Transferase Enzyme Research Articles

Diversity and functional role of bacterial microbiota in spontaneous coffee fermentation in northern Peru using shotgun metagenomics.

Peru is the ninth-largest coffee producer and the largest organic coffee exporter worldwide. Specific modifications in the microbial consortia during fermentation control the flavor of coffee. It is still unclear how fermentation duration affects microbial communities. This study aimed to provide insights into the diversity and functional behavior of the bacterial microbiome during coffee fermentation in northern Peru using shotgun metagenomics. Accordingly, metagenomic DNA was extracted and sequenced from samples of the liquid fraction during the short fermentation process (SFP) in Amazonas (6 and 12h) and long fermentation process (LFP) in Cajamarca (6, 12, 18, 24, and 36h). Our findings indicate that common (e.g., Acetobacter, Lactobacillus, Leuconostoc, and Weissella) and unique (e.g., Acidiphilium and Methylobacterium) acid-tolerant bacteria from the SFP and LFP play crucial roles and have a positive impact on the sensory qualities of coffee. Specifically, the LFP from San Ignacio might be associated with the high sensory quality of coffee based on the release of catalytic, hydrolase, oxidoreductase, transferase, and transporter enzymes in the InterPro and KEGG profiles. Additionally, these bacterial microorganisms metabolize several compounds (e.g., isoleucine, betaine, galactose, tryptophan, arginine, and cobalamin) into volatile compounds, mainly in the LFP, enhancing the flavor and aroma of coffees. This characteristic suggests that the LFP has a stronger effect on coffee quality than does the SFP on the basis of bacterial diversity and functional prediction. These findings provide new perspectives on the potential biotechnological uses of autochthonous microorganisms to produce superior-quality coffee beans from northern Peru.

Therapeutic potency of kaempferol against Naja haje venom induced neurotoxicity, inflammation, biological activities, and antioxidant system damage: a pre-clinical antivenom evaluation.

Naja haje envenoming manifests organ system disorders leading to severe fatalities due to the venom's toxins. The neutralizing capacity of kaempferol has been reported against some medically significant snake venoms with exception of N. haje venom (NhV). This current study assessed the neutralizing profile of kaempferol against toxicities induced by NhV using in vitro and in vivo methods. In in vitro, NhV induced wide spectrum of toxic biological activities with substantial increase in hemorrhagic, anticoagulating, and hemolytic effects. Likewise in the in vivo study, the venom caused hematological disorders by inducing acute anemia, thrombocytopenia, and leucopenia in envenomed rats. Also, the venom caused detrimental effect on the antioxidant defense system of the brain through significant (P < 0.05) elevation of malondialdehyde (MDA), nitrite (NIT), and suppression of reduce glutathione (GSH) antioxidant, superoxide dismutase (SOD), catalase (CAT), and glutathione transferase (GST) enzymes. Additionally, the levels of neurotoxicity biomarkers, monoamine oxidase (MAO) and acetylcholinesterase (AChE), and pro-inflammatory cytokines, tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), were significantly (P < 0.0.5) enhanced by NhV in the brain of envenomed rats. Severe pathohistological effects were observed in brain tissues of the envenomed rats. However, kaempferol substantially (P < 0.05) inhibited the NhV-induced biological activities, normalized the hematological disorders, enhanced antioxidants system functions, and significantly (P < 0.05) suppressed the levels of neurotoxicity biomarkers and pro-inflammatory cytokines. Severe structural alterations observed in the brain tissues were ameliorated after kaempferol treatment. Further exploration of kaempferol could lead to the development of an alternative therapy for treatment of N. haje envenoming.

TPCA-1 compound, inhibiting testis-specific serine/threonine protein kinase 3 for potential male sterile in Bombyx mori.

