Development of an isotope dilution mass spectrometry assay for the quantification of insulin based on signature peptide analysis.

An isotope dilution mass spectrometry (IDMS) method that involves peptide-based protein analysis was developed to accurately quantify insulin. In this study, a signature peptide (GFFYTPK) obtained from tryptic digestion of insulin was selected as a surrogate for insulin. Then, the optimal conditions for signature peptide analysis through mass spectrometry detection and enzymatic digestion were determined. The analytical performance of this method was assessed and validated using porcine insulin-certified reference material. The linear range of the insulin calibration curve ranged from 0.05 ~ 2 mass ratios, with recoveries ranging from 96.15 to approximately 101.15%. The limit of detection was 0.19 ng/mL, and the limit of quantification was 0.63 ng/mL. The quantitative results corresponded well with a certified value that was obtained from measuring a porcine insulin reference material with amino acid-based IDMS. In addition, the target peptide GFFYTPK can be found in other species of insulin. This method was also applied for the quantification of human insulin-certified reference material. Finally, we applied the method to quantify the concentrations of simulated serum insulin. These findings suggested that this signature peptide-based IDMS approach can accurately quantify insulin levels, can assign a certified value to insulin reference materials, and has the potential to quantify serum insulin with traceable measurements.

Insulin and aging - a disappointing relationship.

Experimental studies in animal models of aging such as nematodes, fruit flies or mice have observed that decreased levels of insulin or insulin signaling promotes longevity. In humans, hyperinsulinemia and concomitant insulin resistance are associated with an elevated risk of age-related diseases suggestive of a shortened healthspan. Age-related disorders include neurodegenerative diseases, hypertension, cardiovascular disease, and type 2 diabetes. High ambient insulin concentrations promote increased lipogenesis and fat storage, heightened protein synthesis and accumulation of non-functional polypeptides due to limited turnover capacity. Moreover, there is impaired autophagy activity, and less endothelial NO synthase activity. These changes are associated with mitochondrial dysfunction and oxidative stress. The cellular stress induced by anabolic activity of insulin initiates an adaptive response aiming at maintaining homeostasis, characterized by activation of the transcription factor Nrf2, of AMP activated kinase, and an unfolded protein response. This protective response is more potent in the long-lived human species than in short-lived models of aging research resulting in a stronger pro-aging impact of insulin in nematodes and fruit flies. In humans, resistance to insulin-induced cell stress decreases with age, because of an increase of insulin and insulin resistance levels but less Nrf2 activation. These detrimental changes might be contained by adopting a lifestyle that promotes low insulin/insulin resistance levels and enhances an adaptive response to cellular stress, as observed with dietary restriction or exercise.

Simultaneous monitoring of multiple hormones from human islets of Langerhans using solid-phase extraction-mass spectrometry.

Islets of Langerhans release peptide hormones in controlled amounts and patterns to ensure proper maintenance of blood glucose levels. The overall release of the hormones is shaped by external factors and by autocrine and paracrine interactions occurring within the islets. To better understand what controls the secretion of islet-secreted peptides, and how these processes go awry in diabetes, methods to monitor the release of multiple hormones simultaneously are needed. While antibody-based assays are typically used, they are most often applied to quantification of a single hormone. Mass spectrometry (MS), on the other hand, is well suited for quantifying multiple hormones simultaneously but typically requires time-consuming separation steps with biological samples. In this report, response surface methodology was used to identify a set of optimal solid-phase extraction (SPE) conditions for the islet-secreted peptides, insulin, C-peptide, glucagon, and somatostatin. The optimized SPE method was used with multiple reaction monitoring and isotopically labeled standards to quantify secretion levels. Calibrations were linear from 0.5 to 50 nM with < 15% RSD peak area ratios. A microfluidic system was used to perfuse 30 human islets with different glucose conditions, and fractions were collected every 2 min for SPE-MS analysis. Results showed the release dynamics of the individual peptides, as well as patterns, such as positively and negatively correlated release and oscillations. This rapid SPE-MS method is expected to be useful for examining other peptide and small-molecule secretions from islets and could be applied to a number of other biological systems for investigating cellular communication.

Comparative analysis of biochemical, hormonal, and mineral compositions of preovulatory and cystic ovarian follicles in buffalo during the non-breeding season.

