultrastructural features along with its capability to support chondrocyte proliferation, specific marker expression and ECM synthesis; 2) SELP microarchitecture, cytocompatibility and mechanical properties. Our results demonstrated that both materials hold unique properties suitable to be exploited to produce a tailored microenvironment to support cell growth and differentiation providing a proof of concept concerning the in vitro biological and mechanical efficacy of the construct. The SELP hydrogel displayed a very interesting physical behavior due to its high degree of resistance to mechanical stress, which is generally associated with physiological mechanical load during locomotion. Intriguingly, the shear-thinning behavior of the hydrogel may also make it suitable to be applied and spread over non-homogeneous surfaces, therefore, we hypothesize that the hybrid biomaterial proposed may be a real asset in the treatment of cartilage defects and injuries.

Mansonella sp. and associated Wolbachia endosymbionts in ring-tailed coatis (Nasua nasua) in periurban areas from Midwestern Brazil.

Coatis (Nasua nasua) are wild carnivorous well adapted to anthropized environments especially important because they act as reservoirs hosts for many arthropod-borne zoonotic pathogens. Information about filarioids from coatis and associated Wolbachia spp. in Brazil is scant. To investigate the diversity of filarial nematodes, blood samples (n = 100 animals) were obtained from two urban areas in midwestern Brazil and analyzed using blood smears and buffy coats and cPCR assays based on the cox1, 12S rRNA, 18S rRNA, hsp70 and myoHC genes for nematodes and 16S rRNA for Wolbachia. When analyzing coati blood smears and buffy coats, 30% and 80% of the samples presented at least one microfilaria, respectively. Twenty-five cox1 sequences were obtained showing 89% nucleotide identity with Mansonella ozzardi. Phylogenetic analyses clustered cox1 sequences herein obtained within the Mansonella spp. clade. Sequences of both myoHC and two hsp70 genes showed 99.8% nucleotide identity with Mansonella sp. and clustered into a clade within Mansonella sp., previously detected in coatis from Brazil. Two blood samples were positive for Wolbachia, with a 99% nucleotide identity with Wolbachia previously found in Mansonella perstans, Mansonella ozzardi and Mansonella atelensis and in ectoparasites of the genus Pseudolynchia, Melophagus and Cimex. The study showed a high prevalence of Mansonella sp. in the coati population examined, suggesting that this animal species play a role as reservoirs of a novel, yet to be described, species within the Onchocercidae family.

Resultados de 5 anos da cirurgia bariátrica em pacientes com obesidade e hipertensão. um estudo clínico randomizado / 5-year outcomes of bariatric surgery in patients with obesity and hypertension. a randomized clinical trial