Protein kinases are a type of transferase enzyme that catalyze the phosphorylation of protein substrates, including receptor proteins. Testis-specific serine/threonine kinases (TSSKs) are a highly conserved group of protein kinases found in various organisms. They play an essential role in male reproduction by influencing sperm development and function. In this study, we report on the characterization of BmTSSK3, a TSSK from the silkworm, Bombyx mori. We found that BmTSSK3 is specifically expressed in the testis and localized to the sperm flagella, particularly in the sperm tail cyst. Furthermore, we developed BmTSSK3 inhibitors through molecular docking and binding assays. Small molecules 5-(4-Fluorophenyl)-2-ureidothiophene-3-carboxamide (TPCA-1)and Imidurea were identified to bind to BmTSSK3. Using site-specific mutation technology, we identified amino acid residues R134 and S184 as crucial binding sites for small molecules. RNA interference assay and Western blot analysis showed that knockdown of BmTSSK3 significantly decreased histone 3 phosphorylation. To confirm the inhibitory effect of these small molecules, we treated silkworm testes with TPCA-1 and observed a strong inhibitory effect. TPCA-1 is an inhibitor of BmTSSK3, which raises its potential as a future candidate for male sterility of the silkworm. Thus, this study may offer a novel strategy for sterile silkworms as well as insects. © 2024 Society of Chemical Industry.

Overall mutational scanning unveils the essential active residues for the mechanistic action of MCR-1

Polymyxins, including colistin and polymyxin B, serve as crucial last-resort antibiotics for managing infections caused by carbapenem-resistant Enterobacterales (CRE). However, the rapid spread of the mobilized colistin resistance gene (mcr-1) challenged the efficacy of treatment by polymyxins. The mcr-1 gene encoded a transmembrane phosphoethanolamine (PEA) transferase enzyme, MCR-1. MCR-1 could catalyze the transfer of PEA moiety of phosphatidylethanolamine (PE) to the 1’ (or 4’)-phosphate group of the lipid A. Despite the determination of several structures of the soluble domain of MCR-1, the structural and biochemical mechanisms of integral MCR-1 remain less understood. In this study, we utilized an alanine scanning mutagenesis approach to systematically investigate the functional attributes of distinct regions within MCR-1. We identified fifteen critical residues that are indispensable for the enzymatic activity of MCR-1 and are pivotal for its ability to confer resistance to colistin. Furthermore, molecular docking of MCR-1 complexed with the phosphoethanolamine (PE) substrate revealed the presence of a channel-shaped cavity, a characteristic feature shared with other phosphoethanolamine transferases. Despite MCR-1 exhibiting a low sequence identity with both MCR homologues and other phosphoethanolamine (PEA) transferases, several conserved sites were identified, including Y97, M105, K333, H395, L477, and H478, suggesting a potentially shared catalytic mechanism among them for modifying LPS-lipid A. Overall, these findings provide a deep understanding of the catalytic mechanism of MCR-1 for colistin resistance. Moreover, these findings provide a robust structural and functional foundation, enabling the rational design of targeted inhibitors and restoring colistin activity against serious infections with carbapenem-resistant Enterobacterales (CRE).

MurA-catalyzed synthesis of 5-enolpyruvylshikimate-3-phosphate confers glyphosate tolerance in bryophytes

Glyphosate is a broad-spectrum herbicide that kills most vascular plant weeds but is ineffective against many bryophytes. Glyphosate competitively inhibits the enolpyruvyl transferase enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). EPSPS catalyzes the production of 5-enolpyruvylshikimate-3-phosphate (EPSP)-an intermediate in the shikimate pathway-from shikimate-3-phosphate (S3P) and phosphoenolpyruvate (PEP) substrates. Here, we show that Marchantia polymorpha mutants with loss-of-function mutations in a closely related enolpyruvyl transferase, MpMurA, were more sensitive to glyphosate than wild-type controls. Overexpression of MpMurA in M. polymorpha increased glyphosate tolerance, and heterologous expression of the M. polymorpha MurA enzyme in Arabidopsis thaliana conferred glyphosate resistance. Furthermore, we demonstrate that MpMurA catalyzes the production of EPSP from S3P and PEP substrates. These data demonstrate that MpMurA contributes to glyphosate tolerance in M. polymorpha. We speculate that the existence of two independent mechanisms for EPSP synthesis-one canonical and EPSPS-dependent, and the other MurA-dependent-may account for glyphosate tolerance in bryophytes. Alternatively, MurA in bryophytes may bind glyphosate, thereby leaving more unbound EPSPS enzymes available, allowing aromatic amino acid biosynthesis to continue.

Mitochondrial Phosphopantetheinylation is Required for Oxidative Function.

