Comparison of protocols for genomic DNA extraction from ‘velame pimenta’ (Croton linearifolius), a native species to the Caatinga, Brazil

The Caatinga biome occupies some 12% of the Brazilian territory, which is present in at least nine states. The species that constitute its biodiversity have the potential to be used as natural resources, among them are approximately 700 species of the genus Croton. As an example of this potential, the Croton linearifolius specie is used by local communities as a natural insecticide. Associated with the economic potential of the Caatinga species, one must stress the risk of extinction or genetic erosion due to the growing deforestation of natural areas of this biome. These factors make it relevant in genetic studies in order to guide conservation strategies. Considering the lack of molecular studies involving C. linearifolius, we compared the efficiency of six protocols for genomic DNA extraction previously described in literature. The DNA extraction buffers [based on the use of Cetyl trimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), mannitol and sorbitol] were different in their efficiency to obtaining the genomic DNA of C. linearifloius. In general, protocols using CTAB buffer were more efficient. The use of liquid nitrogen in the maceration process was also evaluated and its use was considered a no necessary factor in obtaining DNA in adequate quantity and quality for PCR platform procedures.Keywords: DNA Isolation, molecular markers, native species of CaatingaAfrican Journal of Biotechnology Vol. 12(30), pp. 4761-476

Long-Term weight loss and metabolic syndrome remission after bariatric surgery: The effect of sex, age, metabolic parameters and surgical technique-a 4-year follow-up study

Introduction: Bariatric surgery is an effective treatment for morbid obesity and its metabolic related comorbidities. However, the literature reports inconsistent results regarding weight loss (WL) and the resolution of comorbidities associated with obesity. Objective: We aim to evaluate long-Term differences in WL between different surgical techniques and the impact of each surgical technique on metabolic parameters (type 2 diabetes mellitus [T2DM], dyslipidemia,hypertension, and metabolic syndrome). We also aim to evaluate the effect of baseline clinical characteristics in WL and in the evolution of metabolic syndrome (MetS) components. Our hypothesis is that different types of surgery have different effects on WL and the prevalence of comorbidities over time. Methods: We retrospectively evaluated WL and metabolic parameter remission (T2DM, dyslipidemia, hypertension, and MetS) during 4 years in 1,837 morbidly obese patients (females, 85%; age, 42.5 ± 10.6 years; BMI, 44.0 ± 5.8) who underwent bariatric surgery (Roux-en-Y gastric bypass [RYGB], laparoscopic sleeve gastrectomy [LSG], and laparoscopic adjustable gastric band [LAGB]). Results: The mean percentage of WL for RYGB, LSG, and LAGB was, respectively, 32.9 ± 8.7, 29.8 ± 9.8, and 16.2 ± 9.6 at 12 months and 30.6 ± 9.1, 22.7 ± 10.0, and 15.8 ± 10.8 at 48 months (p < 0.001), even after adjustment for baseline weight, BMI, age, and sex (p < 0.001). Women had more WL during the first 36 months (p = 0.013 and 0.007 at 12 and 36 months, respectively) and older patients had less WL compared to younger ones (p <0.001), except at 48 months. Patients with T2DM had less WL than those without diabetes after adjustment (sex, age, and surgical technique) during the same period. Patients with hypertension had less WL at 12 months (p = 0.009) and MetS at 24 months (p = 0.020) compared to those without these comorbidities. There was no significant difference regarding the presence of dyslipidemia in WL. The RYGB group showed better results for MetS resolution. Conclusion:During the 4-year follow-up, RYGB was the surgical procedure that caused the highest WL and MetS resolution

Bariatric Surgery Impact on Cardiovascular Risk Factors: Is Age a Factor to Consider?

