Computational fluid dynamic analysis of physical forces playing a role in brain organoid cultures in two different multiplex platforms.

BACKGROUND: Organoid cultivation in suspension culture requires agitation at low shear stress to allow for nutrient diffusion, which preserves tissue structure. Multiplex systems for organoid cultivation have been proposed, but whether they meet similar shear stress parameters as the regularly used spinner flask and its correlation with the successful generation of brain organoids has not been determined. RESULTS: Here we used computational fluid dynamics (CFD) to simulate two multiplex culture conditions: steering plates on an orbital shaker and the use of a previously described bioreactor. The bioreactor had low speed and high shear stress regions that may affect cell aggregate growth, depending on volume, whereas the computed variables of the steering plates were closer to those of the spinning flask. CONCLUSION: Our protocol improves the initial steps of the standard brain organoid formation, and the produced organoids displayed regionalized brain structures, including retinal pigmented cells. Overall, we conclude that suspension culture on orbital steering plates is a cost-effective practical alternative to previously described platforms for the cultivation of brain organoids for research and multiplex testing.

Interspecies Isobaric Labeling-Based Quantitative Proteomics Reveals Protein Changes in the Ovary of Aedes aegypti Coinfected With ZIKV and Wolbachia.

Zika is a vector-borne disease caused by an arbovirus (ZIKV) and overwhelmingly transmitted by Ae. aegypti. This disease is linked to adverse fetal outcomes, mostly microcephaly in newborns, and other clinical aspects such as acute febrile illness and neurologic complications, for example, Guillain-Barré syndrome. One of the most promising strategies to mitigate arbovirus transmission involves releasing Ae. aegypti mosquitoes carrying the maternally inherited endosymbiont bacteria Wolbachia pipientis. The presence of Wolbachia is associated with a reduced susceptibility to arboviruses and a fitness cost in mosquito life-history traits such as fecundity and fertility. However, the mechanisms by which Wolbachia influences metabolic pathways leading to differences in egg production remains poorly known. To investigate the impact of coinfections on the reproductive tract of the mosquito, we applied an isobaric labeling-based quantitative proteomic strategy to investigate the influence of Wolbachia wMel and ZIKV infection in Ae. aegypti ovaries. To the best of our knowledge, this is the most complete proteome of Ae. aegypti ovaries reported so far, with a total of 3913 proteins identified, were also able to quantify 1044 Wolbachia proteins in complex sample tissue of Ae. aegypti ovary. Furthermore, from a total of 480 mosquito proteins modulated in our study, we discuss proteins and pathways altered in Ae. aegypti during ZIKV infections, Wolbachia infections, coinfection Wolbachia/ZIKV, and compared with no infection, focusing on immune and reproductive aspects of Ae. aegypti. The modified aspects mainly were related to the immune priming enhancement by Wolbachia presence and the modulation of the Juvenile Hormone pathway caused by both microorganism's infection.

Identification of Potentially Therapeutic Immunogenic Peptides From Paracoccidioides lutzii Species.

Paracoccidioidomycosis (PCM) is an endemic mycosis in Latin America caused by the thermodimorphic fungi of the genus Paracoccidioides spp. Paracoccidioides lutzii (PL) is one of the 5 species that constitute the Paracoccidioides genus. PL expresses low amounts of glycoprotein (Gp) 43 (PLGp43) and PLGp43 displays few epitopes in common with the P. brasiliensis (PB) immunodominant antigen PBGp43, which is commonly used for serological diagnosis of PCM. This difference in structure between the glycoproteins markedly reduces the efficiency of serological diagnosis in patients infected with PL. We previously demonstrated that peptide 10 (P10) from the PBGp43 induces protective immune responses in in vitro and in vivo models of PB PCM. Since, P10 has proven to be a promising therapeutic to combat PB, we sought to identify peptides in PL that could similarly be applied for the treatment of PCM. PL yeast cell proteins were isolated from PL: dendritic cell co-cultures and subjected to immunoproteomics. This approach identified 18 PL peptides that demonstrated in silico predictions for immunogenicity. Eight of the most promising peptides were synthesized and applied to lymphocytes obtained from peptide-immunized or PL-infected mice as well as to in vitro cultures with peptides or dendritic cells pulsed the peptides. The peptides LBR5, LBR6 and LBR8 efficiently promoted CD4+ and CD8+ T cell proliferation and dendritic cells pulsed with LBR1, LBR3, LBR7 or LBR8 stimulated CD4+ T cell proliferation. We observed increases of IFN-γ in the supernatants from primed T cells for the conditions with peptides without or with dendritic cells, although IL-2 levels only increased in response to LBR8. These novel immunogenic peptides derived from PL will be employed to develop new peptide vaccine approaches and the proteins from which they are derived can be used to develop new diagnostic assays for PL and possibly other Paracoccidioides spp. These findings identify and characterize new peptides with a promising therapeutic profile for future against this important neglected systemic mycosis.

