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Thromb Res ; 196: 291-296, 2020 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-32949961


INTRODUCTION: Direct thrombin inhibitor, dabigatran and factor Xa inhibitors, apixaban, edoxaban, and rivaroxaban (DOACs/NOACs), are currently the first-choice drugs in some indications. Life-threatening bleeding occurring during DOACs treatment may benefit from the use of reversal agents, however there are some concerns regarding potential rebound thrombotic events. In this systematic review we aimed to estimate the incidence of thrombotic events in patients treated with idarucizumab or andexanet alfa. METHODS: This systematic review included all prospective and retrospective studies, enrolling patients that received specific antidotes (idarucizumab, andexanet alfa and cirapantag) for anticoagulation reversal, published until October 2019 in CENTRAL, MEDLINE and PsycINFO. Studies in healthy individuals and those with less than 10 patients were excluded. The primary outcome was the incidence of thrombotic events and the secondary outcome was all-cause mortality. Studies screening and data extraction was performed in duplicate by reviewers. A random-effects meta-analysis was performed using the Freeman-Tukey transformation of the data. The results are expressed in percentages, with 95%-confidence intervals (CI), limited between 0 and 100% due to the data transformation. RESULTS: Overall 16 studies with 1774 patients were included (13 studies enrolling 1384 patients that received idarucizumab; 3 studies enrolling 390 patients that received andexanet alfa; cirapantag studies were not found). The pooled incidence rate of thrombotic events in the patients treated with specific antidote was 5.5% (95% CI 2.0-10.1%) until 30-90 days. The incidence of all-cause mortality was 13.3% (95% CI 9.6-17.5%). CONCLUSIONS: In patients requiring idarucizumab or andexanet alfa to reach haemostasis, our data shows that there were 5.5% thrombotic events. The causality of harm associated to antidotes remains to be established due to the design of studies without a control group.

Rev Col Bras Cir ; 47: e20202524, 2020 Sep 04.
Artigo em Inglês, Português | MEDLINE | ID: mdl-32901705


OBJECTIVE: to identify predictive factors for lethality and complications of deep fascial space infections of the neck (DFSIN), establishing an early and aggressive treatment in the neck before the progression to descending mediastinitis. METHODS: we retrospectively analyzed 133 cases of DFSIN treated at Discipline of Head and Neck Surgery of the Medicine School of Santa Casa de Misericórdia de São Paulo. We accessed demographic characteristics, associated diseases, clinical presentation, laboratorial tests, length of hospital stay, number of involved anatomic neck spaces, intra-operative and microbiology findings. We analyzed these data using logistic regression to predict DFSIN lethality and life threatening complications (mediastinitis, septic shock, pneumonia, pleural empyema, skin necrosis). RESULTS: lethality and complication ratios were 9% and 50.3%, respectively. The logistic regression model showed that patients with septic shock were more likely to have progression to death (p < 0.001) and, the presence of more than two involved neck spaces (p < 0.001) and older individuals (p = 0.017) were more likely to have complicated deep neck infections. Descending necrotizing mediatinitis increased the lethality ratio by 50%, and was associated to necrotizing fasciitis (p=0.012) and pleural empyema (p<0.001). CONCLUSION: septic shock is a lethal predictive factor and age as well as more than two involved neck spaces are the predictive factors for complications. Necrotizing fasciitis is an important factor for complications and death. Therefore, its surgical treatment must be more aggressive. Descending mediastinitis has a high lethal rate and the successful treatment is based on early diagnosis and aggressive surgical approach.

Fasciite Necrosante/mortalidade , Infecções , Pescoço , Adulto , Empiema Pleural , Fáscia , Humanos , Infecções/complicações , Infecções/etiologia , Pessoa de Meia-Idade , Estudos Retrospectivos
Sci Rep ; 10(1): 12384, 2020 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-32709946


Some lineages of ants, termites, and beetles independently evolved a symbiotic association with lignocellulolytic fungi cultivated for food, in a lifestyle known as fungiculture. Fungus-growing insects' symbiosis also hosts a bacterial community thought to integrate their physiology. Similarities in taxonomic composition support the microbiota of fungus-growing insects as convergent, despite differences in fungus-rearing by these insects. Here, by comparing fungus-growing insects to several hosts ranging diverse dietary patterns, we investigate whether the microbiota taxonomic and functional profiles are characteristic of the fungiculture environment. Compared to other hosts, the microbiota associated with fungus-growing insects presents a distinctive taxonomic profile, dominated by Gammaproteobacteria at class level and by Pseudomonas at genera level. Even with a functional profile presenting similarities with the gut microbiota of herbivorous and omnivorous hosts, some differentially abundant features codified by the microbiota of fungus-growing insects suggest these communities occupying microhabitats that are characteristic of fungiculture. These features include metabolic pathways involved in lignocellulose breakdown, detoxification of plant secondary metabolites, metabolism of simple sugars, fungal cell wall deconstruction, biofilm formation, antimicrobials biosynthesis, and metabolism of diverse nutrients. Our results suggest that the microbiota could be functionally adapted to the fungiculture environment, codifying metabolic pathways potentially relevant to the fungus-growing insects' ecosystems functioning.

