The selective extraction of dietary polyamines from chicken breast using the application of a lab-on-a-chip electromembrane and dispersive liquid-liquid microextraction followed by gas chromatography-mass spectrometry.

Excessive dietary polyamines (PAs), including putrescine (PUT), spermine (SPM), and spermidine (SPD), have become a worldwide concern due to their carcinogenicity and reduced shelf life. A modern miniaturized on-chip electromembrane extraction (EME) has been applied to extract these compounds from chicken breast samples. This method is based fundamentally on ionic compounds' electrostatic attraction, diffusion, and solubility in the acceptor phase. The chemical structure of polyamines enables their efficient extraction using an electric driving force on a microchip device. HCl solution (0.1 mol L-1) was applied as an aqueous acceptor solvent. Dispersive liquid-liquid microextraction was performed after EME to facilitate joining three-phase EME to GC-MS and improve the merit figures. The total ranges of 3.77-7.89 µg g-1, 3.48-7.02 µg g-1, and 0.78-2.20 µg g-1 were acquired as PUT, SPM and SPD concentrations in chicken breast, respectively. The results demonstrate that the level of PAs in fresh chicken breast samples is not concerning, but it may reduce the quality of chicken meat over time. This novel analytical technique has several advantages: high recovery, substantial quickness, remarkable selectivity, and good enrichment factors. This emerging method could be generalized to other studies to analyze different foodstuffs.

Volatile N-nitrosamines in processed meat products: An approach for monitoring dietary exposure, assessing human risk, and evaluating variable correlations by principal component analysis and heat map.

Several epidemiological studies have reported a positive association between the consumption of processed meats containing N-nitrosamines (NAs) and the incidence of hepatocellular and colon cancer. The health risk assessment in this investigation was based on the concentration of six volatile N-nitrosamines (VNAs) (N-nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosomethylethylamine, N-nitrosopiperidine, N-nitrosodibutylamine, and N-nitrosodi-n-propylamine) found in processed meat products (sausage and kielbasa) in the Iranian market. Direct supported liquid membrane two-phase hollow fiber electromembrane extraction coupled to gas chromatography/mass spectrometry was used to analyse six VNAs. The mean concentration of the six VNAs in sausages and kielbasa was 38.677 ± 27.56 and 48.383 ± 35.76 µg/kg, respectively. The 95th percentile for the chronic daily intake of total VNAs for children (3-14 years) and adults (15-70 years) were calculated to be 5.06 × 10-4 and 1.09 × 10-4 mg/kg bw/day, respectively. The cancer risk assessment showed that the risk associated with NDEA was the highest among the other VNAs studied in Iranian processed meat, with a 95th percentile for the child and adult groups. Based on an incremental lifetime cancer risk (ILCR) value of ≤10-4 for the carcinogenic effects of exposure to a total of six VNAs, it indicates low concern for all age groups.

Synthesis, design, biological evaluation, and computational analysis of some novel uracil-azole derivatives as cytotoxic agents.

The design and synthesis of novel cytotoxic agents is still an interesting topic for medicinal chemistry researchers due to the unwanted side effects of anticancer drugs. In this study, a novel series of uracil-azole hybrids were designed and synthesized. The cytotoxic activity, along with computational studies: molecular docking, molecular dynamic simulation, density functional theory, and ADME properties were also, evaluated. The compounds were synthesized by using 3-methyl-6-chlorouracil as the starting material. Cytotoxicity was determined using MTT assay in the breast carcinoma cell line (MCF-7) and Hepatocellular carcinoma cell line (HEPG-2). These derivatives demonstrated powerful inhibitory activity against breast and hepatocellular carcinoma cell lines in comparison to Cisplatin as positive control. Among these compounds, 4j displayed the best selectivity profile and good activity with IC50 values of 16.18 ± 1.02 and 7.56 ± 5.28 µM against MCF-7 and HEPG-2 cell lines respectively. Structure-activity relationships revealed that the variation in the cytotoxic potency of the synthesized compounds was affected by various substitutions of benzyl moiety. The docking output showed that 4j bind well in the active site of EGFR and formed a stable complex with the EGFR protein. DFT was used to investigate the reactivity descriptors of 4a and 4j. The outputs demonstrated that these uracil-azole hybrids can be considered as potential cytotoxic agents.

