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1.
Alcohol-related chronic exocrine pancreatic insufficiency: diagnosis and therapeutic management. A proposal for treatment by the Italian Association for the Study of the Pancreas (AISP) and the Italian Society of Alcohology (SIA).
Pezzilli, R, Caputo, F, Testino, G, Patussi, V, Greco, G, Macciò, L, Rossin, MR, Mioni, D, Balbinot, P, Gandin, C, et al
Minerva medica. 2019;(5):425-438
Abstract
Current estimates of the prevalence of chronic pancreatitis, one of the most common causes of exocrine pancreatic insufficiency, are in the range of 3-10 per 100,000 people in many parts of the world. Alcohol consumption is a very important risk factor for exocrine pancreatic insufficiency and is involved in nearly half of all cases. The main hypothesis regarding the role of chronic alcohol consumption in pancreatitis is that there must be additional environmental or genetic risk factors involved for ongoing damage to occur. Treatment of patients with alcohol-related exocrine pancreatic insufficiency is complex, as the patient has two concomitant pathologies, alcohol-use disorder (AUD) and exocrine pancreatic insufficiency/chronic pancreatitis. Alcohol abstinence is the starting point for treatment, although even this along with the most advanced therapies allow only a slowdown in progression rather than restoration of function. This position paper of the Italian Association for the Study of the Pancreas and the Italian Society of Alcohology provides an overview of the pathogenesis of alcohol-related pancreatitis and discuss diagnostic issues. Treatment options for both exocrine pancreatic insufficiency/chronic pancreatitis (with a focus on pancreatic enzyme replacement therapy) and AUD (acamprosate, disulfiram, oral naltrexone, long-acting injectable naltrexone, sodium oxybate, nalmefene, baclofen, and psychosocial interventions) are also reviewed.
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2.
Engineering grass biomass for sustainable and enhanced bioethanol production.
Mohapatra, S, Mishra, SS, Bhalla, P, Thatoi, H
Planta. 2019;(2):395-412
Abstract
Bioethanol from lignocellulosic biomass is a promising step for the future energy requirements. Grass is a potential lignocellulosic biomass which can be utilised for biorefinery-based bioethanol production. Grass biomass is a suitable feedstock for bioethanol production due to its all the year around production, requirement of less fertile land and noninterference with food system. However, the processes involved, i.e. pretreatment, enzymatic hydrolysis and fermentation for bioethanol production from grass biomass, are both time consuming and costly. Developing the grass biomass in planta for enhanced bioethanol production is a promising step for maximum utilisation of this valuable feedstock and, thus, is the focus of the present review. Modern breeding techniques and transgenic processes are attractive methods which can be utilised for development of the feedstock. However, the outcomes are not always predictable and the time period required for obtaining a robust variety is generation dependent. Sophisticated genome editing technologies such as synthetic genetic circuits (SGC) or clustered regularly interspaced short palindromic repeats (CRISPR) systems are advantageous for induction of desired traits/heritable mutations in a foreseeable genome location in the 1st mutant generation. Although, its application in grass biomass for bioethanol is limited, these sophisticated techniques are anticipated to exhibit more flexibility in engineering the expression pattern for qualitative and qualitative traits. Nevertheless, the fundamentals rendered by the genetics of the transgenic crops will remain the basis of such developments for obtaining biorefinery-based bioethanol concepts from grass biomass. Grasses which are abundant and widespread in nature epitomise attractive lignocellulosic feedstocks for bioethanol production. The complexity offered by the grass cell wall in terms of lignin recalcitrance and its binding to polysaccharides forms a barricade for its commercialization as a biofuel feedstock. Inspired by the possibilities for rewiring the genetic makeup of grass biomass for reduced lignin and lignin-polysaccharide linkages along with increase in carbohydrates, innovative approaches for in planta modifications are forging ahead. In this review, we highlight the progress made in the field of transgenic grasses for bioethanol production and focus our understanding on improvements of simple breeding techniques and post-harvest techniques for development in shortening of lignin-carbohydrate and carbohydrate-carbohydrate linkages. Further, we discuss about the designer lignins which are aimed for qualitable lignins and also emphasise on remodelling of polysaccharides and mixed-linkage glucans for enhancing carbohydrate content and in planta saccharification efficiency. As a final point, we discuss the role of synthetic genetic circuits and CRISPR systems in targeted improvement of cell wall components without compromising the plant growth and health. It is anticipated that this review can provide a rational approach towards a better understanding of application of in planta genetic engineering aspects for designing synthetic genetic circuits which can promote grass feedstocks for biorefinery-based bioethanol concepts.
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3.
Relation of plasma carnitine and aminotransferases to alcohol dose and time of dependence.