Biomacromolecules ; 21(8): 3081-3091, 2020 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-32573205


Tissue engineering and stem cell research greatly benefit from cell encapsulation within hydrogels as it promotes cell expansion and differentiation. Affinity-triggered hydrogels, an appealing solution for mild cell encapsulation, rely on selective interactions between the ligand and target and also on the multivalent presentation of these two components. Although these hydrogels represent a versatile option to generate dynamic, tunable, and highly functional materials, the design of hydrogel properties based on affinity and multivalency remains challenging and unstudied. Here, the avidin-biotin affinity pair, with the highest reported affinity constant, is used to address this challenge. It is demonstrated that the binding between the affinity hydrogel components is influenced by the multivalent display selected. In addition, the natural multivalency of the interaction must be obeyed to yield robust multicomponent synthetic protein hydrogels. The hydrogel's resistance to erosion depends on the right stoichiometric match between the hydrogel components. The developed affinity-triggered hydrogels are biocompatible and support encapsulation of induced pluripotent stem cells and their successful differentiation into a neural cell line. This principle can be generalized to other affinity pairs using multimeric proteins, yielding biomaterials with controlled performance.

Braz J Microbiol ; 51(3): 989-997, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32333271


Fungi in the genus Trichoderma are notorious producers of secondary metabolites with diverse applications, such as antibacterial, antifungal, and plant growth-promoting properties. Peptaibols are linear peptides produced by such fungi, with more than 440 compounds described to date, including tricholongins, longibrachins, trichobrachins, and trichovirins. Peptaibols are synthesized by non-ribosomal peptide synthetases and they have several biological activities. Our research group isolated four peptaibols (6DP2, 6DP3, 6DP4, and 6DP5) with antifungal activity against the plant pathogen Colletotrichum gloeosporioides and the proteasome (a cancer chemotherapy target) from Trichoderma sp. P8BDA1F1, an endophytic fungus from Begonia venosa. The ethyl acetate extract of this endophyte showed activity of 6.01% and 75% against C. gloeosporioides and the proteasome, respectively. The isolated compounds were identified by MS/MS and compared to literature data, suggesting the presence of trilongins BI, BII, BIII, and BIV, which are peptaibols containing 20 amino acid residues. The minimum inhibitory concentration against C. gloeosporioides was 40 µM for trilongin BI, 320 µM for trilongin BII, 160 µM for trilongin BIII, and 310 µM for trilongin BIV. BI-BIV trilongins inhibited proteasome ChTL activity, with IC50 values of 6.5 ± 2.7; 4.7 ± 1.8; 6.3 ± 2.2; and 2.7 ± 0.5 µM, respectively. The compounds were tested ex vivo against the intracellular amastigotes of Leishmania (L.) infantum but showed no selectivity. It is the first report of trilongins BI-BIV with antifungal activity against C. gloeosporioides and the proteasome target.

São Paulo; s.n; 2020. 31 p. figuras, tabelas.
Tese em Português | LILACS, Inca | ID: biblio-1102466