This study is a comparative analysis of the biochemical, hormonal, and mineral compositions of follicular fluid in preovulatory and cystic follicles of water buffalo (Bubalus bubalis). In total, reproductive tracts from 215 buffalo along with intact ovaries were collected randomly from an abattoir. The incidence of cystic conditions found in this study was 3.72% (8/215), involving the right ovary in 62.5% of instances and the left ovary in 37.5% of instances during the non-breeding season. Follicular fluid was aspirated from preovulatory follicles (12-15 mm diameter, oestrogen-active, follicular phase or stage IV corpus luteum on one of the two ovaries, n = 10) and cystic follicles (at least 20 mm diameter, no corpus luteum on any one of the two ovaries, n = 8). The follicular fluid samples were assayed for biochemical components (uric acid, creatinine, blood urea nitrogen, cholesterol, total protein, glucose, ascorbic acid, and alkaline phosphatase), hormones (progesterone, estradiol, and insulin), and minerals (calcium, magnesium, phosphorus, copper, zinc, and cobalt). Cystic follicles had greater (P < 0.05) concentrations of creatinine, blood urea nitrogen, cholesterol, progesterone, copper, zinc, and cobalt, and lesser (P < 0.05) concentrations of uric acid, glucose, ascorbic acid, estradiol, insulin, calcium, magnesium, and phosphorus compared with preovulatory follicles. These results indicated the marked differences in follicular fluid composition between preovulatory and cystic follicles in buffalo. Some of the changes were indicative of oxidative stress and disturbed steroidogenesis, two important mechanisms shown to be associated with cystic ovarian disease in various species. Further studies are warranted to investigate whether these differences are directly or indirectly involved in the formation of cystic follicles or are mere manifestations of the condition.

Effects of Exercise Training on Inflammatory and Cardiometabolic Risk Biomarkers in Patients With Type 2 Diabetes Mellitus: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.

BACKGROUND: The interaction between type 2 diabetes mellitus (T2DM) and cardiometabolic morbidity and mortality stems from the progressive nature of inflammation underpinning both diseases. Exercise training is considered an effective treatment strategy for T2DM and cardiometabolic diseases. OBJECTIVE: The current systematic review and meta-analysis investigated the effects of exercise training on inflammatory and cardiometabolic risk biomarkers in patients with T2DM. DATA SOURCES: Electronic databases (PubMed/Medline, Embase, Cochrane Library, CINAHL, Google Scholar, Scopus, and Web of Science) were searched for randomized controlled trials (RCTs) from inception to January 2022. We used random effects models to estimate weighted mean differences with 95% confidence intervals. STUDY SELECTION: Twenty-five RCTs were included (N = 1257 participants; mean age = 52 years). Included studies had moderate to good overall methodological quality (TESTEX = 9 (range 7-13). RESULTS: Meta-analysis indicated that exercise training significantly increased adiponectin and decreased fasting insulin, homeostatic model assessment for insulin resistance, tumor necrosis factor-α, interleukin-6, and C-reactive protein (ps ≤ 0.05). Subgroup analysis by type of training indicated that aerobic exercise had the most consistent beneficial effects as compared to other types of exercise training; however, there was high heterogeneity among studies. CONCLUSION: Different types of exercise training increase adiponectin levels and decrease pro-inflammatory cytokines such as TNF-α, IL-6, and CRP, as well as fasting insulin and insulin resistance markers in patients with T2DM. However, these effects were not beneficial for more commonly measured cardiometabolic risk factors (i.e., lipid profiles). Additional relevant clinical trials are required to confirm these results. TRIAL REGISTRATION: This systematic review and meta-analysis was prospectively registered in the PROSPERO database (CRD42022307396).

Diabetes Mellitus gestacional: perfil epidemiológico de maternidade de alto risco / Gestational Diabetes Mellitus: epidemiological profile of high- risk maternity / Diabetes Mellitus gestacional: perfil epidemiológico de una maternidad de alto riesgo

O Diabetes Mellitus Gestacional é definido como doença que se caracteriza pelos altos níveis de glicemia sanguínea, diagnosticada durante a gestação. Este adoecimento pode acarretar várias complicações maternas e fetais, muitas vezes, necessitando de internamento precoce e cuidados avançados. Objetivou-se caracterizar o perfil epidemiológico de gestantes com diabetes mellitus gestacionais atendidas em serviço de referência. Trata-se de estudo descritivo, documental, retrospectivo, de caráter quantitativo, realizado com gestantes atendidas na maternidade do Hospital Regional do Sudoeste ­ PR, Francisco Beltrão. A amostra foi constituída por 216 gestantes, cujos dados foram coletados dos prontuários das pacientes. Incluíram-se as gestantes atendidas e diagnosticadas com diabetes mellitus gestacional no período de 2020 e com pelo menos um exame de glicose em jejum ou um teste de tolerância oral à glicose para comprovação diagnóstica. Foram exclusas as gestantes dos anos de 2019 e 2021 e oito transferências. A amostra teve maior porcentual do Diabetes mellitus gestacional (90,7%), com prevalência na raça branca (69,9%), faixa etária de 15- 35 anos (68,5%). Ademais,65,7% realizaram controle com dieta e 32,4 % necessitaram realizar o uso de insulina e 51,9%delas eram obesas. A presente pesquisateve considerável relevância, pois permitiu obter perfil epidemiológico de gestantes diagnosticadas com diabetes mellitus, trazendo benefícios, como identificação precocemente da doença, de modo a evitar complicações para gestantes e bebês. PALAVRAS-CHAVE: Gestacional; Diabetes; Prevalência; Maternidade.

