Em diferentes mopdelos, o aumento dos níveis séricos de glicose pode ativar cascatas de sinalização envolvendo receptores semelhantes a Toll, o fator de transcrição NF-kB, moléculas da familia NLRP, ASC e caspase-1, que culminam na produção, ativação e liberação de moléculas inflamatórias. Investigamos o perfil inflamatório sérico e placentário em gestantes com distúrbios glicêmicos e a possível participação da via do inflamassoma NLRP1 e NRLP3 para o estabelecimento desse perfil. Para avaliar o papel da hiperglicemia no estabelecimento desse perfil, a via do inflamassoma foi também estudada em explantes de vilos coriônicos tratados com 100, 200, 300 e 400 mg/dL de glicose. Nossos resultados mostraram aumento na ativação de NF-kB e da expressão de NLRP1, NLRP3, ASC e Caspase-1 nas placentas de gestantes hiperglicemicas e nas culturas tratadas com doses de 300 r 400 mg/dL de glicose. Esses dados sugerem que a ativação da via do inflamassoma na porção fetal da placenta contribui para o processo inflamatório nessas pacientes. / In different models, the increase in serum glucose levels can activate signaling cascades involving Toll-like receptors, the transcription factor NF-kB, NLRP proteins, ASC and caspase-1, which culminate in the production, activation and release of inflammatory molecules . We investigated the serum and placental inflammatory profile in pregnant women with glucose disorders and the possible participation of via the NLRP1 and NRLP3 inflammasome to establish this profile. To evaluate the role of hyperglycemia, molecules of the inflammasome pathway was also studied in explants of chorionic villi treated with 100, 200, 300 and 400 mg / dL glucose. Our results showed an increase in NF-kB activation and expression of NLRP1, NLRP3, ASC and Caspase-1 in placentas of pregnant hyperglycemic women and in cultures treated with 300 and 400 mg / dL glucose. These data suggest that the inflammasome activation of the fetal portion of the placenta contributes to the inflammatory process in these patients.

The central nervous system (CNS) has a crucial role in the maintenance of energy homeostasis by orchestrating a plethora of signals from peripheral organs about the state of energy stores and the current energy intake needed to match energy expenditure. These signals converge into the hypothalamic regions and its complex local circuitry. CNS-derived cholecystokinin (CCK) is acting at central level to modulate energy balance by regulating the neuronal activity of hypothalamic neuronal populations that regulate food intake, energy storage and consumption. Moreover, our recent published work identifies CCK neurons as key integrators of the neuroendocrine negative feedback of glucocorticoids to the PVN. Glucose sensing neurons of the Ventromedial Hypothalamus (VMH) are integrating energy signals and are essential for mounting a counter-regulatory response and glucose homeostasis. VMH is also important in energy expenditure by regulating body weight and thermogenesis. CCK neurons are present in high density in the VMH.The source of endogenous CCK that acts on distinct neuronal components has not been elucidated. The research so far does not address the purpose of CCK neurons in the hypothalamus and their potential role in the network dynamics regarding energy homeostasis. In this study, we untangle the role of CCK neurons in the VMH nucleus by employing stereotactic intracranial delivery of adeno-associated viruses that result in cell-type specific chemogenetic inhibition or ablation of these neurons. Acute silencing of their neurotransmission with the cre-dependent AAV expression of the chemogenetic tool of Designer Receptors Exclusively Activated by Designer Drugs (DREADDs) increases their daily food intake due to increased meal numbers and eating frequency without meal size or meal duration being affected. CCK ablation by a newly generated double-recombinase-mediated Diphtheria Toxin Receptor (DTR) mouse line or AAV-DTA-mediated ablation resulted in hyperphagia, obesity and hyperglycaemia. We conclude that CCK^VMH neurons are implicated in the regulation of food intake, body weight and glucose homeostasis in the adult brain.

