Fungal Inactivation by Mexican Oregano (Lippia berlandieri Schauer) Essential Oil Added to Amaranth,Chitosan, or Starch Edible Films

ABSTRACT: Edible films can incorporate antimicrobial agents to provide microbiological stability, since they can be used as carriers of a wide number of additives that can extend product shelf life and reduce the risk of pathogenic bacteria growth on food surfaces. Addition of antimicrobial agents to edible films offers advantages such as the use of low antimicrobial concentrations and low diffusion rates. The aim of this study was to evaluate inhibition of Aspergillus niger and Penicillium spp. by selected concentrations of Mexican oregano (Lippia berlandieri Schauer) essential oil added to amaranth, chitosan, or starch edible films. Oregano essential oil was characterized by gas chromatography-mass spectrometry (GC/MS) analysis. Amaranth, chitosan, and starch edible films were formulated with essential oil concentrations of 0%, 0.25%, 0.50%, 0.75%, 1%, 2%, and 4%. Mold radial growth was evaluated inoculating spores in 2 ways: edible films were placed over inoculated agar, Film/Inoculum mode (F/I), or the edible films were first placed in the agar and then films were inoculated, Inoculum/Film mode (I/F). The modified Gompertz model adequately described growth curves. There was no significant difference (P > 0.05) in growth parameters between the 2 modes of inoculation. Antifungal effectiveness of edible films was starch > chitosan > amaranth. In starch edible films, both studied molds were inhibited with 0.50% of essential oil. Edible films added with Mexican oregano essential oil could improve the quality of foods by controlling surface growth of molds.     

Glucuronidation of zearalenone,zeranol and four metabolites in vitro: Formation of glucuronides by various microsomes and human UDP‐glucuronosyltransferase isoforms

Glucuronidation constitutes an important pathway in the phase II metabolism of the mycotoxin zearalenone (ZEN) and the growth promotor α‐zearalanol (α‐ZAL, zeranol), but the enzymology of their formation is yet unknown. In the present study, ZEN, α‐ZAL and four of their major phase I metabolites were glucuronidated in vitro using hepatic microsomes from steer, pig, rat and human, intestinal microsomes from humans, and eleven recombinant human UDP‐glucuronosyltransferases (UGTs). After assigning chemical structures to the various glucuronides by using previously published information, the enzymatic activities of the various microsomes and UGT isoforms were determined together with the patterns of glucuronides generated. All six compounds were good substrates for all microsomes studied. With very few exceptions, glucuronidation occurred preferentially at the sterically unhindered phenolic 14‐hydroxyl group. UGT1A1, 1A3 and 1A8 had the highest activities and gave rise to the phenolic glucuronide, whereas glucuronidation of the aliphatic hydroxyl group was mostly mediated by UGT2B7 with low activity. Based on these in vitro data, ZEN, α‐ZAL and their metabolites must be expected to be readily glucuronidated both in the liver and intestine as well as in other extrahepatic organs of humans and various animal species.     

Agar-immobilized basil–lactic acid bacteria bioproducts as goat milk taste-masking agents and natural preservatives for the production of unripened goat cheese

Goat milk cheeses have become popular recently; however, many consumers do not choose these products because they have specific sensory properties that are not acceptable to all consumers and the shelf life of the cheese is short. The concept of this work was to increase overall acceptability and shelf life of unripened goat milk cheese by using Ocimum basilicum and lactic acid bacteria (Lactobacillus plantarum LUHS135, Lactobacillus paracasei LUHS244, Pediococcus pentosaceus LUHS100, Pediococcus acidilactici LUHS29, and Lactobacillus brevis LUHS140) bioproducts (basil-LAB) immobilized in agar. A basil-LAB bioproduct could be a promising multifunctional ingredient for cheese manufacturing because it has a low pH, high LAB count, and high total phenolic compound content (after fermentation pH decreased by 25.4%, LAB count averaged 7.2 log₁₀ cfu/g, and total phenolic compound content increased by 30.9%). Use of different LAB in the preparation of basil-LAB bioproducts had a significant influence on cheese pH and hardness, and compared with cheese samples prepared with nonfermented basil, cheese samples prepared with basil-LAB bioproducts had, on average, higher pH (by 2.6%) and lower hardness (by 36.0%), similar to the control cheese (without basil). Overall acceptability of cheese was significantly influenced by the basil-LAB bioproduct immobilization process; in all cases, cheese samples prepared with fermented and immobilized basil-LAB bioproduct had better acceptability (5 points). After 120 h of storage, cheese samples prepared with basil-LAB bioproducts fermented with LUHS135, LUHS244 and LUHS140, no enterobacteria were found, and we detected strong negative and moderate negative correlations, respectively, of LAB count with enterobacteria count and yeast/mold count (r = ?0.7939 and r = ?0.4495, respectively). Finally, immobilization increased LAB viability in fresh goat milk cheese, which led to a reduction in enterobacteria and mold/yeast contamination during storage and an increase in overall acceptability compared with nonimmobilized basil-LAB. Therefore, basil-LAB bioproducts fermented with LUHS135, LUHS244, and LUHS140 strains can be recommended for preparing fresh goat milk cheese with extended shelf life and high acceptability.     