Comprehensive Quantitative Proteome Analysis of Aedes aegypti Identifies Proteins and Pathways Involved in Wolbachia pipientis and Zika Virus Interference Phenomenon.

Zika virus (ZIKV) is a global public health emergency due to its association with microcephaly, Guillain-Barré syndrome, neuropathy, and myelitis in children and adults. A total of 87 countries have had evidence of autochthonous mosquito-borne transmission of ZIKV, distributed across four continents, and no antivirus therapy or vaccines are available. Therefore, several strategies have been developed to target the main mosquito vector, Aedes aegypti, to reduce the burden of different arboviruses. Among such strategies, the use of the maternally-inherited endosymbiont Wolbachia pipientis has been applied successfully to reduce virus susceptibility and decrease transmission. However, the mechanisms by which Wolbachia orchestrate resistance to ZIKV infection remain to be elucidated. In this study, we apply isobaric labeling quantitative mass spectrometry (MS)-based proteomics to quantify proteins and identify pathways altered during ZIKV infection; Wolbachia infection; co-infection with Wolbachia/ZIKV in the A. aegypti heads and salivary glands. We show that Wolbachia regulates proteins involved in reactive oxygen species production, regulates humoral immune response, and antioxidant production. The reduction of ZIKV polyprotein in the presence of Wolbachia in mosquitoes was determined by MS and corroborates the idea that Wolbachia helps to block ZIKV infections in A. aegypti. The present study offers a rich resource of data that may help to elucidate mechanisms by which Wolbachia orchestrate resistance to ZIKV infection in A. aegypti, and represents a step further on the development of new targeted methods to detect and quantify ZIKV and Wolbachia directly in complex tissues.

Quantitative profiling of axonal guidance proteins during the differentiation of human neurospheres.

Axon guidance is required for the establishment of brain circuits. Whether much of the molecular basis of axon guidance is known from animal models, the molecular machinery coordinating axon growth and pathfinding in humans remains to be elucidated. The use of induced pluripotent stem cells (iPSC) from human donors has revolutionized in vitro studies of the human brain. iPSC can be differentiated into neuronal stem cells which can be used to generate neural tissue-like cultures, known as neurospheres, that reproduce, in many aspects, the cell types and molecules present in the brain. Here, we analyzed quantitative changes in the proteome of neurospheres during differentiation. Relative quantification was performed at early time points during differentiation using iTRAQ-based labeling and LC-MS/MS analysis. We identified 6438 proteins, from which 433 were downregulated and 479 were upregulated during differentiation. We show that human neurospheres have a molecular profile that correlates to the fetal brain. During differentiation, upregulated pathways are related to neuronal development and differentiation, cell adhesion, and axonal guidance whereas cell proliferation pathways were downregulated. We developed a functional assay to check for neurite outgrowth in neurospheres and confirmed that neurite outgrowth potential is increased after 10 days of differentiation and is enhanced by increasing cyclic AMP levels. The proteins identified here represent a resource to monitor neurosphere differentiation and coupled to the neurite outgrowth assay can be used to functionally explore neurological disorders using human neurospheres as a model.

Rectus sheath hematoma in patients receiving subcutaneous enoxaparin: A case series of five patients.

