High-throughput platforms for the screening of new therapeutic targets for neurodegenerative diseases

Despite the recent progress in the understanding of neurodegenerative disorders, a lack of solid fundamental knowledge on the etiology of many of the major neurodegenerative diseases has made it difficult to obtain effective therapies to treat these conditions. Scientists have been looking to carry out more-human-relevant studies, with strong statistical power, to overcome the limitations of preclinical animal models that have contributed to the failure of numerous therapeutics in clinical trials. Here, we identify currently existing platforms to mimic central nervous system tissues, healthy and diseased, mainly focusing on cell-based platforms and discussing their strengths and limitations in the context of the high-throughput screening of new therapeutic targets and drugs.This work had the financial support of Fundação para a Ciência e Tecnologia ( FCT ) through National Funds and, when applicable, co-financed by the FEDER through the PT2020 Partnership Agreement under the 4293 Unit I&D. D.N. Rocha acknowledges FCT for her PhD grant

Head-to-head comparison of two online nomograms for prostate biopsy outcome prediction

LEVEL OF EVIDENCE: 2b. What's known on the subject? and What does the study add? In recent years, several nomograms were developed in an effort to decrease the number of unnecessary prostate biopsies. The European SWOP-PRI and the North American PCPT are among the most popular. However, evidence on the relative predictive accuracy is lacking. A head-to-head comparison on the diagnostic accuracy of two previously validated prostate cancer risk predictors on biopsy confirmed the superiority of these tools over PSA alone. Moreover, in the studied population, the European SWOP-PRI proved to be more accurate than the North American PCPT-CRC. OBJECTIVE: To compare the diagnostic accuracy of two previously validated prostate cancer risk predictors on biopsy. PATIENTS AND METHODS: In total, 390 consecutive patients submitted to 10-core systematic transrectal prostate biopsy at our institution were included in this retrospective study. External validation of a European (European Randomized Study of Screening for Prostate Cancer derived Prostate Risk Indicator; SWOP-PRI) and a North American (Prostate Cancer Prevention Trial Cancer Risk Calculator; PCPT-CRC) nomogram was performed. The predictive accuracy of these online available nomograms was calculated based on the area under the curve derived from receiver-operator characteristic curves and then compared using the DeLong method. RESULTS: Both tools were confirmed to be superior to prostate-specific antigen alone. Moreover, the SWOP-PRI (77.9%) displays a 7.96% increase in the predictive accuracy compared to the PCPT-CRC (69.9%) in a statistically significant fashion (P=0.002). CONCLUSIONS: The results obtained in the present study confirm the utility of nomograms with respect to biopsy outcome prediction in patients with suspicion of prostate cancer. In the current sample of patients, the European-based nomogram appears to be more accurate than the North American nonogram, which lacks information regarding prostate volume and prostatic ultrasonographic lesions. • To our knowledge, this is the first study to compare the accuracy of these popular risk calculators in a specific population

Contact sensitizer nickel sulfate activates the transcription factors NF-kB and AP-1 and increases the expression of nitric oxide synthase in a skin dendritic cell line

Nuclear factor kappa B (NF-kB) and activating protein-1 (AP-1) transcription factors are ubiquitously expressed signaling molecules known to regulate the transcription of a large number of genes involved in immune responses, namely the inducible isoform of nitric oxide synthase (iNOS). In this study, we demonstrate that a fetal skin-derived dendritic cell line (FSDC) produces nitric oxide (NO) in response to the contact sensitizer nickel sulfate (NiSO(4)) and increases the expression of the iNOS protein, as determined by immunofluorescence and Western blot analysis. The sensitizer NiSO(4) increased cytoplasmic iNOS expression by 31.9 +/- 10.3% and nitrite production, as assayed by the Griess reaction, by 27.6 +/- 9.5%. Electrophoretic mobility shift assay (EMSA), showed that 30 min of FSDC exposure to NiSO(4) activates the transcription factor NF-kB by 58.2 +/- 7.0% and 2 h of FSDC exposure to NiSO(4) activates the transcription factor AP-1 by 26.0 +/- 1.4%. Together, these results indicate that NiSO(4) activates the NF-kB and AP-1 pathways and induces iNOS expression in skin dendritic cells

Involvement of JAK2 and MAPK on type II nitric oxide synthase expression in skin-derived dendritic cells