Orientador: Débora Cristina Damasceno / Banca: Yuri Karen Sinzato / Banca: Daniela Cristina Ceccatto Gerardin / Banca: Luciano Freitas Felicio / Banca: Maria Martha Bernardi / Resumo: Não disponível / Abstract: The aim of the present study was to evaluate the effect of maternal mild hyperglycemia on maternal behavior, as well as on offspring development, behavior, reproductive function, and development of glucose intolerance in adulthood. At birth, litters were assigned either to Control (subcutaneous(sc)- citrate buffer) or STZ group (streptozotocin(STZ)-100mg/kg-sc.). On PND 90 rats were mated. Glucose tolerance test (GTT) and insulin tolerance test (ITT) were performed during pregnancy. Pregnancy duration, litter size and sex ratio were assessed. Newborns were classified according to birth weight as small (SPA), adequate (APA), or large for pregnancy age (LPA). Maternal behavior was analyzed on PND 5 and 10. Offspring physical and sexual development was evaluated. Offspring body weight, length, and anogenital distance was measured and general activity was assessed on open field. Sexual behavior studies were conducted on both male and female offspring. Levels of reproductive hormones and estrous cycle duration were evaluated in female offspring. Female offspring were mated and GTT and ITT performed during pregnancy. STZ neonatal administration caused mild hyperglcyemia during pregnancy and changed some aspects of maternal care. The hyperglycemic intrauterine millieu impaired physical development and increased imobility on the open field in the offspring although the latter effect appeared at different ages for males (adulthood) and females (infancy). However, offspring sexual behavior was not impaired and, during pregnancy, STZ female offspring did not become glucose intolerant. Therefore, it can be concluded that mild hyperglycemia during pregnancy caused changes in maternal behavior, as well as offspring development and general activity, but failed to induce glucose... (Complete abstract click electronic access below) / Doutor

En la actualidad es difícil encontrar que se monitoree la glucosa antes, durante y al finalizar un proceso quirúrgico para comprobar el estado de estrés del animal y el nivel de dolor que experimenta; ambos episodios conllevan a cuadros de hiperglucemia; esta se presenta como respuesta en el paciente tras haber sufrido un trauma o lesión, que en este caso lo causa la cirugía, generando complicaciones como: Deshidratación, lenta recuperación post-quirúrgica, daños neurológicos y en diversos tejidos del organismo. El objetivo de este estudio fue determinar el porcentaje de pacientes con hiperglucemia del curso de Cirugía de Animales Menores de la Universidad Ricardo palma y la clínica veterinaria Hernández en la ciudad de Lima. Se evaluaron 25 pacientes caninos de sexo macho sometidos a castración, entre Agosto del 2014 y Julio del 2015; a los cuales se les evaluó la glucosa en diferentes etapas de la cirugía con un glucómetro de acuerdo a un tipo de cirujano. Los resultados fueron analizados mediante los programas SPSS y Microsoft Excel. Los resultados promedio indicaron que de las 100 mediciones de glucosas obtenidas un 18% se encontraba en hiperglucemia, siendo el máximo valor registrado el de 212 mg/dl; de los cuales el grupo de Cirujano 1 (profesores) no mostro pacientes en estado hiperglucémico; sin embargo en el grupo de Cirujano 3 (alumnos) el 87.5 % de los pacientes se encontraban en hiperglucemia durante la etapa 3 de la cirugía. Los resultados por American Society of Anesthesiologists (ASA) mostraron un mayor porcentaje de hiperglucemia en los ASA 2. Se comprobó la existencia de hiperglucemia en los procedimientos quirúrgicos de la Sala de Cirugía de Animales Menores y Clínica veterinaria recomendándose realizar un monitoreo más exhaustivo de constantes vitales en los diferentes procedimientos quirúrgicos y clasificaciones ASA, empleando las diferentes fichas de monitoreo incluyendo la medición de dolor.At present it is difficult to find that glucose is monitored before, during and after a surgical procedure to check the status of animal stress and the level of pain experienced; both episodes lead to hyperglycemia boxes; This response occurs in the patient after a trauma or injury, which in this case is caused by surgery, causing complications such as dehydration, slow post-surgical recovery, neurological damage and various body tissues. The aim of this study was to determine the percentage of patients with hyperglycemia course of Small Animal Surgery University of Ricardo Palma and Hernandez veterinary clinic in the city of Lima. 25 canine patients undergoing castration of male sex were evaluated between August 2014 and July 2015; to which glucose were evaluated at different stages of surgery with a meter according to a type of surgery. The results were analyzed using SPSS and Microsoft Excel programs. The average results showed that of the 100 glucose measurements obtained 18% was in hyperglycemia, the maximum recorded the 212 mg / dl value; of which one surgeon group (teachers) showed patients in hyperglycemic state; however in the group of Surgeon 3 (students) 87.5% of patients were in stage 3 hyperglycemia during surgery. The results for American Society of Anesthesiologists (ASA) showed a higher percentage of hyperglycemia in ASA 2. the existence of hyperglycemia was found in surgical procedures for the Board of Surgery and Small Animal Veterinary Clinic recommending a more comprehensive monitoring of constant vital in different surgical procedures and ASA classifications using different tabs monitoring including measurement of pain.