The Measurement of Hazardous Biogenic Amines in Non-Alcoholic Beers: Efficient and Applicable Miniaturized Electro-Membrane Extraction Joined to Gas Chromatography-Mass Spectrometry.

The determination of biogenic amines (BAs) as serious food contaminants and chemical indicators of unwanted microbial contamination or deficient processing conditions in non-alcoholic beers is of great interest for the beverage industries. In the present investigation, the combination of hollow fiber-electro-membrane extraction (HF-EME) and dispersive liquid-liquid microextraction (DLLME) followed by gas chromatography-mass spectrometry (GC/MS) was applied for the analysis of histamine, putrescine, tyramine, cadaverine in non-alcoholic beers. EME is fundamentally based on the electrostatic attraction, diffusion and solvability of analytes in a selected acceptor phase. This membrane-based extraction technique promoted selectivity and the enrichment factor. The DLLME process reduced the volumes of organic solvents and make the coupling of HF-EME to the CG/MS conceivable. The leading variables, which have a great effect on extraction recovery, were optimized. The relative standard deviation was achieved between 4.9 and 7.0%. The recoveries were between 94% and 98%. The limit of detection and limit of quantification were found to be 0.92-0.98 ng mL-1 and 3.03-3.23 ng mL-1, respectively. The enrichment factor was calculated in the range 36-41. The achievements revealed that putrescine and tyramine, with concentrations of 3.87 and 2.33 µg g-1, were at the highest concentration in non-alcoholic beers. This offered method with great benefits could help beverage industries to monitor the concentration of BAs in beers and control them.

Testicular histopathology in rats co-exposed to heat and psychological stressors.

Objectives: The present study aimed to investigate the effect of co-exposure to heat and psychological stressors on testicular tissue as one of the major male reproductive organs in rats. Methods: Forty adult male Wistar rats were divided into four groups of 10 animals each. The first group was exposed to heat stress (Temperature Humidity Index: 57.75 ± 3), the second group was exposed to three psychological stressors including strobe light ultrasonic sound, and tilting cage, and the third group was co-exposed to both heat and psychological stress daily. The order of exposure to various psychological stressors was randomized. Following the last day of the 40 -day exposure, the rats were euthanized and their testicular tissues were fixed in Bouin's solution. Moreover, a tissue processor, a microtome as well as hematoxylin, and eosin staining were used for tissue preparation. Results: The co-exposure to psychological and heat stress can cause negative testis histopathological changes including spermatid retention, degeneration of round spermatids and spermatocytes, degeneration and depletion of elongated Spermatid, Sertoli cell status, tubular degeneration/atrophy, Leydig cell atrophy, tubular dilatation, re-tubular dilation, and sclerosis status in a number of rats. Moreover, the histopathological changes were higher in the group exposed to heat stress compared with those exposed to psychological stress. Conclusions: Although exposure to either stressor alone can affect testicular tissue as part of the reproductive system, co-exposure to both stressors may result in an increased risk of adverse effects on testicular tissue.

New and efficient direct-SLM two-phase hollow fiber electromembrane extraction coupled to GC/MS for the analysis of nitrosamines in different types of sausage: Investigation of meat type, meat percent and cooking methods.

The potent two-phase hollow fiber electromembrane extraction technique coupled to gas chromatography mass spectrometry (HF-EME/GC-MS) was proposed for the determination of six types of carcinogen nitrosamines in sausages samples. Two steps of sample digestion were accomplished for the complete removal of fat globules and efficient release of target analytes. The extraction principle was based on electro-migration of target analytes via specific fiber to extraction solvent. 2-Nitrophenyl octyl ether (NPOE) was dexterously employed as both supported liquid membrane and extraction solvent, which is compatible with GC-MS. After the extraction process, the NPOE containing nitrosamines was directly injected to GC-MS without extra steps requirement to reduce analysis time. The consequences revealed that N-nitrosodiethylamine (NDEA) as the most potent carcinogen has the highest concentration in fried and oven-cooked sausages with 70% of red meat. The meat type and amount and also cooking process could significantly effect on nitrosamines formation.

