Au cours des dernières décennies, les modifications comportementales ont conduit à une forte augmentation de la prévalence des maladies métaboliques comme l’obésité et le diabète de type 2. L’hyperglycémie chronique associée à ces maladies a des effets délétères sur de nombreux tissus, entraînant de graves complications (glucotoxicité). Parmi les différents mécanismes impliqués dans les effets toxiques du glucose, la O-N-acétylglucosaminylation (O-GlcNAc) des protéines joue un rôle très important. La O-GlcNAcylation est une modification post-traductionnelle réversible qui régule l'activité des protéines cytosoliques et nucléaires en fonction de la disponibilité en glucose. Deux enzymes seulement, l’O-GlcNAc transférase (OGT) et l’O-GlcNAcase (OGA), ajoutent ou retirent le groupement GlcNAc sur les protéines, respectivement. Cette modification dépend étroitement de la disponibilité en glucose et de son flux à travers la voie de biosynthèse des hexosamines (HBP). Les maladies métaboliques comme le diabète et l'obésité se caractérisent par ailleurs par une inflammation chronique à bas bruit, qui participe également aux complications observées dans ces pathologies. Les perturbations métaboliques associées à ces maladies (augmentation des acides gras libres et du glucose) favorisent les processus pro-inflammatoires dans le macrophage. Cependant, les relations entre processus inflammatoires et O-GlcNAcylation des protéines restent peu explorées. Le but de ce travail était d’évaluer l’implication de la voie de O-GlcNAcylation dans le macrophage. Les études ont été réalisées en utilisant la lignée de macrophages murins RAW264.7 ou des macrophages, différenciés à partir de moelle osseuse de souris, des macrophages péritonéaux de souris ou des macrophages différenciés à partir à de monocytes humains. La O-GlcNAcylation des protéines a été mesurée dans les macrophages RAW264.7 à l’aide d’un biosenseur BRET adressé dans différents compartiments cellulaires. Nous avons observé que l'activation du Toll-like receptor (TLR) 4 par le LPS (lipopolysaccharide) augmente le signal de BRET à la membrane plasmique, dans le cytosol et le noyau des cellules RAW264.7. Une augmentation de la O-GlcNAcylation induite par le LPS était également observée par western blotting dans les cellules RAW264.7 et dans les macrophages primaires de souris et humains. Cette augmentation de O-GlcNAcylation était en particulier détectée sur la sous-unité p65 du facteur de transcription NFκB, suggérant un rôle de cette modification dans les effets pro-inflammatoires du LPS. En accord avec cette notion, l’inhibition de l’OGA, (responsable de la dé-GlcNAcylation des protéines) à l’aide d’un inhibiteur spécifique (Thiamet G), potentialise les effets du LPS sur l’expression des ARNm de la cytokine pro-inflammatoire IL1β. Nous avons en outre généré un modèle de souris OGT-KO inductible au tamoxifène. L’invalidation de l’OGT dans les macrophages primaires de ces souris réduit l’effet du LPS sur l’expression des ARNm de l’IL1β d’un facteur 2, suggérant que l’induction de la O-GlcNAcylation est au moins en partie impliquée dans la transmission des effets pro-inflammatoires du LPS. Afin d’élucider les mécanismes responsables de l’augmentation de O-GlcNAcylation induite par le LPS, nous avons étudié, dans les cellules RAW264.7, l’effet du LPS sur les enzymes impliquées dans la voie O-GlcNAc. Nous avons observé que le traitement au LPS n’a pas d’effet sur l’expression (ARNm et protéine) et sur les activités enzymatiques de l’OGT et de l’OGA. En revanche, le LPS induit une forte augmentation de l’expression (ARNm et protéine) de la glutamine : fructose-6-phosphate amidotransférase (GFAT), enzyme limitante de la voie HBP. (...) / In the last decades, changes in lifestyle have led to a dramatic increased prevalence of pathologies such as obesity