BNIP3 promotes HIF-1α-driven melanoma growth by curbing intracellular iron homeostasis.

BNIP3 is a mitophagy receptor with context-dependent roles in cancer, but whether and how it modulates melanoma growth in vivo remains unknown. Here, we found that elevated BNIP3 levels correlated with poorer melanoma patient's survival and depletion of BNIP3 in B16-F10 melanoma cells compromised tumor growth in vivo. BNIP3 depletion halted mitophagy and enforced a PHD2-mediated downregulation of HIF-1α and its glycolytic program both in vitro and in vivo. Mechanistically, we found that BNIP3-deprived melanoma cells displayed increased intracellular iron levels caused by heightened NCOA4-mediated ferritinophagy, which fostered PHD2-mediated HIF-1α destabilization. These effects were not phenocopied by ATG5 or NIX silencing. Restoring HIF-1α levels in BNIP3-depleted melanoma cells rescued their metabolic phenotype and tumor growth in vivo, but did not affect NCOA4 turnover, underscoring that these BNIP3 effects are not secondary to HIF-1α. These results unravel an unexpected role of BNIP3 as upstream regulator of the pro-tumorigenic HIF-1α glycolytic program in melanoma cells.

Chromatographic Methods Developed for the Quantification of Quercetin Extracted from Natural Sources: Systematic Review of Published Studies from 2018 to 2022.

Quercetin (QUE) is the most widely used flavonoid for therapeutic purposes. To improve the available knowledge about the properties of some natural products, determining the amount of QUE is crucial. The main objective of this systematic review is to identify the analytical methods validated for detecting and quantifying QUE in different matrices and characterize their sensitivity. A search was conducted until 30 June 2023 in the PubMed database for experimental studies that addressed the validation of chromatographic analytical methods to detect and quantify QUE from consumable natural products. Only studies published between 2018 and 2022, written in English, were included. The risk of bias was assessed by emphasizing methods of comparison according to previously published studies. Descriptive statistics were used to depict the obtained results. The studies were analyzed based on the type of QUE source, chromatographic method, and validation parameters. A total of 17 studies were included in this review. Plants were the most commonly analyzed source of QUE. Among the detection methods, spectrophotometry proved to be the most widely used, surpassing mass spectrometry (MS). After analyzing the bias, all the included studies mentioned/presented, totally or partially, at least four of the eight parameters.

In Vitro Cytotoxic Activity of African Plants: A Review.

In African countries, cancer not only is a growing problem, but also a challenge because available funding and resources are limited. Therefore, African medicinal plants play a significant role in folk medicine and some of them are traditionally used for the treatment of cancer. The high mortality rate and adverse effects associated with cancer treatments have encouraged the search for novel plant-based drugs, thus, some African plants have been studied in recent years as a source of molecules with proven cytotoxicity. This review aims to discuss the cytotoxic activity, in vitro, of African plant crude extracts against cancer cell lines. For the period covered by this review (2017−2021) twenty-three articles were found and analyzed, which included a total of 105 plants, where the main cell lines used were those of breast cancer (MCF-7 and MDA-MBA-231) and colorectal cancer (HCT-116 and Caco-2), which are among the most prevalent cancers in Africa. In these studies, the plant crude extracts were obtained using different solvents, such as ethanol, methanol, or water, with variable results and IC50 values ranging from <20 µg/mL to >200 µg/mL. Water is the preferred solvent for most healers in African countries, however, in some studies, the aqueous extracts were the least potent. Apoptosis and the induction of cell cycle arrest may explain the cytotoxic activity seen in many of the plant extracts studied. Considering that the criteria of cytotoxicity activity for the crude extracts, as established by the American National Cancer Institute (NCI), is an IC50 < 30 µg/mL, we conclude that many extracts from the African flora could be a promising source of cytotoxic agents.

Physicochemical Properties and Antioxidant Activity of Portuguese Craft Beers and Raw Materials.