Phase stability and hydrogen desorption in a quinary equimolar mixture of light-metals borohydrides

The present study aims at investigating, for the first time, a quinary mixture of light-metals borohydrides. The goal is to design combinations of borohydrides with multiple cations in equimolar ratio, following the concept of high entropy alloys. The equimolar composition of the LiBH₄NaBH₄KBH₄Mg(BH₄)₂Ca(BH₄)₂ system was synthetized by ball milling. The obtained phases were analysed by X-ray diffraction and in-situ Synchrotron Radiation Powder X-ray Diffraction, in order to establish the amount of cations incorporated in the obtained crystalline phases and to study the thermal behaviour of the mixture. HP-DSC and DTA were also used to define the phase transformations and thermal decomposition reactions, leading to the release of hydrogen, that was detected by MS. The existence of a quinary liquid borohydride phase is reported for the first time. Effects of the presence of multi-cations compounds or a liquid phase on the hydrogen desorption reactions are described.     

Exploration of Somatostatin Binding Mechanism to Somatostatin Receptor Subtype 4

Somatostatin (also named as growth hormone-inhibiting hormone or somatotropin release-inhibiting factor) is a regulatory peptide important for the proper functioning of the endocrine system, local inflammatory reactions, mood and motor coordination, and behavioral responses to stress. Somatostatin exerts its effects via binding to G-protein-coupled somatostatin receptors of which the fourth subtype (SSTR4) is a particularly important receptor mediating analgesic, anti-inflammatory, and anti-depressant effects without endocrine actions. Thus, SSTR4 agonists are promising drug candidates. Although the knowledge of the atomic resolution-binding modes of SST would be essential for drug development, experimental elucidation of the structures of SSTR4 and its complexes is still awaiting. In the present study, structures of the somatostatin–SSTR4 complex were produced using an unbiased, blind docking approach. Beyond the static structures, the binding mechanism of SST was also elucidated in the explicit water molecular dynamics (MD) calculations, and key binding modes (external, intermediate, and internal) were distinguished. The most important residues on both receptor and SST sides were identified. An energetic comparison of SST binding to SSTR4 and 2 offered a residue-level explanation of receptor subtype selectivity. The calculated structures show good agreement with available experimental results and indicate that somatostatin binding is realized via prerequisite binding modes and an induced fit mechanism. The identified binding modes and the corresponding key residues provide useful information for future drug design targeting SSTR4.     

Peripheral Ion Channel Gene Screening in Painful- and Painless-Diabetic Neuropathy

Neuropathic pain is common in diabetic peripheral neuropathy (DN), probably caused by pathogenic ion channel gene variants. Therefore, we performed molecular inversion probes-next generation sequencing of 5 transient receptor potential cation channels, 8 potassium channels and 2 calcium-activated chloride channel genes in 222 painful- and 304 painless-DN patients. Twelve painful-DN (5.4%) patients showed potentially pathogenic variants (five nonsense/frameshift, seven missense, one out-of-frame deletion) in ANO3 (n = 3), HCN1 (n = 1), KCNK18 (n = 2), TRPA1 (n = 3), TRPM8 (n = 3) and TRPV4 (n = 1) and fourteen painless-DN patients (4.6%—three nonsense/frameshift, nine missense, one out-of-frame deletion) in ANO1 (n = 1), KCNK18 (n = 3), KCNQ3 (n = 1), TRPA1 (n = 2), TRPM8 (n = 1), TRPV1 (n = 3) and TRPV4 (n = 3). Missense variants were present in both conditions, presumably with loss- or gain-of-functions. KCNK18 nonsense/frameshift variants were found in painless/painful-DN, making a causal role in pain less likely. Surprisingly, premature stop-codons with likely nonsense-mediated RNA-decay were more frequent in painful-DN. Although limited in number, painful-DN patients with ion channel gene variants reported higher maximal pain during the night and day. Moreover, painful-DN patients with TRP variants had abnormal thermal thresholds and more severe pain during the night and day. Our results suggest a role of ion channel gene variants in neuropathic pain, but functional validation is required.     

