Weak purifying selection in allelic diversity of the ADSL gene in indigenous and local chicken breeds and red junglefowl in Thailand.

High levels of purine and uric acid, which are associated with health issues such as gout and cardiovascular disease, are found in the meat of fast-growing broiler chickens, which raises concerns about the quality of chicken meat and the health of the consumers who consume it. High genetic homogeneity and uniformity, particularly in genes involved in the synthesis of inosine monophosphate (IMP) and subsequent process of purine synthesis, which are associated with the meat quality, are exhibited in commercial broiler chickens owing to intensive inbreeding programs. Adenosine succinate lyase (ADSL) is a key enzyme involved in de novo purine biosynthetic pathway and its genetic polymorphisms affect IMP metabolism and purine content. In this study, we investigated the polymorphism of the ADSL gene in indigenous and local chicken breeds and red junglefowl in Thailand, using metabarcoding and genetic diversity analyses. Five alleles with 73 single nucleotide polymorphisms in exon 2, including missense and silent mutations, which may act on the synthesis efficiency of IMP and purine. Their protein structures revealed changes in amino acid composition that may affect ADSL enzyme activity. Weak purifying selection in these ADSL alleles was observed in the chicken population studied, implying that the variants have minor fitness impacts and a greater probability of fixation of beneficial mutations than strong purifying selection. A potential selective sweep was observed in Mae Hong Son chickens, whose purine content was lower than that in other breeds. This suggests a potential correlation between variations of the ADSL gene and reduced purine content and an impact of ADSL expression on the quality of chicken meat. However, further studies are required to validate its potential availability as a genetic marker for selecting useful traits that are beneficial to human health and well-being.

Multienzyme-Mediated Dual-Channel Magnetic Relaxation Switching Taste Biosensor (D-MRSTB) for Simultaneous Detection of Umami Compounds and Synergistic Enhancement in Food.

Umami substances play a significant role in the evaluation of food quality, and their synergistic enhancement is of great importance in improving and intensifying food flavors and tastes. Current biosensors available for umami detection still confront challenges in simultaneous quantification of multiple umami substances and umami intensities. In this study, an innovative dual-channel magnetic relaxation switching taste biosensor (D-MRSTB) was developed for the quantitative detection of representative umami substances. The multienzyme signal of D-MRSTB specifically catalyzes the umami substances of interest to generate hydrogen peroxide (H2O2), which is then used to oxidate Fe2+ to Fe3+. Such a valence-state transition of paramagnetic ions was utilized as a magnetic relaxation signaling switch to influence the transverse magnetic relaxation time (T2) within the reaction milieu, thus achieving simultaneous detection of monosodium glutamate (MSG) and inosine 5'-monophosphate (IMP). The biosensor showed good linearity (R2 > 0.99) in the concentration range of 50-1000 and 10-1000 µmol/L, with limits of detection (LOD) of 0.61 and 0.09 µmol/L for MSG and IMP, respectively. Furthermore, the biosensor accurately characterized the synergistic effect of the mixed solution of IMP and MSG, where ΔT2 showed a good linear relationship with the equivalent umami concentration (EUC) of the mixed solution (R2 = 0.998). Moreover, the D-MRSTB successfully achieved the quantitative detection of umami compounds in real samples. This sensing technology provides a powerful tool for achieving the detection of synergistic enhancement among umami compounds and demonstrates its potential for application in the food industry.

PKM2 promotes myoblast growth and inosine monophosphate-specific deposition in Jingyuan chicken.

Inosine monophosphate (IMP) is widely regarded as an important indicator for evaluating the flavour of poultry meat. However, little is known about the molecular mechanisms affecting the specific deposition of IMP. In this study, we functionally verified PKM2 (Pyruvate kinase M2), a candidate gene related to IMP synthesis, in order to reveal the important role of PKM2 in meat flavour and muscle development of Jingyuan chickens. The results showed that the IMP content in breast muscle of Jingyuan chickens was negatively correlated with PKM2 mRNA expression (r = -0.1710), while the IMP content in leg muscle was significantly positively correlated with PKM2 mRNA expression (r = 0.7350) (P < 0.05). During myogenesis, PKM2 promoted the proliferation rate of myoblasts and the expression of proliferation marker genes, inhibited the apoptosis rate and the expression of apoptosis marker genes, and decreased the expression of differentiation marker genes. Up-regulation of PKM2 enhanced the expression of key genes in the purine metabolic pathway and the de novo synthesis pathway of IMP, and suppressed the expression of key genes in the salvage pathway. ELISA assays showed that PKM2 decreased IMP and hypoxanthine (HX) contents, while adenosine triphosphate (ATP) and uric acid (UA) contents were clearly elevated. In summary, these studies revealed that PKM2 regulates myogenesis and specific deposition of IMP, which can be used to improve the quality of Jingyuan chicken meat.