Gestational Diabetes Mellitus is defined as a disease characterized by high levels of blood glucose, which is diagnosed for the first time during pregnancy. It can cause several maternal and fetal complications, often requiring early hospitalization and advanced care. The aim of thestudy was to characterize the epidemiological profile of pregnant women with gestational diabetes mellitus seen at a reference service. This is a descriptive, documentary, retrospective, quantitative study, carried out with pregnant women attended at the maternity hospital of the Hospital Regional do Sudoeste - PR in the city of Francisco Beltrão. The sample consisted of216 pregnant women, and data were collected from the patients' medical records. The study included all pregnant women who were attended and diagnosed with GDM in the period described, and who had at least one fasting glucose test or an oral glucose tolerance test for diagnostic confirmation. All pregnant women from the year 2019 and 2021 were excluded fromthe study. The sample had a higher percentage of GDM 90.7% according to race 69.9% werewhite, aged 15-35 years 68 , 5%, while 65.7% performed control with diet and 32.4% neededto use insulin and 51.9% of them were obese. This research had great results because it had an epidemiological profile of pregnant women diagnosed with Diabetes Mellitus, bringing benefitsand thus being able to identify gestational Diabetes mellitus early, aiming to avoid complications for the pregnant woman and the baby.

La diabetes mellitus gestacional se define como una enfermedad caracterizada por niveles elevados de glucosa en sangre, diagnosticada durante el embarazo. Esta enfermedad puede dar lugar a varias complicaciones maternas y fetales, que a menudo requieren una hospitalización temprana y cuidados avanzados. El objetivo es caracterizar el perfil epidemiológico de las gestantes con diabetes mellitus atendidas en el servicio de referencia. Se trata de un estudio descriptivo, documental, retrospectivo, de carácter cuantitativo, realizado con gestantes atendidas en la maternidad del Hospital Regional del Sudoeste - PR, Francisco Beltrão. La muestra estaba formada por 216 mujeres embarazadas, cuyos datos se recogieron de las historias clínicas de las pacientes. Se incluyeron las mujeres embarazadas atendidas y diagnosticadas de diabetes mellitus gestacional en el periodo 2020 y con al menos una prueba de glucosa en ayunas o una prueba de tolerancia oral a la glucosa para su diagnóstico. Se excluyeron las embarazadas de los años 2019 y 2021 y ocho traslados. La muestra tuvo un mayor porcentaje de Diabetes mellitus gestacional (90,7%), con prevalencia en la raza blanca (69,9%), grupo de edad 15-35 años (68,5%). Además, el 65,7% se controlaba con la dieta y el 32,4% necesitaba utilizar insulina y el 51,9% era obeso. La presente investigación tiene una relevancia considerable, ya que permite obtener el perfil epidemiológico de las gestantes diagnosticadas con diabetes mellitus, lo que conlleva beneficios, como la identificación precoz de la enfermedad, a fin de evitar complicaciones para las gestantes y los bebés.

Gain of Function of Malate Dehydrogenase 2 and Familial Hyperglycemia.

CONTEXT: Genes causing familial forms of diabetes mellitus are only partially known. OBJECTIVE: We set out to identify the genetic cause of hyperglycemia in multigenerational families with an apparent autosomal dominant form of adult-onset diabetes not due to mutations in known monogenic diabetes genes. METHODS: Existing whole-exome sequencing (WES) data were used to identify exonic variants segregating with diabetes in 60 families from the United States and Italy. Functional studies were carried out in vitro (transduced MIN6-K8 cells) and in vivo (Caenorhabditis elegans) to assess the diabetogenic potential of 2 variants in the malate dehydrogenase 2 (MDH2) gene linked with hyperglycemia in 2 of the families. RESULTS: A very rare mutation (p.Arg52Cys) in MDH2 strongly segregated with hyperglycemia in 1 family from the United States. An infrequent MDH2 missense variant (p.Val160Met) also showed disease cosegregation in a family from Italy, although with reduced penetrance. In silico, both Arg52Cys and Val160Met were shown to affect MDH2 protein structure and function. In transfected HepG2 cells, both variants significantly increased MDH2 enzymatic activity, thereby decreasing the NAD+/NADH ratio-a change known to affect insulin signaling and secretion. Stable expression of human wild-type MDH2 in MIN6-K8 cell lines enhanced glucose- and GLP-1-stimulated insulin secretion. This effect was blunted by the Cys52 or Met160 substitutions. Nematodes carrying equivalent changes at the orthologous positions of the mdh-2 gene showed impaired glucose-stimulated insulin secretion. CONCLUSION: Our findings suggest a central role of MDH2 in human glucose homeostasis and indicate that gain of function variants in this gene may be involved in the etiology of familial forms of diabetes.