INTRODUÇÃO: O carcinoma de pênis (CaPe) é uma doença devastadora do ponto de vista físico e psicológico. Sua etiopatogenia é pouco conhecida e o tratamento limitado. Existem evidências do papel de células do sistema imune, como macrófagos e linfócitos no prognóstico desses tumores. Com o avanço do conhecimento da imunoterapia, esta possibilidade se abre para os casos avançados de CaPe. Um dos biomarcadores preditivos para o seu uso é a expressão da proteína ligante (PD-L1) ao receptor programado de morte celular (PD-1). No momento, pouco se sabe sobre o padrão de imunoexpressão deste biomarcador no CaPe e sua relação com as características clinico-patológicos destes pacientes. OBJETIVO: Avaliar a presença, por imunohistoquímica de PD-L1 em CaPe, a relação de linfócitos nos microambiente tumoral e a relação com variáveis clínico-patológicas em pacientes com CaPe. METODOLOGIA: Por imunohistoquímica foi avaliada a presença de PD-L1 em células tumorais e células inflamatórias: macrófagos, CD4, CD8 e CD20 em biopsias de 49 pacientes com diagnostico prévio de CaPe. Os dados foram analisados utilizando o programa R versão 3.5. O teste exato de Fisher foi empregado para comparação estatística entre a expressão de PD-L1 em células tumorais e macrófagos, CD4, CD8 e CD20 e nas variáveis clínico-patológicas. RESULTADOS: Em relação às características clínico-patológicas, 55,1% dos pacientes eram casados, 61,2% foram submetidos a penectomia parcial, 71.4% não necessitou de esvaziamento linfonodal, sendo 49% localizado na glande. A histologia usual para esse tumor estava presente em 64,6% dos pacientes. Em relação ao processo de invasão da neoplasia a outros tecidos, 81,6% apresentaram invasão ao corpo esponjoso, 75,5% não apresentou invasão à uretra, 89,8% não apresentou invasão vascular. As células tumorais do CaPe expressaram PD-L1 em 19% dos casos. Esta se associou positivamente com o predomínio de células CD4 e CD20 no microambiente tumoral. CONCLUSÃO: Por imunohistoquímica PD-L1 foi expresso em cerca de um quinto dos casos e se associou a uma resposta imune anérgica. Embora expresso na minoria dos casos, a capacidade da célula neoplásica ter o inibidor de check-point com o mecanismo de evasão da resposta imune levanta a possibilidade de se testar a imunoterapia como estratégia terapêutica em casos selecionados (AU)

INTRODUCTION: Penile carcinoma (PeCa) is a physically and psychologically devastating disease. Knowledge of its etiopathogenesis and its treatment is limited. There is evidence of the role of immune system cells such as macrophages and lymphocytes in the prognosis of these tumors. With the advance of immunotherapy knowledge, this possibility opens up for advanced cases of PeCa. One of the predictive biomarkers for its use is the expression of the binding protein (PD-L1) to the programmed cell death receptor (PD-1). At the moment, little is known about the immunoexpression pattern of this biomarker on PeCa and its relationship with the clinical and pathological characteristics of these patients. OBJECTIVE: To evaluate the presence, by immunohistochemistry of PD-L1 in PeCa, the relationship of macrophages and lymphocytes in the tumor microenvironment and the relationship with clinical-pathological variables in patients with PeCa. METHODOLOGY: The presence of PD-L1 in tumor cells and inflammatory cells: macrophages, CD4, CD8 and CD20 cells in biopsies of 49 patients with the previous diagnosis of PeCa were evaluated. The data were analyzed using the R version 3.5 program. Fisher's exact test was used for statistical comparison between the expression of PD-L1 in tumor cells and macrophages, CD4, CD8, and CD20 cells and in the clinical-pathological variables. RESULTS: Regarding the clinical and pathological characteristics, 55.1% of the patients were married, 61.2% underwent partial penectomy; 71.4% did not require lymph node dissection and 49% of which were located in the glans. The usual histology for this tumor was present in 64.6% of the patients. Regarding the process of invasion of the neoplasm to other tissues, 81.6% presented invasion to the spongy body of penis; 75.5% did not present invasion to the urethra and 89.8% did not show vascular invasion. Tumor cells from CaPe expressed PD-L1 in 19% of cases. This was positively associated with the predominance of CD4 and CD20 cells in the tumor microenvironment. CONCLUSION: By immunohistochemistry PD-L1 was expressed in about one-fifth of the cases and was associated with an anergic immune response. Although expressed in a minority of cases, the ability of the neoplastic cell to have the checkpoint inhibitor with the immune response evasion mechanisms raises the possibility of testing immunotherapy as a therapeutic strategy in selected cases.

Humanos , Feminino , Idoso , Neoplasias Penianas , Pênis/cirurgia , Imuno-Histoquímica , Imunoterapia
Med Eng Phys ; 73: 77-84, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31477429


Recent studies highlight the ability of inductive architectures to deliver therapeutic magnetic stimuli to target tissues and to be embedded into small-scale intracorporeal medical devices. However, to date, current micro-scale biomagnetic devices require very high electric current excitations (usually exceeding 1 A) to ensure the delivery of efficient magnetic flux densities. This is a critical problem as advanced implantable devices demand self-powering, stand-alone and long-term operation. This work provides, for the first time, a novel small-scale magnetic stimulation system that requires up to 50-fold lower electric current excitations than required by relevant biomagnetic technology recently proposed. Computational models were developed to analyse the magnetic stimuli distributions and densities delivered to cellular tissues during in vitro experiments, such that the feasibility of this novel stimulator can be firstly evaluated on cell culture tests. The results demonstrate that this new stimulative technology is able to deliver osteogenic stimuli (0.1-7 mT range) by current excitations in the 0.06-4.3 mA range. Moreover, it allows coil designs with heights lower than 1 mm without significant loss of magnetic stimuli capability. Finally, suitable core diameters and stimulator-stimulator distances allow to define heterogeneity or quasi-homogeneity stimuli distributions. These results support the design of high-sophisticated biomagnetic devices for a wide range of therapeutic applications.