There is an increase in the popularity of craft beer, which is produced by small, independent, and traditional breweries. Since craft beer popularity is rising in Portugal this research focused on assessing physicochemical parameters, total phenolic content (TPC) and the antioxidant capacity of Portuguese craft beers and raw materials used in beer production. In this experimental study, 19 beer samples were analyzed. Parameters such as pH, Total Acidity, Reducing Sugar Content and TPC were evaluated. For the determination of antioxidant activity, DPPH scavenging activity and metal chelating activity (MCA) were analyzed in all samples. Craft beers demonstrated a high phenolic content (ranging from 343.78 mg GAE/L to 2172.49 mg GAE/L), significantly different from industrial beers. Craft beers demonstrated a higher inhibition of DPPH radicals and higher MCA than the raw materials. DPPH inhibition ranged from 36.5% to 96.0% for malt and 64.7% to 79.6% in hops samples. MCA also varied between the different samples, with results of 12.0% to 24.8% in malt samples and 3.8% to 23.5% in hops. Raw materials can potentially influence the antioxidant activity of the resulting beer. Positive correlations between TPC and physicochemical properties can be useful to help consumers choose beers with added value for health.

Crosstalk between glial and glioblastoma cells triggers the "go-or-grow" phenotype of tumor cells.

BACKGROUND: Glioblastoma (GBM), the most malignant primary brain tumor, leads to poor and unpredictable clinical outcomes. Recent studies showed the tumor microenvironment has a critical role in regulating tumor growth by establishing a complex network of interactions with tumor cells. In this context, we investigated how GBM cells modulate resident glial cells, particularly their paracrine activity, and how this modulation can influence back on the malignant phenotype of GBM cells. METHODS: Conditioned media (CM) of primary mouse glial cultures unexposed (unprimed) or exposed (primed) to the secretome of GL261 GBM cells were analyzed by proteomic analysis. Additionally, these CM were used in GBM cells to evaluate their impact in glioma cell viability, migration capacity and activation of tumor-related intracellular pathways. RESULTS: The proteomic analysis revealed that the pre-exposure of glial cells to CM from GBM cells led to the upregulation of several proteins related to inflammatory response, cell adhesion and extracellular structure organization within the secretome of primed glial cells. At the functional levels, CM derived from unprimed glial cells favored an increase in GBM cell migration capacity, while CM from primed glial cells promoted cells viability. These effects on GBM cells were accompanied by activation of particular intracellular cancer-related pathways, mainly the MAPK/ERK pathway, which is a known regulator of cell proliferation. CONCLUSIONS: Together, our results suggest that glial cells can impact on the pathophysiology of GBM tumors, and that the secretome of GBM cells is able to modulate the secretome of neighboring glial cells, in a way that regulates the "go-or-grow" phenotypic switch of GBM cells.

Phenotypic impact of deregulated expression of class I histone deacetylases in urothelial cell carcinoma of the bladder.

Deregulated expression of histone deacetylases (HDACs) has been implicated in tumorigenesis. Herein, we investigated class I HDACs expression in bladder urothelial cell carcinoma (BUCC), its prognostic value and biological significance. Significantly increased transcript levels of all HDACs were found in BUCC compared to 20 normal mucosas, and these were higher in lower grade and stage tumors. Increased HDAC3 levels were associated with improved patient survival. SiRNA experiments showed decrease cell viability and motility, and increased apoptosis. We concluded that class I HDACs play an important role in bladder carcinogenesis through deregulation of proliferation, migration and apoptosis, constituting putative therapeutic targets.

Epigenetic regulation of MDR1 gene through post-translational histone modifications in prostate cancer.