Novel Human Insulin Isoforms and Cα-Peptide Product in Islets of Langerhans and Choroid Plexus.

Human insulin (INS) gene diverged from the ancestral genes of invertebrate and mammalian species millions of years ago. We previously found that mouse insulin gene (Ins2) isoforms are expressed in brain choroid plexus (ChP) epithelium cells, where insulin secretion is regulated by serotonin and not by glucose. We further compared human INS isoform expression in postmortem ChP and islets of Langerhans. We uncovered novel INS upstream open reading frame isoforms and their protein products. In addition, we found a novel alternatively spliced isoform that translates to a 74-amino acid (AA) proinsulin containing a shorter 19-AA C-peptide sequence, herein designated Cα-peptide. The middle portion of the conventional C-peptide contains ß-sheet (GQVEL) and hairpin (GGGPG) motifs that are not present in Cα-peptide. Islet amyloid polypeptide (IAPP) is not expressed in ChP, and its amyloid formation was inhibited in vitro more efficiently by Cα-peptide than by C-peptide. Of clinical relevance, the ratio of the 74-AA proinsulin to proconvertase-processed Cα-peptide was significantly increased in islets from type 2 diabetes mellitus autopsy donors. Intriguingly, 100 years after the discovery of insulin, we found that INS isoforms are present in ChP from insulin-deficient autopsy donors.

Glucose Tolerance after Pancreatectomy: A Prospective Observational Follow-Up Study of Pancreaticoduodenectomy and Distal Pancreatectomy.

BACKGROUND: Effects of pancreatectomy on glucose tolerance have not been clarified, and evidence regarding the difference in postoperative glucose tolerance between pancreaticoduodenectomy (PD) and distal pancreatectomy (DP) is lacking. STUDY DESIGN: This prospective, single-center observational study analyzed 40 patients undergoing PD and 29 patients undergoing DP (Clinical trial registry number UMIN000008122). Glucose tolerance, including insulin secretion (Δ C-peptide immunoreactivity, ΔCPR) and insulin resistance (homeostasis model assessment of insulin resistance, HOMA-IR) were assessed before and 1 month after pancreatectomy using the oral glucose tolerance test (OGTT) and glucagon stimulation test. We assessed long-term hemoglobin A1c (HbA1c) levels in patients, with a follow-up time of 3 years. RESULTS: Percentages of patients diagnosed with abnormal OGTT decreased after PD (from 12 [30%] to 7 [17.5%] of 40 patients, p = 0.096); however, they increased after DP (from 4 [13.8%] to 8 [27.6%] of 29 patients, p = 0.103), although the changes were not statistically significant. ΔCPR decreased after both PD (from 3.2 to 1.0 ng/mL, p < 0.001) and DP (from 3.3 to 1.8 ng/mL, p < 0.001). HOMA-IR decreased after PD (from 1.10 to 0.68, p < 0.001), but did not change after DP (1.10 and 1.07, p = 0.42). Median HbA1c level was higher after DP than after PD for up to 3 years, but the differences were not statistically significant. CONCLUSIONS: In comparisons of pre- and 1 month post-pancreatectomy data, glucose tolerance showed improvement after PD, whereas it worsened after DP. Insulin secretion decreased after both PD and DP. Insulin resistance improved after PD, but did not change after DP. Further studies are warranted to clarify mechanisms of improved insulin resistance after PD.

Cadmium exposure induces cardiac glucometabolic dysregulation and lipid accumulation independent of pyruvate dehydrogenase activity.