Kępka, A, Zwierz, P, Chojnowska, S, Ochocińska, A, Skorupa, E, Szczepański, M, Szajda, SD, Waszkiewicz, N
Alcohol (Fayetteville, N.Y.). 2019;:62-69
Abstract
BACKGROUND Serum aspartate, alanine aminotransferases (AST, ALT), and plasma carnitine are all indirect biomarkers of alcohol abuse. Carnitine transfers long-chain fatty acids from cytoplasm to mitochondria for β-oxidation. The aim of the study was to determine the relationship between daily alcohol intake, time of alcohol dependence, plasma carnitine, and serum aminotransferases. PATIENTS We studied 26 men who were addicted for 2-30 years, consuming ethanol from 75 to 700 g/day (alcoholic group), as well as 17 healthy men (control group). RESULTS In alcoholics, compared to the controls, we found: a significant increase in serum: AST (p = 0.0014), ALT (p = 0.0071), AST/ALT ratio (p < 0.000); significantly lower plasma free carnitine (FC) (p = 0.0316) and total carnitine (TC) (p = 0.0349); and a significant negative correlation between FC (r = -0.6200; R2 = 0.3844; p = 0.0007), TC (r = -0.4365; R2 = 0.1905; p = 0.0258), and time of alcohol dependence, suggesting carnitine as an indirect marker of alcohol abuse. We did not find any significant correlation between FC, TC, and levels of alcohol or aminotransferase activity. CONCLUSION In the alcoholic group, there was an increase in serum activity of AST, ALT, and AST/ALT ratio that confirms liver injury. In addition, we found low plasma FC and TC, which may indicate damage to mitochondrial β-oxidation caused by alcohol metabolites. The significantly higher plasma FC and TC in patients consuming the most, compared to patients consuming smaller doses of alcohol, may be caused by a lower carnitine demand of injured liver cells, decreased urinary carnitine excretion by impaired renal tubules, and leakage of carnitine into the blood from damaged muscles by the higher quantities of alcohol. The negative correlation between carnitine concentration and time of alcohol dependence may suggest the potential use of carnitine for treatment of alcohol abuse.
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4.
The intriguing effect of ethanol and nicotine on acetylcholine-sensitive potassium current IKAch: Insight from a quantitative model.
Šimurda, J, Šimurdová, M, Bébarová, M
PloS one. 2019;(10):e0223448
Abstract
Recent experimental work has revealed unusual features of the effect of certain drugs on cardiac inwardly rectifying potassium currents, including the constitutively active and acetylcholine-induced components of acetylcholine-sensitive current (IKAch). These unusual features have included alternating susceptibility of the current components to activation and inhibition induced by ethanol or nicotine applied at various concentrations, and significant correlation between the drug effect and the current magnitude measured under drug-free conditions. To explain these complex drug effects, we have developed a new type of quantitative model to offer a possible interpretation of the effect of ethanol and nicotine on the IKAch channels. The model is based on a description of IKAch as a sum of particular currents related to the populations of channels formed by identical assemblies of different α-subunits. Assuming two different channel populations in agreement with the two reported functional IKAch-channels (GIRK1/4 and GIRK4), the model was able to simulate all the above-mentioned characteristic features of drug-channel interactions and also the dispersion of the current measured in different cells. The formulation of our model equations allows the model to be incorporated easily into the existing integrative models of electrical activity of cardiac cells involving quantitative description of IKAch. We suppose that the model could also help make sense of certain observations related to the channels that do not show inward rectification. This new ionic channel model, based on a concept we call population type, may allow for the interpretation of complex interactions of drugs with ionic channels of various types, which cannot be done using the ionic channel models available so far.
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5.
Optimization of fermentation-relevant factors: A strategy to reduce ethanol in red wine by sequential culture of native yeasts.