Condutividade Elétrica , Terapia de Campo Magnético/instrumentação , Próteses e Implantes
An Acad Bras Cienc ; 91(3): e20180583, 2019 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-31365652


Isolation and screening of new fungal strains from extreme and understudied environments, such as caves, is a promising approach to find higher yields enzyme producers. Cellulolytic fungal strains isolated from a Brazilian cave were evaluated for their enzymatic production after submerged (SmF) and solid-state fermentation (SSF). After SmF, three strains were selected for their high enzymatic activities: Aspergillus ustus for endoglucanase (4.76 U/mg), Talaromyces bruneus for ß-glucosidase (11.71 U/mg) and Aspergillus sp. (CBMAI 1926) for total cellulase (1.70 U/mg). After SSF, these strains, showed better yields compared to the reference strain Aspergillus niger 3T5B8. Aspergillus sp. (CBMAI 1926) stood out as a new species that expressed activity of total cellulases (0.10 U/mg) and low protein concentration (0.44 mg/mL). In conclusion, these isolated strains have a more efficient and promising cellulolytic enzyme complex that can be used in fermentation and saccharification processes with a lower protein concentration and a higher enzymatic activity than the reference strain. Therefore, beside the new genetic material characterized, our study highlights the benefits of cave extreme environments exploitation to find new potentially valuable strains.

Cavernas , Celulases/metabolismo , Ambientes Extremos , Fungos/metabolismo , Brasil , Fungos/classificação
Rev. psicol. polit ; 19(45): 351-369, maio-ago. 2019. ilus
Artigo em Português | LILACS-Express | LILACS | ID: biblio-1020838


Considerando nossas apostas, experiências e pesquisas em saúde, partilhamos parte do que temos sentido, experimentado e percebido a partir de nossos campos de pesquisa, por meio do encontro com distintas existências, coletivos e lugares, usando a cartografia como modo de aproximação. Traremos os movimentos produzidos por coletivos das artístico-político-culturais cidades de São Paulo e Belém que, para além do espaço e das temporalidades, evidenciam ressonâncias e conexões quanto às apostas, aos atravessamentos e aos modos intensivos de compor as existências no contemporâneo, dos quais a saúde em sua institucionalidade tem permanecido distante; movimentos que dizem de um ocupar-se de si e do outro.

Considering our issues, our experiences and our research in health care, we hereby share part of what we have sensed, experienced and perceived in our fields of research, through our encounters with different forms of being, different collectives and places, according to a cartographic approach. We will present movements engendered by artistic, politic and cultural collectives from the cities of São Paulo and Belém which, beyond spatial and temporal contingencies, point out resonances and connections concerning the issues, the troubles and the intensiveness of the means by which ways of life are composed nowadays, processes from which health care has remained aloof. Such movements relate to some form of occupying oneself with oneself and each other.

Teniendo en cuenta nuestras cuestiones, nuestras experiencias y nuestros estudios en salud, compartimos aquí parte de lo que hemos sentido, experimentado y percibido en nuestros campos de investigación, através de encuentros con diferentes existencias, colectivos y lugares, según una aproximación cartográfica. Traemos movimientos producidos por colectivos artísticos, políticos y culturales de las ciudades de São Paulo y Belem que, a despecho de las particularidades de espacio y tiempo, senalan resonancias y conexiones respecto a las cuestiones, los obstáculos y la intensividad de los modos por los cuales se componen las existencias en lo contemporáneo, de los cuales la salud en su institucionalidad se ha quedado alejada. Estos movimientos hablan de un ocuparse de sí mismo y del otro.

Compte tenu de nos paris, nos expériences et nos recherches en santé, nous partageons une partie de ce que nous avons ressenti, expérimenté et perçu dans nos recherches sur la terrain, à travers la rencontre avec différentes existences, collectifs et endroits, en utilisant la cartographie comme méthode d'approche. Nous apporterons les mouvements produits par des collectifs artistiques, politiques e culturelles des villes de São Paulo et de Belém, lesquels, au-delà de l'espace et de la temporalité, dévoilent des résonances et des connexions concernant les paris, les enjeux et les modes intensives par lesquels les existences se constituent dans le contemporain, dont la santé, en son institutionnalité, reste écarté; mouvements qui disent d'un s 'occuper de soi de l'autre.