and type 2 diabetes. Chronic hyperglycaemia associated with these diseases has deleterious effects on many tissues, resulting in serious complications (glucotoxicity). Among the different mechanisms involved in the toxic effects of glucose, O-N-acetylglucosaminylation (O-GlcNAc) of proteins plays an important role. O-GlcNAcylation is a reversible post-translational modification that regulates the activities of cytosolic and nuclear proteins. Only two enzymes, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA), control the level of O-linked N-acetyl glucosamine (O- GlcNAc) on proteins. OGT is the enzyme that O-GlcNAcylates proteins, whereas OGA removes O-GlcNAc from proteins. This post-translational modification tightly depends on glucose availability and its flux through the hexosamines biosynthesis pathway (HBP). Metabolic diseases such as diabetes and obesity are also characterized by chronic low level inflammation, which contributes to the complications observed in these pathologies. The metabolic disturbances associated with these diseases (increased free fatty acids and glucose) promote pro-inflammatory processes in the macrophage. However, the relationships between inflammatory processes and O-GlcNAcylation of proteins remain poorly explored. The aim of this work was to evaluate the involvement of the O-GlcNAcylation pathway in the macrophage. The studies were carried out using the RAW264.7 mouse macrophage cell line or primary macrophages differentiated from mouse bone marrow (BMDM), peritoneal mouse macrophages, or human monocytes derived macrophages (hMDM). O-GlcNAcylation of proteins was measured in RAW264.7 macrophages using a BRET biosensor targeted to different cell compartments. We have observed that activation of Toll-like receptor (TLR) 4 by LPS (lipopolysaccharide) increases the BRET signal at the plasma membrane, in the cytosol, and nucleus of RAW264.7 cells. An increase in LPS-induced O-GlcNAcylation was also observed by western blotting in RAW264.7 cells and primary mouse and human macrophages. This increase in O-GlcNAcylation was in particular detected on the p65 subunit of the NFκB transcription factor, suggesting a role for this modification in the pro-inflammatory effects of LPS. In agreement with this notion, inhibition of OGA, (responsible for de-GlcNAcylation of proteins) using a specific inhibitor (Thiamet G) potentiates the effects of LPS on mRNA expression of the pro-inflammatory cytokine IL1β. In addition, we generated a tamoxifen-inducible OTG-KO mouse model. Invalidation of OGT in primary macrophages of these mice reduces the effect of LPS on the expression of IL1β mRNAs by a factor of 2, suggesting that the induction of O-GlcNAcylation is at least in part involved in the transmission of the pro-inflammatory effects of LPS. In order to elucidate the mechanisms responsible for LPS-induced increase in O-GlcNAcylation, we studied the effect of LPS on the enzymes involved in the O-GlcNAc pathway in RAW264.7 cells. We observed that LPS treatment had no effect on the expression (mRNA and protein) and on the enzymatic activities of OGT and OGA. On the other hand, LPS induced a strong increase in the expression (mRNA and protein) of glutamine: fructose-6-phosphate amidotransferase (GFAT), the rate limiting enzyme of the HBP pathway. Inhibition of GFAT with 6-Diazo-5-oxo-L-norleucine (DON) inhibited LPS-induced increase in O-GlcNAcylation and LPS effect on the expression of IL1β mRNA, but not NOS2 expression, confirming a partial dependence of the pro-inflammatory effects of LPS on the O-GlcNAc pathway. In conclusion, our results indicate that activation of the O-GlcNAcylation pathway could be considered as an integral part of the signal induced by TLR4, and suggest that this pathway is involved in some of the pro-inflammatory effects of LPS in the macrophage.

