Modulation of inflammation by anti-TNF α mAb-dendrimer nanoparticles loaded in tyramine-modified gellan gum hydrogels in a cartilage-on-a-chip model.

Rheumatoid arthritis (RA) is an autoimmune and chronic inflammatory disease characterized by joint inflammation. Since the inflammatory condition plays an important role in the disease process, it is important to develop and test new therapeutic approaches that specifically target and treat joint inflammation. In this study, a human 3D inflammatory cartilage-on-a-chip model was established to test the therapeutic efficacy of anti-TNFα mAb-CS/PAMAM dendrimer NPs loaded-Tyramine-Gellan Gum in the treatment of inflammation. The results showed that the proposed therapeutic approach applied to the human monocyte cell line (THP-1) and human chondrogenic primary cells (hCH) cell-based inflammation system revealed an anti-inflammatory capacity that increased over 14 days. It was also possible to observe that Coll type II was highly expressed by inflamed hCH upon the culture with anti-TNF α mAb-CS/PAMAM dendrimer NPs, indicating that the hCH cells were able maintain their biological function. The developed preclinical model allowed us to provide more robust data on the potential therapeutic effect of anti-TNF α mAb-CS/PAMAM dendrimer NPs loaded-Ty-GG hydrogel in a physiologically relevant model.

Gastrointestinal Toxicity of Pelvic Radiotherapy: Are We Letting Women Down?

For all cancers there are four areas of importance: prevention, early diagnosis, optimising therapy and living with and beyond. For women diagnosed with gynaecological cancers, progress in these first three areas has been immense. However, living with and beyond has largely been ignored as a significant issue. As a group, patients treated for gynaecological cancer are more often young and more often suffer the most difficult long-term issues. Despite the growing number of long-term survivors, little has been done to ensure appropriate assessment and treatment of side-effects of cancer therapies, especially when radiotherapy has been used. For many affected patients their symptoms become part of everyday life, 'normality' is adjusted and these changes are tolerated even when severely limiting activities. Data show that even expert clinicians frequently do not appreciate the true impact of these problems and the focus of treatment and of follow-up remains fixed on 5-year survival and cancer recurrence, respectively. Many clinicians are unaware of what experts can do for toxicity and do not know where to refer their patients. However, rapid identification of patients with significant symptoms can lead to earlier diagnosis of treatable pathologies and improvement in patients' quality of life. In addition, the underlying pathophysiology of radiation-induced damage is potentially amenable to disease-modifying therapies. This review focuses on the factors that contribute to patients developing pelvic radiation disease, what can be done to mitigate the toxicity of treatment and highlights the challenges that must be addressed to reduce the gastrointestinal toxicity of pelvic radiotherapy.

Investigation of genetic susceptibility to Mycobacterium tuberculosis (VDR and IL10 genes) in a population with a high level of substructure in the Brazilian Amazon region.

OBJECTIVES: Tuberculosis (TB) is an infectious and contagious disease that has been very influential in human history and presents high rates of mortality. The objective of this study was to investigate the association of VDR, IL10, and SLC11A1 gene polymorphisms with susceptibility to the presence of Mycobacterium tuberculosis infection. METHODS: A total of 135 patients with confirmed TB and 141 healthy individuals were included in the analysis. Blood samples were collected for DNA extraction. Genotyping of the polymorphisms in the VDR and IL10 genes was performed by real-time PCR, and genotyping of the polymorphisms in the SLC11A1 gene by conventional PCR, followed by visualization in polyacrylamide gel. The genomic ancestry was obtained using an autosomal panel with 48 insertion/deletion ancestry-informative markers. RESULTS: Polymorphisms TaqI (TT, p=0.004), FokI (CC and CC+CT, p=0.012 and p=0.003, respectively), and BsmI (GG, p=0.008) in the VDR gene, as well as A-592C (GC+AG, p=0.001) in the IL10 gene, were significantly associated with susceptibility to TB In addition, high production of VDR combined with low production of IL10 showed protection for the TB group (p=0.035). CONCLUSIONS: The VDR polymorphisms may confer an increased risk and the IL10 haplotype may be a protection factor for the presence of M. tuberculosis infection in the Brazilian population.