Otolaryngol Head Neck Surg ; 161(6): 954-959, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31359817


OBJECTIVE: Tertiary hyperparathyroidism, an autonomous hyperproduction of parathyroid hormone (PTH), has a challenge in its treatment. This study asked whether the intraoperative PTH and calcium drop can confirm the resection of all parathyroid tissues. STUDY DESIGN: Case series with planned data collection. SETTING: Tertiary referral medical center. SUBJECTS AND METHODS: The study assessed patients with tertiary hyperparathyroidism who were treated at the Hospital of the State University of Campinas from 2007 to 2015. All patients underwent total parathyroidectomy with autotransplantation of parathyroid fragments. PTH and calcium were collected during the preoperative period; at 10, 20, and 240 minutes after resection of the glands; and at 1 year after the procedure. Data were analyzed by analysis of variance and logistic regression analysis with statistical values of P < .05. RESULTS: Thirty-five patients were assessed: 17 women (48.57%) and 18 men (51.43%). The percentage of PTH drop was statistically significant at all times, unlike the calcium analysis, but only PTH collected at 20 minutes was able to confirm the removal of all parathyroid tissues (P = .029). Based on the receiver operating characteristic curve, the 71.2% drop obtained high sensitivity and specificity (P = .028). CONCLUSIONS: Treatment success can be predicted by analyzing the decrease of intraoperative PTH and not by calcium. The 71.2% PTH drop at 20 minutes after parathyroidectomy had high sensitivity and specificity to predict surgical cure.

Cálcio/sangue , Hiperparatireoidismo/cirurgia , Hormônio Paratireóideo/sangue , Paratireoidectomia , Adulto , Idoso , Estudos de Coortes , Feminino , Humanos , Hiperparatireoidismo/sangue , Masculino , Pessoa de Meia-Idade , Monitorização Intraoperatória , Valor Preditivo dos Testes , Curva ROC , Resultado do Tratamento
Sci Rep ; 9(1): 5001, 2019 03 21.
Artigo em Inglês | MEDLINE | ID: mdl-30899061


Cosurface electrode architectures are able to deliver personalized electric stimuli to target tissues. As such, this technology holds potential for a variety of innovative biomedical devices. However, to date, no detailed analyses have been conducted to evaluate the impact of stimulator architecture and geometry on stimuli features. This work characterizes, for the first time, the electric stimuli delivered to bone cellular tissues during in vitro experiments, when using three capacitive architectures: stripped, interdigitated and circular patterns. Computational models are presented that predict the influence of cell confluence, cosurface architecture, electrodes geometry, gap size between electrodes and power excitation on the stimuli delivered to cellular layers. The results demonstrate that these stimulators are able to deliver osteoconductive stimuli. Significant differences in stimuli distributions were observed for different stimulator designs and different external excitations. The thickness specification was found to be of utmost importance. In vitro experiments using an osteoblastic cell line highlight that cosurface stimulation at a low frequency can enhance osteoconductive responses, with some electrode-specific differences being found. A major feature of this type of work is that it enables future detailed analyses of stimuli distribution throughout more complex biological structures, such as tissues and organs, towards sophisticated biodevice personalization.

MycoKeys ; (46): 97-118, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30814906


Escovopsis (Ascomycota: Hypocreales, Hypocreaceae) is the only known parasite of the mutualistic fungi cultivated by fungus-growing ants (Formicidae: Myrmicinae: Attini: Attina, the "attines"). Despite its ecological role, the taxonomy and systematics of Escovopsis have been poorly addressed. Here, based on morphological and phylogenetic analyses with three molecular markers (internal transcribed spacer, large subunit ribosomal RNA and the translation elongation factor 1-alpha), we describe Escovopsisclavatus and E.multiformis as new species isolated from fungus gardens of Apterostigma ant species. Our analysis shows that E.clavatus and E.multiformis belong to the most derived Escovopsis clade, whose main character is the presence of conidiophores with vesicles. Nevertheless, the most outstanding feature of both new species is the presence of a swollen region in the central hypha of the conidiophore named swollen cell, which is absent in all previously described Escovopsis species. The less derived Escovopsis clades lack vesicles and their phylogenetic position within the Hypocreaceae still remains unclear. Considering the high genetic diversity in Escovopsis, the description of these new species adds barely two pieces to a huge taxonomic puzzle; however, this discovery is an important piece for building the systematics of this group of fungi.