Introduction: Despite the abundance of data addressing the influence of patient's age on surgery-related complications, its impact on cardiometabolic outcomes following bariatric surgery has been overlooked. Methods: Retrospective unicentric study of 1,728 obese patients who underwent bariatric surgery between January 2010 and June 2015. Patients were divided in 3 age groups, according to their age at surgery: ?40 (n = 751), 40-59 (n = 879), and 60 years (n = 98). Parameters with cardiometabolic impact, such as body anthropometric measures, lipid profile, and glycemic status, before and 24 months after surgery, were compared between these groups. A multiple linear regression was performed, adjusting differences between groups for sex, surgery type, and body mass index variation. Results: The group ?40 years presented more weight loss (-35.4 ± 9.0 kg, p ? 0.001), greater BMI reduction (-15.8 ± 6.1 kg/m2, p ? 0.001), and larger changes in waist (-34 ± 13.8 cm, p ? 0.001) and hip circumferences (-28.7 ± 11.9 cm, p ? 0.05). The group of 60 years presented the heaviest reduction in fasting glucose (-17.7 ± 32.8 mg/dL, p ? 0.001) and HbA1c (0.7 ± 1.0, p ? 0.001), and also had a tendency to have the biggest changes in systolic blood pressure (-14.7 ± 18.7 mm Hg, p = 0.071). Conclusion: Patients with 60 years benefit the most from bariatric surgery regarding cardiometabolic parameters, presenting heavier reductions in fasting glucose, as well as HbA1c and a tendency towards a higher decrease in systolic blood pressure. No clinically significant differences in lipid profile were observed between groups. (c) 202

Ectopic Cushing's Syndrome Unveiling a Metastatic Parotid Carcinoma

Introduction. Adrenocorticotropic hormone (ACTH) ectopic production is a rare cause of Cushing syndrome (CS). The most commonly associated tumours are small-cell lung carcinoma along with bronchial and thymic carcinoids. To date, only 5 cases have been published in the literature featuring ectopic ACTH secretion from metastatic acinic cell carcinoma (ACC) of the parotid gland. We hereby describe a very uncommon case of ectopic CS (ECS) unveiling a metastatic parotid ACC. Case Presentation. A 46-year-old man with hypertension and dyslipidemia diagnosed 4-months before, as well as new-onset diabetes mellitus unveiled 1-month earlier, was referred to emergency department for hypokalemia. Hormonal study and dynamic biochemical tests performed indicated ECS. Imaging and cytological findings pointed toward a likely primary right parotid malignancy with liver metastases. Somatostatin receptor scintigraphy has shown an increased uptake in the parotid gland and mild expression in liver metastasis. The patient underwent right parotidectomy, and histopathologic examination confirmed ACC. Meanwhile, hypercortisolism was managed with metyrapone, ketoconazole, and lanreotide. Despite chemotherapy onset, a rapid disease progression and clinical course deterioration was observed. Conclusion. The present report highlights a rare ECS, exposing a metastatic parotid ACC, with an aggressive and challenging clinical course, representing the first case whose diagnosis of ECS came prior to ACC

Effect of Weight Loss after Bariatric Surgery on Thyroid-Stimulating Hormone Levels in Patients with Morbid Obesity and Normal Thyroid Function

Background: Several studies have reported that morbid obesity is associated with increased thyroid-stimulating hormone (TSH) levels. However, it is not clear what is the impact of bariatric surgery on postoperative thyroid function. The aim of this study was to evaluate the effect of weight loss after bariatric surgery on TSH levels in euthyroid patients with morbid obesity. Methods: We performed a retrospective observational study of 949 euthyroid patients (86.1% female; age 42.0 ± 10.3 years, BMI 44.3 ± 5.7 kg/m2) with morbid obesity submitted to bariatric surgery (laparoscopic adjustable gastric band, Roux-en-Y gastric bypass, or sleeve gastrectomy). Patients were subdivided in two groups: normal TSH group (TSH <2.5 mU/L) and high-normal TSH group (TSH ≥2.5 mU/L). The impact of anthropometric parameters, comorbidities, TSH, free thyroxine (FT4), free triiodothyronine (FT3), type of surgery, and excessive body weight loss (EBWL) on TSH variation 12 months after surgery was evaluated. Results: The high-normal TSH group (24.3% of patients) included more women, presented a higher BMI, higher systolic blood pressure, and higher FT3 levels. There was a significant decrease of TSH 12 months after surgery that was more marked in the high-normal TSH group (normal TSH group: 1.57 ± 0.49 to 1.53 ± 0.69 mIU/L, p = 0.063; high-normal TSH group: 3.23 ± 0.59 to 2.38 ± 0.86 mIU/L, p < 0.001). In a multivariate analysis, after adjusting for relevant covariates, EBWL, baseline BMI, and baseline FT3 were significantly associated with TSH decrease 12 months after bariatric surgery. Conclusion: Bariatric surgery promotes a decrease of TSH that is significantly greater in patients with high-normal TSH and is independently associated with EBWL after surgery