A Low-fat Diet Containing Heated Soybean Oil Promotes Hyperglycemia in C57BL/6J Mice Introduction: The metabolic effects of consuming mildly oxidized lipids as compared to highly oxidized lipids are not well documented. Consumption of highly oxidized polyunsaturated fatty acids, even in a low-fat diet, may be a threat to one’s health. Previous studies in our lab have shown that mice consuming soybean oil heated for 3 hours, compared to unheated, gain less body weight, but more fat pad mass, and 3T3-L1 adipocytes treated with soybean oil heated for ≥ 6 hours have abrogated triglyceride accumulation. Another study showed that rats fed highly oxidized oil (heated > 24 hrs), as compared to fresh soybean oil and fish oil, had lower fat pad mass and weight gain but developed glucose intolerance. This suggested that the extent of lipid oxidation determined the subsequent metabolic risk. Objective: Our aim, in the current study, is to investigate if a low-fat diet with soybean oil heated for increasing time points (3h, 6h and 9h) alter fat mass and glucose tolerance. Methods: Six week old, male, C57BL/6J mice were randomly divided into six groups (n=8/group). Three groups were fed a low fat diet with soybean oil heated for 3 (3hr-HO), 6 (6hr-HO), or 9 (9hr-HO) hours for 16 weeks. Another three groups were pair-fed to each of the 3hr-HO, 6hr-HO and 9hr-HO groups with a low-fat diet containing unheated oil (PF-UHO). Food consumption was recorded every 3-4 days, and body weights were recorded weekly. Soy oil in the diets was analyzed for products of oxidation. At 16 weeks, blood glucose levels were measured after a 6 hour fast; fat pad and liver weights were recorded, and blood was collected by cardiac puncture for serum insulin analysis. Results: Final weight gain was not significantly different between all HO groups as a percent of their respective PF groups (p >0.05). The feeding efficiency for 3hr, 6hr, and 9hr-HO groups as a percent of PF was 92.75, 113.02, and 111.28, respectively. Mean weights of all fat pads for HO groups decreased with heating time as a percent of PF, although these differences were not statistically significant. Blood glucose was lowest in the 3hr-HO group and significantly increased from 3hr-HO group to 6hr-HO group (p=0.021) as a percent of PF. Serum insulin levels decreased for the HO groups as heating time increased, although these differences were not statistically significant. Conclusion: Consuming a diet with increasing amounts of oxidized lipids decreased fat pad mass and insulin levels, while increasing fasting glucose levels. This paradoxical relationship between increased glucose in the presence of decreased insulin in c57BL/6J mice could be due to either reduced insulin secretion or increased insulin resistance. Further research in our lab will aim to analyze triglyceride accumulation in the liver and muscle cells of these mice to determine if oxidized lipids promote ectopic fat deposition.