Antonie Van Leeuwenhoek ; 112(2): 305-317, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30206787


Parasite-host associations are widespread in nature and the fungus-growing ants are considered model organisms to study such interactions. These insects cultivate basidiomycetous fungi for food, which are threatened by mycotrophic fungi in the genus Escovopsis. Although recently described from colonies of the lower attine ant Mycetophylax morschi, the biology and pathogenicity of Escovopsis kreiselii are unknown. Herein, we evaluated the interaction of E. kreiselii with fungi cultivated by M. morschi (native hosts) and with a fungus cultivated by another attine ant species (non-native host). In addition, we examined the physical interactions between hypha of E. kreiselii and hypha from its native hosts using scanning electron microscopy. Escovopsis kreiselii inhibited the growth of fungal cultivars by 24% or more (with exception of one isolate), when compared to the fungal cultivars growing alone. Escovopsis kreiselii is attracted towards its native hosts through chemotaxis and inhibition occurs when there is physical contact with the hyphae of the fungal cultivar. As reported for Escovopsis parasites associated with leafcutter ants (higher attines), E. kreiselii growth increased in the presence of its native hosts, even before contact between both fungi occurred. In interactions with the fungal cultivar that is not naturally infected by E. kreiselii (non-native host), it caused inhibition but not at the same magnitude as in native hosts. Multiple lines of evidence suggest that E. kreiselii is an antagonist of the fungus cultivated by M. morschi and can chemically recognize such fungus.

Agaricales/crescimento & desenvolvimento , Formigas/microbiologia , Especificidade de Hospedeiro , Interações Hospedeiro-Parasita , Hypocreales/classificação , Hypocreales/patogenicidade , Animais , Brasil , Hypocreales/genética
Biotechnol J ; 14(4): e1800461, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30320457


The development of bioprocesses capable of producing large numbers of human induced pluripotent stem cells (hiPSC) in a robust and safe manner is critical for the application of these cells in biotechnological and medical applications. Scalable expansion of hiPSC is often performed using polystyrene microcarriers, which have to be removed from the cell suspension using a separation step that causes loss of viable cells. In this study, application of novel xeno-free dissolvable microcarriers (DM) for an efficient and integrated expansion and harvesting of hiPSC is demonstrated. After an initial screening under static conditions, hiPSC culture using DM is performed in dynamic culture, using spinner-flasks. A maximum 4.0 ± 0.8-fold expansion is achieved after 5 days of culture. These results are validated with a second cell line and the culture is successfully adapted to fully xeno-free conditions. Afterwards, cell recovery is made within the spinner flask, being obtained a 92 ± 4% harvesting yield, which is significantly higher than the one obtained for the conventional filtration-based method (45 ± 3%). Importantly, the expanded and harvested hiPSC maintain their pluripotency and multilineage differentiation potential. The results here described represent a significant improvement of the downstream processing after microcarrier-based hiPSC expansion, leading to a more cost-effective and efficient bioprocess.

Biotecnologia/métodos , Técnicas de Cultura de Células/métodos , Células-Tronco Pluripotentes Induzidas/citologia , Diferenciação Celular/genética , Proliferação de Células/genética , Humanos
Microb Ecol ; 77(1): 12-24, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29916010


Extreme environments such as the Antarctic can lead to the discovery of new microbial taxa, as well as to new microbial-derived natural products. Considering that little is known yet about the diversity and the genetic resources present in these habitats, the main objective of this study was to evaluate the fungal communities from extreme environments collected at Aldmiralty Bay (Antarctica). A total of 891 and 226 isolates was obtained from soil and marine sediment samples, respectively. The most abundant isolates from soil samples were representatives of the genera Leucosporidium, Pseudogymnoascus, and a non-identified Ascomycota NIA6. Metschnikowia sp. was the most abundant taxon from marine samples, followed by isolates from the genera Penicillium and Pseudogymnoascus. Many of the genera were exclusive in marine sediment or terrestrial samples. However, representatives of eight genera were found in both types of samples. Data from non-metric multidimensional scaling showed that each sampling site is unique in their physical-chemical composition and fungal community. Biotechnological potential in relation to enzymatic production at low/moderate temperatures was also investigated. Ligninolytic enzymes were produced by few isolates from root-associated soil. Among the fungi isolated from marine sediments, 16 yeasts and nine fungi showed lipase activity and three yeasts and six filamentous fungi protease activity. The present study permitted increasing our knowledge on the diversity of fungi that inhabit the Antarctic, finding genera that have never been reported in this environment before and discovering putative new species of fungi.

