Effect of inulin supplementation in male mice fed with high fat diet on biochemical profile and α-amylase gene expression

Purpose: To evaluate the preventive and therapeutic effects of inulin supplementation in Naval Medical Research Institute (NMRI) male mice fed with high fat diet. Methods: NMRI male mice (n = 36) were divided into three groups. Control (C1), obese (O1) and experimental mice (E1) were fed during 8 weeks as follows: C1 with normal rodent pellet, O1 with high fat diet, and E1 with high fat diet plus 20 % inulin. C2, O2, and E2 were fed as follows: C2 with normal rodent pellets for 12 weeks; O2 with high fat diet during 8 weeks and switched to normal rodent pellet during next 4 weeks; and E2 with high fat diet over a period of 8 weeks and switched to normal rodent pellet plus 20 % inulin for 4 weeks. Body weight, serum glucose, triglycerides, total cholesterol, high density lipoprotein (HDL), low density lipoprotein (LDL), and hepatic α-amylase gene expression were measured. Results: Groups receiving high fat diet showed higher weight (30.71 ± 0.66 g in O2, p < 0.001), nonfasting blood glucose levels (257.69 ± 5.10 mg/dl in O2, p < 0.001), TG (282.15 ± 1.83 mg/dl in O2, (p < 0.001)), and cholesterol levels (335.72 ± 2.23 mg/dl in O2, (p < 0.001)), compared with control. In C2 group, mean body weight was 25.71 ± 0.54 g, non-fasting blood level 161.54 ± 4.48 mg/dl, TG level 214.29 ± 5.54 mg/dl, and cholesterol level 164.29 ±4.57 mg/dl. Compared to obese group, mice receiving inulin showed lower blood glucose levels (223.10 ± 8.7 mg/dl in E2, p < 0.001), body weight (27.86 ± 0.57 g in E2, p < 0.001), TG (232.14 ± 4.02 mg/dl in E2, p < 0.001) and cholesterol (249.97 ± 2.28 in E2, p < 0.001). A slight decrease in hepatic α-amylase gene expression was observed only in E1. Conclusion: Besides its sweetening properties, inulin may also find use as a potential anti-obesity compound.

Characteristics of microRNAs and their potential relevance for the diagnosis and therapy of the acute respiratory distress syndrome: from bench to bedside

Acute respiratory distress syndrome (ARDS) is a complex disease associated with high morbidity and mortality. Biomarkers and specific pharmacologic treatment of the syndrome are lacking. MicroRNAs (miRNAs) are small (∼19–22 nucleotides) noncoding RNA molecules whose function is the regulation of gene expression. Their uncommon biochemical characteristics (eg, their resistance to degradation because of extreme temperature and pH fluctuations, freeze-thaw cycles, long storage times in frozen conditions, and RNAse digestion) and their presence in a wide range of different biological fluids and the relatively low number of individual miRNAs make these molecules good biomarkers in different clinical conditions. In addition, miRNAs are suitable therapeutic targets as their expression can be modulated by different available strategies. The aim of the present review is to offer clinicians a global perspective of miRNA, covering their structure and nomenclature, biogenesis, effects on gene expression, regulation of expression, and features as disease biomarkers and therapeutic targets, with special attention to ARDS. Because of the early stage of research on miRNAs applied to ARDS, attention has been focused on how knowledge sourced from basic and translational research could inspire future clinical studies.

Neuroprotective role of nutraceutical and phytochemical components

Neurodegenerative diseases (NDs) are characterized by a multifactorial etiology, in which oxidative stress and inflammation are the main causative factors. For this reason, increasing attention is being paid to the characterization and the identification of nutraceuticals and phytochemicals with intrinsic pleiotropic activity. Moreover, in a Circular Economy perspective, these natural compounds can be obtained also from renewable resources derived from the food industry by-products and can be used for both preventive and therapeutic purposes. The aim of this PhD program was to identify nutraceuticals and phytochemicals, both as extracts and pure compounds, and obtained from both plant and renewable sources, which due to their antioxidant and anti-inflammatory properties, were able to counteract cellular and molecular alterations that characterize NDs. Their neuroprotective potential has been evaluated in an in vitro model of neuroinflammation (the LPS-activated BV-2 microglial cell line), and/or in an in vitro model of neuronal oxidative stress (the neuron-like SH-SY5Y cell line differentiated with retinoic acid and exposed to H2O2). Four different projects, although deeply linked by the aforementioned common goal, have been discussed in this thesis: 1_ Impact of phenolic profile of different cherry cultivars on the potential neuroprotective effect in SH-SY5Y cells. 2_Anti-inflammatory activities of Spilanthol-rich essential oil from Acmella oleracea (L.). 3_Study of the anti-inflammatory activity of novel tacrine derivatives with lipids extracted from cashew nutshell liquid. 4_Coffee Silverskin (CSS) and Spent Coffee Grounds (SCG): coffee industry by-products as a promising source of neuroprotective agents. In general, it is, therefore, possible to conclude that the natural compounds studied in this thesis have been proven, due to their antioxidant and/or anti-inflammatory properties, to be valid preventive and therapeutic strategies for the treatment of NDs, to improve the life quality of these patients and of the general population by preventing and combating the onset of these deleterious diseases.