Molecular characterization of adenosine monophosphate deaminase 1 and the correlation analysis between its mRNA expression levels and inosine monophosphate content in large yellow croaker (Larimichthys crocea).

Declining flesh quality has drawn considerable attention in the farmed large yellow croaker (LYC; Larimichthys crocea) industry. Inosine monophosphate (IMP) is the primary flavor substance in aquatic animals. Adenosine monophosphate deaminase 1 (AMPD1) plays a critical role in IMP formation by catalyzing the deamination of AMP to IMP in the purine nucleotide cycle. To further evaluate the correlation between ampd1 mRNA expression levels and IMP content in the LYC muscle tissue, the relevant open reading frame (ORF) of L. crocea (Lcampd1) was cloned, and the IMP content and Lcampd1 mRNA expression in the muscles of LYCs of different sizes were examined. The ORF cDNA of Lcampd1 was 2211 bp in length and encoded a polypeptide of 736 amino acids (AAs). The deduced protein, LcAMPD1, possesses conserved AMPD active regions (SLSTDDP) and shows high homology with AMPD proteins of other teleost fishes. The genomic DNA sequence of Lcampd1 exhibits a high degree of evolutionary conservation in terms of structural organization among species. Phylogenetic analysis of the deduced AA sequence revealed that teleost fish and mammalian AMPD1 were separate from each other and formed a cluster with AMPD3, suggesting that AMPD1 and AMPD3 arose by duplication of a common primordial gene. In healthy LYC, Lcampd1 mRNA was expressed only in the muscle tissue. The IMP content in the muscle of LYCs with different average body weights was measured by high-performance liquid chromatography; the results showed that the IMP content in the muscle of LYCs with greater body weight was significantly higher than that in LYC with lower body weight. Moreover, a similar trend in Lcampd1 expression was observed in these muscle tissues. The Pearson correlation analysis further showed that the Lcampd1 mRNA expression was positively correlated with IMP content in the muscles of different-sized LYCs. These results suggest the potential function of Lcampd1 in determining the IMP content in LYC and provide a theoretical basis for flesh quality improvement, as well as a scientific basis for the development of the molecular breeding of LYC.

Structure, cooperativity and inhibition of the inosine 5'-monophosphate-specific phosphatase from Saccharomyces cerevisiae.

The nucleoside inosine is a main intermediate of purine nucleotide catabolism in Saccharomyces cerevisiae and is produced via the dephosphorylation of inosine monophosphate (IMP) by IMP-specific 5'-nucleotidase 1 (ISN1), which is present in many eukaryotic organisms. Upon transition of yeast from oxidative to fermentative growth, ISN1 is important for intermediate inosine accumulation as purine storage, but details of ISN1 regulation are unknown. We characterized structural and kinetic behavior of ISN1 from S. cerevisiae (ScISN1) and showed that tetrameric ScISN1 is negatively regulated by inosine and adenosine triphosphate (ATP). Regulation involves an inosine-binding allosteric site along with IMP-induced local and global conformational changes in the monomer and a tetrameric re-arrangement, respectively. A proposed interaction network propagates local conformational changes in the active site to the intersubunit interface, modulating the allosteric features of ScISN1. Via ATP and inosine, ScISN1 activity is likely fine-tuned to regulate IMP and inosine homeostasis. These regulatory and catalytic features of ScISN1 contrast with those of the structurally homologous ISN1 from Plasmodium falciparum, indicating that ISN1 enzymes may serve different biological purposes in different organisms.

Physicochemical properties as indicators of sensory characteristics of jidori and broiler chicken meat.