A doença arterial coronária (DAC) tem sido a maior causa de morte por doenças nos paises ocidentais. Existem vários fatores responsáveis pela iniciação e progressão desta doença - fatores ambiental ou genético. Muitos fatores de risco para a DAC estão relacionados a alterações do metabolismo lipídico, como acúmulo de lipoproteína de baixa densidade (LDL) no plasma seguida da deposição da lipoproteína na parede arterial. Recentemente, foi demonstrado que uma emulsão rica em colesterol que se assemelha a composição lipídica da LDL liga-se aos receptores que captam a lipopoteína da circulação e internalizam-na no citoplasma. A emulsão, denominada LDE, é feita sem proteína mas quando injetada na circulação sangüínea adquire várias apolipoproteínas (apo) como apo E que pode ser reconhecida pelo receptor de LDL. A apo E tem mais afinidade pelo receptor do que a apo B, a apo que liga a LDL nativa ao receptor. No presente estudo, para esclarecer o processo metabólico que a LDL enfrenta no plasma e o processo de captação da lipoproteína pelos vasos, a LDE marcada com colesterol livre-3H (CL) e oleato de colesterol-14C (CE) foi injetada em 10 pacientes portadores de DAC (57 ± 2,2 anos) submetidos à cirurgia de revascularização miocárdica. Amostras de sangue foram coletadas em intervalos de tempo pré-determinados. A radioatividade presente nas alíquotas de plasma foi determinada por cintilação líquida e a taxa fracionai de remoção (TFR) calculada por análise compartimental. Os fragmentos dos enxertos de aorta, artérias radial e torácica interna, veia safena e pericárdio removidos durante o procedimento cirúrgico foram coletados para extração lipídica, separação por cromatografia de camada delgada e quantificação radioativa. A remoção plasmática do CE da LDE foi similar a do CL da LDE (0,0617 ± 0,0087 vs 0,0528 ± 0,0123, p = 0,5635, respectivamente). A captação do CL da LDE foi maior do que a do CE da LDE na aorta (21% vs 3,1%, p = 0,0049), artéria torácica interna (10,3% vs 2%, p = 0,0007) e veia safena (8% vs 2%, p = 0,0326). Nos fragmentos de artéria radial (14,4% vs 4,3%) e de pericardio (2,2% vs 0,3%), a captação do CL tendeu ser maior do que a captação do CE, porém não foi estatisticamente confirmado. A taxa de esterificação foi maior nos fragmentos de aorta, de toráxica interna e de pericárdio do que nos fragmentos de veia safena (p < 0,001). Concluindo, a LDE foi captada pelos vasos e pericardio em quantidades concideráveis e a captação do CL pelos tecidos foi maior do que a do CE. Ainda, a taxa de esterificação do colesterol livre foi mais intensa nos fragmentos de aorta e torácica interna do que nos fragments de veia safena. / Coronary artery disease (CAD) is the main mortality cause in western countries. There are many factors responsible for the onset and progression of the disease - either environmental or genetic. Many risk factors in CAD are related with disorders of lipid metabolism, such as accumulation of low-density lipoprotein (LDL) in the plasma with deposition of the lipoprotein in the arterial wall. Recently, it was shown that a cholesterol-rich emulsion that mimics the lipid composition of LDL binds to the receptors that take-up the lipoprotein from the circulation and internalizes it into the cytoplasm. The emulsion, denominated LDE, is made without protein but when injected into the bloodstream it picks-up several apolipoproteins (apo) such as apo E that can be recognized by the LDL receptor. Apo E has even more affinity for the receptor than apo B, the apo that binds native LDL to the receptors. In the current study, aiming to clarity the metabolic processes that LDL undergoes in the plasma and the process of lipoprotein uptake by the vessels, LDE labeled with 3H- Cholesterol (CL) and 14C-Cholesteryl Oleate (CE) was injected into 10 CAD patients (57 ± 2,2 yr.) scheduled to be submitted to myocardial revascularization surgery. Blood samples were collected over 24 hour at pre-established intervals. Radioactivity present in plasma aliquots was determined in a scintillation solution and the fractional clearance rate (FCR) was calculated by compartimental analysis. The gratt\'s fragments of aortic, radial, internal thoracic arteries, safenous vein and pericardium discarded during the surgical procedure were collected for lipid extraction, separation by thin layer chromatography and radioactive counting. The removal from plasma of the LDE CE was similar to that of the LDE CL (0,0617 ± 0,0087 vs 0,0528 ± 0,0123, p = 0,5635, respectively). The uptake of LDE CL was greater than that of LDE CE in aorta (21% vs 3,1%, p = 0,0049), internal toracic artery (10,3% vs 2%, p = 0,0007) and safenous vein (8% vs 2%, p = 0,0326). In the radial artery (14,4% vs 4,3%) and pericardium (2,2% vs 0,3%) fragments, the CL uptake also tended to be greater than that of CE, but this was not statistically confirmed. The esterification rate was greater in the aorta, internal thoracic artery and pericardium fragments than in safenous vein fragments (p < 0,001). In conclusion, LDE was taken-up by vessels and pericardium at considerable amounts and LDE CL uptake by those tissues was greater than that of CE. In addition, the cholesterol esterification rate was more intense in the aorta and internal thoracic artery than in venous fragments.