INTRODUÇÃO: Dados de estudos randomizados avaliando os efeitos a longo prazo da cirurgia bariátrica no controle e remissão da hipertensão (HTN) são escassos. O seguimento de curto e médio prazo do Estudo GATEWAY (1 e 3 anos) mostrou melhora significativa e remissão da hipertensão após a cirurgia bariátrica. Aqui, apresentamos os dados de acompanhamento de 5 anos deste estudo. MÉTODOS: Conduzimos um ensaio clínico randomizado, em um centro único, fase III, paralelo e aberto. Pacientes com obesidade graus 1 e 2, tomando no mínimo 2 medicamentos anti-hipertensivos em doses máximas para hipertensão, foram alocados aleatoriamente para Bypass gástrico em Y de Roux (RYGB) combinado com tratamento clínico (TC) ou TC (estilo de vida/aconselhamento nutricional) isoladamente. O desfecho primário foi a redução de pelo menos 30% do total de anti-hipertensivos, mantendo níveis controlados de pressão arterial (PA) <140/90mmHg. A análise principal seguiu o princípio de intenção de tratar modificada e os valores ausentes foram imputados com a técnica de imputação múltipla. Os desfechos secundários incluíram vários biomarcadores. RESULTADOS: Foram incluídos 100 pacientes (76% mulheres, idade: 43,8±9,2 anos, índice de massa corporal, IMC: 36,9±2,7Kg/ m2). Após 5 anos, o IMC foi de 27,8±3,8kg/m2 para RYGB e 36,4±3,9kg/m2 para TC (P<0,001). A análise de sensibilidade considerando apenas os casos com seguimento concluído (n=69) revelou resultados consistentes. Os desfechos secundários nos grupos RYGB e TC, aos 60 meses, foram: circunferência abdominal 88,05 ± 9,84cm e 109,44 ± 9,76cm (P<0,001) ; hemoglobina glicada 5,4 ± 0,43% e 5,96 ± 0,9% (P<0,001); insulina 6,41 ± 2,46mU/L e 24,65 ± 16,57mU/L (P<0,001); LDL colesterol 93.61 ± 31.06mg/dL e 109.94 ± 37.58mg/dL (P=0,022); triglicerídeos 86,76 ± 38,45mg/dL e 140,33 ± 59,11 mg/dL (P<0,001); proteína C reativa de alta sensibilidade 0,81 ± 0,76mg/L e 6,13 ± 6,22mg/L (P=0,005), respectivamente. CONCLUSÕES: A cirurgia bariátrica é uma estratégia eficaz e duradoura no controle da PA com menor número ou nenhum medicamento anti-hipertensivo e na melhora do perfil metabólico e inflamatório em pacientes com obesidade graus 1 ou 2 e hipertensão.

Efficacy and safety of ultra-low-dose estradiol and norethisterone in postmenopausal Brazilian women.

OBJECTIVE: This study aimed to evaluate the efficacy and safety of oral ultra-low-dose continuous combination of 17ß-estradiol (17ß-E2) and norethisterone acetate (NETA) in postmenopausal Brazilian women. METHODS: Postmenopausal women (age 45-60 years) with amenorrhea >12 months and intact uterus, with moderate to severe vasomotor symptoms, were included. The vasomotor symptoms and endometrial bleeding were evaluated by a daily diary for 24 weeks, and the women were assessed at baseline and endpoint. RESULTS: A total of 118 women were included. The group treated with 0.5 mg 17ß-E2/0.1 mg NETA (n = 58) showed a percentage reduction of 77.1% in the frequency of vasomotor symptoms versus 49.9% in the placebo group (n = 60) (p = 0.0001). The severity score showed a reduction in the treatment group when compared to the placebo (p < 0.0001). The adverse events were comparable between the groups; however, in the 0.5 mg 17ß-E2/0.1 mg NETA group there were more complaints of vaginal bleeding; despite that, in most cycles in both treatment groups, more than 80% of women experienced amenorrhea. CONCLUSIONS: The combination of 0.5 mg 17ß-E2/0.1 mg NETA in a continuous combination regimen was shown to be effective in reducing the frequency and severity of vasomotor symptoms in Brazilian postmenopausal women.

Molecular detection of Babesia spp. and Rickettsia spp. in coatis (Nasua nasua) and associated ticks from midwestern Brazil.

Procyonids are reservoirs of many zoonotic infectious diseases, including tick-borne pathogens. The role of coatis (Nasua nasua) in the epidemiology of piroplasmids and Rickettsia has not been fully addressed in Brazil. To molecularly study these agents in coatis and associated ticks, animals were sampled in two urban areas in Midwestern Brazil. Blood (n = 163) and tick (n = 248) DNA samples were screened by PCR assays targeting the 18S rRNA and gltA genes of piroplasmids and Rickettsia spp., respectively. Positive samples were further molecularly tested targeting cox-1, cox-3, ß-tubulin, cytB, and hsp70 (piroplasmid) and ompA, ompB, and htrA 17-kDa (Rickettsia spp.) genes, sequenced and phylogenetically analyzed. All coatis' blood samples were negative for piroplasmids, whereas five pools of ticks (2%) were positive for two different sequences of Babesia spp.. The first from Amblyomma sculptum nymphs was close (i.e., ≥ 99% nucleotide identity) to a Babesia sp. previously found in capybaras (Hydrochoerus hydrochaeris); the second from Amblyomma dubitatum nymphs and Amblyomma spp. larvae was identical (100% nucleotide identity) to a Babesia sp. detected in opossums (Didelphis albiventris) and associated ticks. Four samples (0.8%) were positive by PCR to two different Rickettsia spp. sequences, being the first from Amblyomma sp. larva identical to Rickettsia belli and the second from A. dubitatum nymph identical to Rickettsia species from Spotted Fever Group (SFG). The detection of piroplasmids and SFG Rickettsia sp. highlights the importance of Amblyomma spp. in the maintenance of tick-borne agents in urban parks where humans and wild and domestic animals are living in sympatry.