Σκοπός. Yπεργλυκαιμία του stress (ΥΣ) χαρακτηρίζεται η παρουσία υπεργλυκαιμίας σε μη διαβητικούς ασθενείς σε παρουσία διαφόρων παραγόντων στρες όπως τραύμα, έγκαυμα, χειρουργείο, έμφραγμα μυοκαρδίου και σήψη. Σκοπός της μελέτης είναι η διερεύνηση της υπεργλυκαιμίας του stress σε βαριά σηπτικούς ασθενείς και η επίδρασης της στην παραγωγή προ και αντι- φλεγμονωδών κυτταροκινών όπως IL-6, IL-10, TNF-alpha and TGF-beta 1 και στην τελική έκβαση ασθενών με βαριά σήψη. Μέθοδος. Μελετήσαμε 265 σηπτικούς ασθενείς που εισήχθηκαν σε 3 παθολογικές κλινικές της ΝΔ Ελλάδος στη διάρκεια ενός έτους. Οι ασθενείς χωρίστηκαν σε 3 ομάδες ανάλογα με το γλυκαιμικό τους προφίλ κατά το πρώτο 24ωρο της εισαγωγής τους: ασθενείς με στρες υπεργλυκαιμία (ομάδα ΥΣ,ν=45), με σακχαρώδη διαβήτη (ομάδα ΣΔ,ν=67),και με φυσιολογικές τιμές σακχάρου (ομάδα ΦΓ,ν=153). Ως ΥΣ ορίστηκε η παρουσία γλυκόζης νηστείας 126mg/dl ή τυχαίας τιμής 200 mg/dl σε ≥2 μετρήσεις. Η βαρύτητα της σήψης εκτιμήθηκε με SOFA score. Σε 62 από τους ασθενείς μετρήθηκαν επιπλέον οι κυτταροκίνες TNF-alpha , IL-6 , IL-10 and TGFb-1 μέσα στο πρώτο 24ωρο της εισαγωγής. Αποτελέσματα. Ποσοστό 39.4% των βαριά σηπτικών ασθενών είχαν υπεργλυκαιμία, ενώ 15.3% είχε υπεργλυκαιμία του στρες προκαλούμενη από σήψη. Δεν παρατηρήθηκε κληρονομικό ιστορικό σακχαρώδη διαβήτη στην ομάδα ΥΣ. Υψηλότερο ποσοστό ασθενών με υπεργλυκαιμία του στρες απεβίωσε συγκριτικά με αυτούς με φυσιολογικές τιμές γλυκόζης (42.5 % vs 13.6 %) και αυτούς με σακχαρώδη διαβήτη 42.5 % vs 24.6%). Η ομάδα ΣΔ είχε χειρότερη πρόγνωση από την ομάδα ΦΓ (24.6% vs 13.6 %). Θετική συσχέτιση παρατηρήθηκε μεταξύ των τιμών γλυκόζης αίματος νηστείας των ομάδων ΣΔ και ΥΣ και τη βαρύτητα της σήψης όπως εκφράζεται από το SOFA score. Η ομάδα ΥΣ είχε υψηλότερο SOFA score και επίπεδα IL-6 και IL-10 από τις ομάδες ΣΔ και ΦΓ. Είχε επίσης υψηλότερα επίπεδα TNF-alpha από την ομάδα ΣΔ αλλά όχι από την ομάδα ΦΓ. Δεν παρατηρήθηκε διαφορά στα επίπεδα TGFb-1μεταξύ των 3 ομάδων. Οι επιζώντες είχαν υψηλότερες τιμές IL-10 από αυτούς που απεβίωσαν, δεν παρατηρήθηκε διαφορά για IL-6, TNF-alpha, το λόγο IL-10/TNF-alpha και TGFb-1 μεταξύ των 3 ομάδων. Οι τιμές της IL-10, οι μέσες τιμές νηστείας γλυκόζης και η ηλικία ανευρέθηκαν ως προγνωστικοί παράγοντες σχετιζόμενοι με την πρόγνωση. Συμπεράσματα: Η υπεργλυκαιμία συμπεριλαμβανόμενης και της υπεργλυκαιμίας που παρουσιάζεται στα πλαίσια του στρες παρατηρείται συχνά σε ασθενείς με βαριά σήψη. Η υπεργλυκαιμία του στρες φαίνεται να μη σχετίζεται με ανθρωπομετρικά χαρακτηριστικά και να σχετίζεται με βαρύτερη νόσο. Η υπεργλυκαιμία του στρες συνοδεύεται με αυξημένη παραγωγή κυταροκινών και με κακή έκβαση σε ασθενείς με βαριά σήψη. / Aims /hypothesis. Stress hyperglycemia is a medical term referring to elevation of blood glucose levels in the absence of diabetes due to various kinds of stress, like trauma, burn injury, surgery, myocardial infraction and sepsis. The aim of our study was to investigate the clinical and laboratory characteristics of severe septic patients with baseline hyperglycemia and the impact of hyperglycemia on the final outcome. e also studied whether stress hyperglycemia affects the production of the main pro- and anti-inflammatory cytokines and the 28 days hospital mortality in patients with severe sepsis. Methods-Patients: A total of 265 patients admitted with severe sepsis in three major Hospitals in South-Western Greece, during a 1-year period, were included in the study. Patients were divided in three groups according to their glycemic profile at admission: patients with stress hyperglycemia (group SH, n=45), diabetes mellitus (group DM, n=67) and normal glucose level (group NG, n=153). Hyperglycemia was defined as an admission or in-hospital fasting glucose level of ≥126 mg/dl, or a random blood glucose level of ≥ 200mg/dl on ≥ 2 determinations. The serum levels of the cytokines TNF-alpha, IL-6, IL-10 and TGFb-1 were measured in 62 patients with severe sepsis within 24 hours after admission. Results: 39.4% of septic patients had baseline hyperglycemia with 15.3% having sepsis-induced stress hyperglycemia. No family history was noted in the SH group. A higher percentage of septic patients with stress hyperglycemia died compared to patients with normal glucose levels (42.5 % vs. 13.6 %) and diabetics (42.5 % vs. 24.6%). Group DM had also a poorer prognosis than group NG (24.6% vs. 13.6 %). A positive correlation was detected between the fasting blood glucose levels of groups DM and SH and the severity of sepsis indicated by SOFA score. Group SH had higher SOFA score and levels of IL-6 and IL-10 than group DM and group NG. It also had higher levels of TNF-alpha than group DM but not group NG. There was no difference in the levels of TGFb-1 among the 3 groups. Survivors had higher levels of IL-10 than no survivors, no difference was detected for IL-6, TNF-alpha, IL-10/TNF-alpha ratio and TGFb-1. Interleukin-10 values, mean fasting glucose values and age were found as prognostic factors associated with outcome. Conclusions: Baseline hyperglycemia, including stress induced hyperglycemia is common in patients with severe sepsis. Stress induced hyperglycemia seems not to be related with anthropometric characteristics and is related to a more severe disease. Stress hyperglycemia is associated with increased cytokine production and an adverse clinical outcome in patients with severe sepsis.