Response Surface Methodology of Quantitative of Heterocyclic Aromatic Amines in Fried Fish Using Efficient Microextraction Method Coupled with High-Performance Liquid Chromatography: Central Composite Design.

Meat and meat products are indispensable part of our diet. Heat processing of these tasty foods such as fried fish causes to form heterocyclic aromatic amines (HAAs). The sources of heating have directly affected on the level and type of HAAs. In this research, 2-amino-1-methyl-6-phenylimidazo [4'5-b] pyridine (PhIP), 2-amino-3-methylimidazo [4,5-f]quinolone (IQ), 2-amino-3,4-dimethylimidazo [4,5-f] quinoline (MeIQ) and 2-amino-3,4-dimethylimidazo [4,5-f] quinoxaline (MeIQx) were determined using an efficient analytical methodology coupled with high-performance liquid chromatography. The effective parameters were optimized by central composite design. The results of this survey demonstrated that rang of relative standard deviation were between 4.5 and 8.2, extraction recoveries were obtained 86-97% and limits of detection were between 0.40 and 0.63 for 4 HAAs. The amounts of HAAs found in 20 different fried fish samples were between 0 and 4.8 ng g-1. PhIP with 1.57 ng g-1 and MeIQ with 2.08 ng g-1 have the lowest and highest average level of HAAs, respectively.

Heterocyclic aromatic amines in doner kebab: Quantitation using an efficient microextraction technique coupled with reversed-phase high-performance liquid chromatography.

The safety of doner kebab as a traditional Middle East tasty food can threaten via the formation of dangerous compounds such as heterocyclic aromatic amines during heat process. In this regard, the current investigation was devoted to measuring of 4 HAAs (2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), and 2-amino-3-methylimidazo[4,5-f]quinoline (IQ)) in doner kebab samples with an innovative microextraction technique combined with high-performance liquid chromatography. The limit of detection was in the range of 4.8 and 5.3 ng/g, while relative standard deviations were between 6.5% and 8.3%, and recoveries were calculated in the range of 89%-97%. The most and the least total mean values of HAA levels were 13.30 ng/g for MeIQx and 5.0 ng/g for IQ. The proposed method showed a high capability to extract trace amount of HAAs from a complex matrix such as doner kebab. Also, this technique is easy, high sensitive, selective, accurate and efficient.

Adjunctive Memantine Treatment of Schizophrenia: A Double-Blind, Randomized Placebo-Controlled Study.

PURPOSE/BACKGROUND: This double-blind, placebo-controlled clinical trial was designed to assess the efficacy and safety of memantine augmentation to standard regimen of antipsychotic treatment on psychotic symptoms and cognitive function in individuals with chronic schizophrenia for 8 weeks. METHODS/PROCEDURES: Forty stabilized individuals with chronic schizophrenia were randomized in a 1:1 ratio to memantine (20 mg/d) and control (placebo) groups, along with their antipsychotic regimen for 8 weeks. The efficacy of treatment was assessed by the Positive and Negative Syndrome Scale (PANSS) and Brief Assessment of Cognition Scale, and the safety was measured by the Abnormal Involuntary Movement Scale and Barnes Akathisia Rating Scale at baseline and at weeks 4 and 8. FINDINGS/RESULTS: No significant differences were observed in demographic or clinical variables between both groups at baseline. During the study, all subscales and total scores of PANSS decreased significantly within both groups, except the subscale score in memantine, which was found to be positive. Reduction in general subscale and total scores of PANSS was significantly higher in the control group compared with the memantine group. All subscale scores of the Brief Assessment of Cognition Scale increased significantly only in the memantine group. The increase in the Verbal Memory, Working Memory, Verbal Fluency Letter, and Verbal Fluency Total subscale scores was significantly higher in the memantine group than in the control group. There was no significant difference in the Abnormal Involuntary Movement Scale and Barnes Akathisia Rating Scale scores between the 2 groups during the study. IMPLICATIONS/CONCLUSIONS: This study showed that adjunctive memantine to antipsychotic regimen improved the verbal memory, learning, verbal letter fluency, and working memory without improvement on psychotic symptoms in individuals with chronic schizophrenia.

Heterocyclic aromatic amines in cooked food: A review on formation, health risk-toxicology and their analytical techniques.