Co-infection by multiple vector-borne agents in wild ring-tailed coatis (Nasua nasua) from Iguaçu National Park, southern Brazil.

The present study aimed to detect molecularly the presence of co-infections by vector-borne agents (VBA) in ring-tailed coatis' (Nasua nasua) blood samples from Iguaçu National Park (INP), southern Brazil, and assess the phylogenetic positioning of the detected agents. DNA blood samples were submitted to molecular screening and characterization for Anaplasmataceae agents, Piroplasmids, Hepatozoon sp., hemotropic mycoplasmas, and Bartonella spp. In total, 42 (85.7%) coatis were positive for hemotropic Mycoplasma sp., 12 (24.5%) for Bartonella machadoae, 7 (14.3%) for Anaplasma sp. closely related to 'Candidatus Anaplasma brasiliensis', and 3 (6%) for Hepatozoon procyonis. The most prevalent co-infections observed was from bacterial VBA: while 18.3% were co-infected by hemotropic Mycoplasma sp. and Bartonella sp., 12.2% were co-infected by Anaplasma sp. and hemotropic Mycoplasma sp. Only two animals (4%) presented co-infections by three VBA (Bartonella sp., Anaplasma sp. and hemotropic Mycoplasma sp.). The coati is a wild carnivore found in INP, mainly in areas visited by tourists. These animals are frequently seen searching for food in garbage dumps or in tourists' belongings. The present study expands the host specificity range of B. machadoae, which has been isolated only from rodents until the present moment. Since the zoonotic potential and transmission routes of the detected VBA are not yet known, surveillance in this area is much needed.

Adverse reactions to the injection of face and neck aesthetic filling materials: a systematic review.

BACKGROUND: Adverse reactions, caused during the inflammation and healing process, or even later, can be induced by the injection of dermal filler and can present a variety of clinical and histological characteristics. In this study we aimed to review the adverse reactions associated with the injection of aesthetic filling materials in the face and neck. MATERIAL AND METHODS: The review was reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses checklist. Studies published that mentioned adverse reactions in patients with aesthetic filling materials in the face or neck were included. Risk of bias was assessed using the Joanna Briggs Institute appraisal tool. After a 2-step selection process, 74 studies were included: 51 case reports, 18 serial cases, and five cohorts. RESULTS: A total of 303 patients from 20 countries were assessed. Lesions were more prevalent in the lip (18%), nasolabial folds (13%), cheeks (13%), chin (10%), submental (8%), glabella (7%), and forehead (6%). Histopathological analysis revealed a foreign body granuloma in 87.1% of the patients, 3% inflammatory granuloma, 3% lipogranuloma, 2.3% xanthelasma-like reaction, 1% fibrotic reaction, 0.7% amorphous tissues, 0.7% xanthelasma, 0.3% sclerosing lipogranuloma, 0.3% siliconoma, and 0.3% foreign body granuloma with scleromyxedema. In addition, two patients displayed keratoacanthoma and two others displayed sarcoidosis after cutaneous filling. The most commonly used materials were silicone fillers (19.7%), hyaluronic acid (15.5%), and hydroxyethyl methacrylate/ethyl methacrylate suspended in hyaluronic acid acrylic hydrogel (5.6%). All patients were treated, and only 12 had prolonged complications. CONCLUSIONS: There is evidence that adverse reaction can be caused by different fillers in specific sites on the face. Although foreign body granuloma was the most common, other adverse lesions were diagnosed, exacerbating systemic diseases. In this way, we reinforce the importance of previous systemic evaluations and histopathological analyses for the correct diagnosis of lesions.