To improve the eating quality of chicken, the physicochemical properties were examined, which serve as indicators for breeding. Thigh meat was collected from 384 chickens from seven and nine types of the jidori (free-range local traditional pedigree chickens) or broiler chickens, respectively. The principal component analysis of the physicochemical values of the jidori and broilers were arranged as different groups in the score plot. The results of multiple regression analysis of the sensory characteristics and physicochemical properties of thigh meat indicated that the tenderness decreased with the higher crude protein content and cooking loss in the jidori and increased with lower cooking loss and higher moisture and ether extract content in the broiler. The juiciness of the jidori decreased as the moisture content decreased, and that of the broiler decreased as the cooking loss and crude protein content increased. The umami of both the jidori and broiler was improved by increasing the 5'-inosinic acid content. Therefore, it was suggested that the physicochemical properties, which serve as indicators of the eating quality of chicken, differed between the jidori and broiler, and that the general biochemical components, cooking loss and 5'-inosinic acid content may serve as breeding indicators.

A c-di-GMP signaling module controls responses to iron in Pseudomonas aeruginosa.

Cyclic dimeric guanosine monophosphate (c-di-GMP) serves as a bacterial second messenger that modulates various processes including biofilm formation, motility, and host-microbe symbiosis. Numerous studies have conducted comprehensive analysis of c-di-GMP. However, the mechanisms by which certain environmental signals such as iron control intracellular c-di-GMP levels are unclear. Here, we show that iron regulates c-di-GMP levels in Pseudomonas aeruginosa by modulating the interaction between an iron-sensing protein, IsmP, and a diguanylate cyclase, ImcA. Binding of iron to the CHASE4 domain of IsmP inhibits the IsmP-ImcA interaction, which leads to increased c-di-GMP synthesis by ImcA, thus promoting biofilm formation and reducing bacterial motility. Structural characterization of the apo-CHASE4 domain and its binding to iron allows us to pinpoint residues defining its specificity. In addition, the cryo-electron microscopy structure of ImcA in complex with a c-di-GMP analog (GMPCPP) suggests a unique conformation in which the compound binds to the catalytic pockets and to the membrane-proximal side located at the cytoplasm. Thus, our results indicate that a CHASE4 domain directly senses iron and modulates the crosstalk between c-di-GMP metabolic enzymes.

Ribonucleotides differentially modulate oral glutamate detection thresholds.

The savory or umami taste of the amino acid glutamate is synergistically enhanced by the addition of the purines inosine 5'-monophosphate (IMP) and guanosine 5'-monophosphate (GMP) disodium salt. We hypothesized that the addition of purinergic ribonucleotides, along with the pyrimidine ribonucleotides, would decrease the absolute detection threshold of (increase sensitivity to) l-glutamic acid potassium salt (MPG). To test this, we measured both the absolute detection threshold of MPG alone and with a background level (3 mM) of 5 different 5'-ribonucleotides. The addition of the 3 purines IMP, GMP, and adenosine 5'-monophosphate (AMP) lowered the MPG threshold in all participants (P < 0.001), indicating they are positive modulators or enhancers of glutamate taste. The average detection threshold of MPG was 2.08 mM, and with the addition of IMP, the threshold was decreased by approximately 1.5 orders of magnitude to 0.046 mM. In contrast to the purines, the pyrimidines uridine 5'-monophosphate (UMP) and cytidine 5'-monophosphate (CMP) yielded different results. CMP reliably raised glutamate thresholds in 10 of 17 subjects, suggesting it is a negative modulator or diminisher of glutamate taste for them. The rank order of effects on increasing sensitivity to glutamate was IMP > GMP> AMP >> UMP// CMP. These data confirm that ribonucleotides are modulators of glutamate taste, with purines enhancing sensitivity and pyrimidines displaying variable and even negative modulatory effects. Our ability to detect the co-occurrence of glutamate and purines is meaningful as both are relatively high in evolutionarily important sources of nutrition, such as insects and fermented foods.

Inosine monophosphate dehydrogenase intranuclear inclusions are markers of aging and neuronal stress in the human substantia nigra.