Maturano, YP, Mestre, MV, Kuchen, B, Toro, ME, Mercado, LA, Vazquez, F, Combina, M
International journal of food microbiology. 2019;:40-48
Abstract
Current consumer preferences are determined by well-structured, full-bodied wines with a rich flavor and with reduced alcohol levels. One of the strategies for obtaining wines with reduced ethanol content is sequential inoculation of non-Saccharomyces and Saccharomyces cerevisiae yeasts. However, different factors affect the production of metabolites like ethanol, glycerol and acetic acid by inoculated yeasts. In order to obtain low alcohol wines without quality loss, the aims of our study were: i) to determine optimum conditions (fermentation temperature and time of permanence and initial inoculum size of the non-Saccharomyces population at the beginning of the process, prior to inoculation with S. cerevisiae); ii) to validate the optimized factors; and iii) to assess sensory quality of the wines obtained after validation. Two combinations of yeasts were used in this study: Hanseniaspora uvarum BHu9/S. cerevisiae BSc114 and Candida membranaefaciens BCm71/S. cerevisiae BSc114. Optimization of three fermentation factors that affect to non-Saccharomyces yeasts prior to S. cerevisiae inoculation was carried out using a Box-Behnken experimental design. Applying the models constructed by Response Surface Methodology, the lowest ethanol production by H. uvarum BHu9/S. cerevisiae BSc114 co-culture was obtained when H. uvarum BHu9 was inoculated 48 h 37 min prior to S. cerevisiae inoculation, at a fermentation temperature of 25 °C and at an initial inoculum size of 5 × 106 cells/mL. Lowest alcohol production with C. membranaefaciens BCm71/S. cerevisiae BSc114 was observed when C. membranaefaciens BCm71 was inoculated 24 h 15 min prior to S. cerevisiae at a fermentation temperature of 24.94 °C and at an initial inoculum size of 2.72 × 106 cells/mL. The optimized conditions of the two co-cultures were subsequently submitted to lab-scale validation. Both proposed strategies yielded ethanol levels that were significantly lower than control cultures (S. cerevisiae). Wines fermented with non-Saccharomyces/Saccharomyces co-cultures under optimized conditions were also associated with higher aromatic complexity characterized by the presence of red fruit aromas, whereas wines obtained with S. cerevisiae BSc114 were described by parameters linked with high ethanol levels.
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6.
Semi-Quantitative Impacts of Major Step on Antigen in Pre-Treatment Required for Immunohistochemistry.
Feng, X, Wang, W, Liu, H
Clinical laboratory. 2019;(9)
Abstract
BACKGROUND We investigated the impact of immunohistochemistry (IHC) pre-treatment steps on antigens. METHODS Salmonella typhimurium was selected as the observed antigen. The antigen was subjected to IHC pre-treatment steps involving a series of reagents, including 10% formaldehyde, ethanol, and xylene. Antigenicity was then measured by agglutination reaction. RESULTS The agglutination titer for S. typhimurium was higher in the untreated control group than in the experimental group, indicating that pre-treatment inhibited antigen activity. The inhibitory effect of ethanol was greater than that of 10% formaldehyde and xylene. Unexpectedly, partial antigen recovery can be achieved from a preparation of paraffin section after hydration. CONCLUSIONS S. Antigens may be strongly inhibited (inhibition: 70.8%) by IHC pre-treatment steps, especially by alcohol treatment. There is an experimental foundation for antigen retrieval in IHC.
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7.
Destabilization of Insulin Hexamer in Water-Ethanol Binary Mixture.
Mukherjee, S, Deshmukh, AA, Mondal, S, Gopal, B, Bagchi, B
The journal of physical chemistry. B. 2019;(49):10365-10375
Abstract
We report combined experimental and simulation studies which reveal that the structural integrity of insulin hexamer, the storehouse of the important hormone in our body, is compromised by the interactions with ethanol. X-ray crystal structures suggest that ethanol replaces water molecules inside the insulin hexamer cavity. At the maximum physiologically tolerable concentration of ethanol (∼0.6% v/v), molecular dynamics simulations show that ethanol molecules get exchanged between the bulk and the cavity with a free energy cost of ∼5 kcal mol-1. However, biological time scales are orders of magnitude longer than that achievable by molecular dynamics simulations. Hence, to accelerate the process we investigate insulin hexamer in ∼30% v/v ethanol concentration. We find that the entrance and exit of ethanol from the hexamer cavity lead to the modification of atomic contacts in the protein. This causes large-scale fluctuations that force the protein out of its native state free energy minimum. Structural perturbations are also observed at lower ethanol concentration. The computational findings are consistent with dynamic light scattering experiments that suggest an abrupt reduction in the population of insulin hexamers at a critical ethanol concentration. The structural changes triggered by interaction of ethanol with the insulin hexamer are likely to represent a general dynamic event of amphiphilic cosolvent induced changes in macromolecular assemblies with the consequent effects on cellular homeostasis.
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8.
Substance P in Solution: Trans-to-Cis Configurational Changes of Penultimate Prolines Initiate Non-enzymatic Peptide Bond Cleavages.