Profile of Brazilian climacteric women: results from the Brazilian Menopause Study.

OBJECTIVE: This study aims to understand the epidemiological characteristics of Brazilian menopausal women, and their view on menopause hormone therapy (MHT). METHODS: A national cross-sectional study with 1500 women between 45 and 65 years old was carried out through questionnaires. RESULTS: The overall median age of participants was 52 [47-56] years, and 55 [52-59] years for the postmenopausal subgroup. Menstrual irregularity started at median age 46 [44-49] years. Median menopause age was 48 [45-51] years with no differences between socioeconomic classes. The prevalence of any climacteric symptoms was 87.9% and hot flashes started at median age 47 [45-50] years. Among women in menopause/menopausal transition, 52.1% received any medical prescription, and MHT was recommended for 22.3%. Among those who started MHT, 45.4% were still using the treatment and the median duration of use was 8 months, but different among socioeconomic classes (24 months for class A against 3 months for class D/E). CONCLUSIONS: In this first Brazilian national population-based study on menopause and MHT, it was observed that, in spite of being symptomatic when entering menopause around 48 years of age, only a small part of Brazilian women started MHT and the median duration of treatment was less than 1 year, but the duration was higher for higher socioeconomic class.

Longitudinal dynamics and health impact of Hepatozoon procyonis (Apicomplexa: Hepatozoidae) on naturally infected ring-tailed coatis Nasua nasua (Carnivora: Procyonidae) from Midwestern Brazil.

This study aimed to morphologically and molecularly detect Hepatozoon procyonis in ring-tailed coatis' (Nasua nasua) blood and associated ticks from central-western Brazil, Campo Grande, Mato Grosso do Sul state and also evaluate the impact of the protozoa in blood parameters and coati´s health. Samplings were performed in a conservation area Parque Estadual do Prosa (PEP) and in a Brazilian Air Force Private Area namely Vila da Base Aérea (VBA), between March 2018 and April 2019. We collected 165 blood samples, 61 from recaptured coatis. Peripheral blood smears were stained with Romanovsky-type stain for H. procyonis parasitemia assessment. DNA extracted from blood samples and ticks (Amblyomma spp.) were submitted to a nested PCR (nPCR) assay based on the 18S rRNA gene for Hepatozoon spp. Out of 104 individuals sampled, 80 (77%) were positive for H. procyonis in at least one capture. Overall, 67/165 (40.6%) blood smears showed H. procyonis gametocytes (PEP: 41/63 - 65%; VBA: 26/102 - 25.5%). Parasitemia based on 500 assessed leucocytes ranged from 1 (0.2%) to 50 (10%) and 1 (0.2%) to 25 (5%), from animals sampled in PEP and VBA, respectively. Fluctuation on the parasitemia was observed during recaptures. nPCR results showed higher positivity when compared to blood smears, i.e. 112/165 (68%) positive blood samples [PEP: 41/63 (65%), VBA: 26/102 (25.5%)]. In total, 63/248 (25.4%) tick DNA samples were positive at nPCR for Hepatozoon sp., including 32/87 (37%) pools (1 to 10 larvae) of Amblyomma larvae, 21/105 (20%) pools (1 to 5 nymphs) of Amblyomma sculptum nymphs, 9/43 (21%) pools (1 to 5 nymphs) of Amblyomma dubitatumnymphs, and 1/12 (8%) A. sculptum adult female. The partial 18S rRNA sequence from one coati's blood sample and one representative of each positive tick species randomly selected from each area for sequencing (1,000 bp) showed 100% identity with sequences of H. procyonis from GenBank previously detected in coatis. Regarding H. procyonis infection, no statistical differences were obtained when comparing males vs. females (p-value 0.67), immature animals vs. adults (p-value 0.31), rainy vs. dry season (p-value 0.51) and sampling location (p-value 0.42). No noticeable alteration in blood parameters or heath status was observed in parasite animals. H. procyonis circulates in a high prevalence in coatis from central-western Brazil. Parasitemia fluctuates among different coatis' recaptures and apparently the infection has no influence in coatis' hematological and clinical parameters.