We explored mechanisms involved in the age-dependent degeneration of human substantia nigra (SN) dopamine (DA) neurons. Owing to its important metabolic functions in post-mitotic neurons, we investigated the developmental and age-associated changes in the purine biosynthetic enzyme inosine monophosphate dehydrogenase (IMPDH). Tissue microarrays prepared from post-mortem samples of SN from 85 neurologically intact participants humans spanning the age spectrum were immunostained for IMPDH combined with other proteins. SN DA neurons contained two types of IMPDH structures: cytoplasmic IMPDH filaments and intranuclear IMPDH inclusions. The former were not age-restricted and may represent functional units involved in sustaining purine nucleotide supply in these highly metabolically active cells. The latter showed age-associated changes, including crystallization, features reminiscent of pathological inclusion bodies, and spatial associations with Marinesco bodies; structures previously associated with SN neuron dysfunction and death. We postulate dichotomous roles for these two subcellularly distinct IMPDH structures and propose a nucleus-based model for a novel mechanism of SN senescence that is independent of previously known neurodegeneration-associated proteins.

Amplified drying in South Asian summer monsoon precipitation due to anthropogenic sulfate aerosols.

A declining trend in Indian summer monsoon precipitation (ISMP) in the latter half of the 20th century is a scientifically challenging and societally relevant research issue. Heavy aerosol loading over India is one of the key factors in modulating the ISMP. Using the state-of-the-state-of-the-art chemistry-climate model, ECHAM6-HAMMOZ, the impacts of South Asian anthropogenic sulfate aerosols on the Indian summer monsoon precipitation were investigated against: (1) 2010 La Niña (excess monsoon), (2) 2015 El Niño (deficit monsoon) in comparison to (3) normal monsoon 2016. Sensitivity simulations were designed with 48% enhancement in South Asian SO2 emissions based on a trend estimated from Ozone Monitoring Instrument (OMI) satellite observations during 2006-2017. The model simulations showed that sulfate aerosols reduce ISMP by 27.5%-43.3 %, while simulations without sulfate loading enhanced ISMP by 23% in 2010 La Niña and reduction by 35% in 2015 El Niño. This paper reports that sulfate aerosols loading over India reduce precipitation by aerosol-induced direct and indirect effects by inducing atmospheric cooling, weakening in the convection, and reduction in moisture transport to Indian landmass. This paper emphasizes the necessity of alternate use of energy to reduce sulfate aerosol emissions to solve water issues in South Asia.

Integrating proteomics and metabolomics to elucidate the molecular network regulating of inosine monophosphate-specific deposition in Jingyuan chicken.

Inosine monophosphate (IMP) plays a significant role in meat taste, yet the molecular mechanisms controlling IMP deposition in muscle tissues still require elucidation. The present study systematically and comprehensively explores the molecular network governing IMP deposition in different regions of Jingyuan chicken muscle. Two muscle groups, the breast and leg, were examined as test materials. Using nontargeted metabolomic sequencing, we screened and identified 20 metabolites that regulate IMP-specific deposition. We maintained regular author and institution formatting, used clear, objective, and value-neutral language, and avoided biased or emotional language. We followed a consistent footnote style and formatting features and used precise word choice with technical terms where appropriate. Out of these, 5 were identified as significant contributors to the regulation of IMP deposition. We explained technical term abbreviations when first used and ensured a logical flow of information with causal connections between statements. The results indicate that PGM1, a key enzyme involved in synthesis, is higher in the breast muscle compared to the leg muscle, which may provide an explanation for the increased deposition of IMP in the breast muscle. We aimed for a clear structure with logical progression, avoided filler words, and ensured grammatical correctness. The activity of key enzymes (PKM2, AK1, AMPD1) involved in this process was higher in the breast muscle than in the leg muscle. In the case of IMP degradation metabolism, the activity of its participating enzyme (PurH) was lower in the breast muscle than in the leg muscle. These findings suggest that the increased deposition of IMP in Jingyuan chickens' breast muscle may result from elevated metabolism and reduced catabolism of key metabolites. In summary, a metaomic strategy was utilized to assess the molecular network regulation mechanism of IMP-specific deposition in various segments of Jingyuan chicken. These findings provide insight into genetic improvement and molecular breeding of meat quality traits for top-notch broilers.

Inosine 5'- monophosphate derived umami taste intensity of beef determination by electrochemistry and chromatography.