This thesis describes biochemical and molecular methods diagnostic for a spectrum of steroid metabolic diseases. Deficiency of any enzyme in the steroid hormone biosynthetic pathways leads to disorders including congenital adrenal hyperplasia, while some cause disorders of sex development (DSD). A gas chromatography mass spectrometry-based analytical technique called urinary steroid profiling (USP) has been shown to be a useful diagnostic test for these diseases and for steroid-secreting adrenocortical tumours. To test the hypothesis that this approach would be effective in our local population, we interpreted 482 USP results using reference intervals set up from 371 local healthy subjects. Characteristic steroid metabolite excretion patterns were found in 39 patients, including 21 patients with 21-hydroxylase deficiency (21OHD) where there were grossly increased 17-hydroxyprogesterone metabolites, 12 patients with 5α-reductase 2 deficiency (5ARD) with extremely low 5α- to 5β-reduced steroid metabolite ratios, and five patients with adrenocortical carcinoma with markedly raised tetrahydro-11-deoxycortisol and 3,16,20-pregnenetriols levels. / The genetic basis of 21OHD in various populations is mainly due to conversion between the CYP21A2 and the CYP21A1P genes but this has not yet been explored in our population. By using DNA sequencing and multiplex ligation-dependent probe amplification, 74 mutations were found in 35 patients with 21OHD. Gross deletion/conversion of the CYP21A2 gene accounted for 27%. c.290-13A/C>G was the most common point mutation (27%), followed by p.Ile172Asn (17.6%). One novel mutation c.1367delA was also detected. Their prevalence in our patients differs from those in other populations. / The most common cause of 46,XY DSD in Western populations is androgen insensitivity syndrome (AIS) but this has not been verified locally. A prospective study was conducted where 64 patients were recruited for comprehensive hormonal profiling and targeted molecular analysis. In this study, a genetic diagnosis was established in 22 patients, with 5ARD being the most common disease, followed by AIS. Traditionally the diagnosis of 5ARD relies on measuring dihydrotestosterone. However, with our experience in diagnosing this condition based on USP and mutational analysis of the SRD5A2 gene, two new diagnostic algorithms for 46,XY DSD were proposed where dihydrotestosterone is not required. / In vitro study is the preferred method for characterising the function of novel genetic variants. However, clinical laboratories rarely have the facilities and resources for it. In silico prediction programmes appear to be practical alternatives but their performance on testing non-synonymous variants in genes related to steroid metabolism has not been verified. Three web-based in silico prediction programmes, namely Sorting Intolerant From Tolerant, PolyPhen-2 and Pathogenic-Or-Not-Pipeline, were tested by analysing 797 published non-synonymous genetic variants in 12 genes related to steroid metabolism. The results of in vitro functional study and/or clinical phenotype were used as gold standards. The performance of these three programmes were: sensitivity (76.6%, 84.1%, 70.0%), specificity (56.6%, 56.3%, 89.4%) and accuracy (70.1%, 75.2%, 76.8%), respectively. / In conclusion, USP is a valuable biochemical phenotyping technique that helps to select patients for subsequent genetic confirmation. Since the mutation spectrum of 21OHD and the aetiological basis of 46,XY DSD in our population differ from the others, laboratory diagnostic algorithms and molecular analytical strategies must be adjusted accordingly. / Chan, On Kei Angel. / Thesis (M.D.) Chinese University of Hong Kong, 2014. / Includes bibliographical references (leaves 250-269). / Appendixes includes Chinese.