CONTEXT: Suppressed glucose metabolism, elevated fatty acid metabolism and lipid deposition within myocardial cells are the key pathological features of diabetic cardiomyopathy. Studies have associated cadmium exposure with metabolic disturbances. OBJECTIVE: To examine the effects of cadmium exposure on cardiac glucose homeostasis and lipid accumulation in male Wistar rats. METHODS: Male Wistar rats were treated for 21 days as (n = 5): Control, cadmium chloride Cd5 (5 mg/kg, p.o.), cadmium chloride Cd30 (30 mg/kg, p.o). RESULTS: The fasting serum insulin level in this study decreased significantly. Pyruvate and hexokinase activity reduced significantly in the Cd5 group while no significant change in lactate and glycogen levels. The activity of pyruvate dehydrogenase enzyme significantly increased with an increasing dosage of cadmium. The free fatty acid, total cholesterol and triglyceride levels in the heart increased significantly with increasing dosage of cadmium when compared with the control. Lipoprotein lipase activity in the heart showed no difference in the Cd5 group but a reduction in the activity in the Cd30 group was observed. CONCLUSION: This study indicates that cadmium exposure interferes with cardiac substrate handling resulting in impaired glucometabolic regulation and lipid accumulation which could reduce cardiac efficiency.

Target-triggered and controlled release plasmon-enhanced fluorescent AIE probe for conformational monitoring of insulin fibrillation.

In this work, we constructed a target-triggered and controlled-release plasmon-enhanced fluorescent AIE probe to realize the purpose of conformational monitoring of insulin fibrillation. We synthesized a novel water-soluble anthracene derivative, 4,4',4'',4'''-(anthracene-9,10-diylbis(ethene-2,1,1-triyl))tetrakis(N,N,N-trimethylbenzenaminium) iodide (BDVAI), with AIE properties, high biocompatibility and good self-assembly effect. Gold nanocages (AuNCs) were selected as the substrate for PEF, and the inner space of hollow AuNCs was filled with BDVAI. Thiol-modified DNA chains were bonded to the surface of AuNCs by Au-S bonds, and an insulin aptamer was combined with the sulfhydryl chain to seal the AuNCs. This PEF-AIE sensor produces different fluorescence signals when interacting with native insulin and fibrillar insulin; thus, monitoring conformational changes in insulin can be realized by detecting fluorescence intensity changes during insulin fibrillation. Based on this design, this system realized sensitive detection of fibrillar insulin with a detection limit of 23.6 pM. This AIE molecular-based PEF fluorescence enhancement system improves the optical properties of fluorescent substances, which is of great significance in improving the detection sensitivity of amyloid fibrils conformational changes and providing a reliable basis for further understanding the pathogenesis of amyloidosis.

Improved Protein and PTM Characterization with a Practical Electron-Based Fragmentation on Q-TOF Instruments.

Electron-based dissociation (ExD) produces uncluttered mass spectra of intact proteins while preserving labile post-translational modifications. However, technical challenges have limited this option to only a few high-end mass spectrometers. We have developed an efficient ExD cell that can be retrofitted in less than an hour into current LC/Q-TOF instruments. Supporting software has been developed to acquire, process, and annotate peptide and protein ExD fragmentation spectra. In addition to producing complementary fragmentation, ExD spectra enable many isobaric leucine/isoleucine and isoaspartate/aspartate pairs to be distinguished by side-chain fragmentation. The ExD cell preserves phosphorylation and glycosylation modifications. It also fragments longer peptides more efficiently to reveal signaling cross-talk between multiple post-translational modifications on the same protein chain and cleaves disulfide bonds in cystine knotted proteins and intact antibodies. The ability of the ExD cell to combine collisional activation with electron fragmentation enables more complete sequence coverage by disrupting intramolecular electrostatic interactions that can hold fragments of large peptides and proteins together. These enhanced capabilities made possible by the ExD cell expand the size of peptides and proteins that can be analyzed as well as the analytical certainty of characterizing their post-translational modifications.

Metabolic parameters in cord blood of neonate of mothers with obese and non-obese PCOS and controls: retrospective cohort study.