4'-phosphopantetheinyl (4'PP) groups are essential co-factors added to target proteins by p hospho p antetheinyl transferase (PPTase) enzymes. Although mitochondrial 4'PP-modified proteins have been described for decades, a mitochondrially-localized PPTase has never been found in mammals. We discovered that the cytoplasmic PPTase a mino a dipate s emialdehyde d ehydrogenase p hospho p antetheinyl t ransferase (AASDHPPT) is required for mitochondrial respiration and oxidative metabolism. Loss of AASDHPPT results in failed 4'-PP modification of the mitochondrial acyl carrier protein and blunted activity of the mitochondrial fatty acid synthesis (mtFAS) pathway. We found that in addition to its cytoplasmic localization, AASDHPPT localizes to the mitochondrial matrix via an N-terminal mitochondrial targeting sequence contained within the first 13 amino acids of the protein. Our data show that this novel mitochondrial localization of AASDHPPT is required to support mtFAS activity and oxidative function. We further identify two variants of uncertain significance in AASDHPPT that are likely pathogenic in humans due to loss of mtFAS activity.

Chemical synthesis, in vitro testing and in silico Nampt-based molecular docking of novel aniline aromatic ring-substituted 2-Aminothiazole analogues.

The heterocyclic 2-Aminothiazoles scaffolds are used in a wide range of therapeutic applications against various diseases for its antioxidant, anti-inflammatory, antimicrobial and anticancer actions. In the present study, we synthesized novel aniline aromatic ring-substituted 2-Aminothiazole derivatives. Molecular docking was performed using Glide module of the Schrödinger Suite to fit compounds JG-49, JG-62, and KBA-18 against the Nicotinamide phosphoribosyl transferase (Nampt) enzyme, an intracellular regulator of Nicotinamide adenine dinucleotide (NAD) redox cofactor involved in energy metabolism and epigenetics and are implicated in aging and metabolic diseases. The three compounds viz. JG-49, JG-62, and KBA-18 showed an increase in Nampt enzymatic activity in vitro. All three substituted derivatives of 2-Aminothiazole showed no cytotoxicity with the mouse C2C12 myoblasts cultures assessed with the MTT cell viability assay. Moreover, the wound closure of the mouse C2C12 myoblasts in vitro displayed no significant difference between the treatment groups of the 2-Aminothiazole derivatives compared with the control naïve and DMSO treated myoblasts cultures, except for the 2-Aminothiazole substituted derivatives JG-62 and KBA-18, which showed a significant increase in the wound closure compared with the control cells at different concentrations. Taken together, we demonstrated that 2-Aminothiazole substituted derivatives provide enhanced Nampt activity, wound closure, and no cytotoxic effects in vitro. Further studies will allow to improve the substitution of 2-Aminothiazole derivatives and test their potential therapeutic applications.

8700 Mono(ADP-Ribosyl)ation of Ribosomal Proteins Promotes Differentiation During Adipogenesis

Abstract Disclosure: X. Tan: None. S. Challa: None. C.V. Camacho: None. T.S. Nandu: None. M.S. Stokes: None. W.L. Kraus: Other; Self; W.L.K. is a founder and stockholder for Ribon Therapeutics, Inc; a founder, member of the SAB, member of the BOD, and a stockholder for and ARase Therapeutics, Inc. He is also coholder of U.S. Patent. ADP-ribosylation, a crucial post-translational modification, involves the covalent attachment of ADP-ribose units to amino acids on proteins through ADP-ribosyl transferase (ART) enzymes, utilizing NAD+ synthesized by nicotinamide mononucleotide adenylyl transferases (NMNATs). We previously showed that NMNAT-2, a cytoplasmic NAD+ synthase, increases during adipogenesis, leading to reduced nuclear NAD+ levels through compartmentalized NMN depletion and enhanced differentiation of preadipocytes. However, the implications of increased cytoplasmic NAD+ synthesis by NMNAT-2 in adipocytes remain unknown. Our current findings demonstrate elevated ribosome MARylation during 3T3-L1 preadipocyte differentiation, driven by NMNAT-2 induction. Inhibiting NAD+ biosynthesis by knockdown of NMNAT-2 or FK866 results in reduced ribosome MARylation, with PARP16 identified as the primary ART mediating MARylation. Knockdown of NMNAT-2 or PARP16 impaired global protein synthesis and preadipocyte differentiation. Treatment with the PARP16 inhibitor DB008 hindered ribosomal MARylation and preadipocyte differentiation. Immunofluorescence and subcellular fractionation analysis demonstrated that MARylated ribosomes are mostly associated with the endoplasmic reticulum. Using TMT mass spectrometry, we identified numerous high-confidence ADP-ribosylation sites on glutamate and aspartate residues in both undifferentiated and differentiated cells. We examined five proteins with specific sites of MARylation that are consistently more modified in the differentiated state. Among these, ADP-ribosylation of RPSA significantly promote preadipocyte differentiation; mutation of the site inhibits this effect. Ongoing investigations involve studying the impact of Parp16 knockout on high fat diet-induced obesity in mice to further determine the biological role of ribosome protein MARylation during adipogenesis. This work is supported by a grant from the NIH/NIDDK (R01 DK069710) and the DOD OCRP (OC200311). Presentation: 6/1/2024