Influence of apocynin on cardiac remodeling in rats with streptozotocin-induced diabetes mellitus.

BACKGROUND: Increased reactive oxygen species (ROS) generation in diabetes mellitus (DM) is an important mechanism leading to diabetic cardiomyopathy. Apocynin, a drug isolated from the herb Picrorhiza kurroa, is considered an antioxidant agent by inhibiting NADPH oxidase activity and improving ROS scavenging. This study analyzed the influence of apocynin on cardiac remodeling in diabetic rats. METHODS: Six-month-old male Wistar rats were assigned into 4 groups: control (CTL, n = 15), control + apocynin (CTL + APO, n = 20), diabetes (DM, n = 20), and diabetes + apocynin (DM + APO, n = 20). DM was induced by streptozotocin. Seven days later, apocynin (16 mg/kg/day) or vehicle was initiated and maintained for 8 weeks. Left ventricular (LV) histological sections were used to analyze interstitial collagen fraction. NADPH oxidase activity was evaluated in LV samples. Comparisons between groups were performed by ANOVA for a 2 × 2 factorial design followed by the Bonferroni post hoc test. RESULTS: Body weight (BW) was lower and glycemia higher in diabetic animals. Echocardiogram showed increased left atrial diameter, LV diastolic diameter, and LV mass indexed by BW in both diabetic groups; apocynin did not affect these indices. LV systolic function was impaired in DM groups and unchanged by apocynin. Isovolumic relaxation time was increased in DM groups; transmitral E/A ratio was higher in DM + APO compared to DM. Myocardial functional evaluation through papillary muscle preparations showed impaired contractile and relaxation function in both DM groups at baseline conditions. After positive inotropic stimulation, developed tension (DT) was lower in DM than CTL. In DM + APO, DT had values between those in DM and CTL + APO and did not significantly differ from either group. Myocardial interstitial collagen fraction was higher in DM than CTL and did not differ between DM + APO and CTL + APO. Serum activity of antioxidant enzymes glutathione peroxidase, superoxide dismutase (SOD), and catalase was lower in DM than CTL; apocynin restored catalase and SOD levels in DM + APO. Myocardial NADPH oxidase activity did not differ between groups. CONCLUSION: Apocynin restores serum antioxidant enzyme activity despite unchanged myocardial NADPH oxidase activity in diabetic rats.

Ativação de células de memória na produção de anticorpos e na expressão de células IgM positivas no baço de tilápias-do-nilo / Activation of memory cells in the antibody production and the expression of IgM positive cells in Nile tilapia spleen

O presente trabalho avaliou o papel do baço no armazenamento e na reativação das linhagens de células B, representadas por células IgM positivas imunomarcadas no tecido esplênico, bem como a funcionalidade dessas células, sobre a cinética dos linfócitos e na produção sistêmica de anticorpos em tilápias-do-nilo (Oreochromis niloticus). Foram separados dois grupos: grupo memória, constituído por peixes previamente imunizados com hemácia de carneiro a 2,5%, para a geração da memória imune, e o grupo naive, que recebeu o mesmo volume de solução salina a 0,65%. Após 32 dias, os dois grupos foram submetidos a uma nova dose do antígeno na mesma concentração, volume e via de inoculação. A reativação dos clones de memória foi evidenciada pelo aumento do número de células IgM positivas no baço do grupo memória no dia zero/pré-imune. Além disso, o mesmo grupo apresentou aumento dos títulos de anticorpos séricos no 14º dia e no número absoluto de linfócitos no 21º dia em relação ao grupo naive. Esses resultados sugerem que o baço não seja apenas um local de armazenamento, mas também de reativação de células B de memória em tilápia-do-nilo.(AU)

This work aimed to evaluate the role of the spleen in storage and reactivation of the memory B cells, represented by IgM positive cells and the systemic production of sheep antibodies anti-red cell in Nile tilapia (Oreochromis niloticus). Two groups were established: the memory group, containing fish previously immunized with a 2,5% sheep anti-red cell, to generate the immune memory; and the naive group, containing fish that received a 0,65% saline solution. After 32 days, both groups were subjected to a new dose of the same antigen at the same concentration, volume, and inoculation via. The memory clones reactivation was correlated to the increase of the IgM positive cells in the spleen in the memory group at 0 day. The memory group showed an increase in the absolute number of lymphocytes at 21 days and an increase in the antibodies at 14 days after inoculation when compared to the naive group. The results suggest that the spleen may be a storage and reactivation place of memory B cells in Nile tilapia.(AU)

Apocynin influence on oxidative stress and cardiac remodeling of spontaneously hypertensive rats with diabetes mellitus.