Introdução: Beribéri é uma doença causada pela deficiência de tiamina (vitamina B1) que, apesar de facilmente tratável, pode levar a óbito. Há pelo menos setenta anos não se tinha referência de surtos de beribéri no Brasil. Objetivo: Descrever o perfil epidemiológico dos casos de beribéri e óbitos notificados no Estado do Maranhão. Método: Estudo descritivo-retrospectivo que analisou 1.207 casos notificados de beribéri e 40 óbitos ocorridos no Maranhão no período de 2006 a 2008. As informações foram obtidas do banco de dados da Vigilância Epidemiológica e fichas de notificação. Resultados: Os casos distribuíram-se em 434 (36,0%) em 2006, 551 (45,6%) em 2007 e 222 (18,4%) em 2008. Afetou dois terços das Unidades Regionais de Saúde e 26,3% dos municípios localizados nas regiões centro-oeste, noroeste e sudoeste do Estado. As notificações foram maiores nos meses de março a agosto, com pico em junho. O coeficiente de incidência no período foi de 4,32/10.000 hab. Homens foram mais acometidos (81,9%), com concentração de casos na faixa etária de 20-40 anos (57,0%). Predominou a ocorrência do beribéri seco (84,6%) e o tempo decorrido entre os primeiros sintomas e a notificação foi inferior a três meses para dois terços dos casos. A hospitalização ocorreu para 50% dos casos em 2006, 30% em 2007 e 15% em 2008. O consumo de álcool foi referido por 53,2% dos acometidos em 2006, com proporção discretamente menor nos anos subseqüentes, e o hábito de fumar por cerca de um terço dos acometidos. Sintomas mais comuns foram: diminuição da força, dormência e edema das pernas, dificuldade para caminhar e dor na panturrilha. Em relação à escolaridade, constatou-se que mais da metade (57,2%) tinha quatro anos ou menos de estudo, dois terços (66,2%) desempenhava atividade laboral pesada e 72,9% tinham renda familiar inferior a um salário mínimo. Foram registrados 40 óbitos (3,3% do total de casos notificados) apenas em 2006, concentrados no mês de junho (61,9%) e distribuídos em 21 municípios (9,7%). A taxa de mortalidade para o Estado foi de 0,45/10.000 hab. Praticamente a totalidade dos óbitos ocorreu no sexo masculino (97,5%), sendo 72,5% na faixa etária de 20-30 anos. Para dois terços, o tempo decorrido entre os primeiros sintomas e a notificação foi inferior a três meses e a hospitalização ocorreu para 42,5%. Hábito de consumir álcool e fumar foi elevado entre aqueles que foram a óbito, 75,0% e 66,7%, respectivamente. Conclusão: O estudo se destaca por abranger análise de todas as notificações de beribéri do estado do Maranhão. Constatou-se que os casos e óbitos apresentaram distribuição espacial e temporal relacionado à realidade do estado. A ausência de óbitos em 2007 e 2008 e redução dos casos em 2008 sugerem efeitos positivos das ações de enfretamento governamentais implementadas. Esta avaliação descritiva é importante para nortear as atividades e áreas a serem priorizadas no planejamento das ações de combate, tanto no nível central (federal, estadual e municipal), como para os profissionais da atenção básica e especializada, no direcionamento de suas práticas. Os resultados contribuem também para o delineamento de outras pesquisas que possam auxiliar no estudo da determinação do beribéri no Brasil. / Introduction: Beriberi is a disease caused by a deficiency of thiamine (vitamin B1) which, although easily treatable, may lead to death. There had been no register of beriberi outbreaks in Brazil for at least seventy years. Objective: To describe the epidemiological profile of reported beriberi cases in the State of Maranhão. Method: A descriptive retrospective study of 1,207 reported beriberi cases and 40 deaths in the State of Maranhão from 2006 to 2008. The information was obtained from the database of Epidemiological Surveillance and the reporting forms. Results: The cases were distributed in 434 (36.0%) in 2006, 551 (45.6%) in 2007 and 222 (18.4%) in 2008. It affected two thirds of the Regional Health Units and 26.3% of the municipalities located in the Central-Western, Northwestern and Southwestern of the State. The notifications were higher in the months from March to August, peaking in June. The incidence rate in the period was 4.32 / 10,000 inhabitants. Men were more affected (81.9%), with more concentration of cases in the age group 20-40 years (57.0%). Dry beriberi was predominant (84.6%) and the elapsed time