Degradation of carbendazim in aqueous solution by different settings of photochemical and electrochemical oxidation processes.

The present study analyzed the performance of photochemical and electrochemical techniques in the degradation and mineralization of the pesticide carbendazim (CBZ). Direct photolysis (DP), heterogeneous photocatalysis (HP), photoelectrocatalysis (PEC), and electrochemical oxidation (EO) were tested, and the influence of UV radiation, current density (j), and supporting electrolyte concentration were evaluated. The results suggest that CBZ is only degraded by DP when UV-C254nm is used. For HP, the CBZ degradation was observed both when UV-A365nm or UV-C254nm were used, which is related to the reactive oxygen species (ROS) formed by the photocatalytic activity (photon-ROS). Neither DP nor HP were able to mineralize CBZ, demonstrating its resistance to photomediated processes. For EO, regardless of the j, there were higher CBZ degradation and mineralization than those observed when using DP and HP. The increase in the supporting electrolyte concentration (Na2SO4) did not affect the levels of degradation and mineralization of CBZ. Concerning the PEC, a CBZ mineralization of 52.2% was accomplished. These findings demonstrate that the EO is the main pathway for CBZ mineralization, suggesting an additional effect of the electro-ROS on the photon-ROS and UV-C254nm. The values of mineralization, kinetics, and half-life show that PEC UV-C254nm with a j of 15 mA cm-2 was the best setting for the degradation and mineralization of CBZ. However, when the values of specific energy consumption were considered for industrial applications, the use of EO with a j of 3 mA cm-2 and 4 g L-1 of Na2SO2 becomes more attractive. The assessment of by-products formed after this best cost-efficient treatment setting revealed the presence of aromatic and aliphatic compounds from CBZ degradation. Acute phytotoxicity results showed that the presence of sodium sulfate can be a representative factor regarding the toxicity of samples treated in electrochemical systems.

Psychological symptomatology in a prison population: an exploratory study of age, psychopathological history and time in prison.

OBJECTIVES: Identify psychological symptoms relating to age, psychopathological history and time in prison in women and men incarcerated in a prison. MATERIAL AND METHOD: Cross-sectional study. The sample was made up of 100 inmates, 50 men and 50 women and the symptom Checklist-90-Revised (SCL-90-R) was used to assess psychopathological symptoms. RESULTS: This study shows that inmates with a psychopathological history prior to entry to prison, younger inmates (18-29 years) and inmates who have been a short time in prison present more psychopathological symptoms. DISCUSSION: The results found suggest the implementation of a protocol for psychological care of prisoners in general, but highlights a particular interest in the care of cases with people with a psychopathological history prior to entering prison, in those who are younger and those who have been in prison for a short time.

COVID-19 symptoms: a case-control study, Portugal, March-April 2020.

COVID-19, although a respiratory illness, has been clinically associated with non-respiratory symptoms. We conducted a negative case-control study to identify the symptoms associated with SARS-CoV-2-positive results in Portugal. Twelve symptoms and signs included in the clinical notification of COVID-19 were selected as predictors, and the dependent variable was the RT-PCR test result. The χ2 tests were used to compare notified cases on sex, age group, health region and presence of comorbidities. The best-fit prediction model was selected using a backward stepwise method with an unconditional logistic regression. General and gastrointestinal symptoms were strongly associated with a positive test (P < 0.001). In this sense, the inclusion of general symptoms such as myalgia, headache and fatigue, as well as diarrhoea, together with actual clinical criteria for suspected cases, already updated and included in COVID-19 case definition, can lead to increased identification of cases and represent an effective strength for transmission control.

Vaginal Health: Insights, Views & Attitudes survey in Latin America (VIVA-LATAM): focus on Brazil.