BACKGROUND: Multidrug resistance 1 (MDR1) gene encodes for an ATP binding cassette transporter--P-glycoprotein (P-gp)-- involved in chemoresistance to taxanes. MDR1 promoter methylation is frequent in prostate carcinoma (PCa), suggesting an epigenetic regulation but no functional correlation has been established. We aimed to elucidate the epigenetic mechanisms involved in MDR1 deregulation in PCa. RESULTS: MDR1 promoter methylation and P-gp expression were assessed in 121 PCa, 39 high-grade prostatic intraepithelial neoplasia (HGPIN), 28 benign prostatic hyperplasia (BPH) and 10 morphologically normal prostate tissue (NPT) samples, using quantitative methylation specific PCR and immunohistochemistry, respectively. PCa cell lines were exposed to a DNA methyltransferases inhibitor 5-aza-2'deoxycytidine (DAC) and histone deacetylases inhibitor trichostatin A (TSA). Methylation and histone posttranscriptional modifications status were characterized and correlated with mRNA and protein expression. MDR1 promoter methylation levels and frequency significantly increased from NPTs, to HGPIN and to PCa. Conversely, decreased or absent P-gp immunoexpression was observed in HGPIN and PCa, inversely correlating with methylation levels. Exposure to DAC alone did not alter significantly methylation levels, although increased expression was apparent. However, P-gp mRNA and protein re-expression were higher in cell lines exposed to TSA alone or combined with DAC. Accordingly, histone active marks H3Ac, H3K4me2, H3K4me3, H3K9Ac, and H4Ac were increased at the MDR1 promoter after exposure to TSA alone or combined with DAC. CONCLUSION: Our data suggests that, in prostate carcinogenesis, MDR1 downregulation is mainly due to histone post-translational modifications. This occurs concomitantly with aberrant promoter methylation, substantiating the association with P-gp decreased expression.

Relation between weight gain during pregnancy and postpartum reclassification in gestational diabetes.

PURPOSE: Gestational diabetes mellitus (GDM) is the most common metabolic disease in pregnancy. It is known that GDM is a precursor to type 2 diabetes (T2D). There is evidence that excessive gestational weight variation (GWV) increases the risk of GDM. So, in this study, we aimed to evaluate the association between GWV and the persistence of diabetes in postpartum reclassification. METHODS: A retrospective observational study including pregnant women based on data from the Portuguese National Registry of Gestational Diabetes. Six-to-eight weeks after delivery, all women included underwent a reclassification test. We performed unadjusted and adjusted logistic regression models to evaluate the associations between GWV and diabetes diagnosis at the reclassification test. A subgroup analysis according to the pre-gestational BMI was also performed. RESULTS: We included 10,389 pregnant women, of which 19.6% had GDM in a previous pregnancy. The median of GWV was 10.0 [6.4, 14.0] kg and was found to be higher for those with a normal BMI. At the DM reclassification test, 1% of the women were diagnosed with T2D. We found a negative association between GWV and postpartum diabetes mellitus (DM). We also present a subgroup analysis, and these associations were only significant for the group with a normal pre-gestational BMI. CONCLUSION: Our results showed that women with normal pre-gestational BMI and lower GWV were more likely to have a diagnosis of DM in the postpartum reclassification test. This study helps to fill the gap in the effect of GWG on the persistence of diabetes in postpartum reclassification.

Validation of an HPLC-DAD Method for Quercetin Quantification in Nanoparticles.

The evaluation of the efficacy of incorporation of quercetin in nanoparticles is crucial, both for the development and quality control of pharmaceutical formulations. The validation of analytical methods for the precise quantification of quercetin is useful for the evaluation of various potential quercetin delivery systems and quercetin pharmacokinetics. This work aimed to validate a high-performance liquid chromatography with diode array detection (HPLC-DAD) method for quercetin detection and quantification in nanoparticles. Different mobile phase conditions and detection wavelengths (254 and 368 nm) were tested, and the major validation parameters were assessed (precision, accuracy, linearity, sensitivity, stability, and selectivity). The best peak resolution was obtained when quercetin was analyzed at 368 nm with a mobile phase of 1.5% acetic acid and a water/acetonitrile/methanol ratio of 55:40:5. Under these conditions, quercetin also eluted rapidly (retention time of 3.6 min). The method proved to be linear (R2 > 0.995), specific, and repeatable (variation coefficient between 2.4% and 6.7%) and presented intermediate precision (variation coefficient between 7.2% and 9.4%). The accuracy of the analysis ranged between 88.6% and 110.7%, and detection and quantification limits were 0.046 and 0.14 µg/mL, respectively. Quercetin solutions were more stable when stored at 4 °C than at room temperature or -20 °C. This validated method satisfied more parameters of bias assessment than most recent methods for quercetin determination and presented itself as more sensitive and efficient than general spectrophotometric methods. The method was successfully used for the analysis of quercetin incorporation in nanoparticles and will be evaluated in the future for its adequacy for the determination of quercetin in more complex matrices.