Heterocyclic aromatic amines (HAAs) are defined as a major class of poisonous compounds formed from proteinaceous foods during heat processing and flavour-forming. These toxicants have detrimental effects on the human body and eventually bring about mutagenicity and carcinogenicity. Owing to the presence of HAAs at a marginal level in the intricate tissue of food and their interactivity, an effective sample preparation should be employed to extract these controversial chemicals from the food matrix. For separation and detection of HAAs, advanced extraction methods and instrumental techniques have been applied. According to the type of sample preparation, sensitivity of the experiment, laboratory conditions and available facilities, the choice of analytical equipment will be different. In this review, various cooked food samples containing HAAs and heat processing of them have been listed. Also, sample pre-treatment and analytical techniques that have been applied to determine HAAs are discussed.

Alterations in oxidative stress markers and its correlation with clinical findings in schizophrenic patients consuming perphenazine, clozapine and risperidone.

BACKGROUND AND OBJECTIVE: Today, the role of oxidative stress in development of schizophrenia has gained attention. Also, some atypical antipsychotic agents showed antioxidant properties. Therefore, in this study, we evaluated the level of oxidative stress parameters between patients treated with perphenazine, clozapine and risperidone and their relationship with schizophrenia symptoms' severity. MATERIALS AND METHODS: This was a descriptive study on 100 patients with chronic schizophrenia. Patient selection was done based on the DSM-IV-TR criteria for at least 3 months regular use of clozapine or risperidone or perphenazine and a minimum period of 2 years of schizophrenia. Ten ml of patient's blood samples were used to assess serum levels of glutathione (GSH), protein carbonyl, lipid peroxidation (LPO), superoxide dismutase (SOD) and Ferric Reducing Ability of Plasma (FRAP). Also, the severity of symptoms was assessed with the positive and negative syndrome scale (PANSS scale). P-values of less than 0.05 were considered significant. RESULTS: The results showed a significant difference between clozapine and risperidone with perphenazine in all subscales of PANSS. Also, there was a positive correlation between MDA and PANSS all subscales in risperidone and perphenazine groups and a negative correlation between MDA and PANSS in all subscales in the clozapine group. Serum level of GSH and negative symptoms in patients receiving clozapine showed a negative correlation. The results also represented that clozapine significantly increased SOD levels in comparison to perphenazine and risperidone and reduced LPO in comparison to perphenazine and risperidone, While the protein carbonyl level did not show a significant difference between three groups (p-value = 0.8). CONCLUSION: This study showed that clozapine, as an atypical antipsychotic agent, has significant antioxidant effects compared to risperidone and perphenazine. Especially, it increased SOD and GSH levels and reduced LPO in patients with schizophrenia. Therefore, clozapine's antioxidant effect may be contributive to improving negative symptoms of schizophrenic patients.

Cytotoxic activity of abietane diterpenoids from roots of Salvia sahendica by HPLC-based activity profiling

ABSTRACT Screening of medicinal plants from Iranian flora against human cancer cell-lines have shown that an hexane extract from roots of Salvia sahendica Boiss. & Buhse, Lamiaceae, is active against human cervical cancer (HeLa) and colorectal adenocarcinoma (Caco-2) cell-lines at the test concentration of 100 µg/ml (100% inhibition). Cytotoxicity of the extract was localized with the aid of HPLC-time-based activity profiling adapted to the tetrazolium colorimetric bioassay. Four abietane-type diterpenoids in active time-windows were identified as cytotoxic compounds namely: sahandone (1), sahandol (2), 12-deoxy-salvipisone (3) and sahandinone (4). Compound 1 showed the highest toxicity against HeLa cells (IC50 = 5.6 ± 0.1 µg/ml), which was comparable with betulinic acid (IC50 = 4.3 ± 1.2 µg/ml), the positive control. Compound 2 was active against the HeLa cells (IC50 = 8.9 ± 0.7 µg/ml) but not the Caco-2 cell-line. Compounds 1, 3 and 4 exhibited moderate activity (IC50 = 22.9-41.4 µg/ml) against the Caco-2 cells. This study reveals that the HeLa cells are more sensitive to all tested compounds than the Caco-2 cells. In silico molecular docking study showed a rigid binding of the compounds to tyrosine kinase pp60src, and proved their cytotoxic activity.