Fungos/classificação , Fungos/isolamento & purificação , Sedimentos Geológicos/microbiologia , Água do Mar/microbiologia , Microbiologia do Solo , Regiões Antárticas , Ascomicetos/classificação , Ascomicetos/isolamento & purificação , Basidiomycota/classificação , Basidiomycota/isolamento & purificação , Baías , Biodiversidade , Temperatura Baixa , Extremófilos , Fungos/enzimologia , Fungos/genética , Ilhas , Lipase/metabolismo , Peptídeo Hidrolases/metabolismo , Filogenia , Especificidade da Espécie
Rev. bras. med. fam. comunidade ; 13(40): 1-6, jan.-dez. 2018.
Artigo em Português | LILACS-Express | LILACS | ID: biblio-969326


Objetivo: Estimar a confiabilidade e a validade da codificação de motivos de consulta e problemas por estudantes utilizando a Classificação Internacional da Atenção Primária, 2ª edição (CIAP-2). Métodos: Para cada encontro supervisionado durante todo um semestre, três professores médicos de família e comunidade registraram os motivos de consulta e problemas em um questionário usando texto livre. Dois de quatro estudantes de medicina e um professor codificaram cada motivo de consulta ou problema usando a CIAP-2. No começo do estudo, houve duas seções de padronização com três horas de duração, até os professores julgarem que os estudantes estavam prontos para a codificação. Após todos os motivos de consulta e problemas terem sido codificados independentemente, os sete codificadores resolveram os códigos definitivos por consenso. Definiu-se confiabilidade como concordância entre estudantes, e validade como a concordância destes com os códigos definitivos; essa concordância foi estimada com o AC1 de Gwet. Resultados: Após a exclusão dos encontros codificados antes da última sessão de padronização, a amostra consistiu em 149 encontros consecutivos, somando 262 motivos de consulta e 226 problemas. A codificação teve confiabilidade moderada a substancial (AC1 0,805; IC 95% 0,767­0,843) e validade substancial (AC1 0,864; IC 95% 0,833­0,891). C

Objective: To estimate how reliably and validly can medical students encode reasons for encounter and diagnoses using the International Classification of Primary Care, revised 2nd edition (ICPC-2-R). Methods: For every encounter they supervised during an entire semester, three family and community physician teachers entered the reasons for encounter and diagnoses in free text into a form. Two of four medical students and one teacher encoded each reason for encounter or diagnosis using the ICPC-2-R. In the beginning of the study, two three-hour workshops were held, until the teachers were confident the students were ready for the encoding. After all the reasons for encounter and the diagnoses had been independently encoded, the seven encoders resolved the definitive codes by consensus. We defined reliability as agreement between students and validity as their agreement with the definitive codes, and used Gwet's AC1 to estimate this agreement. Results: After exclusion of encounters encoded before the last workshop, the sample consisted of 149 consecutive encounters, comprising 262 reasons for encounter and 226 diagnoses. The encoding had moderate to substantial reliability (AC1 , 0.805; 95% CI, 0.767­0.843) and substantial validity (AC1 , 0.864; 95% CI, 0.833­0.891). Conclusion: Medical students can encode reasons for encounter and diagnoses with the ICPC-2-R if they are adequately trained.

Objetivo: Estimar la confiabilidad y la validez de la codificación de motivos de consulta y problemas de salud por estudiantes utilizando la Clasificación Internacional de Atención Primaria, 2ª edición (CIAP-2). Métodos: Para cada encuentro supervisado durante todo un semestre, tres profesores médicos de familia y comunidad registraron los motivos de consulta y los problemas de salud en un formulario usando texto libre. Dos de cuatro estudiantes de medicina y un profesor codificaron cada motivo de consulta o problema de salud utilizando la CIAP-2. En el comienzo del estudio, se llevaron a cabo dos sesiones de estandarización de tres horas, hasta que los profesores estuvieron seguros de que los estudiantes estaban listos para la codificación. Después de que todos los motivos de consulta y problemas de salud fueran codificados independientemente, los siete codificadores resolvieron los códigos definitivos por consenso. Se definió confiabilidad como concordancia entre los estudiantes y validez como la concordancia de éstos con los códigos definitivos; se estimó esta concordancia con el AC1 de Gwet. Resultados: Después de la exclusión de los encuentros codificados antes de la última sesión de estandarización, la muestra consistió en 149 encuentros consecutivos, que comprendían 262 motivos de consulta y 226 problemas de salud. La codificación tuvo una confiabilidad moderada a sustancial (AC1 0,805; IC 95% 0,767­0,843) y validez sustancial (AC1 0,864; IC 95% 0,833­0,891). Conclusión: Los estudiantes de medicina pueden codificar los motivos de consulta y los problemas de salud con la CIAP-2 si fueran adecuadamente capacitados.