The umami sensation contributes to beef taste and acceptability. Inosine 5'- monophosphate (IMP), the most abundant 5'-ribonucleotide in meat, is known to impart an umami taste without the undesired side effects commonly associated with glutamate. Nevertheless, the investigation of IMP's role in beef flavor has thus far been overlooked. Traditional methods for detecting IMP have relied on liquid chromatography coupled with ultraviolet spectroscopy or mass spectrometry techniques. However, these methods are not practical for production settings due to the complexity and resource demands of sophisticated laboratory techniques. Alternative methods like cyclic voltammetry might offer more practical solutions for rapidly detecting IMP. The objectives of this study were to evaluate the efficiency of using electrochemistry and chromatography on differentiating beef strips spiked with different IMP contents. The IMP threshold was 0.30 mM determined by a trained panel using the Best Estimates Threshold method. Beef strip steaks of USDA Prime, Choice, and Select were spiked at 0.30 and 0.60 mM of IMP, based on green weight and an estimated moisture content of 65%. In this study, differences in the IMP content of steaks were not detected by liquid chromatography-mass spectrometry. However, the cyclic voltammetry approach differentiated IMP concentrations at 0.50 mM or above in aqueous solutions and subsequentially meat extracts from the buffered blank solutions. In conclusion, cyclic voltammetry holds potential as a rapid and effective approach for detecting IMP in beef and other meat products, offering promising applications for future research.

Preparation of sunflower seed-derived umami protein hydrolysates and their synergistic effect with monosodium glutamate and disodium inosine-5'-monophosphate.

Sunflower seeds are rich in protein and can be an excellent raw material for the production of umami peptides. In this study, sunflower seed meal, which was defatted at a low temperature, was taken as the raw material, and proteins were separated, followed by hydrolyzation for 4 h by flavourzyme® to obtain hydrolysates with strong umami intensity. These hydrolysates were deamidated using glutaminase to increase the umami intensity. The highest umami value of 11.48 was recorded for hydrolysates deamidated for 6 h, and the umami intensity was determined. The umami hydrolysates mixed with 8.92 mmol IMP + 8.02 mmol MSG showed the highest umami value of 25.21. Different concentrations of ethanol were used for further separation of hydrolysates, and the highest umami value of 13.54 was observed for 20% ethanol fraction. The results of this study provide utilization method for sunflower seed meal protein and a theoretical basis for the preparation of umami peptides. PRACTICAL APPLICATION: A large number of sunflower seed meals after oil production are used as feed for livestock and poultry. Sunflower seed meal is rich in protein, and umami amino acid composition in sunflower seed meal is up to 25%-30%, which is potentially an excellent raw material for the production of umami peptides. The umami flavor and synergistic effect of obtained hydrolysates, with MSG and IMP, were analyzed in the present study. We intend to provide a novel way for utilization of protein from sunflower seed meal along with a theoretical basis for the preparation of umami peptides.

IMPDH1-associated autosomal dominant retinitis pigmentosa: natural history of novel variant Lys314Gln and a comprehensive literature search.

BACKGROUND: Inosine monophosphate dehydrogenase (IMPDH) is a key regulatory enzyme in the de novo synthesis of the purine base guanine. Mutations in the inosine monophosphate dehydrogenase 1 gene (IMPDH1) are causative for RP10 autosomal dominant retinitis pigmentosa (adRP). This study reports a novel variant in a family with IMPDH1-associated retinopathy. We also performed a comprehensive review of all reported IMPDH1 disease causing variants with their associated phenotype. MATERIALS AND METHODS: Multimodal imaging and functional studies documented the phenotype including best-corrected visual acuity (BCVA), fundus photograph, fundus autofluorescence (FAF), full field electroretinogram (ffERG), optical coherence tomography (OCT) and visual field (VF) data were collected. A literature search was performed in the PubMed and LOVD repositories. RESULTS: We report 3 cases from a 2-generation family with a novel heterozygous likely pathogenic variant p. (Lys314Gln) (exon 10). The ophthalmic phenotype showed diffuse outer retinal atrophy with mild pigmentary changes with sparse pigmentary changes. FAF showed early macular involvement with macular hyperautofluorescence (hyperAF) surrounded by hypoAF. Foveal ellipsoid zone island can be found in the youngest patient but not in the older ones. The literature review identified a further 56 heterozygous, 1 compound heterozygous, and 2 homozygous variant. The heterozygous group included 43 missense, 3 in-frame, 1 nonsense, 2 frameshift, 1 synonymous, and 6 intronic variants. Exon 10 was noted as a hotspot harboring 18 variants. CONCLUSIONS: We report a novel IMPDH1 variant. IMPDH1-associated retinopathy presents most frequently in the first decade of life with early macular involvement.