between first symptoms and reporting was less than three months in two thirds of cases. Hospitalization occurred for 50.0% of cases in 2006, 30.0% in 2007 and 15.0% in 2008. Alcohol consumption was reported by 53.2% of patients in 2006 and slightly lower proportion in subsequent years, and smoking was reported for about one-third of patients. Main symptoms were loss of strength, numbness and swelling of the legs, difficulty in walking and calf pain. Regarding education it was found that more than half (57.2%) had four or fewer years of study, two thirds (66.2%) performed heavy labor activity and 72.9% had a family income lower than minimum wage. Forty deaths were recorded (3.3% of all reported cases) only in 2006, concentrated in the month of June (61.9%) and distributed in 21 municipalities (9.7%). The mortality rate for the State was 0.45 per 10,000 inhabitants. Almost all the deaths occurred in males (97.5%), and 72.5% aged 20-30 years. For two-thirds of the patients who died, the elapsed time between first symptoms and reporting was less than three months and hospitalization occurred in 42.5%. Alcohol consumption and smoking were higher among those who died, 75.0% and 66.7% respectively. Conclusion: This study stands out for including analysis of all reported beriberi cases in the State of Maranhão. It was found that cases and deaths showed spatial and temporal distribution related to the social reality of the State. The absence of deaths in 2007 and 2008 and reduction of cases in 2008 suggest positive effects of the implemented governmental actions. This descriptive evaluation is important to guide the activities and areas to be prioritized in the planning of actions at the central level (federal, state and municipal), as well as for guiding the practice of primary and specialized healthcare professionals. The results also contribute to the design of other research that may support the study of beriberi in Brazil.

Background: The association of asthma and obesity in youth is well studied. However, much of this research has been observational and the temporal relationship between the two factors remains the subject of debate. Moreover, there are mixed findings regarding the role of physical activity in the association between asthma and obesity in youth. Methods: A systematic review of existing literature of the relationship between asthma and obesity in youth, while examining the role of physical activity as a mediator, was conducted using PubMed and Medline databases. Studies conducted on children and adolescents aged 0 to 18 years, published in English during the period of 2000 to 2014 were included. A comprehensive search yielded 143 studies in PubMed and 133 studies in Medline databases. Of these, 76 studies met the review’s eligibility criteria. The Strengthening the Reporting of Observational Studies in Epidemiology guidelines were consulted to evaluate quality of selected citations. Results: The association between asthma and obesity in youth is well explored. However, there are varying hypotheses and conclusions regarding this relationship, of which temporality remains to be elucidated. Existing evidence was identified to support the mediation of this association by physical activity. Negative self- or parental- perception of exercise ability due to asthma symptoms secondary to physical exertion was identified as a determinant of physical activity in asthmatic youth. Also, control of asthma symptoms was found to directly impact daily physical activity levels. Greater odds of increased weight were observed among asthmatic children compared to non-asthmatic peers. However, annual weight gain was found to be similar among asthmatic children with well controlled symptoms to that of their healthy peers. Lastly, the review found that adverse respiratory symptoms secondary to exercise are experienced by 90% of asthmatic youth, resulting in activity limitation. According to this finding, it is reasonable that the incidence of obesity frequently observed to be greater in asthmatic youth than healthy peers is mediated by decreased expenditure of energy in asthmatic children due to reduced physical activity. Conclusions: It is likely that physical activity mediates the relation between asthma and obesity, though further research is required to determine the temporal relation and degree of this association.