Adequate magnesium level as an associated factor of pre-diabetes and diabetes mellitus remission in patients with obesity submitted to bariatric surgery

Bariatric surgery (BS) can lead to remission of type 2 diabetes mellitus (T2DM), however, the evidence on the influence of preoperative serum magnesium levels on this reversal is scarce. To study the influence of preoperative serum magnesium levels on the pre-T2DM and T2DM remission one year after BS. Retrospective study carried out among 1656 patients with obesity who underwent BS in the Centro Hospitalar Universitário São João. T2DM and pre-T2DM remission were defined as being normal glycaemic measures of at least one year's after BS and without pharmacological therapy. To assess the association between preoperative serum magnesium levels and pre- and T2DM remission, logistic regression models, crude and adjusted for sex, age and body mass index were computed. Patients with normoglycaemia presented hypomagnesaemia less often than those patients with pre-T2DM and T2DM (17.0% vs. 21.3% vs. 39.9%) (p < 0.001). One year after BS, 62.9% of patients with pre-T2DM or T2DM before BS showed remission. Adequate magnesium levels were positively associated with T2DM and pre-T2DM remission, one year after BS (OR 1.79; 95% CI 1.34-2.38), independently of sex, age, and body mass index. Adequate preoperative serum magnesium levels showed to be an important clinical parameter for pre-T2DM and T2DM remission.Isabel Maia holds a PhD Grant (ref:SFRH/BD/117371/2016/PT), which is co-funded by the Foundation for Science and Technology (FCT) and the POCH/FSE Program

Oral glucose tolerance testing at 1 h and 2 h: relationship with glucose and cardiometabolic parameters and agreement for pre-diabetes diagnosis in patients with morbid obesity

Background: One hour plasma glucose concentration (1hPG) during an oral glucose tolerance test (OGTT) may be an alternative to 2-h plasma glucose concentration (2hPG) in the identification of individuals at increased risk of hyperglycaemia, although its role is not fully understood. Aim: We aim to investigate the relationship of these measures with other glucose parameters, as well as their relationship with cardiometabolic risk markers and the level of agreement for prediabetes mellitus diagnosis, in a sample of patients with morbid obesity. Methods: We retrospectively evaluated 656 patients with morbid obesity without diagnosed diabetes. To define prediabetes with 2hPG, 2022 American Diabetes Association guidelines criteria were used, while for 1hPG, glucose ≥ 155 mg/dL was considered. Cohen’s Kappa coefficient was used to assess the agreement between both measures of prediabetes mellitus diagnosis. Results: A Cohen’s Kappa coefficient of 0.405 (p < 0.001) was obtained. The 1hPG were positively correlated with homeostatic model assessment for insulin resistance (HOMA-IR) (ρ = 0.281, p < 0.001), fasting plasma glucose (FPG) (ρ = 0.581, p < 0.001), glycated haemoglobin (Hb1AC) (ρ = 0.347, p < 0.001) and were negatively correlated with homeostatic model assessment for cell-β function (HOMA-β) (ρ = −0.092, p = 0.018). 2hPG were also correlated with the same parameters, except for HOMA-β. Conclusion: A fair agreement between 1 and 2hPG was verified. 1hPG criteria may be a useful indicator of β-cell dysfunction and insulin resistance in patients with morbid obesity without diabetes diagnosis. © 2022, The Author(s).Funding text 1: We would like to acknowledge the support of all the endocrinologists, surgeons and nutritionists of the Multidisciplinary Group for Surgical Management of Obesity.; Funding text 2: Isabel Maia holds a PhD Grant (ref: SFRH/BD/117371/2016) co-funded by the Foundation for Science and Technology—FCT (The Portuguese Ministry of Science, Technology and Higher Education) and the POCH/FSE programme