BACKGROUND: Polycystic ovary syndrome (PCOS) is characterized by reproductive disorder and increased risk of metabolic syndrome. This study aimed to assess the metabolic parameters in the cord blood of neonate of mothers with obese PCOS and comparison with non-obese PCOS and controls. METHODS: This retrospective cohort study was conducted in Arash and Kamali Hospital in 2017-2018. The biochemical test was conducted on 78 neonates from obese PCOS mothers, 78 neonates from non-obese PCOS mothers, and 78 neonates from healthy mothers. Finally, cord blood lipid profile and insulin and blood sugar were determined by specific kits. Correlations between variables were compared with chi-square, Mann-Whitney's U, Kruskal-Wallis H tests and regression model by SPSS 23 and P < 0.05 was considered significant. RESULTS: Triglycerides (TG) and high-density lipoprotein cholesterol (HDL) were higher in cord blood of newborn of obese PCOS women than non-obese PCOS and controls (P = 0.02, P < 0.001, respectively). Also, the mean insulin was higher in cord blood of neonate of non-obese PCOS women than in obese PCOS and controls (12.26 ± 12.79 vs. 11.11 ± 16.51 vs. 6.21 ± 10.66, P = 0.01). But in the study, there was no significant difference between the mean of umbilical cord low-density lipoprotein cholesterol (LDL), total cholesterol and blood sugar in three groups. The logistic regression model showed that metabolic parameters were related to PCOS. CONCLUSIONS: In the present study, there was a significant difference between the mean of umbilical cord HDL, cholesterol, and the insulin level in the three groups. But, there was no significant association between the mean of blood sugar, LDL, and TG in the groups. The metabolic disorder in PCOS might affect cord blood lipid and insulin and adulthood health.

[Pancreatic proinsulinoma]. / Proinsulinoma podzheludochnoi zhelezy.

OBJECTIVE: To report own experience in the treatment of patients with proinsulinoma. MATERIAL AND METHODS: There were 10 patients with increased proinsulin production and normal insulin level since 2017. Most of them were young women. RESULTS: Fasting hypoglycemia in all patients was severe (up to 0.7 mmol/l). Clinical picture consisted of typical symptoms similar to those in insulinoma. The main difference in the course of proinsulinoma was the absence of weight gain in 7 patients and rapid weight loss (from 210 to 90 kg within 9 months) in 1 patient. All patients with proinsulinoma underwent surgery. In most cases, minimally aggressive surgery was performed. CONCLUSION: Proinsulinoma is an extremely rare endocrine-active neuroendocrine pancreatic tumor. Differential features of proinsulinoma are the absence of weight gain and normal insulin levels in the presence of hypoglycemia. Surgery is the only radical method of treatment.

Dietary insulin index and insulin load in relation to glioma: findings from a case-control study.

BACKGROUND: Although hyperinsulinemia is assumed to be involved in brain carcinogenesis, data on the link between dietary insulin index (DII) and dietary insulin load (DIL) and risk of glioma are lacking. OBJECTIVE: The current study aimed to investigate the relation between DII and DIL and risk of glioma in a case-control study among Iranian adults. METHODS: In this hospital-based case-control study, 128 pathologically confirmed cases of glioma and 256 age and sex-matched controls were enrolled. Dietary intakes of study participants were assessed using a validated Block-format 123-item semi-quantitative FFQ. DII and DIL were computed using a published food insulin index (FII) data. RESULTS: A significant positive association was found between DIL and glioma (OR: 3.56; 95% CI: 1.85-6.58, P < 0.001); such that after controlling for potential confounders, participants in the highest quartile of DIL had 2.95 times greater odds of glioma than those in the lowest quartile (OR: 2.95; 95% CI: 1.40-6.24, Ptrend = 0.006). Furthermore, we observed a significant positive association between DII and glioma (OR: 2.65; 95% CI: 1.43-4.93, Ptrend = 0.001). This association remained significant even after considering energy intake (OR: 2.66; 95% CI: 1.43-4.95, Ptrend = 0.001). However, when further potential confounders were taken into account, this relationship became non-significant (OR: 1.87; 95% CI: 0.92-3.80, Ptrend = 0.03), despite a significant trend of increased odds ratios (P = 0.03). CONCLUSIONS: In conclusion, we found a significant positive association between DIL and odds of glioma. DII was not significantly associated with odds of glioma after controlling for confounders.

The effect and safety of Berberine on polycystic ovary syndrome: a systematic review.

The aim of this systematic review is to assess the effect of Berberine (BBR) on women's health to provide greater insights about its effect on women with polycystic syndrome for both patients and health care providers. Electronic databases such as PubMed, Web of Science, Scopus, Cochrane Central Register of Controlled Trials (CENTRAL) were systematically searched from the base to July 1th, 2019 to identify clinical trials and randomised controlled trials that had explored the effect of BBR on the polycystic syndrome. With regard to the weight and composition body, BBR did not have any significant effect on reducing body weight and conflicting findings had been reported about waist circumference (WC) and body mass index (BMI). However, BBR led to a significant decrease in waist to hip ratio (WHR), profile hormonal insulin resistance (IR), and insulin resistance (HOMA-IR). Further, androstenedione dropped significantly following treatment with BBB. However, BBB did not have a significant effect on follicle stimulating hormone (FSH) and luteinizing hormone (LH).