Post-acute sequelae of COVID-19 is characterized by diminished peripheral CD8+ß7 integrin+ T cells and anti-SARS-CoV-2 IgA response.

Several millions of individuals are estimated to develop post-acute sequelae SARS-CoV-2 condition (PASC) that persists for months after infection. Here we evaluate the immune response in convalescent individuals with PASC compared to convalescent asymptomatic and uninfected participants, six months following their COVID-19 diagnosis. Both convalescent asymptomatic and PASC cases are characterised by higher CD8+ T cell percentages, however, the proportion of blood CD8+ T cells expressing the mucosal homing receptor ß7 is low in PASC patients. CD8 T cells show increased expression of PD-1, perforin and granzyme B in PASC, and the plasma levels of type I and type III (mucosal) interferons are elevated. The humoral response is characterized by higher levels of IgA against the N and S viral proteins, particularly in those individuals who had severe acute disease. Our results also show that consistently elevated levels of IL-6, IL-8/CXCL8 and IP-10/CXCL10 during acute disease increase the risk to develop PASC. In summary, our study indicates that PASC is defined by persisting immunological dysfunction as late as six months following SARS-CoV-2 infection, including alterations in mucosal immune parameters, redistribution of mucosal CD8+ß7Integrin+ T cells and IgA, indicative of potential viral persistence and mucosal involvement in the etiopathology of PASC.

Quercetin-biapigenin nanoparticles are effective to penetrate the blood-brain barrier.

Search for efficient therapeutic agents for central nervous system (CNS) disorders has been extensive. Nevertheless, blood-brain barrier (BBB) is an obstacle that prevents the majority of compounds to act in these diseases. It is, thus, of extreme relevance the BBB overcome, in order to deliver a drugs therapeutically active concentration to the action site, with the least losses and interaction with other organs, tissues, or cells. The present study aimed to investigate the potential protective effect of quercetin-biapigenin encapsulated into poly(Ɛ-polycaprolactone) (PCL) nanoparticles against t-BOOH-induced oxidative stress in several brain cell lines, as well as evaluate the permeability of those active molecules through an in vitro BBB model. The three cell lines under study (BV-2, hcmec/D3, and U87) presented different reactions to t-BOOH. In general, quercetin-biapigenin PCL-loaded nanoparticles were able to minimize compound toxicity they convey, regardless the cell line. Quercetin-biapigenin PCL-loaded nanoparticles (Papp of approximately 80 × 10-6 cm/s) revealed to be more permeable than free compounds (Papp of approximately 50 × 10-6 cm/s). As of our knowledge, this is the first report of quercetin-biapigenin PCL-loaded nanoparticle activity in brain cells. It is also the first determining its permeability through BBB, as an effective nanocarrier for brain delivery.

Subacute Thyroiditis After Severe Acute Respiratory Syndrome Coronavirus 2 Vaxzevria Vaccination in a Patient With Thyroid Autoimmunity.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has been challenging the scientific community to promptly treat the patients and mitigate its spreading. The rapid development of vaccination against SARS-CoV-2 is being highly effective, but it is still lacking knowledge about its side effects. Epidemiological studies point toward virus infection as causative agents of subacute thyroiditis. More than 20 cases of thyroiditis after SARS-CoV-2 have also been described. Here, we aim to broad the spectrum of SARS-CoV-2 vaccination thyroid-associated disorders with the description of a new case of subacute thyroiditis associated with thyroid autoimmunity. The temporal association with the inoculation of the vaccine and the absence of other plausible etiological agents makes it highly possible that this thyroiditis was caused by Vaxzevria vaccine. It remains to be established whether the presence of thyroid autoimmunity can facilitate this condition, as this is one of the few described cases associated with autoimmunity.