OBJECTIVE: A previous survey investigated postmenopausal vaginal atrophy in a sample of women across Latin America. To help implement a tailored approach to improve postmenopausal care and outcomes in Brazil, we consider results from the survey for this country. METHODS: A total of 2509 postmenopausal women resident in Argentina, Brazil, Chile, Colombia, or Mexico completed an online questionnaire. The Brazilian cohort comprised 504 women. RESULTS: Over half of the Brazilian cohort (56%) reported experiencing symptoms of vaginal atrophy; most described them as moderate or severe (76%), and almost half (48%) experienced symptoms for at least 1 year. Three-quarters of the Brazilian cohort (75%) were unaware of the chronic nature of the condition. Upon experiencing symptoms of vaginal atrophy, 92% had visited a health-care provider to discuss treatment options. Overall, 56% were aware of some form of local hormone therapy and 40% of those affected by vaginal atrophy had used such treatment. CONCLUSION: Postmenopausal women in Brazil are likely to benefit from increased awareness of the symptoms of vaginal atrophy. Health-care providers can potentially improve outcomes by helping women to understand the chronic nature of the condition and available treatment options. Women may be open to education pre menopause, before symptoms occur.

Effect of organic compost and inorganic nitrogen fertigation on spinach growth, phytochemical accumulation and antioxidant activity.

The aim of this study was to evaluate the effect of the addition of organic compost in combination with the inorganic nitrogen fertigation on growth, phytochemical accumulation, and antioxidant activity of spinach (Spinacia oleracea L. cv. Manatee). Soil blocked spinach seedlings (six seedlings per block), three blocks per pot (316 plants m-2) were transplanted after 18 days after emergence into to 12 L pots. The treatments were: unfertilized soil, organic compost, organic compost +75 kg of N ha-1, applied as ammonium sulfate; and organic compost +75 kg N ha-1, applied as ammonium nitrate. The addition of organic compost to unfertilized soil increased the fresh yield. The application of inorganic N from the two sources in relation to organic compost treatment increased spinach fresh yield from 2.3 to 4.81 kg m-2 and shoot dry weight from 0.60 to 1,31 g plant-1. Levels of carotenoids also increased with inorganic N addition, producing higher values in plants grown with organic compost + ammonium nitrate (31.14 mg/100 g fresh weight). However, the addition of N led to a decrease in leaf-blade total phenols from 75 to 56 mg gallic acid equivalents/100mg fresh weight. The addition of inorganic N led to a dramatic decrease in leaf-blade ferric reducing antioxidant activity. This effect was higher with ammonium sulfate application. The application of organic compost and inorganic nitrogen had no influence on the petiole's phytochemical accumulation and antioxidant activity.

Evaluation of cell damage and modulation of cytokines TNF-α, IL-6 and IL-10 in macrophages exposed to PpIX-mediated photodynamic therapy / Avaliação do dano celular e da modulação das citocinas TNF-α, IL-6 e IL-10 em macrófagos expostos à terapia fotodinâmica mediada pela PpIX

Abstract Little is known regarding whether photodynamic therapy (PDT)-induced cell death can substantially compromise macrophages (MΦ), which are important cells in PDT-induced immune responses. Here, parameters of PDT-mediated MΦ cytotoxicity and cytokine production in response to protoporphyrin IX (PpIX) were evaluated. Peritoneal MΦ from BALB/c mice were stimulated in vitro with PDT, light, PpIX, or lipopolysaccharide (LPS). After that, cell viability, lipid peroxidation, Nitric Oxide (NO), DNA damage, TNF-α, IL-6 and IL-10 were evaluated. Short PDT exposure reduced cell viability by 10-30%. There was a two-fold increase in NO and DNA degradation, despite the non-increase in lipoperoxidation. PDT increased TNF-α and IL-10, particularly in the presence of LPS, and decreased the production of IL-6 to 10-fold. PDT causes cellular stress, induces NO radicals and leads to DNA degradation, generating a cytotoxic microenvironment. Furthermore, PDT modulates pro- and anti-inflammatory cytokines in MΦ.