The Role of Arginase and some Parameters in Diabetic Type 2 Patients with and without Retinopathy

Background: Oxidative stress plays a major role in the pathogenesis of diabetes mellitus by damaging cellular organelles and enzymes in blood such as arginase1, insulin and glutathione s-transferase; increasing lipid peroxidation such as malondialdehyde and increasing insulin resistance which can lead to diabetic complications such as diabetic retinopathy. Objectives: To explore the relationship of oxidative stress to the development of diabetic retinopathy by measuring the levels of Arginase1, the activity of glutathione s-transferase enzyme, and the levels of malondialdehyde as a secondary product of lipid peroxidation (biomarker for oxidative stress). Patients and MethodsThis study was conducted from November 2022 to January 2023 at the Ibn Al-Haitham Teaching Eye Hospital in Baghdad, the University of Baghdad / Department of Chemistry and the National Diabetes Centre for Treatment and Research at Al-Mustansyrriah University. This study was conducted on 120 subjects distributed as follows: 40 non-diabetic obese controls, 40 type 2 diabetics with no retinopathy, and 40 type 2 diabetic retinopathy patients, between 30 and 65 years of age. All groups were subjected to tests: measuring fasting blood glucose (FBG), HbA1c, lipid profile (cholesterol, triglycerides, HDL- cholesterol, LDL- cholesterol, and VLDL cholesterol), serum total arginase1, malondialdehyde glutathione s -transferase, body mass index (BMI), and waist-to-hip ratio. Results: Mean arginase1 levels were significantly higher in diabetic patients than in diabetic retinopathy and control groups (p ≤ 0.05). The mean xidative stress marker malondialdehyde concentration was significantly higher in diabetic retinopathy patients than in type 2 diabetics and the control group (P ≤ 0.05). The mean glutathione s-transferase activity in diabetic retinopathy patients was significantly higher than in the control group and type 2 diabetics (P ≤ 0.05). Conclusion: There is a relationship between oxidative stress and the development of diabetic retinopathy, where the levels of arginase1 and malondialdehyde increased and the activity of glutathione s –transferase enzyme increased as a result of oxidative stress and inflammation associated with complications of type 2 diabetes.

Thiopurine S-methyl Transferase (TPMT) Enzyme Level in Healthy Sudanese Population

Background: Thiopurine drugs have limited use due to their toxicity, related to the enzyme thiopurine S-methyl transferase (TPMT) activity, which varies between individuals. This is the first study in Sudan, which aimed to assess the TPMT phenotypic status of healthy Sudanese volunteers. Methods: A total of 177 healthy volunteers from Sudan were included in the study. TPMT enzymatic activities were measured using the ELISA serum protocol. We used SPSS to analyze the data and determined enzyme level categories and normal range with Z scores and quartile tests. The Sudan Medical Specialization Board (SMSB) Ethical Committee approved the study. Results: There were 117 males and 60 females among the volunteers, with ages ranging from 16 to 70 years and a mean age ± SD of 28.0 ±1 0.2, median = 24. Most candidates were from the Afro-Asiatic linguistic group (64.5%), followed by Nilo-Saharan (18.6%) and Niger-Kordofanian (16.9%). The TPMT enzyme level ranged between 0.17 and 9.5 ng/ml, with a mean of 2.26 ± 0.75 ng/ml. The quartile classification included very low enzyme (&lt;0.76 ng/ml) seen in 4 candidates (2.3%), intermediate low (0.76-1.4 ng/ml) seen in 34 (19.2%), the normal range (1.5 – 3.75 ng/ml) seen in 119 (67.2%), and high enzyme activity (&gt;3.76 ng/ml) seen in 20 (11.3%). No significant correlations between age, sex, and ethnic groups were recorded. Conclusion: The normal TPMT enzyme activity is between 1.5 and 3.76 ng/ml. A higher prevalence of TPMT deficiency was recorded and compared with international studies. Pretreatment screening using serum ELISA test for TPMT enzyme activity should be used to predict the risk of toxicity.