Clinical Characteristics and Mortality-Associated Factors in COVID-19 Critical Patients in a Portuguese ICU.

INTRODUCTION: Severe COVID-19 is associated with serious complications and poor outcomes. Older age and underlying comorbidities are known risk factors for severe COVID-19, but a better understanding of baseline characteristics and outcomes of patients with severe COVID-19 is urgently needed. METHODS: This study was a retrospective case series of 227 consecutive patients with laboratory-confirmed COVID-19 admitted to the intensive care unit (ICU) at our institution between March 2020 and December 2021. Demographic and clinical data were collected. RESULTS: The median age of patients was 65 years, and 180 (79.3%) were male. Cardiovascular comorbidities were frequent and included hypertension (n=148; 65.2%), dyslipidemia (n=116; 51.1%), obesity (n=114; 50.2%), and diabetes mellitus (n=80; 35.2%). About 20% of the patients had the chronic respiratory disease, with sleep apnea being the most common. Immunosuppression was identified in 13% of the patients, with autoimmunity, post-transplantation, and neoplasms being the most represented causes. Most patients were admitted to the ICU at six to 15 days after symptom onset, corresponding to stages IIb (pulmonary involvement/hypoxia) and III (hyperinflammatory). All patients received systemic steroids, with an average treatment duration of 22 days. Several ventilatory support strategies were used; 80 patients were supported entirely noninvasively with high flow nasal oxygenation and noninvasive ventilation, while 164 patients were invasively ventilated. Most intubations (65%) occurred in the first 24 hours after admission, and the mean duration of mechanical ventilation was 14 days. The reintubation rate was 10%, occurring on average two to three days after planned extubation. Thirty-two tracheostomies were performed. Bacterial co-infection was treated in 75% of patients, and Aspergillus co-infection complicating COVID-19 pneumonia was diagnosed in eight patients. Median ICU and hospital stays were 15 and 25 days, respectively, and the 28-day mortality rate was 38%. Patients over 75 years experienced a higher mortality rate (56%). Increased age and multimorbidity, particularly comprising cardiovascular disease and associated risk factors, were significantly more common in patients who died within 28 days after ICU admission. CONCLUSIONS: A large proportion of critically ill COVID-19 patients required prolonged mechanical ventilation. ICU/hospital stay and mortality were particularly elevated in older patients and patients with cardiovascular risk factors. Considerable discrepancy existed between the proportion of patients with microbiological documentation of bacterial infections and those receiving antimicrobials. Improved methods for adequate microbiological diagnosis are needed and stewardship programs should be reinforced.

Virchow node of an unusual aetiology - the experience of a clinical case.

BACKGROUND: The cause of adult adenomegalies may be defiant. On the other hand, ectopic thyroid is a rare condition that happens in every 1:100000 to 300,000 of healthy individuals. Here, we present a case report that joins these two clinical rare and defiant challenges. CLINICAL CASE: Forty-seven-year-old woman, with known thyroid nodules for several years. She had no other relevant personal or familiar history. At our appointment she had no complaints. At the physical examination she had a palpable right thyroid nodule (previously known). The routine blood analysis showed normal thyroid function. The routine cervical ultrasonography showed no dimensional progression of the known thyroid nodules and identified a 31x18mm nodule at the left supraclavicular fossa. The patient underwent a cervical, thoracic, and abdominal computed tomography that exhibited no relevant findings, such as abdominal malignancies. The cytology of the nodule showed characteristics that were "compatible with a benign follicular nodule in ectopic thyroid tissue". CONCLUSION: This is a rare case in which we incidentally found a follicular nodule in ectopic thyroid tissue in the left supraclavicular fossa. Given the rarity of the situation, clinical sense is the mainstay of treatment and follow-up.