In this report, we demonstrate that a fetal mouse skin-derived dendritic cell line produces nitric oxide (NO) in response to the endotoxin [lipopolysaccharide (LPS)] and to cytokines [tumor necrosis factor-alpha (TNF-alpha) and granulocyte-macrophage colony-stimulating factor (GM-CSF)]. Expression of the inducible isoform of NO synthase (iNOS) was confirmed by immunofluorescence with an antibody against iNOS. The tyrosine kinase inhibitor genistein decreased LPS- and GM-CSF-induced nitrite (NO(-2)) production. The effect of LPS and cytokines on NO(-2) production was inhibited by the Janus kinase 2 (JAK2) inhibitor tyrphostin B42. The p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB-203580 also reduced the NO(-2) production evoked by LPS, TNF-alpha, or GM-CSF, but it was not as effective as tyrphostin B42. Inhibition of MAPK kinase with PD-098059 also slightly reduced the effect of TNF-alpha or GM-CSF on NO(-2) production. Immunocytochemistry studies revealed that the transcription factor nuclear factor-kappaB was translocated from the cytoplasm into the nuclei of fetal skin-derived dendritic cells (FSDC) stimulated with LPS, and this translocation was inhibited by tyrphostin B42. Our results show that JAK2 plays a major role in the induction of iNOS in FSDC

Differential activation of nuclear factor kappa B subunits in a skin dendritic cell line in response to the strong sensitizer 2,4-dinitrofluorobenzene

Dendritic cell (DC) maturation is essential for the initiation of T-dependent immune responses. Nuclear factor kappa B (NF-kappaB) transcription factors are ubiquitously expressed signalling molecules, known to regulate the transcription of a large number of genes involved in immune responses, including cytokines and cell surface molecules. In this work, we studied the time-dependent activation of five members of the NF-kappaB family, p50, p52, p65, RelB and cRel, in a mouse skin DC line in response to stimulation with the strong sensitizer, 2,4-dinitrofluorobenzene (DNFB). Western blot assay revealed that exposure of fetal skin DC (FSDC) to DNFB induced the degradation of the inhibitor of NF-kappaB (IkappaB). Three out of its five members, i.e. p50, p52, and RelB, were similarly activated upon DNFB stimulation, with subsequent translocation of these subunits from the cytosol to the nucleus, but with different kinetics. In contrast, p65 expression was diminished in both the nucleus and the cytosol. The electrophoretic mobility shift assay (EMSA) showed that exposure of FSDC to DNFB induced DNA binding to NF-kappaB. Together, these results show that DNFB differentially activates the various members of the NF-kappaB family in skin DC

Muscle strength and mortality while on a liver transplant waiting list

OBJETIVO: Avaliar a força de músculos respiratórios e de mão em pacientes na lista de espera para o transplante de fígado e associá-los a mortalidade. MATERIAIS E MÉTODOS: Foram estudados retrospectivamente 132 pacientes submetidos à avaliação fisioterapêutica de rotina e que esperavam o transplante de fígado. A força dos músculos ventilatórios foi avaliada por meio das pressões inspiratória e expiratória máximas e a força do membro superior por meio de dinamometria. Os pacientes foram divididos em dois grupos: grupo A, com 51 pacientes (14 mulheres, 50,1±12,3 anos) que morreram enquanto estavam na lista de espera e grupo B, com 81 pacientes (31 mulheres, 45,0±3,8 anos) que sobreviveram até o transplante de fígado. Foi utilizado o teste de t de Student com nível de significância de 5%. RESULTADOS: Os valores médios da pressão inspiratória máxima (PImax) dos grupos A e B foram 65,7±28,0 e 77,5±33,8mmHg (p=0,04), respectivamente, e as pressões expiratórias máximas foram 72,9±32,9 e 84,4±33,1mmHg (p=0,07), respectivamente. Os valores médios da força da mão esquerda dos grupos A e B foram 18,5±8,1 e 21,5±10,5kgf (p=0,08), respectivamente, e da força da mão direita foram 20,2±9,7 e 23,5±12,5kgf (p=0,10), respectivamente. CONCLUSÕES: A PImax é menor nos pacientes que morreram enquanto aguardavam o transplante. No mesmo grupo, foi observado que a pressão expiratória máxima e a força da mão direita e esquerda foram menores, apesar de não apresentarem diferenças estatisticamente significante.OBJECTIVE: To evaluate respiratory muscle strength and hand strength in patients on a liver transplant waiting list and to associate these with mortality. METHODS: one hundred and thirty-two patients who underwent routine physical therapy evaluation while waiting for liver transplantation were studied retrospectively. Respiratory muscle strength was assessed by measuring the maximum inspiratory pressure (MIP) and maximum expiratory pressure (MEP), and upper-limb strength was evaluated by dynamometry. The patients were divided into two groups: group A, consisting of 51 patients (14 females, 50.1±12.3 years) who died while on the waiting list; and group B, consisting of 81 patients (31 females, 45.0±3.8 years) who survived until the time of liver transplant. Student’s t test was used with a 5% significance level. RESULTS: The mean MIP values for groups A and B were 65.7±28.0 and 77.5±33.8mmHg (p=0.04), respectively, and the mean MEP values were 72.9±32.9 and 84.4±33.1mmHg (p=0.07), respectively. The mean values for left-hand strength in groups A and B were 18.5±8.1 and 21.5±10.5kgf (p=0.08), and the mean values for right-hand strength were 20.2±9.7 and 23.5±12.5kgf (p=0.10), respectively. CONCLUSIONS: MIP was lower in the patients who died while waiting for liver transplantation. In the same group, it was observed that the MEP values and right and left-hand strength were numerically lower, although they did not reach statistically significant differences