Cardioprotective action of apocynin in isoproterenol-induced cardiac damage is mediated through Nrf-2/HO-1 signaling pathway.

This investigation evaluated the therapeutic benefit of apocynin in isoproterenol (ISO)-induced cardiac damage in rats. ISO-administered male Wistar rats were treated with apocynin for 2 weeks. Blood plasma and left ventricle of heart tissues were collected and analyzed for oxidative stress-related parameters such as malondialdehyde (MDA), advanced oxidation protein product (AOPP), and nitric oxide (NO). The activities of endogenous antioxidant enzymes such as superoxide dismutase (SOD) and catalase were also measured. The gene expressions of oxidative stress-related proteins such as Nrf-2, HO-1, and HO-2 in cardiac tissues were also measured. In silico studies like molecular docking and molecular dynamics were also performed to detect how apocynin interacts with NADPH and nitric oxide synthase at the molecular level. This investigation revealed significant elevation of serum transferase enzymes and creatinine kinase-Muscle Brain (CK-MB) activities in ISO-administered rats compared to the control. Apocynin effectively normalized the serum transferases and CK-MB activities in the blood of ISO-stressed rats. Moreover, ISO-induced elevations of MDA, NO, and AOPP levels were also suppressed by apocynin treatment. Consistently, apocynin restored the reduced SOD and catalase activities in ISO-administered rats. This restoration of enzyme activity might be due to the increased expression of Nrf-2 and HO-1 and reduced expression of iNOS and TNF-α in ISO-administered rats. Histological analysis revealed that apocynin treatment ameliorated the mononuclear cell adherence and fibrosis in the cardiac tissue of ISO-administered rats. Computational studies also support the experimental findings. This study demonstrates that apocynin prevents ISO-induced cardiac injury not only by preventing inflammation but also by empowering the antioxidant defense system.

Unveiling the multitargeted potential of deprodone and control comparison with linezolid against hydrolase and transferase enzymes of methicillin-resistant Staphylococcus aureus

Staphylococcus aureus (S. aureus), commonly found on the skin and nose, causes minor skin conditions to life-threatening diseases, including boils or impetigo, pneumonia, and bloodstream infections. MRSA (Methicillin-Resistant S. aureus) is a strain resistant to many antibiotics and poses a significant challenge in clinical settings. Nowadays, the alternative drug Linezolid is used, and it is not clear when MRSA starts resistance to it, necessitating the need for more alternative drugs with the least chance of developing resistance. This study aims to identify a multitargeted drug candidate with better efficacy than Linezolid. We have taken three hydrolase and transferase proteins from S. aureus, performed the multitargeted docking studies with human-approved drugs, and compared them with the control drug Linezolid. The docking and MM\\GBSA scores ranging from −6.79 to −5.78 Kcal/mol and − 37.47 to 30.16 Kcal/mol, respectively, that revealed Deprodone (used for inflammatory skin disorders, bowel disease, and fatty acid metabolism disorders) can be a far better and multitargeted drug candidate than Linezolid. We extended our studies to include extensive pharmacokinetics and molecular interaction fingerprints for interaction pattern studies. Also, the DFT computations optimised the drug, and we extended our studies for MD Simulation in water for 100 ns, which showed the complexes among the identified drug with proteins are entirely stable with acceptable deviation, fluctuations and many intermolecular interactions that make them stable. We also performed the MM\\GBSA studies on MD simulation's all 1000 frames to understand the complex energy level. All the results reveal promising interactions between Deprodone and the targeted enzymes, suggesting its potential as a multitargeted therapeutic agent—however, experimental studies need to validate Deprodone against MRSA.