Candida albicans repetitive elements display epigenetic diversity and plasticity

Transcriptionally silent heterochromatin is associated with repetitive DNA. It is poorly understood whether and how heterochromatin differs between different organisms and whether its structure can be remodelled in response to environmental signals. Here, we address this question by analysing the chromatin state associated with DNA repeats in the human fungal pathogen Candida albicans. Our analyses indicate that, contrary to model systems, each type of repetitive element is assembled into a distinct chromatin state. Classical Sir2-dependent hypoacetylated and hypomethylated chromatin is associated with the rDNA locus while telomeric regions are assembled into a weak heterochromatin that is only mildly hypoacetylated and hypomethylated. Major Repeat Sequences, a class of tandem repeats, are assembled into an intermediate chromatin state bearing features of both euchromatin and heterochromatin. Marker gene silencing assays and genome-wide RNA sequencing reveals that C. albicans heterochromatin represses expression of repeat-associated coding and non-coding RNAs. We find that telomeric heterochromatin is dynamic and remodelled upon an environmental change. Weak heterochromatin is associated with telomeres at 30?°C, while robust heterochromatin is assembled over these regions at 39?°C, a temperature mimicking moderate fever in the host. Thus in C. albicans, differential chromatin states controls gene expression and epigenetic plasticity is linked to adaptation

Beta Cell Function as a Baseline Predictor of Weight Loss After Bariatric Surgery

Background: Obesity is a multifactorial disease, which is strongly associated to other metabolic disorders. Bariatric surgery is the most effective treatment of morbid obesity. The role of beta cell function in weight loss after bariatric surgery is uncertain. Aim: To evaluate the association between beta cell function and percentage of total body weight loss (TBWL%) 1, 2, 3, and 4 years after bariatric surgery in patients with morbid obesity. Methods: Retrospective longitudinal study in patients with morbid obesity followed in our center between January 2010 and July 2018. Patients were excluded if they had diabetes at baseline or missing data on the needed parameters. We evaluated baseline Homeostatic Model Assessment of IR, Homeostatic Model Assessment of ß-cell function (HOMA-beta), Quantitative Insulin Sensitivity Check Index, and Matsuda and DeFronzo index, and TBWL% at years 1 to 4. Linear regression models were used to evaluate the association of indexes of insulin resistance with TBWL% (unadjusted and adjusted for age, sex, BMI, and type of surgery). Results: There were 1,561 patients included in this analysis. HOMA-beta was negatively associated with TBWL% at second, third, and fourth years post-surgery (ß = -1.04 [-1.82 to -0.26], p<0.01; ß = -1.16 [-2.13 to -0.19], p=0.02; ß = -1.29 [-2.64 to 0.06], p=0.061, respectively). This was not observed in the first year post-surgery nor for the other indexes. Glycemia at baseline was positively associated to EWL% at second and third years post-surgery. Conclusion: ß-cell function at baseline seems to be associated to long-term weight loss, explicitly after the first year post bariatric surgery. This might be a helpful predictor of weight loss in clinical practice.The authors would like to thank all the CRIO group members for following these patients: John Rodrigues Preto; Eduardo Jorge Lima da Costa; Hugo Miguel Santos Sousa; André Manuel Costa Pinho; Carla Cristina Oliveira Rodrigues Teixeira Galego; Maria Flora Ferreira Sampaio Carvalho Correia; Cidália Fátima Castro Carção Gil; Diva Bizarro Figueiredo Melim; Eduardo Gil Ferreira Rodrigues Pinto; Marco António Costa Silva; Cristina Sarmento Pontes Martins; Luis Miguel Gonçalves Pereira; Inês Vasconcelos Sousa Magalhães; Isabel Maria Boavista Vieira Marques Brandão; Sertório Manuel Freitas Andrade, and Patrícia Maria Lopes Nunes. The authors would also like to thank the patients and the hospital for their support. The authors would like to thank to Associação dos Amigos do Serviço de Endocrinologia do Hospital de S. João