Conant, CR, Fuller, DR, El-Baba, TJ, Zhang, Z, Russell, DH, Clemmer, DE
Journal of the American Society for Mass Spectrometry. 2019;(6):919-931
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Abstract
We report ion mobility spectrometry and mass spectrometry studies of the non-enzymatic step-by-step degradation of substance P (subP), an 11-residue neuropeptide, with the sequence Arg1-Pro2-Lys3-Pro4-Gln5-Gln6-Phe7-Phe8-Gly9-Leu10-Met11-NH2, in ethanol. At elevated solution temperatures (55 to 75 °C), several reactions are observed, including a protonation event, i.e., [subP+2H]2+ + H+ → [subP+3H]3+, that appears to be regulated by a configurational change and two sequential bond cleavages (the Pro2-Lys3 peptide bond is cleaved to form the smaller nonapeptide Lys3-Met11-NH2 [subP(3-11)], and subsequently, subP(3-11) is cleaved at the Pro4-Gln5 peptide bond to yield the heptapeptide Gln5-Met11-NH2 [subP(5-11)]). Each of the product peptides [subP(3-11) and subP(5-11)] is accompanied by a complementary diketopiperazine (DKP): cyclo-Arg1-Pro2 (cRP) for the first cleavage, and cyclo-Lys3-Pro4 (cKP) for the second. Insight about the mechanism of degradation is obtained by comparing kinetics calculations of trial model mechanisms with experimental data. The best model of our experimental data indicates that the initial cleavage of subP is regulated by a conformational change, likely a trans→cis isomerization of the Arg1-Pro2 peptide bond. The subP(3-11) product has a long lifetime (t1/2 ~ 30 h at 55 °C) and appears to transition through several structural intermediates prior to dissociation, suggesting that subP(3-11) is initially formed with a Lys3-trans-Pro4 peptide bond configuration and that slow trans→cis isomerization regulates the second bond cleavage event as well. From these data and our model mechanisms, we obtain transition state thermochemistry ranging from ΔH‡ = 41 to 85 kJ mol-1 and ΔS‡ = - 43 to - 157 J mol-1 K-1 for each step in the reaction. Graphical Abstract.
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Modelling of Molasses Fermentation for Bioethanol Production: A Comparative Investigation of Monod and Andrews Models Accuracy Assessment.
Zentou, H, Zainal Abidin, Z, Yunus, R, Awang Biak, DR, Zouanti, M, Hassani, A
Biomolecules. 2019;(8)
Abstract
Modelling has recently become a key tool to promote the bioethanol industry and to optimise the fermentation process to be easily integrated into the industrial sector. In this context, this study aims at investigating the applicability of two mathematical models (Andrews and Monod) for molasses fermentation. The kinetics parameters for Monod and Andrews were estimated from experimental data using Matlab and OriginLab software. The models were simulated and compared with another set of experimental data that was not used for parameters' estimation. The results of modelling showed that μmax = 0.179 1/h and Ks = 11.37 g.L-1 for the Monod model, whereas μmax = 0.508 1/h, Ks = 47.53 g.L-1 and Ki = 181.01 g.L-1 for the Andrews model, which are too close to the values reported in previous studies. The validation of both models showed that the Monod model is more suitable for batch fermentation modelling at a low concentration, where the highest R squared was observed at S0 = 75 g.L-1 with an R squared equal to 0.99956, 0.99954, and 0.99859 for the biomass, substrate, and product concentrations, respectively. In contrast, the Andrews model was more accurate at a high initial substrate concentration and the model data showed a good agreement compared to the experimental data of batch fermentation at S0 = 225 g.L-1, which was reflected in a high R squared with values 0.99795, 0.99903, and 0.99962 for the biomass, substrate, and product concentrations respectively.
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Development of Robust Yeast Strains for Lignocellulosic Biorefineries Based on Genome-Wide Studies.
Zhang, MM, Chen, HQ, Ye, PL, Wattanachaisaereekul, S, Bai, FW, Zhao, XQ
Progress in molecular and subcellular biology. 2019;:61-83
Abstract
Lignocellulosic biomass has been widely studied as the renewable feedstock for the production of biofuels and biochemicals. Budding yeast Saccharomyces cerevisiae is commonly used as a cell factory for bioconversion of lignocellulosic biomass. However, economic bioproduction using fermentable sugars released from lignocellulosic feedstocks is still challenging. Due to impaired cell viability and fermentation performance by various inhibitors that are present in the cellulosic hydrolysates, robust yeast strains resistant to various stress environments are highly desired. Here, we summarize recent progress on yeast strain development for the production of biofuels and biochemical using lignocellulosic biomass. Genome-wide studies which have contributed to the elucidation of mechanisms of yeast stress tolerance are reviewed. Key gene targets recently identified based on multiomics analysis such as transcriptomic, proteomic, and metabolomics studies are summarized. Physiological genomic studies based on zinc sulfate supplementation are highlighted, and novel zinc-responsive genes involved in yeast stress tolerance are focused. The dependence of host genetic background of yeast stress tolerance and roles of histones and their modifications are emphasized. The development of robust yeast strains based on multiomics analysis benefits economic bioconversion of lignocellulosic biomass.