Physicians must acknowledge the potential risk of RSH with enoxaparin. Switching home anticoagulation by enoxaparin upon hospital admission is common, but it may put patients at higher risk for RSH. Management guidelines are needed in this setting.

THE USE OF THE INTERNET BY THE PATIENT AFTER BARIATRIC SURGERY: CONTRIBUTIONS AND OBSTACLES FOR THE FOLLOW-UP OF MULTIDISCIPLINARY MONITORING.

BACKGROUND: Bariatric surgery is presented as the last treatment option for obesity. It requires from all candidates a multidisciplinary evaluation and monitoring throughout treatment. The non-adherence to follow-up with health care teams is related to weight regain. It's possible that the use of internet influences the doctor-patient relationship and patients replace medical care or information provided by health professionals for information from the internet. AIM: Identify and analyze the pattern of internet use by patients after bariatric surgery and check the influence of such use in attending medical appointments with the multidisciplinary team. METHOD: Electronic questionnaire available on the Internet was used to verify patient's patterns of Internet use and its influence on in attending multidisciplinary care after surgery. RESULTS: Of the 103 participants, 95% were female, 64% married, 59% with children and 54% with higher education. The mean age was 35.69 years and the mean duration of performing surgery, 11.74 months. The surgical technique that prevailed was Roux-en-Y gastric by-pass (90.3%), the local monitoring concentrated in the private care (93.2%). In the preoperative, most participants consulted more than three times with the surgeon (n=81), nutritionists (n=70), psychologist (n=70). After the surgery, p most patients maintained monitoring with the surgeon and nutritionist. Concerning the internet use, 51.5% accessed the internet in search of information about health and bariatric surgery every day. Facebook and search tools were the most used sites.Conclusion - Data showed the influence of the information contained on the Internet and the adherence to multidisciplinary monitoring. This fact requires the team to consider the use of the Internet as a variable that may interfere and must be handled during follow-up. It is suggested that an active participation of professionals on their websites and social networks and the diversification of services and interventions to stimulate follow-up after surgery.

Posterior ankle impingement syndrome: a diagnosis rheumatologists should not forget. Two case reports.

The ankle is a common site of painful symptoms in athletes and nonathletes. Posterior ankle pain can be the end result of several pathologies, and a diagnostic challenge for rheumatologists. The posterior ankle impingement syndrome, also known as os trigonum syndrome and posterior tibiotalar compression syndrome, is a clinical disorder characterized by acute or chronic posterior ankle pain triggered by forced plantar flexion, which causes chronic repetitive microtrauma. Pathology of the os trigonum-talar process is the most common cause of this syndrome, but there are other causes, such as tenosynovitis of the flexor hallucis longus, ankle osteochondritis, subtalar joint disease, and fracture. Diagnosis is based on clinical history and physical examination, and complemented by findings on plain radiography (RX), ultrasound (US), scintigraphy, computed tomography (CT), and magnetic resonance imaging (MRI). It is worth noting that RX has low cost and good sensitivity, US can provide guidance to therapeutic infiltrations, and MRI allows the assessment of surrounding soft tissues.

THE USE OF THE INTERNET BY THE PATIENT AFTER BARIATRIC SURGERY: CONTRIBUTIONS AND OBSTACLES FOR THE FOLLOW-UP OF MULTIDISCIPLINARY MONITORING / O USO DA INTERNET PELO PACIENTE APÓS CIRURGIA BARIÁTRICA: CONTRIBUIÇÕES E ENTRAVES PARA O SEGUIMENTO DO ACOMPANHAMENTO MULTIPROFISSIONAL