Insulina modula migração celular e a secreção de citocinas na vigência da infecção pulmonar por Paracoccidioides brasiliensis em camundongos diabéticos e não-diabéticos / Insulin modulates cell migration and cytokines secretion during pulmonary infection caused by Paracoccidioides brasiliensis in diabetic and non-diabetic mice

Diabetes mellitus (DM) compreende um conjunto de doenças metabólicas de grande importância e incidência mundial. Nele, o DM do tipo 1 é caracterizado pela destruição de células pancreáticas produtoras de insulina, e dentre seus sintomas, a disfunção imunológica relacionada à falta de insulina foi observada por diversos estudos, descrevendo pacientes diabéticos como mais susceptíveis a infecções e complicações decorrentes destas. Paracoccidioidomicose (PCM) é uma enfermidade sistêmica causada por fungos da espécie Paracoccidioides sp., bastante importante no Brasil e endêmica em toda a América Latina. Este trabalho utiliza um modelo de carência relativa de insulina (DM experimental) para estudar a intervenção da insulina em um modelo de micose pulmonar causada por P. brasiliensis, analisando o processo de migração celular (expressão de moléculas de adesão por imunohistoquímica e fenótipo dos leucócitos do pulmão por citometria de fluxo), os mecanismos moleculares (produção/liberação de citocinas por cytometric bead array), intracelulares (vias de sinalização por Western blot), e a atividade fagocítica e microbicida dos macrófagos alveolares. Em resultados observamos que, comparados aos não-diabéticos, camundongos tornados diabéticos apresentam maior susceptibilidade evidenciada por menor atividade fagocítica e reduzidas secreções de interferon-γ e de interleucina-12 na fase inicial da inflamação, que leva a uma resposta menos efetiva com menor expressão de molécula de adesão de células vasculares, reduzidas populações de linfócitos TCD4+, TCD8+, células natural killer, culminando em inflamação crônica resultante da proliferação aumentada do fungo nos pulmões (aumento de interferon-γ e fator necrótico tumoral-ß). Vemos ainda que o tratamento de insulina em animais diabéticos restaurou as secreções de citocinas pró-inflamatórias e a atividade fagocítica de macrófagos em 24 horas de infecção, e aumentou a celularidade, a expressão de moléculas de adesão de células vasculares-1 e restaurou as populações de linfócitos B, de células natural killer e de células coestimuladas por CD80, além de reduzir a inflamação crônica no pulmão. Estes dados em conjunto nos permitem inferir que a insulina modulou o ambiente inflamatório de animais tornados diabéticos de formas diferentes em estágios iniciais e tardios da infecção pelo isolado Pb18 do Paracoccidioides brasiliensis

Diabetes mellitus comprehends a group of metabolic diseases of great importance and incidence worldwide. Type 1 diabetes mellitus is characterized by destruction of insulin producing-pancreatic cells and, among its symptoms, an impaired immunological function has been observed in many studies having diabetic patients described as more susceptible to infections and complications resulted of them. Paracoccidioidomycosis is a systemic disease caused by fungi of Paracoccidioides spp. , also of great importance in Brazil and endemic in the whole Latin America. This work uses a model of experimental T1DM to investigate the intervention of insulin in a model of murine PCM induced by Paracoccidioides brasiliensis, analyzing the process of cell migration (adhesion molecules expression, leukocyte phenotyping), molecular mechanisms (production and secretion of cytokines), intracellular mechanisms (signaling pathways) and phagocytic and microbicidal activities in alveolar macrophages. In results, compared to controls, we observed higher susceptibility in diabetic mice to PCM, evidenced by reduced phagocytic activity and reduced levels of interferon-γ and interleukin-12 on initial stages of infection, and a less effective inflammation with lesser expression of adhesion molecules, reduced migration of TCD4+, TCD8+, NK cells and B lymphocytes, resulting in chronic inflammation caused by higher fungal proliferation in lungs (higher interferon-γ and tumours necrosis factor-α levels). In addition, we saw treatment with insulin in diabetic animals restored secretion of pro-inflammatory cytokines and phagocytic activity on early stages and allowed higher cellularity, higher expression of vascular cells adhesion molecule-1 and restored populations of B lymphocytes, NK cells and the expression of costimularoty molecule CD80, also reducing the chronic inflammation in lungs. Taken together, these data lead us to suggest insulin modulated the inflammatory microenvironment in lungs of mice rendered diabetic, in different forms on earlier and later stages of an infection by Pb18 isolate

Role of the Metabolic Profile in Mediating the Relationship Between Body Mass Index and Left Ventricular Mass in Adolescents: Analysis of a Prospective Cohort Study.