Anxiety-like Behavior and GABAAR/BDZ Binding Site Response to Progesterone Withdrawal in a Stress-Vulnerable Strain,the Wistar Kyoto Rats

Stress susceptibility could play a role in developing premenstrual anxiety due to abnormalities in the hypothalamus–pituitary–adrenal (HPA) axis and impairments in the GABAA receptors’ benzodiazepine (BDZ) site. Hence, we studied the stress-vulnerable Wistar Kyoto rat strain (WKY) to evaluate progesterone withdrawal (PW) effects on anxiety, HPA axis response, and to explore indicators of GABAA functionality in the BDZ site. For five days, ovariectomized WKY rats were administered 2.0 mg/kg of progesterone. Twenty-four hours after the last administration, rats were tested in the anxiety-like burying behavior test (BBT) or elevated plus maze test (EPM), and corticosterone was determined. [3H]Flunitrazepam binding autoradiography served as the BDZ binding site index of the GABAA receptor in amygdala nuclei and hippocampus’s dentate gyrus (DG). Finally, different doses of diazepam in PW-WKY rats were tested in the BBT. PW induced anxiety-like behaviors in both BBT and EPM compared with No-PW rats. PW increased corticosterone, but was blunted when combined with PW and BBT. PW increased [3H]Flunitrazepam binding in the DG and central amygdala compared with No-PW rats. Diazepam at a low dose induced an anxiogenic-like response in PW rats, suggesting a paradoxical response to benzodiazepines. Overall, PW induced anxiety-like behavior, a blunted HPA axis response, and higher GABAAR/BZD binding site sensitivity in a stress-vulnerable rat strain. These findings demonstrate the role of stress-susceptibility in GABAAR functionality in a preclinical approximation of PMDD.     

Physical aggression and marital dysfunction: A longitudinal analysis.

Shortly after marriage, 56 couples provided data on physical aggression and other predictors of marital adjustment. At 6-month intervals over the next 4 years, spouses reported on their marital quality and stability. Results indicated that marital dysfunction was more common among aggressive than among nonaggressive couples (70% vs. 38%) and among severely aggressive than among moderately aggressive couples (93% vs. 46%). Aggression remained a reliable predictor of marital outcomes after the authors controlled for stressful events and negative communication. These findings help to refine developmental models of marital dysfunction, which often overlook the role of aggression, and can provide information for prevention programs for marital distress, which typically do not distinguish between aggressive and nonaggressive couples. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Targeting Tumor Acidosis and Regulatory T Cells Unmasks Anti-Metastatic Potential of Local Tumor Ablation in Triple-Negative Breast Cancer

Triple-negative breast cancer (TNBC) is an immunologically heterogenous disease that lacks clinically actionable targets and is more likely to progress to metastatic disease than other types of breast cancer. Tumor ablation has been used to increase response rates to checkpoint inhibitors, which remain low for TNBC patients. We hypothesized that tumor ablation could produce an anti-tumor response without using checkpoint inhibitors if immunosuppression (i.e., Tregs, tumor acidosis) was subdued. Tumors were primed with sodium bicarbonate (200 mM p.o.) to reduce tumor acidosis and low-dose cyclophosphamide (100–200 mg/kg i.p.) to deplete regulatory T cells, as has been shown independently in previous studies. A novel injectable ablative was then used to necrose the tumor, release tumor antigens, and initiate an immune event that could create an abscopal effect. This combination of bicarbonate, cyclophosphamide, and ablation, called “BiCyclA”, was tested in three syngeneic models of TNBC: E0771 (C57BL/6), 67NR (BALB/c), and 4T1-Luc (BALB/c). In E0771 and 67NR, BiCyclA therapy significantly reduced tumor growth and cured 5/7 and 6/10 mice 50 days after treatment respectively. In the metastatic 4T1-Luc tumors, for which surgery and checkpoint inhibitors fail, BiCyclA cured 5/10 mice of primary tumors and lung metastases. Notably, CD4+ and CD8+ T cells were found to be crucial for the anti-metastatic response, and cured mice were able to resist tumor rechallenge, suggesting production of immune memory. Reduction of tumor acidity and regulatory T cells with ablation is a simple yet effective therapy for local and systemic tumor control with broad applicability as it is not limited by expensive supplies.