Braz. j. microbiol ; 49(4): 840-847, Oct.-Dec. 2018. graf
Artigo em Inglês | LILACS | ID: biblio-974284


ABSTRACT Anthracnose is a crop disease usually caused by fungi in the genus Colletotrichum or Gloeosporium. These are considered one of the main pathogens, causing significant economic losses, such as in peppers and guarana. The current forms of control include the use of resistant cultivars, sanitary pruning and fungicides. However, even with the use of some methods of controlling these cultures, the crops are not free of anthracnose. Additionally, excessive application of fungicides increases the resistance of pathogens to agrochemicals and cause harm to human health and the environment. In order to find natural antifungal agents against guarana anthracnose, endophytic fungi were isolated from Amazon guarana. The compounds piliformic acid and cytochalasin D were isolated by chromatographic techniques from two Xylaria spp., guided by assays with Colletotrichum gloeosporioides. The isolated compounds were identified by spectrometric techniques, as NMR and mass spectrometry. This is the first report that piliformic acid and cytochalasin D have antifungal activity against C. gloeosporioides with MIC 2.92 and 2.46 µmol mL-1 respectively. Captan and difenoconazole were included as positive controls (MIC 16.63 and 0.02 µmol mL-1, respectively). Thus, Xylaria species presented a biotechnological potential and production of different active compounds which might be promising against anthracnose disease.

Doenças das Plantas/prevenção & controle , Xylariales/química , Paullinia/microbiologia , Endófitos/química , Fungicidas Industriais/farmacologia , Filogenia , Doenças das Plantas/microbiologia , Espectrometria de Massas , Xylariales/isolamento & purificação , Xylariales/genética , Xylariales/metabolismo , Estrutura Molecular , Colletotrichum/efeitos dos fármacos , Colletotrichum/fisiologia , Endófitos/isolamento & purificação , Endófitos/genética , Endófitos/metabolismo , Fungicidas Industriais/isolamento & purificação , Fungicidas Industriais/química
BMC Microbiol ; 18(1): 130, 2018 10 10.
Artigo em Inglês | MEDLINE | ID: mdl-30305028


BACKGROUND: Fungus gardens of fungus-growing (attine) ants harbor complex microbiomes in addition to the mutualistic fungus they cultivate for food. Fungi in the genus Escovopsioides were recently described as members of this microbiome but their role in the ant-fungus symbiosis is poorly known. In this study, we assessed the phylogenetic diversity of 21 Escovopsioides isolates obtained from fungus gardens of leafcutter ants (genera Atta and Acromyrmex) and non-leafcutter ants (genera Trachymyrmex and Apterostigma) sampled from several regions in Brazil. RESULTS: Regardless of the sample locality or ant genera, phylogenetic analysis showed low genetic diversity among the 20 Escovopsisoides isolates examined, which prompted the identification as Escovopsioides nivea (the only described species in the genus). In contrast, one Escovopsioides isolate obtained from a fungus garden of Apterostigma megacephala was considered a new phylogenetic species. Dual-culture plate assays showed that Escovopsioides isolates inhibited the mycelium growth of Leucoagaricus gongylophorus, the mutualistic fungus cultivated by somes species of leafcutter ants. In addition, Escovopsioides growth experiments in fungus gardens with and without ant workers showed this fungus is detrimental to the ant-fungus symbiosis. CONCLUSIONS: Here, we provide clues for the antagonism of Escovopsioides towards the mutualistic fungus of leafcutter ants.

Antibiose , Formigas/microbiologia , Fungos/classificação , Filogenia , Simbiose , Animais , Brasil , Fungos/isolamento & purificação , Microbiota
Sensors (Basel) ; 18(10)2018 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-30347883


Dependability and standardization are essential to the adoption of Wireless Sensor Networks (WSN) in industrial applications. Standards such as ZigBee, WirelessHART, ISA100.11a and WIA-PA are, nowadays, at the basis of the main process-automation technologies. However, despite the success of these standards, management of WSNs is still an open topic, which clearly is an obstacle to dependability. Existing diagnostic tools are mostly application- or problem-specific, and do not support standard-based multi-network monitoring. This paper proposes a WSN monitoring architecture for process-automation technologies that addresses the mentioned limitations. Specifically, the architecture has low impact on sensor node resources, uses network metrics already available in industrial standards, and takes advantage of widely used management standards to share the monitoring information. The proposed architecture was validated through prototyping, and the obtained performance results are presented and discussed in the final part of the paper. In addition to proposing a monitoring architecture, the paper provides an in-depth insight into metrics, techniques, management protocols, and standards applicable to industrial WSNs.