Development of sensitive and stable electrochemical impedimetric biosensor based on T1R1 receptor and its application to detection of umami substances.

Umami substances can provide a palatable flavour for food. In this study, an electrochemical impedimetric biosensor was developed for detecting umami substances. This biosensor was fabricated by immobilising T1R1 onto AuNPs/reduced graphene oxide/chitosan which was in advance electro-deposited onto a glassy carbon electrode. The evaluation by the electrochemical impedance spectrum method showed that the T1R1 biosensor performed well with low detection limits and wide linear ranges. Under the optimised incubation time (60 s), the electrochemical response was linearly related to the concentrations of the detected targets monosodium glutamate and inosine-5'-monophosphate within their respective linear range of 10-14 to 10-9 M and 10-16 to 10-13 M. The low detection limit of monosodium glutamate and inosine-5'-monophosphate was 10-15 M and 10-16 M, respectively. Moreover, the T1R1 biosensor exhibited high specificity to umami substances even in the real food sample. The developed biosensor still retained 89.24% signal intensity after 6-day storage, exhibiting a desirable storability.

LncRNA UCA1 Participates in De Novo Synthesis of Guanine Nucleotides in Bladder Cancer by Recruiting TWIST1 to Increase IMPDH1/2.

Metabolic dysregulation has been identified as one of the hallmarks of cancer biology. Based on metabolic heterogeneity between bladder cancer tissues and adjacent tissues, we discovered several potential driving factors for the bladder cancer occurrence and development. Metabolic genomics showed purine metabolism pathway was mainly accumulated in bladder cancer. Long noncoding RNA urothelial carcinoma-associated 1 (LncRNA UCA1) is a potential tumor biomarker for bladder cancer diagnosis and prognosis, and it increases bladder cancer cell proliferation, migration, and invasion via the glycolysis pathway. However, whether UCA1 plays a role in purine metabolism in bladder cancer is unknown. Our findings showed that UCA1 could increase the transcription activity of guanine nucleotide de novo synthesis rate limiting enzyme inosine monophosphate dehydrogenase 1 (IMPDH1) and inosine monophosphate dehydrogenase 2 (IMPDH2), triggering in guanine nucleotide metabolic reprogramming. This process was achieved by UCA1 recruiting the transcription factor TWIST1 which binds to the IMPDH1and IMPDH2 promoter region. Increased guanine nucleotide synthesis pathway products stimulate RNA polymerase-dependent production of pre-ribosomal RNA and GTPase activity in bladder cancer cells, hence increasing bladder cancer cell proliferation, migration, and invasion. We have demonstrated that UCA1 regulates IMPDH1/2-mediated guanine nucleotide production via TWIST1, providing additional evidence of metabolic reprogramming.

Enhanced preference for dried bonito dashi by prior experience with dashi and various taste substances in mice.

Dried bonito dashi, a complex mixture of sour, bitter, and umami substances as well as over 400 odorants, is the most widely used Japanese fish broth that enhances palatability of various dishes. Recent studies have suggested that prior experience with dried bonito dashi produces strong enhancement of subsequent intake and preference for dried bonito dashi. The present study investigated taste substances in dried bonito dashi that enhance subsequent dashi preference by its prior exposure. Male C57BL/6N mice were initially exposed for 10 days to (1) dried bonito dashi, (2) a chemical mixture of taste substances identified in dried bonito dashi (artificially reconstituted dashi), or (3) individual chemical solutions such as NaCl, monosodium l-glutamate (MSG), inosine 5'-monophosphate (IMP), lactic acid, histidine, and glucose. Intakes of 0.01-100% dried bonito dashi with water were then measured using ascending concentration series of 2-day two-bottle choice tests. Prior exposure to 1-100% dashi enhanced subsequent dashi preference in a concentration-dependent manner and the greatest effects were attained with 10-100% dashi exposure. Exposure to the reconstituted dashi also enhanced subsequent dashi preference. Among individual chemical solutions, 0.1% IMP produced modest enhancement of subsequent dashi preference, but neither NaCl, MSG, histidine, lactic acid, nor glucose did. These results suggest that IMP is at least a key substance that produces experience-based enhancement of dried bonito dashi preference.