Clinical characteristics and incidence of glucose metabolism disorders during the follow-up of surgically treated insulinomas.

PURPOSE: Insulinomas are pancreatic endocrine tumors characterized by hypoglycemia resulting from hypersecretion of insulin. The long-term impact of surgical treatment of insulinomas, particularly the risk of glucose metabolism disorders, remains largely unknown. METHODS: We retrospectively evaluated all patients with insulinoma submitted to surgery at Centro Hospitalar Universitário de São João (Porto, Portugal) between 1980 and 2016. We evaluated baseline characteristics of patients at presentation, imaging evaluation, surgical treatment, characteristics of the tumors, perioperative complications, disease remission, and long-term follow-up and metabolic outcomes. RESULTS: Twenty-eight patients with insulinomas submitted to surgical treatment were included. Sixty-one percent were female, and the average age was 46.4 years. The most reported symptoms were confusion (72%) and diaphoresis (56%). The most used imaging technique was abdominal CT (72%), and the test with the highest percentage of positive results was endoscopic ultrasound (80%). The most used surgical procedure was partial pancreatic resection (71%). The mean tumor diameter was 2.1 cm and 11% of the tumors had lymph node involvement at diagnosis. Pancreatic fistula was the most common postoperative acute complications (21%). After surgery, patients were followed for a median time of 80 months (25th-75th percentile: 20-148 months). Eight patients (32%) developed glucose metabolism disorders (seven developed diabetes and one prediabetes). One of these patients developed albuminuria, and no macrovascular complications were observed during the follow-up. CONCLUSIONS: Disorders of glucose metabolism are a frequent complication during follow-up of surgically treated insulinomas. The prevention, early diagnosis, and treatment of diabetes should be a priority in the follow-up of these patients.

Interleukin-6 Is a Biomarker for the Development of Fatal Severe Acute Respiratory Syndrome Coronavirus 2 Pneumonia.

Hyper-inflammatory responses induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are a major cause of disease severity and death. Predictive prognosis biomarkers to guide therapeutics are critically lacking. Several studies have indicated a "cytokine storm" with the release of interleukin-1 (IL-1), IL-6, and IL-8, along with tumor necrosis factor alpha (TNFα) and other inflammatory mediators. Here, we proposed to assess the relationship between IL-6 and outcomes of patients with coronavirus disease 2019 (COVID-19). Our cohort consisted of 46 adult patients with PCR-proven SARS-CoV-2 infection admitted in a COVID-19 ward of the Hospital de Braga (HB) from April 7 to May 7, 2020, whose IL-6 levels were followed over time. We found that IL-6 levels were significantly different between the disease stages. Also, we found a significant negative correlation between IL-6 levels during stages IIb and III, peripheral oxygen saturation (SpO2), and partial pressure of oxygen in arterial blood (PaO2), showing that IL-6 correlates with respiratory failure. Compared to the inflammatory markers available in the clinic routine, we found a positive correlation between IL-6 and C-reactive protein (CRP). However, when we assessed the predictive value of these two markers, IL-6 behaves as a better predictor of disease progression. In a binary logistic regression, IL-6 level was the most significant predictor of the non-survivors group, when compared to age and CRP. Herein, we present IL-6 as a relevant tool for prognostic evaluation, mainly as a predictor of outcome.

Minimally Invasive Surgical Treatment of Pilonidal Disease: Mid-Term Retrospective Analysis of a Single Center.