Overexpression of VaSS4 negatively regulates cold tolerance by disturbing ROS balance and decreasing soluble sugar content

Cold stress is one of the major limiting factors for grapevine growth and yield. Sucrose synthetase (SS) is a glycosyl transferase enzyme involved in plant sugar metabolism, which is essential for plant growth and development and response to abiotic stress. However, the function of SS genes in grape cold resistance has not been systematically understood. In the present study, a SS gene from Vitis amurensis named VaSS4 was identified, which was significantly suppressed in response to cold stress. The physicochemical properties, evolutionary relationships, synteny, and selection pressure of VaSS4 were analyzed. Furthermore, VaSS4-overexpressed Arabidopsis and grape calli were obtained to characterize their tolerance to cold stress. Amino acid sequence analysis demonstrated that VaSS4 protein encoded 806 amino acids and contained conserved sucrose synthase domain and glycosyl transferase domain. Overexpression of VaSS4 decreased SS decomposition activity and soluble sugar accumulation under cold stress. Meanwhile, VaSS4 overexpression decreased the reactive oxygen scavenging ability and cold tolerance of Arabidopsis and grape calli. Importantly, overexpression of VaSS4 negatively regulated the expression of cold-related genes in the CBF-dependent pathway compared with WT plants. Overall, these findings not only provide extensive systematic information for Vitis amurensis VaSS4 gene research, but also provide valuable germplasm resources for grape cold resistance breeding.

Ultraviolet‐B Stress Increases Epidermal UV‐Screening Effectiveness and Alters Growth and Cell‐Wall Constituents of the Brown Midrib bmr6 and bmr12 Mutants of Sorghum bicolor

ABSTRACTThe brown midrib bmr6 and bmr12 mutants of sorghum (Sorghum bicolor) have alterations to the phenylpropanoid pathway impairing the activity of cinnamyl alcohol dehydrogenase (CAD) and/or caffeate5/hydroxyferulate O‐methyl transferase (COMT) enzymes, which inhibit lignin synthesis. Interestingly, these phenylpropanoids can also act as sunscreen compounds in plants and potentially attenuate ultraviolet radiation. We examined the effects of ultraviolet‐B (UV‐B; 280–320 nm) exclusion on growth, cell‐wall constituents and UV‐screening abilities of bmr6, bmr12, a double mutant (bmr6 bmr12; dm) and wild‐type (WT) genotypes of S. bicolor. Plants were grown in a UV‐transparent greenhouse under filters that either transmitted 2.8% (Mylar) or 90% (Aclar) of UV‐B. The greenhouse experiment was a 2 × 4 (UV treatment × genotype) complete factorial design. Sorghum grown under reduced UV were 23% taller and had 22% fewer leaves. Among genotypes, the WT plants were 5%–12% taller than the bmr6, bmr12 and dm mutants. The near‐ambient UV‐B treatment group was more effective at UV screening and had a 16% higher UV‐screening effectiveness than those under reduced UV‐B. Sorghum plants with the bmr6 and dm genotypes had 8%–19% higher UV‐shield than the bmr12 and WT. Plants grown under the reduced UV‐B treatment had 5% less hemicellulose and 6% more cellulose in their cell walls. There were no overall treatment effects on bulk soluble phenolics, chlorophyll fluorescence (Fv/Fm) or lignin concentrations. These results are a possible indication that the bmr mutants of S. bicolor have a varied response to UV‐B exclusion due to alterations in the phenylpropanoid pathway leading to redistribution of metabolites.

The Evaluation of the Genetic Variation Types of the Uridine Diphosphate Glucuronosyl Transferase 1A1 Gene by Next-Generation Sequencing and Their Effects on Bilirubin Levels in Obese Children.

Background and Objectives: Obesity is a major nutritional problem with an increasing prevalence among children and adolescents. The uridine-diphosphate-glucuronosyl-transferase1A1 (UGT1A1) gene encodes the UDP-glucuronosyl transferase enzyme, converting the toxic form of bilirubin to a soluble, nontoxic form. There are yet to be studies on the evaluation of the UGT1A1 variant types detected by next-generation sequencing (NGS) and their effects on bilirubin levels in nonsyndromic obese children. Methods: Forty-five children with body mass index (BMI) >95 percentile (p) constituted the obesity group and fourteen healthy children with BMI <85p constituted the control group. Anthropometric, clinical features, and biochemical parameters were evaluated. Furthermore, the UGT1A1 gene was sequenced by NGS. Results: The obese patients had lower total, direct, and indirect bilirubin levels (p = 0.422, 0.026, and 0.568, respectively). In addition, obese patients had more genetic variations in the UGT1A1 gene compared with the control group (62.2% and 50%, respectively). We found that children with variations had higher total direct and indirect bilirubin levels compared with those without variation (p = 0.016, 0.028, and 0.015, respectively). Children diagnosed with obesity in the first two years of their life had fewer genetic variations and lower total bilirubin levels (p = 0.000 and 0.013, respectively). Conclusions: It is assumed that bilirubin can be protective against many chronic diseases. Although bilirubin levels are found to be lower in obese children compared with the control group, some variations in the UGT1A1 gene may be supported by raising bilirubin. We suggest that high bilirubin levels caused by those UGT1A1 variations may be protective against obesity and its many negative effects.