Design and synthesis of E3 ligase RNF5 inhibitors as innovative strategy to trigger mutant CFTR rescue in Cystic Fibrosis

In Cystic Fibrosis (CF) the deletion of phenylalanine 508 (F508del) in the CFTR anion channel is associated to misfolding and defective gating of the mutant protein. Among the known proteins involved in CFTR processing, one of the most promising drug target is the ubiquitin ligase RNF5, which normally promotes F508del-CFTR degradation. In this context, a small molecule RNF5 inhibitor is expected to chemically mimic a condition of RNF5 silencing, thus preventing mutant CFTR degradation and causing its stabilization and plasma membrane trafficking. Hence, by exploiting a virtual screening (VS) campaign, the hit compound inh-2 was discovered as the first-in-class inhibitor of RNF5. Evaluation of inh-2 efficacy on CFTR rescue showed that it efficiently decreases ubiquitination of mutant CFTR and increases chloride current in human primary bronchial epithelia. Based on the promising biological results obtained with inh-2, this thesis reports the structure-based design of potential RNF5 inhibitors having improved potency and efficacy. The optimization of general synthetic strategies gave access to a library of analogues of the 1,2,4-thiadiazol-5-ylidene inh-2 for SAR investigation. The new analogues were tested for their corrector activity in CFBE41o- cells by using the microfluorimetric HS-YFP assay as a primary screen. Then, the effect of putative RNF5 inhibitors on proliferation, apoptosis and the formation of autophagic vacuoles was evaluated. Some of the new analogs significantly increased the basal level of autophagy, reproducing RNF5 silencing effect in cell. Among them, one compound also displayed a greater rescue of the F508del-CFTR trafficking defect than inh-2. Our preliminary results suggest that the 1,2,4-thiadiazolylidene could be a suitable scaffold for the discovery of potential RNF5 inhibitors able to rescue mutant CFTRs. Biological tests are still ongoing to acquire in-depth knowledge about the mechanism of action and therapeutic relevance of this unprecedented pharmacological strategy.

Harnessing the cellular and molecular complexity of high-risk neuroblastoma to investigate novel potential treatment approaches

Neuroblastoma (NB) is the most common type of tumor in infants and the third most common cancer in children. Current clinical practices employ a variety of strategies for NB treatment, ranging from standard chemotherapy to immunotherapy. Due to a lack of knowledge about the molecular mechanisms underlying the disease's onset, aggressive phenotype, and therapeutic resistance, these approaches are ineffective in the majority of instances. MYCN amplification is one of the most well-known genetic alterations associated with high risk in NB. The following work is divided into three sections and aims to provide new insights into the biology of NB and hypothetical new treatment strategies. First, we identified RUNX1T1 as a key gene involved in MYCN-driven NB onset in a transgenic mouse model. Our results suggested that that RUNX1T1 may recruit the Co-REST complex on target genes that regulate the differentiation of NB cells and that the interaction with RCOR3 is essential. Second, we provided insights into the role of MYCN in dysregulating the CDK/RB/E2F pathway controlling the G1/S transition of the cell cycle. We found that RB is dispensable in regulating MYCN amplified NB's cell cycle, providing the rationale for using cyclin/CDK complexes inhibitors in NBs carrying MYCN amplification and relatively high levels of RB1 expression. Third, we generated an M13 bacteriophage platform to target GD2-expressing cells in NB. Here, we generated a recombinant M13 phage capable of binding GD2-expressing cells selectively (M13GD2). Our results showed that M13GD2 chemically conjugated with the photosensitizer ECB04 preserves the retargeting capability, inducing cell death even at picomolar concentrations upon light irradiation. These results provided proof of concept for M13 phage employment in targeted photodynamic therapy for NB, an exciting strategy to overcome resistance to classical immunotherapy.