Background: Sacrococcygeal Pilonidal Disease (PD) is commonly treated with excision and primary closure techniques (EPC). Minimally invasive techniques (MIT), such as EPSiT and Pit-picking, had been recently advocated promising better outcomes. We analyzed mid-term results from our center after introduction of MIT to treat PD. Methods: Patients submitted to MIT (n = 44) with a median follow-up of 37 months were analyzed and compared with patients submitted to EPC (n = 70) with a median follow-up of 5 years. Both groups included patients operated in our department between 2011 and 2016 and have similar demographic and clinical characteristics. We compared operative time and post-operative parameters such as time with pain, dressing time and time to relapse. Results: The post-operative time with pain was significantly lower, whereas the dressing time was significantly longer, in MIT when compared to the EPC group. The relapse rate was similar in both groups but the follow-up is shorter in the MIT group. In addition, the analysis of patients free of disease using Kaplan-Meier curves revealed that relapse tends to occur more precociously in MIT than in EPC patients (p = 0.014). Interestingly, in the subgroup of patients with previous surgery, MIT's relapse rate was significantly lower than in the EPC group (30 vs. 100%, p < 0.001). Conclusions: MIT has the advantage of having a shorter time with pain in the postoperative period, while EPC benefits from a shorter dressing time. In general, the relapse of the disease tends to manifest more precociously in MIT patients. Moreover, in the subgroup of patients with previous surgery, MIT seems to have significantly better results when compared to EPC.

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.

Metformin in overweight and obese women with gestational diabetes: a propensity score-matched study.

PURPOSE: Obesity and gestational diabetes mellitus (GDM) have an independent negative impact in pregnancy outcomes. Excessive gestational weight gain (GWG) represents an additional high-risk condition for adverse outcomes. The aims of this study were to evaluate the potential effect of metformin in GWG in overweight or obese women with GDM, to report our experience and to assess metformin's safety in this population. METHODS: Retrospective observational cohort study involving pregnant women with GDM and pregestational overweight or obesity. Demographic, anthropometric, glycemic control data, obstetric, fetal and neonatal outcomes were evaluated. The sample was divided into two groups according to metformin treatment. A propensity score-matched analysis was performed using age, initial body mass index (BMI), trimester at GDM diagnosis and previous history of GDM or macrosomia as covariates. RESULTS: Of the 457 enrolled in the study, 177 (38.7%) were treated with metformin. Two groups of 130 well matched patients were balanced regarding baseline characteristics. Women in metformin group had significantly less excessive GWG (29.23% vs. 42.31%, OR 0.56, 95% CI 0.34-0.94, p = 0.028) and more adequate GWG (36.92% vs. 23.08%, OR 1.95, 95% CI 1.14-3.35, p = 0.015). No significant differences were found between both groups regarding glycemic control, rate of insulinization, and obstetric, fetal, and neonatal outcomes. CONCLUSIONS: This study highlights metformin as an important and safe tool to prevent excessive GWG and promote adequate GWG in overweight or obese women with GDM, regardless of age, BMI, timing of GDM diagnosis, previous history of GDM or macrosomia.

Chemistry, bioactivities, extraction and analysis of azadirachtin: State-of-the-art.

Azadirachta indica A. Juss. (Neem) is an Indian tree recognized for its activity as pesticide, as well as several pharmacological properties. Among the various compounds already isolated and studied from Neem tree, azadirachtin (AZA) was identified as the main bioactive compound. Azadirachtin can be found at different parts of the Neem plant but assumes its maximum concentration at the seed level. This compound features a quite complex chemical structure, which justifies the 20-plus-year difficulty to identify the synthetic pathway that subsequently permitted to carry out its artificial synthesis. Azadirachtin is widely used as a basis for production of biopesticides; nevertheless, other properties have been recognized for this substance, among which the anticancer and antimalarial activity stand out. The methods available for azadirachtin extraction are diverse, including solid-liquid extraction and extraction with solvents at high or low temperatures. Alcohol based solvents are associated with higher extraction yields and are therefore preferred for the isolation of azadirachtin from plant parts. Clean-up of the extracts is generally required for further purification. The highest azadirachtin levels have been obtained from Neem seeds but concentration values present a large variation between batches. Therefore, in addition to extraction procedures, it is essential to establish routine methods for azadirachtin identification and quantification. Chromatography-based techniques are preferably selected for detection and quantification of azadirachtin in plant matrices. Overall, this process will guarantee a future reproducible, safe and effective use of the extracts in formulations for commercial applications.