Granulocyte-macrophage colony-stimulating factor activates the transcription of nuclear factor kappa B and induces the expression of nitric oxide synthase in a skin dendritic cell line.

Nitric oxide (NO) produced by skin dendritic cells and keratinocytes plays an important role in skin physiology, growth and remodelling. Nitric oxide is also involved in skin inflammatory processes and in modulating antigen presentation (either enhancing or suppressing it). In this study, we found that GM-CSF stimulates the expression of the inducible isoform of nitric oxide synthase (iNOS) in a fetal-skin-derived dendritic cell line (FSDC) and, consequently, increases the nitrite production from 11.9 +/- 3.2 micromol/L (basal level) to 26.9 +/- 4.2 micromol/L. Pyrrolidinedithiocarbamate (PDTC) inhibits nitrite production, with a half maximal inhibitory concentration (IC50) of 19.3 micromol/L and the iNOS protein expression in FSDC. In addition, western blot assays revealed that exposure of FSDC to GM-CSF induces the phosphorylation and degradation of the inhibitor of NF-kappaB (IkB), with subsequent translocation of the p50, p52 and RelB subunits of the transcription nuclear factor kappa B (NF-kappaB) from the cytosol to the nucleus. Electrophoretic mobility shift assays (EMSA) showed that FSDC exposure to GM-CSF activates the transcription factor NF-kappaB. Together, these results show that GM-CSF induces iNOS expression in skin dendritic cells by a mechanism involving activation of the NF-kappaB pathway

Network orchestration: new role of business incubators?

Purpose – The purpose of this paper is to analyze the performance of incubators in the stages of formationand development of incubated business networks, especially in bottom-up and top-down network models.Design/methodology/approach – The research is defined as qualitative and descriptive, with theapplication of multiple case studies, in which two networks of incubated businesses were investigated, onebeing top-down and the other bottom-up, which emerged within the incubation process of two businessincubators (CIETEC and INCIT). To make the study operational, 11 semi-structured interviews were carriedout and the thematic analysis of content was developed.Findings – The results pointed out that in the top-down network the incubator performs a new assignment,the network orchestration, which corresponds to the actions of formation, coordination and governance of thegroup. In the bottom-up network, it was found that the role of the incubator was to expand the value offersusually practiced.Research limitations/implications – As a limitation of the research, the very limitation of case studiesis pointed out that is they do not allow for generalizations.Practical implications – The research contributes to reflections on the effectiveness of the incubator andsheds light on the complementarity of networks in incubation processes, providing gains for incubators,incubated businesses and society.Originality/value – The originality of this document is the new role of the incubator, which isorchestration, and its categorization. The results allow us to understand the effects of providing networks andrelationships for incubated businesses. In addition, this study broadens the focus of traditional analyses of theincubator–incubated duo to consider the incubator–network–incubated trio

The importance of serological assays in diagnosing acute pulmonary histoplasmosis

Histoplasmosis is a systemic mycosis caused by inhalation of Histoplasma capsulatum microconidia. The disease does not normally affect immunocompetent individuals after a single, transient inhalation exposure. However, longer exposure may cause chronic or disseminated acute pulmonary infection. Herein, we report the case of a 24-year-old immunocompetent patient, who presented fever, cough and dyspnea for one month. The chest radiography revealed interstitial infiltrate and diffuse micronodules. The patient reported having had close and prolonged contact with bats. Diagnosis was confirmed by positive double immunodifusion and immunoblotting assays. She was treated with ketoconazole (400 mg) and there was complete resolution of the disease

Surgical treatment of complete penile duplication

Penile duplication is a rare anomaly with an incidence of 1 in 5,500,000. It is almost associated with other malformations like double bladder, presence of the cloaca, imperforate anus, duplication of the recto sigmoid and vertebral deformities. The authors present the surgical technique to resolve a rare case of complete penile duplication in a 4 years old child, without any other malformation