Resumo Pouco se sabe se a morte celular induzida pela terapia fotodinâmica (PDT) compromete os macrófagos (MΦ), envolvidos nas respostas imunes induzidas pela PDT. Neste estudo, foram avaliados parâmetros de citotoxicidade dos MΦ mediada pela PDT e a produção de citocinas, frente à protoporfirina IX (PpIX). MΦ peritoneais de camundongos BALB/c foram estimulados in vitro com PDT, luz, PpIX ou lipopolissacarídeo (LPS). Após isto, a viabilidade celular (VC), a lipoperoxidação, os níveis de óxido nítrico (NO), de DNA degradado, de TNF-α, IL-6 e IL-10 foram avaliados. A exposição curta à PDT reduziu a VC em 10-30%. Os níveis de NO e de DNA degradado duplicaram, sem aumento da lipoperoxidação. Houve aumento de TNF-α e IL-10, sendo maior na presença de LPS. Já a produção de IL-6 reduziu em dez vezes. A PDT induz estresse celular, gera radicais NO e causa dano ao DNA, tornando o microambiente citotóxico. Ainda, modula citocinas pró e anti-inflamatórias em MΦ.

Antiproliferative and photoprotective activities of the extracts and compounds from Calea fruticosa.

In this paper, we complement our previous study on the antiproliferative activity of Calea fruticosa (Asteraceae) by isolating the compounds apigenin-4',7-dimethyl ether (1), budlein A (2), quercetin (3), and cichoriin (4) from the plant's aerial parts. The antiproliferative activity of these compounds was evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method against human tumor cell lines. Compound 3 displayed moderate antiproliferative activity in three cell lines (HCT-116, PC-3, and SF-295, with cell growth inhibition values of 72.97, 74.55, and 68.94%) and high antiproliferative activity (90.86%) in the HL-60 cell line. The in vitro sun protection factor (SPF) of the extracts and compound 4, with and without sunscreen, was determined by a spectrophotometric method. The ethanol extract exhibited the highest SPF (9.67) at a concentration of 0.100 mg/mL, while compound 4, isolated from this extract, showed a SPF of 13.79 at the same concentration. A relative increased efficacy of SPF was observed for the extracts and compound 4 when sunscreen was also used. Compound 4 has not been reported previously from any species within the genus Calea. Compounds 1-4 were obtained from this species for the first time.

A novel de novo heterozygous ALPL nonsense mutation associated with adult hypophosphatasia.

Using genetic, clinical, biochemical, and radiographic assessment and bioinformatic approaches, we present an unusual case of adult HPP caused by a novel de novo heterozygous nonsense mutation in the alkaline phosphatase (ALPL). INTRODUCTION: Hypophosphatasia (HPP) is caused by genetic alterations of the ALPL gene, encoding the tissue-nonspecific isozyme of alkaline phosphatase (TNSALP). Here, the purpose was to perform clinical and molecular investigation in a 36-year-old Caucasian woman suspected to present adult HPP. METHODS: Medical and dental histories were obtained for the proposita and family members, including biochemical, radiographic, and dental assessments. ALPL mutational analysis was performed by the Sanger sequencing method, and the functional impact prediction of the identified mutations was assessed by bioinformatic methods. RESULTS: We identified a novel heterozygous nonsense mutation in the ALPL gene (NM_000478.6:c.768G>A; W[TGG]>*[TGA]) associated with spontaneous vertebral fracture, severe back pain, musculoskeletal pain, low bone density, and short-rooted permanent teeth loss. Functional prediction analysis revealed that the Trp256Ter mutation led to a complete loss of TNSALP crown domain and extensive loss of other functional domains (calcium-binding domain, active site vicinity, and zinc-binding site) and over 60% loss of homodimer interface residues, suggesting that the mutant TNSALP molecules are nonfunctional and form unstable homodimers. Genotyping of the ALPL in the proposita's parents, sister, and niece revealed that in this case, HPP occurred due to a de novo mutation. CONCLUSION: The present study describes a novel genotype-phenotype and structure-function relationship for HPP, contributing to a better molecular comprehension of HPP etiology and pathophysiology.