PURPOSE: The association of asthma and overweight in youth is well studied. However, the temporal relationship between asthma and overweight, the strength of their association, and mediating factors involved in this relationship remain unclear. This review investigates the relationship between asthma and overweight in youth, while examining the role of physical activity as a mediator. METHODS: A systematic review of literature was conducted using PubMed and Medline databases. Studies conducted among youth aged 0-18 years, published in English between 2000-2014 were included. The Strengthening the Reporting of Observational Studies in Epidemiology guidelines were consulted to evaluate quality of selected citations. RESULTS: A comprehensive search yielded 143 studies in PubMed and 133 studies in Medline databases. Of these, 75 studies met the eligibility criteria. The review found varying hypotheses regarding the temporal relationship between asthma and overweight in youth; existing evidence supports the mediation of this association by decreased expenditure of energy due to reduced physical activity. Negative self-perception or parental perception of exercise ability due to asthma symptoms secondary to physical exertion was identified as a determinant of physical activity in asthmatic youth. CONCLUSIONS: Physical activity likely mediates the relationship between asthma and overweight in youth. Temporality of this relationship remains unclear.

No description available.

No description available.

Background C1q TNF related protein 3 (CTRP3) is a relatively novel hormonal factor primarily derived from adipose tissue and has anti-diabetic properties. To determine if CTRP3 could play a role in early childhood development, the purpose of this study was to establish the presence of CTRP3 in breast milk (BM) and to determine whether CTRP3 levels were correlated with pregravid obesity status of the mother. Methods Breast milk was collected from breast-feeding mothers who had a pregravid body mass index (BMI) classification of normal weight (BMI 18–25 kg/m2, n = 23) or obese (BMI > 30 kg/m2, n = 14). Immunoprecipitation followed by immunoblot analysis confirmed the presence of CTRP3 in BM. The concentration of CTRP3 in BM samples was determined by ELISA. Additional bioactive components were also measured by commercially available assays: ghrelin, insulin, leptin, adiponectin, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and glucose. Bioactive components in normal weight and obese mothers were compared using unpaired t-test (parametric) and Mann–Whitney U-test (non-parametric), as appropriate. Results The primary findings of this study are that the adipokine CTRP3 is present in BM and CTRP3 levels are increased with pregravid obesity. Additionally, this study independently confirmed previous work that BM from obese mothers has a higher concentration of insulin and leptin. Further, no differences were observed in BM between obese and normal weight mothers in ghrelin, adiponectin, IL-6, TNF-α, or glucose levels. Conclusion This study identified a novel factor in BM, CTRP3, and showed that BM CTRP3 levels higher in obese mothers. Because of the purported insulin sensitizing effect of CTRP3, it is possible that the elevated levels of CTRP3 in the BM of obese mothers may offset negative effects of elevated leptin and insulin levels in the BM of obese mothers. Future studies will need to be conducted to determine the relevance of CTRP3 in BM and to examine the presence of other adipose tissue-derived hormonal factors.

Abstract available in The FASEB Journal.