ABSTRACTBackground: Bariatric surgery is presented as the last treatment option for obesity. It requires from all candidates a multidisciplinary evaluation and monitoring throughout treatment. The non-adherence to follow-up with health care teams is related to weight regain. It's possible that the use of internet influences the doctor-patient relationship and patients replace medical care or information provided by health professionals for information from the internet.Aim: Identify and analyze the pattern of internet use by patients after bariatric surgery and check the influence of such use in attending medical appointments with the multidisciplinary team.Method: Electronic questionnaire available on the Internet was used to verify patient´s patterns of Internet use and its influence on in attending multidisciplinary care after surgery.Results: Of the 103 participants, 95% were female, 64% married, 59% with children and 54% with higher education. The mean age was 35.69 years and the mean duration of performing surgery, 11.74 months. The surgical technique that prevailed was Roux-en-Y gastric by-pass (90.3%), the local monitoring concentrated in the private care (93.2%). In the preoperative, most participants consulted more than three times with the surgeon (n=81), nutritionists (n=70), psychologist (n=70). After the surgery, p most patients maintained monitoring with the surgeon and nutritionist. Concerning the internet use, 51.5% accessed the internet in search of information about health and bariatric surgery every day. Facebook and search tools were the most used sites.Conclusion - Data showed the influence of the information contained on the Internet and the adherence to multidisciplinary monitoring. This fact requires the team to consider the use of the Internet as a variable that may interfere and must be handled during follow-up. It is suggested that an active participation of professionals on their websites and social networks and the diversification of services and interventions to stimulate follow-up after surgery.

Racional: O candidato à cirurgia bariátrica precisa passar por avaliação e acompanhamento multiprofissional durante todo o processo de tratamento. Perder o seguimento com as equipes de saúde tem relação com reganho de peso e deficiências nutricionais. O uso da internet é considerado como um dos fatores que pode afetar a relação médico-paciente.Objetivo: Identificar e analisar o padrão de uso da internet pelos pacientes no pós-operatório e verificar se influencia no seguimento multiprofissional.Metódo: Utilizou-se um questionário eletrônico, disponibilizado on-line, para verificar o padrão de uso da Internet e seu impacto no acompanhamento multiprofissional após o procedimento cirúrgico.Resultados: Participaram 103 pacientes, em sua maioria mulheres (95%), casados (64%) e com ensino superior (54%). A idade média foi 35,69 anos e o tempo médio desde a operação de 11,74 meses. Prevaleceram o uso da técnica by-pass gástrico em Y-de-Roux (90,3%), e o acompanhamento foi em serviço privado (93,2%). No pré-cirúrgico a maioria teve maior número de consultas com os profissionais de saúde do que no pós-cirúrgico, havendo relatos que indicam substituição do acompanhamento por informações contida na internet e participação em grupos de discussão on-line, principalmente pela perda de peso inicial gerar sensação de segurança e percepção de não haver mais necessidade de auxílio. O padrão de uso da internet que sobressaiu foi o acesso diário (51,5%) para buscar informações sobre saúde e cirurgia bariátrica. O Facebook e as ferramentas de pesquisa foram os locais mais utilizados.Conclusão: Os dados evidenciaram que as informações contidas na Internet influenciam o seguimento multiprofissional, fato este que exige a equipe considerar o uso da rede como variável que interfere e deve ser manejada. Sugere-se a participação ativa dos profissionais em seus sites e redes sociais e a diversificação de intervenções e serviços para estimular o acompanhamento após a operação.

Síndrome do impacto posterior do tornozelo: um diagnóstico que deve ser lembrado pelo reumatologista. Relato de dois casos / Posterior ankle impingement syndrome: a diagnosis rheumatologists should not forget. Two case reports

O tornozelo é sítio frequente de sintomas dolorosos em atletas e não atletas. A dor localizada na região posterior pode ser o resultado final de diversas patologias, sendo um desafio diagnóstico para o reumatologista. A síndrome do impacto (pinçamento) posterior do tornozelo, também denominada síndrome os trigonum e síndrome compressiva tibiotalar posterior, é um distúrbio clínico caracterizado por dor aguda ou crônica na região posterior do tornozelo, desencadeada pela flexão plantar forçada, que promove microtrauma crônico repetitivo. A patologia do processo os trigonum-talar é a causa mais comum dessa síndrome, mas existem outras causas, como tenossinovite do flexor longo do hálux, osteocondrite de tornozelo, doença da articulação subtalar e fratura. O diagnóstico baseia-se na história clínica e exame físico, e complementado por achados na radiografia simples (RX), ultrassom (US), cintilografia, tomografia computadorizada (TC) e ressonância magnética (RM). Destacamos o RX por seu baixo custo e boa sensibilidade, o US pela possibilidade de guiar infiltrações terapêuticas e a RM pela possibilidade de avaliar partes moles adjacentes.