The pursuit of new alternative ways to eradicate Helicobacter pylori continues: Detailed characterization of interactions in the adenylosuccinate synthetase active site.

Purine nucleotide synthesis is realised only through the salvage pathway in pathogenic bacterium Helicobacter pylori. Therefore, the enzymes of this pathway, among them also the adenylosuccinate synthetase (AdSS), present potential new drug targets. This paper describes characterization of His6-tagged AdSS from H. pylori. Thorough analysis of 3D-structures of fully ligated AdSS (in a complex with guanosine diphosphate, 6-phosphoryl-inosine monophosphate, hadacidin and Mg2+) and AdSS in a complex with inosine monophosphate (IMP) only, enabled identification of active site interactions crucial for ligand binding and enzyme activity. Combination of experimental and molecular dynamics (MD) simulations data, particularly emphasized the importance of hydrogen bond Arg135-IMP for enzyme dimerization and active site formation. The synergistic effect of substrates (IMP and guanosine triphosphate) binding was suggested by MD simulations. Several flexible elements of the structure (loops) are stabilized by the presence of IMP alone, however loops comprising residues 287-293 and 40-44 occupy different positions in two solved H. pylori AdSS structures. MD simulations discovered the hydrogen bond network that stabilizes the closed conformation of the residues 40-50 loop, only in the presence of IMP. Presented findings provide a solid basis for the design of new AdSS inhibitors as potential drugs against H. pylori.

Effects of different levels of inosine-5'-monophosphate (5'-IMP) supplementation on the growth performance and meat quality of finishing pigs (75 to 100 kg).

This study aimed to assess the effects of dietary supplementation of inosine-5'-monophosphate (5'-IMP) on energy efficiency, growth performance, carcass characteristics, meat quality, oxidative status, and biochemical profile of blood plasma in finishing pigs. Fifty-four crossbred castrated male pigs were distributed in a randomized block design consisting of nine blocks, with six treatments per block and one animal per treatment per block. Experimental diets were as follows: positive control diet (PC, 3300 kcal ME/kg), negative control diet (NC, 3200 kcal ME/kg), and four diets prepared by supplementing the NC diet with 0.050%, 0.100%, 0.150%, or 0.200% 5'-IMP. Based on regression analysis, supplementation with 0.129% 5'-IMP increased average daily weight gain (1.30 kg). Backfat thickness, pH45minutes and redness of m. Longissimus Lumborum (LL) increased linearly with 5'-IMP supplementation level. Drip loss and pH at 24 h post-slaughter had a quadratic response to 5'-IMP supplementation. It is concluded that 5'-IMP supplementation positively influenced growth performance, carcass characteristics and LL meat quality in finishing barrows.

Targeted deletion of ecto-5'-nucleotidase results in retention of inosine monophosphate content in postmortem muscle of medaka (Oryzias latipes).

Inosine monophosphate (IMP) is an important indicator of meat freshness and contributes to its umami taste. An attractive strategy for enhancing umami is to suppress the IMP-degrading activity and increase the IMP content in the skeletal muscle through genome editing technology using the CRISPR-Cas9 system. However, the molecular mechanisms underlying IMP degradation remain unclear. We cloned two ecto-5'-nucleotidase genes, designated as ecto-5'-nucleotidase-a (nt5ea) and ecto-5'-nucleotidase-b (nt5eb), from medaka (Oryzias latipes), a vertebrate model organism. Expression analysis using embryos showed that nt5ea or nt5eb overexpression remarkably upregulated IMP degradation, and that the IMP-degrading activity was higher in Nt5ea than in Nt5eb. Furthermore, we established frame-shifted or large deletion (lacking nt5ea or nt5eb locus) mutant strains and assayed the effects of gene disruptions on the amount of IMP in skeletal muscle. The nt5ea-deficient medaka showed considerable higher levels of IMP at 48 h postmortem than did the wild-type fish. The nt5eb mutants also exhibited higher IMP contents than that in the wild types, but the increase was less than that in the nt5ea mutants. Our results demonstrated that nt5e is an important regulator of IMP levels in skeletal muscle and that its loss of function was effective in maintaining IMP content.