Effect of DNMT3A R882H Hot Spot Mutations on DDX43 Promoter Methylation in Acute Myeloid Leukemia.

Epigenetic alterations have been observed in many hematological malignancies, including acute myeloid leukemia (AML). Many of these alterations result from mutations in DNA methyl transferase (DNMT) enzymes, disabling them to methylate target genes in a proper way. In this case-control study, we investigated the association between R882H mutation in DNMT3A gene and DDX43 gene methylation in patients with AML. 47 AML patients and 6 controls were included in this study. After DNA extraction, amplification refractory mutation system (ARMS)-PCR was used to evaluate R882H mutations in DNMT3A gene. The high-resolution melting (HRM) method was used to determine the methylation changes of the DDX43 gene promoter. R882H mutation was only found in 10.6% (5 out of 47) of AML patients. The frequency of DDX43 gene methylation was significantly higher in patients without R882H mutations compared to patients with R882H mutations (P < 0.05). The DNMT3A R882H mutation is typically present in a minority of AML patients. Nevertheless, this mutation is associated with a reduced frequency of methylation in the DDX43 promoter region.

Design, Synthesis, DFT, docking Studies, and antimicrobial evaluation of novel benzimidazole containing sulphonamide derivatives

In silico approaches have been employed to design a new series of benzimidazole-containing sulphonamide derivatives and qualified compounds have been synthesized to analyze their potential as antimicrobial agents. Antibacterial screening of all synthesized compounds was done using the broth microdilution method against several human pathogenic bacteria, viz. Gram-positive bacteria [B. cerus (NCIN-2156), B. subtilis (ATCC-6051), S. aureus (NCIM-2079)] and Gram-negative bacteria [P. aeruginosa (NCIM-2036), E. coli (NCIM-2065), and a drug-resistant strain of E. coli (U-621)], and the compounds presented admirable MIC values, ranging between 100–1.56 µg/mL. The combinatorial analysis showed the magnificent inhibitory efficiency of the tested compounds, acquired equipotent to ten-fold more potency compared to original MIC values. An immense synergistic effect was exhibited by the compounds during combination studies with reference drugs chloramphenicol and sulfamethoxazole was presented as fractional inhibitory concentration (∑FIC). Enzyme inhibition studies of all synthesized compounds were done by using peptidyl transferase and dihydropteroate synthase enzymes isolated from E. coli and S. aureus and each of the compound presented the admirable IC50 values, where the lead compound 3 bound to peptidyl transferase (of S. aureus with IC50 363.51 ± 2.54 µM and E. coli IC50 1.04 ± 0.08 µM) & dihydropteroate synthase (of S. aureus IC50 3.51 ± 0.82 µM and E. coli IC50 2.77 ± 0.65 µM), might account for the antimicrobial effect, exhibited excellent inhibition potential. Antifungal screening was also performed employing food poisoning methods against several pathogenic fungal species, viz A. flavus, F. oxysporum, A. niger, and A. brassicae. The obtained result indicated that few compounds can prove to be a potent drug regimen against dreaded MDR strains of microbes. Structural activity relationship (SAR) analysis and docking studies reveal that the presence of electron-withdrawing, polar, and more lipophilic substituents positively favor the antibacterial activity, whereas, electron-withdrawing, more polar, and hydrophilic substituents favor the antifungal activities. A robust coherence has been found in in-silico and in-vitro biological screening results of the compounds.

Crosstalk between long non-coding RNAs and miRNAs regulates transferase activities in response to salt stress in Pistachio (Pistacia vera L.)