PURPOSE: Although increased oxidative stress is a major component of diabetic hypertensive cardiomyopathy, research into the effects of antioxidants on cardiac remodeling remains scarce. The actions of antioxidant apocynin include inhibiting reactive oxygen species (ROS) generation by nicotinamide adenine dinucleotide phosphate (NADPH) oxidases and ROS scavenging. We evaluated the effects of apocynin on cardiac remodeling in spontaneously hypertensive rats (SHR) with diabetes mellitus (DM). METHODS: Male SHR were divided into four groups: control (SHR, n = 16); SHR treated with apocynin (SHR-APO; 16 mg/kg/day, added to drinking water; n = 16); diabetic SHR (SHR-DM, n = 13); and SHR-DM treated with apocynin (SHR-DM-APO, n = 14), for eight weeks. DM was induced by streptozotocin (40 mg/kg, single dose). Statistical analyzes: ANOVA and Tukey or Mann-Whitney. RESULTS: Echocardiogram in diabetic groups showed higher left ventricular and left atrium diameters indexed for body weight, and higher isovolumetric relaxation time than normoglycemic rats; systolic function did not differ between groups. Isolated papillary muscle showed impaired contractile and relaxation function in diabetic groups. Developed tension was lower in SHR-APO than SHR. Myocardial hydroxyproline concentration was higher in SHR-DM than SHR, interstitial collagen fraction was higher in SHR-DM-APO than SHR-APO, and type III collagen protein expression was lower in SHR-DM and SHR-DM-APO than their controls. Type I collagen and lysyl oxidase expression did not differ between groups. Apocynin did not change collagen tissue. Myocardial lipid hydroperoxide concentration was higher in SHR-DM than SHR and SHR-DM-APO. Glutathione peroxidase activity was lower and catalase higher in SHR-DM than SHR. Apocynin attenuated antioxidant enzyme activity changes in SHR-DM-APO. Advanced glycation end-products and NADPH oxidase activity did not differ between groups. CONCLUSION: Apocynin reduces oxidative stress independently of NADPH oxidase activity and does not change ventricular or myocardial function in spontaneously hypertensive rats with diabetes mellitus. The apocynin-induced myocardial functional impairment in SHR shows that apocynin actions need to be clarified during sustained chronic pressure overload.

Transferência passiva de soro hiperimune anti-Streptococcus agalactiae e seu efeito profilático em tilápias-do-nilo infectadas experimentalmente: sobrevivência e títulos de anticorpos / Passive transfer of hyperimmune serum anti Streptococcus agalactiae and its prophylactic effect on Nile tilapia experimentally infected

A bactéria Streptococcus agalactiae é um potente agente causador de surtos por doenças bacterianas em peixes. O estresse provocado pelo manejo zootécnico e pela má qualidade ambiental torna a tilápia susceptível às infecções por essa bactéria. O objetivo do presente trabalho foi avaliar a resistência de tilápias-do-nilo imunizadas com soro hiperimune anti-S. agalactiae, posteriormente desafiadas com cepa homóloga da mesma bactéria. Após determinação da DL 50 de S. agalactiae, 36 tilápias foram distribuídas em quatro aquários, dois para o grupo controle e dois para inoculação celomática para produção de anticorpos anti-S. agalactiae. No 21° e 28° dias, foi coletado sangue para obtenção de soro hiperimune utilizado na transferência passiva. Em seguida, 30 tilápias foram distribuídas em três aquários e submetidas a três tratamentos: GI: controle; GII: imunizadas com o soro inativado; GIII: imunizadas com soro ativo...