Assessing brain connectivity through electroencephalographic signal processing and modeling analysis

Brain functioning relies on the interaction of several neural populations connected through complex connectivity networks, enabling the transmission and integration of information. Recent advances in neuroimaging techniques, such as electroencephalography (EEG), have deepened our understanding of the reciprocal roles played by brain regions during cognitive processes. The underlying idea of this PhD research is that EEG-related functional connectivity (FC) changes in the brain may incorporate important neuromarkers of behavior and cognition, as well as brain disorders, even at subclinical levels. However, a complete understanding of the reliability of the wide range of existing connectivity estimation techniques is still lacking. The first part of this work addresses this limitation by employing Neural Mass Models (NMMs), which simulate EEG activity and offer a unique tool to study interconnected networks of brain regions in controlled conditions. NMMs were employed to test FC estimators like Transfer Entropy and Granger Causality in linear and nonlinear conditions. Results revealed that connectivity estimates reflect information transmission between brain regions, a quantity that can be significantly different from the connectivity strength, and that Granger causality outperforms the other estimators. A second objective of this thesis was to assess brain connectivity and network changes on EEG data reconstructed at the cortical level. Functional brain connectivity has been estimated through Granger Causality, in both temporal and spectral domains, with the following goals: a) detect task-dependent functional connectivity network changes, focusing on internal-external attention competition and fear conditioning and reversal; b) identify resting-state network alterations in a subclinical population with high autistic traits. Connectivity-based neuromarkers, compared to the canonical EEG analysis, can provide deeper insights into brain mechanisms and may drive future diagnostic methods and therapeutic interventions. However, further methodological studies are required to fully understand the accuracy and information captured by FC estimates, especially concerning nonlinear phenomena.

[pediatric Patient Classification System: Improvement Of An Instrument].

Improve the content validity of the instrument for classification of pediatric patients and evaluate its construct validity. A descriptive exploratory study in the measurement of the content validity index, and correlational design for construct validation through exploratory factor analysis. The content validity index for indicators was 0.99 and it was 0.97 for graded situations. Three domains were extracted in the construct validation, namely: patient, family and therapeutic procedures, with 74.97% of explained variance. The instrument showed evidences of content and construct validity. The validation of the instrument occurred under the approach of family-centered care, and allowed incorporating some essential needs of childhood such as playing, interaction and affection in the content of the instrument.

Motivating Medical Students To Learn Basic Science Concepts Using Chronic Myeloid Leukemia As An Integration Theme.

To report on the use of chronic myeloid leukemia as a theme of basic clinical integration for first year medical students to motivate and enable in-depth understanding of the basic sciences of the future physician. During the past thirteen years we have reviewed and updated the curriculum of the medical school of the Universidade Estadual de Campinas. The main objective of the new curriculum is to teach the students how to learn to learn. Since then, a case of chronic myeloid leukemia has been introduced to first year medical students and discussed in horizontal integration with all themes taught during a molecular and cell biology course. Cell structure and components, protein, chromosomes, gene organization, proliferation, cell cycle, apoptosis, signaling and so on are all themes approached during this course. At the end of every topic approached, the students prepare in advance the corresponding topic of clinical cases chosen randomly during the class, which are then presented by them. During the final class, a paper regarding mutations in the abl gene that cause resistance to tyrosine kinase inhibitors is discussed. After each class, three tests are solved in an interactive evaluation. The course has been successful since its beginning, 13 years ago. Great motivation of those who participated in the course was observed. There were less than 20% absences in the classes. At least three (and as many as nine) students every year were interested in starting research training in the field of hematology. At the end of each class, an interactive evaluation was performed and more than 70% of the answers were correct in each evaluation. Moreover, for the final evaluation, the students summarized, in a written report, the molecular and therapeutic basis of chronic myeloid leukemia, with scores ranging from 0 to 10. Considering all 13 years, a median of 78% of the class scored above 5 (min 74%-max 85%), and a median of 67% scored above 7. Chronic myeloid leukemia is an excellent example of a disease that can be used for clinical basic integration as this disorder involves well known protein, cytogenetic and cell function abnormalities, has well-defined diagnostic strategies and a target oriented therapy.

Saturated Fatty Acids Modulate Autophagy's Proteins In The Hypothalamus.