Type 2 diabetes mellitus (T2DM) is often characterized with hyperglycemia, delayed gastric emptying time and a blunted response of gut hormones during feeding that may modulate hunger and satiety. We hypothesized that poor diabetes control is associated with greater hunger suppression, satiation and satiety than good control. We studied 9 T2DM men, after an overnight fast and in response to a 689 kcal mixed meal, twice with or without oral antihyperglycemic agents in a crossover design. Untreated, subjects had higher fasting and postprandial glucose, showed prolonged gastric emptying time and higher thermic effect of food; all factors associated with hunger suppression. Treated, glycemia decreased by 24% and postprandially GLP-1 and PYY3-36 , peptides associated with hunger suppression, were higher than without medication. Thus, no differences were observed in satiation scores between studies. However food intake from a buffet offered 5 hours post meal, an index of satiety, related to hunger scores only in the study with medication.

Critically ill patients are known to experience stress-induced hyperglycemia. Inhibiting the physiological response to increased glycaemic levels in these patients are factors such as increased insulin resistance, increased dextrose input, absolute or relative insulin deficiency, and drug therapy. Although hyperglycemia can be a marker for severity of illness, it can also worsen outcomes, leading to an increased risk of further complications. Recent studies have shown that tight control can reduce mortality up to 43%. Metabolic modelling has been used to study physiological behaviour and/or to control glycaemia for a long time and many successful approximate system models have been developed. Due to the malfunction of medical equipments, clinical measurements obtained usually come with noise. In addition, the few such systems currently available can have errors in excess of 20-30%. Therefore, to fully simulate the clinical data, the system model also needs to couple with a successful noise model. This research has developed a new noise model that better fits the current available statistical description of the noise profile and therefore can be applied to achieve better simulation results. The research also designed a filter algorithm that is capable of reducing the sensor measurement error down to an acceptable value. Achieving such a goal is a significant step towards fully automated adaptive control of hyperglycaemia in critically ill patients and would therefore reduce mortality.

Diabetes mellitus is a systemic condition that remains undiagnosed in a large portion of the population, which presents potential challenges for implant-based rehabilitation. Currently, the effects of diabetes on bone healing are not fully understood. Thus prior to employing a model of diabetes in studies of peri-implant healing, it was important to investigate the temporal effects of hyperglycemia on bone healing. Bone healing of femoral drill-defects was compared between streptozotocin-induced hyperglycemic (DB) and normoglycemic (NDC) rats at 5, 10, 15, and 30 days P.O. Quantitative assessment of bone samples using μCT demonstrated a delay in bone formation occurring up to 10 days in DB animals. Histological assessment confirmed these quantitative findings. Additionally, fluorescently stained bone samples indicated possible defects in mineralization of bone in DB group. In summary hyperglycemia affects bone healing at the early stages of bone formation, concurrent with the osteoconduction phase of bone healing.

Diabetes mellitus is a systemic condition that remains undiagnosed in a large portion of the population, which presents potential challenges for implant-based rehabilitation. Currently, the effects of diabetes on bone healing are not fully understood. Thus prior to employing a model of diabetes in studies of peri-implant healing, it was important to investigate the temporal effects of hyperglycemia on bone healing. Bone healing of femoral drill-defects was compared between streptozotocin-induced hyperglycemic (DB) and normoglycemic (NDC) rats at 5, 10, 15, and 30 days P.O. Quantitative assessment of bone samples using μCT demonstrated a delay in bone formation occurring up to 10 days in DB animals. Histological assessment confirmed these quantitative findings. Additionally, fluorescently stained bone samples indicated possible defects in mineralization of bone in DB group. In summary hyperglycemia affects bone healing at the early stages of bone formation, concurrent with the osteoconduction phase of bone healing.