The Streptococcus agalactiae bacteria is a potent agent which causes outbreaks of bacterial diseases in fish. The stress caused by management and poor environmental quality makes tilapia susceptible to infections, including by bacterium. The aim of this study was to evaluate the resistance of the Nile tilapia immunized with hyperimmune serum against S. agalactiae subsequently challenged with homologous strain of the same bacteria. After determining the DL 50 of S. agalactiae, 36 tilapias were distributed in 4 aquariums, 2 for the control group and 2 for the group via coelomic, inoculated with the DL 50 for anti-S. agalactiae antibodies production. On the 21st and 28th day blood was collected for the obtainment of hiperimmune serum used in passive transference. Then, 30 tilapias were distributed in 3 aquariums submitted to 3 treatments (GI: control; GII: immunized with inactivated-serum; GIII: immunized with non-inactivated serum)...

Análise dos itens da Escala de Aconselhamento Profissional - EAP- pelo Modelo de Rasch / Analysis of items of Escala de Aconselhamento Profissional by the Rasch model

Esta pesquisa teve como objetivo analisar os itens da Escala de Aconselhamento Profissional segundoo modelo de Rasch, enfocando principalmente o funcionamento diferencial dos itens em relação aotipo de escola (particular ou pública). Participaram 615 estudantes do ensino médio (68% do sexofeminino) de instituições do interior do estado de São Paulo, sendo 179 de escola privada e 436 deescola pública, com idades variando entre 14 a 27 anos e média de 16,36. Os resultados mostraramque os itens tiveram, em geral, valores adequados de ajuste. Em relação à análise de DIF, observou-seque 20 itens estavam enviesados, sendo que o favorecimento ao tipo de escola mostrou-serelativamente equilibrado. Sugere-se uma análise mais aprofundada dos itens, para verificar se énecessária a separação da escala em duas versões, ou até mesmo se esse viés é parte do constructo(AU)

N-acetyl transferase 2 and cytochrome P450 2E1 genes and isoniazid-induced hepatotoxicity in Brazilian patients.

SETTING: Isoniazid (INH) is related to the development of hepatotoxicity in some patients. OBJECTIVE: To investigate the role of N-acetyl transferase 2 (NAT2) and cytochrome P450 2E1 (CYP2E1) in the hepatotoxicity of patients treated with INH in an Amazonian Brazilian population. DESIGN: Patients undergoing anti-tuberculosis treatment were investigated. Hepatotoxicity was defined as an increase of more than three times the upper limit of normal in serum alanine aminotransferase levels after treatment. NAT2 genotypes were identified by sequencing, whereas CYP2E1 alleles were detected using polymerase chain reaction based methods. RESULTS: Of the 270 individuals included in the study, 18 (6.7%) developed drug-related hepatotoxicity. A high association was found between slow acetylators and hepatotoxicity, particularly with regard to allele *5. The adjusted risk of developing hepatotoxicity was significant in individuals carrying two slow acetylation alleles (P = 0.036, OR 3.05, 95%CI 1.07-8.64). In all of the CYP2E1 markers examined, wild homozygous genotypes were more prevalent in subjects with hepatotoxicity than in controls; however, the difference was not statistically significant. Joint evaluation of the genes revealed a high risk of developing hepatotoxicity in slow acetylators with CYP2E1 wild alleles (adjusted OR 4.26; 95%CI 1.47-12.37, P = 0.008). CONCLUSIONS: Large-scale screening for NAT2 and CYP2E1 genotypes can prove useful in predicting the risk of adverse effects.

Abdominal aortic aneurysm in association with horseshoe kidney.

The authors report 3 cases of infra-renal abdominal aortic aneurysms resected in the presence of horseshoe kidney. In all these cases the diagnosis of the renal anomaly was done before the aneurysmectomy. In the first case, the diagnosis of horseshoe kidney was done by an aortography. In the second case by a previous operation to remove an urinary calculi. Three years after this surgery the diagnosis of abdominal aortic aneurysm was done when a computed tomography, an intravenous pyelography and an aortography, confirmed the diagnosis of horseshoe kidney. The third case come to our hospital in the fourth day after an exploratory laparotomy done in another hospital, when was observed the aneurysm and the horseshoe kidney. The second case had two anomalous arteries. One of them arose from the aneurysm to the renal isthmus and the other one from the common iliac to the isthmus. The three patients had an uncomplicated postoperative course. Only in the second case we had the necessity to reattache an anomalous artery directly to the graft.