The use of Platelet-rich Fibrin in the management of medication-related osteonecrosis of the jaw: A case series.

Medication related osteonecrosis of the jaw (MRONJ) is characterized by exposed necrotic bone in the maxillofacial region that persists for more than eight weeks in patients taking antiresorptive or antiangiogenic drugs for bone metastasis or osteoporosis. The management of such condition depends on several factors, among which the staging of MRONJ. Though, a specific gold standard treatment has not been established to date. The aim of this case series is to describe the outcome of surgical treatment of MRONJ with the adjunct of Platelet-rich Fibrin (PRF). Eleven patients under therapy with alendronate underwent surgical removal of necrotic bone and debridement, followed by placement of PRF membranes in the bone defect. The outcome of the surgical treatment was successful in all patients, in a follow-up range from 12 to 36 months. In the cases presented, the macroscopic evaluation showed excellent and fast soft tissue healing, with no recurrence of bone exposure and no signs of infections. PRF membranes were also effective for postsurgical pain control. The use of PRF may represent a valuable adjunct in the surgical management of MRONJ.

Evaluation of cell damage and modulation of cytokines TNF-α, IL-6 and IL-10 in macrophages exposed to PpIX-mediated photodynamic therapy.

Little is known regarding whether photodynamic therapy (PDT)-induced cell death can substantially compromise macrophages (MΦ), which are important cells in PDT-induced immune responses. Here, parameters of PDT-mediated MΦ cytotoxicity and cytokine production in response to protoporphyrin IX (PpIX) were evaluated. Peritoneal MΦ from BALB/c mice were stimulated in vitro with PDT, light, PpIX, or lipopolysaccharide (LPS). After that, cell viability, lipid peroxidation, Nitric Oxide (NO), DNA damage, TNF-α, IL-6 and IL-10 were evaluated. Short PDT exposure reduced cell viability by 10-30%. There was a two-fold increase in NO and DNA degradation, despite the non-increase in lipoperoxidation. PDT increased TNF-α and IL-10, particularly in the presence of LPS, and decreased the production of IL-6 to 10-fold. PDT causes cellular stress, induces NO radicals and leads to DNA degradation, generating a cytotoxic microenvironment. Furthermore, PDT modulates pro- and anti-inflammatory cytokines in MΦ.

Sources of food contamination in a closed hydroponic system.

This study investigated potential contamination sources in a commercial, closed hydroponic system. Water, substrate and lettuce (Lactuca sativa) samples were evaluated for microbiological indicator populations, including aerobic plate count (APC), coliform bacteria (CB) and yeast and mould (YM). Listeria spp. detection via cultural enrichment and agglutination was negative for all samples. Peat moss substrate (postharvest) had the highest counts for APC (6·8 log CFU per g), CB (4·5 log MPN per g) and YM (5·1 and 4·8 log CFU per g respectively). Roots embedded in plugs demonstrated counts for all populations nearly as high as the substrate. Among water samples, a seedling water reservoir housing germinated plants yielded the highest count for APC (5·1 log CFU per g) and CB (2·4 log MPN per g) likely due to the large numbers of plugs and their close proximity in the reservoir. Harvested lettuce leaves demonstrated higher APC (4·1 log CFU per g) than preharvest leaves (1·7 log CFU per g) due to the transfer of microbes from the root ball. These data suggest that substrates are a significant potential source of contamination in hydroponic systems and likely facilitate microbial transfer to harvested leaves. There is, therefore, the need to further investigate mitigation of potential contamination events. SIGNIFICANCE AND IMPACT OF THE STUDY: Hydroponic production is known to provide safe, clean produce. This study, however, suggests that the hydroponic substrate (peat moss plug) is a possible source of contamination in the hydroponic system. This finding is important as most harvested hydroponic lettuces are packaged and sold with substrate and root ball intact. This implies a high probability of microbial transfer from the root ball to edible harvested lettuce leaves.