Background We aimed to quantify the role of the plasma metabolic profile in explaining the effect of adiposity on cardiac structure. Methods and Results Body mass index (BMI) was measured at age 11 in the Avon Longitudinal Study of Parents and Children. Left ventricular mass indexed to height2.7 (LVMI) was assessed by echocardiography at age 17. The metabolic profile was quantified via 1H-nuclear magnetic resonance spectroscopy at age 15. Multivariable confounder (maternal age, parity, highest qualification, maternal smoking, prepregnancy BMI, prepregnancy height, household social class, adolescent birthweight, adolescent smoking, fruit and vegetable consumption, and physical activity)-adjusted linear regression estimated the association of BMI with LVMI and mediation by metabolic traits. We considered 156 metabolomic traits individually and jointly as principal components explaining 95% of the variance in the nuclear magnetic resonance platform and assessed whether the principal components for the metabolic traits added to the proportion of the association explained by putative cardiovascular risk factors (systolic and diastolic blood pressures, insulin, triglycerides, low-density lipoprotein cholesterol, and glucose). A 1 kg/m2 higher BMI was associated with a 0.70 g/m2.7 (95% CI, 0.53-0.88 g/m2.7) and 0.66 g/m2.7 (95% CI, 0.53-0.79 g/m2.7) higher LVMI in males (n=437) and females (n=536), respectively. Putative risk factors explained 3% (95% CI, 2%-5%) of this association in males, increasing to 10% (95% CI, 8%-13%) when including metabolic principal components. In females, the standard risk factors explained 3% (95% CI, 2%-5%) of the association and did not increase when including the metabolic principal components. Conclusions The addition of the nuclear magnetic resonance-measured metabolic traits appears to mediate more of the association of BMI on LVMI than the putative risk factors alone in adolescent males, but not females.

Sub/supercritical fluid chromatography employing water-rich modifier enables the purification of biosynthesized human insulin.

There is a paucity of knowledge surrounding the SFC purification of human insulin. The current conventional method of insulin purification involves traditional RP-HPLC that utilises copious amounts of toxic solvents. In this study, we envisaged the development of an environmentally friendly SFC method for biosynthesized human insulin purification. Various commercially available SFC columns derived with silica, 2'ethyl pyridine, diol-HILIC, and the PFP functionalities were evaluated to determine the optimal stationary phase for purification. The PFP column gave the best results with respect to efficiencies of this important biologic that yielded average recoveries of 84%. LC-MS was used to initially detect and quantify the SFC purified standard sample of insulin (purchased) as well as the biosynthesized version. Protein sequencing was employed to verify the amino acid sequencing of the insulins; as such, the standard had a 90% probability to human insulin from the database, whereas the biosynthesized version had a 96% probability. The biological activities of both versions of the SFC purified proteins were assessed in vitro using a MTT assay. The results indicated that the biological activities of both samples were retained subsequent to SFC purification. This study successfully proposes a greener and more efficient method for the purification of insulin derivatives.

The effects of physical activity on physiological markers in breast cancer survivors: A meta-analysis.

BACKGROUND: To systematically evaluate the effects of physical activity on physiological markers in breast cancer survivors. METHODS: A systematic search of the PubMed, Wed of Science, Medline, CNKI and Wanfang Database was performed to identify eligible randomized controlled trials to explore physical activity on physiological markers in breast cancer survivors. STATA version 13.0 (Stata Corp LP, College Station, TX) was used for all statistical analyses. RESULTS: A total of 11 articles with 941 cases were eligible in this meta-analysis. The results of the meta-analysis showed that physical activity could decrease the levels of insulin (SMD = -1.90, 95%CI: -3.2 to -0.60; I = 92.3%, P < .001), insulin-like growth factor 1 (IGF-I) (WMD = -4.67, 95%CI: -23.14 to 13.79; I = 96.2%, P < .001), insulin-like growth factor binding protein-3 (IGFBP-3) (WMD = -20.09, 95%CI: -47.15 to 6.97; I = 93.3%, P < .001). However, compared with the control group, there was not the significant change of insulin-like growth factor 2 (IGF-II), insulin-like growth factor binding protein-1 (IGFBP-1), leptin, adiponectin, glucose, C-reactive protein (CRP), Interleukin-6 (IL-6), Interleukin-10 (IL-10), and tumor necrosis factor alpha (TNF-ɑ) levels after the intervention. CONCLUSIONS: Physical activity could improve the insulin function that might be associated with decreasing the levels of IGF-I, IGFBP-3 and insulin in breast cancer survivors.