The ankle is a common site of painful symptoms in athletes and nonathletes. Posterior ankle pain can be the end result of several pathologies, and a diagnostic challenge for rheumatologists. The posterior ankle impingement syndrome, also known as os trigonum syndrome and posterior tibiotalar compression syndrome, is a clinical disorder characterized by acute or chronic posterior ankle pain triggered by forced plantar flexion, which causes chronic repetitive microtrauma. Pathology of the os trigonum-talar process is the most common cause of this syndrome, but there are other causes, such as tenosynovitis of the flexor hallucis longus, ankle osteochondritis, subtalar joint disease, and fracture. Diagnosis is based on clinical history and physical examination, and complemented by findings on plain radiography (RX), ultrasound (US), scintigraphy, computed tomography (CT), and magnetic resonance imaging (MRI). It is worth noting that RX has low cost and good sensitivity, US can provide guidance to therapeutic infiltrations, and MRI allows the assessment of surrounding soft tissues.

Estudo da correlação entre os níveis de hemoglobina fetal e o prognóstico dos pacientes com anemia falciforme / Study of the correlation between fetal hemoglobin levels and the prognosis of patients with sickle cell anemia

Os níveis de hemoglobina fetal (HbF) foram analisados em 47 pacientes com anemia falciforme, residentes em Fortaleza, capital do Ceará, com a finalidade de fornecer informações sobre a influência da HbF no prognóstico desta doença. A determinação dos valores hematológicos foi realizada em contador automático de células sanguíneas (modelo Sysmex KX-21N, Roche) e os níveis de HbF foram determinados pela técnica da desnaturação alcalina. Dentre os resultados foi demonstrada uma tendência de menor número de pacientes com crises vaso-oclusivas (p=0,0405) e úlceras de perna (p=0,0076) com o aumento dos níveis de HbF. Não houve diferença estatisticamente significativa na comparação entre os níveis de HbF e os valores de hemácias, hemoglobina, hematócrito, leucócitos e plaquetas. Os resultados demonstram que os níveis de hemoglobina fetal podem ser utilizados como fator prognóstico nos pacientes com anemia falciforme, no entanto não deve ser a única ferramenta diagnóstica para prever o quadro clínico destes pacientes.

The fetal hemoglobin (HbF) levels of 47 patients with sickle cell anemia living in Fortaleza, Brazil were analyzed to provide information on the influence of HbF on the prognosis of this disease. Measurement of hematological variables was achieved using a Sysmex KX-21N cell counter ( Roche) and the levels of HbF were determined by the alkali denaturation technique. With increased levels of HbF, there was a trend that fewer patients had painful episodes (p = 0.0405) and leg ulcers (p = 0.0076). There were no statistically significant differences between the levels of HbF and the concentrations of red blood cells, hemoglobin, hematocrit, leukocytes and platelets. The results show that levels of fetal hemoglobin can be used as a prognostic factor in patients with sickle cell anemia however they should not be the only diagnostic tool to predict the clinical status of these patients.

Características de imagem na ressonância magnética de gossipiboma intracraniano: relato de caso e revisão da literatura / Magnetic resonance imaging findings of intracranial gossypiboma: a case report and literature review

Vários trabalhos têm descrito as características de gossipibomas por meio de radiografias convencionais e tomografia computadorizada, porém, seus achados na ressonância magnética são menos conhecidos. O objetivo deste estudo foi descrever as características de imagem de gossipibomas, ilustrando um caso de uma paciente com quadro clínico de febre e confusão mental com antecedente de cirurgia encefálica.

Many reports in the literature have described findings of gossypibomas at conventional radiography and computed tomography. However, magnetic resonance imaging findings are still to be completely known. The purpose of the present study was to describe imaging findings of gossypibomas in a patient with previous history of brain surgery, presenting fever and mental confusion.