Long non-coding RNA (lncRNA) molecules are important regulators of a broad spectrum of biological processes, including adaptation to diverse stresses in plants. However, how plant lncRNAs function during salt stress conditions is largely unknown. In our study, we identified 48 conserved mature Pistacia vera L. (P. vera) miRNAs among pistachio miRNA precursors and used RNA sequencing data to predict and validate the interaction relationship between lncRNAs and miRNAs. We indicated that the regulatory networks of pve-miRNA-lncRNA-mRNAs were more complex in the salt-tolerant (Ghazvini) compared to the salt-sensitive (Sarakhs) pistachio cultivar under NaCl treatment. We also found that 7 and 8 salt-responsive lncRNAs play a role as competing endogenous target mimics (eTMs) for 7 pve-miRNA families in Ghazvini (mir164, mir169, mir397, and mir827) and Sarakhs (mir156, mir159, mir169, and mir482), respectively. The GO-term enrichment analysis showed that protein-coding target genes of pve-miRNAs that regulated with target mimic relationship mainly enriched in transferase activity include: 7-deoxyloganetin glucosyltransferase, Arginine N-methyltransferase, Glutathione transferase, Flavonol sulfotransferase, and Beta-glucuronosyltransferase. Their activity regulates alkaloids and polysulfated flavonols biosynthesis, sequesters sodium, improves antioxidant capacity and controls epigenetic elements in Ghazvini under salt treatment. In Sarakhs, the analysis of GO-term enrichment in pve-miRNAs protein-coding target genes of lncRNA-related mimic relationship enriched in response to wounding, cell death, and jasmonic acid biosynthetic process. Compared to Sarakhs, Ghazvini had a more effective ceRNA network for regulating transferase enzymes and specific transcription factors including MYB, WRKY, and ABR1 in response to salt stress, which led to more salt tolerance in the Ghazvini cultivar. The present study provides new information about the complex lncRNA-miRNA-mRNA regulatory networks in response to plant stress conditions in pistachio. Furthermore, our findings can be used for different breeding programs and the introduction of salt-tolerant pistachio rootstocks.

Metabolic response of endangered goats fed spineless cactus associated with Tifton-85 hay or Maniçoba hay

The aim of this study was to evaluate the effects of replacement of Tifton-85 hay by maniçoba hay, in spineless cactus-based diets, on markers of energy, protein, mineral metabolism and serum enzyme activity of Moxotó goats. Sixteen uncastrated male Moxotó breed goats, with 20 months old and initial body weight of 21.7 ± 3.92 kg were used in a randomized blocks design, using the initial weight as the criterion for the formation of blocks, with two treatments and eight replicates per treatment. The experimental treatments consisted of two diets: diets containing Miúda spineless cactus associated with Tifton-85 hay (1) or maniçoba hay (2), with roughage:concentrate ratio of 70:30. Blood samples were collected fortnightly (baseline, 15 days, 30 days and 45 days) during the feedlot period. There was a higher serum concentration of total protein and fructosamine (P &lt; 0.05) in goats that received maniçoba hay. However, there was higher serum gamma-glutamyl transferase (GGT) enzyme activity in goats that ingested Tifton-85 hay (P &lt; 0.05). Concerning to the profile of blood metabolites in the different fortnightly collections, it was possible to verify a significant effect for urea, fructosamine, GGT enzyme activity, K and Cl levels (P &lt; 0.05). In conclusion, Tifton-85 hay and maniçoba hay combined with spineless cactus does not cause metabolic disorders in feedlot native goats.

Sphingolipid biosynthesis is essential for metabolic rewiring during TH17 cell differentiation.

T helper 17 (TH17) cells are implicated in autoimmune diseases, and several metabolic processes are shown to be important for their development and function. In this study, we report an essential role for sphingolipids synthesized through the de novo pathway in TH17 cell development. Deficiency of SPTLC1, a major subunit of serine palmitoyl transferase enzyme complex that catalyzes the first and rate-limiting step of de novo sphingolipid synthesis, impaired glycolysis in differentiating TH17 cells by increasing intracellular reactive oxygen species (ROS) through enhancement of nicotinamide adenine dinucleotide phosphate oxidase 2 activity. Increased ROS leads to impaired activation of mammalian target of rapamycin C1 and reduced expression of hypoxia-inducible factor 1-alpha and c-Myc-induced glycolytic genes. SPTLCI deficiency protected mice from developing experimental autoimmune encephalomyelitis and experimental T cell transfer colitis. Our results thus show a critical role for de novo sphingolipid biosynthetic pathway in shaping adaptive immune responses with implications in autoimmune diseases.