Autophagy is an important process that regulates cellular homeostasis by degrading dysfunctional proteins, organelles and lipids. In this study, the hypothesis that obesity could lead to impairment in hypothalamic autophagy in mice was evaluated by examining the hypothalamic distribution and content of autophagic proteins in animal with obesity induced by 8 or 16 weeks high fat diet to induce obesity and in response to intracerebroventricular injections of palmitic acid. The results showed that chronic exposure to a high fat diet leads to an increased expression of inflammatory markers and downregulation of autophagic proteins. In obese mice, autophagic induction leads to the downregulation of proteins, such as JNK and Bax, which are involved in the stress pathways. In neuron cell- line, palmitate has a direct effect on autophagy even without inflammatory activity. Understanding the cellular and molecular bases of overnutrition is essential for identifying new diagnostic and therapeutic targets for obesity.

Óxido nítrico: propriedades e potenciais usos terapêuticos

Nitric oxide ( NO) is a substance that acts as a second-messenger and is associated with a number of important physiological functions such as regulation of the vascular tonus, immune modulation and neurotransmission. As a physiological mediator, alteration of its concentration level may cause pathophysiological disfunctions such as hypertension, septic shock and impotence. Possible therapeutic approaches are being developed to control NO levels in vivo. We review herein the main physical and chemical properties of NO, its biological functions and available chemical interventions to reduce and increment its physiological concentration levels. Recent developments in the field are also highlighted.

Indicações para ceratoplastia penetrante no Hospital das Clínicas-UNICAMP

PURPOSE: To determine the main causes of penetrating keratoplasty indications at Hospital das Clínicas-UNICAMP (January, 1999 to December, 2003). METHODS: A non-comparative, retrospective series of case studies. The authors reviewed the files of 857 patients who underwent penetrating keratoplasty at Hospital das Clínicas-UNICAMP between 1999-2003 and classified them into different categories according to diagnostic indication for surgery. RESULTS: The age range was between 0-88 years (average 44 years ±1.2). The main causes of penetrating keratoplasty were: keratoconus in 427 cases (49.82%); 152 cases (17.74%) of corneal ulceration (perforated or not); corneal graft failure in 87 cases (10.15%); bullous keratopathy, 72 cases (8.40%); Fuchs dystrophy in 59 cases (6.88%); trachoma complications in 28 cases (3.27%); other causes, 32 (3.74%). In children under 10 years of age, the main cause of penetrating keratoplasty indications was infectious ulcer (77.78%) and between 11-50 years of age, keratoconus was the main cause (71.65%). CONCLUSION: This study was composed of a young population, and the main causes of penetrating keratoplasty were keratoconus and therapeutic keratoplasty.

Educação do corpo e vida ao ar livre : natureza e educação fisica em São Paulo (1930-1945)

Universidade Estadual de Campinas . Faculdade de Educação Física

Práticas corporais alternativas e educação física : entre a formação e a intervenção

Universidade Estadual de Campinas. Faculdade de Educação Física

Linguagem corporal de expressão da criatividade e seu (des)envolvimento na educação fisica

Universidade Estadual de Campinas. Faculdade de Educação Física

Ressecções eletiva e de urgência para tratamento de neoplasia maligna do cólon em hospital universitário: estudo de 66 casos

O câncer de cólon é uma doença de alta prevalência e mortalidade, cujo tratamento baseia-se na ressecção cirúrgica. A possibilidade de cura aumenta com o diagnóstico precoce, daí a importância dos programas de rastreamento populacional do câncer colorretal. O presente estudo analisou, retrospectivamente, 66 pacientes submetidos a ressecções do cólon por neoplasia em um período de 58 meses no Hospital Universitário da Universidade de São Paulo. Os pacientes foram divididos em dois grupos: grupo 1, submetidos a cirurgia eletiva (28 pacientes), e grupo 2, submetidos a cirurgia de urgência (38 pacientes). Os grupos foram comparados com relação às variáveis sexo, idade, apresentação clínica, aspectos da técnica cirúrgica, sítio anatômico da lesão, estádio patológico, taxas de complicações, permanência hospitalar pós-operatória e óbitos na internação. Verificou-se no presente estudo que a idade entre os grupos foi semelhante. Houve uma predominância do sexo masculino entre os pacientes operados de urgência. No grupo de cirurgia eletiva, o principal sintoma foi a hematoquezia, enquanto os operados na urgência, tinham como principal queixa dor abdominal. A grande maioria dos pacientes, no momento da cirurgia, apresentava-se sintomática há meses. Os pacientes operados na urgência apresentaram mais tumores pT4 e os operados eletivamente apresentaram mais neoplasias em estádio I. Em ambos os grupos, o caráter oncológico dos procedimentos foi preservado, bem como foi alto o índice de anastomoses primárias (81,8%). As taxas de complicações pós-operatórias, o tempo de permanência hospitalar pós-operatório e a mortalidade foram semelhantes.