Analysis of functional candidate genes related to ubiquitination process for meat quality in commercial pigs

During the conversion from muscle to meat, the degradation of myofibrillar proteins impacts on the water holding capacity of meat. Besides the calpain/calpastatin proteolytic system, the activity of ubiquitin proteasome system also causes the degradation of intermediate filament proteins and integrins that leads to an increase of drip loss formation in the muscle cells. The current study sought to evaluate the effects on meat quality of the four functional candidate genes, UBXN1, UBE3B, TRIP12 and ZRANB1, that related to the ubiquitination processes. Nine novel polymorphisms were identified in the transcribed and 3´-UTR regions of these genes. Seven of these novel SNPs were genotyped in a total of about 570 animals derived from the two populations, German Landrace (GL) and the commercial crossbreed of Pietrain × (German Large White × German Landrace) (PiF1). The SNPs of the four candidate genes exhibited strong associations with the indicators of water holding capacity, including muscle conductivity (UBXN1, UBE3B, TRIP12); drip loss (UBXN1, UBE3B), pH values (UBXN1, TRIP12 and ZRANB1) and meat redness (UBE3B). The SNPs of the four candidate genes had also significant association with carcass traits such as loin eye area, loin fat depth and meat to fat ratio. Moreover, the variation of transcript abundances of UBXN1, ZRANB1 and TRIP12 were significantly associated with the respective polymorphisms. At the same time, their transcript abundances were correlated with muscle conductivity, pH and drip loss, respectively. UBE3B transcript abundance was associated with meat redness. The integration of association and expression data imply the existence of causal polymorphisms in the cis-regulatory regions of these candidate genes, which are in incomplete linkage disequilibrium with the detected SNPs, and which primarily affect their transcript abundance and, in consequence, traits related to water holding capacity. Thus the study revealed the consistent triangular relationship among genotype - phenotype - transcript abundance across the four candidate genes. In fact, the detected SNPs were in linkage phase with alleles of causal sites increasing the transcript abundances, and enhancing the purge loss in the case of UBXN1 and UBE3B, whereas decreasing drip loss formation in the case of TRIP12 and ZRANB1 genes. Moreover, interactions observed among these genes of the ubiquitination system and the ryanodine 1 receptor (RYR1) indicate options for further improvement of meat quality, in particular in RYR1 heterozygous animals, by considering genotypes at these loci. The results of this study provide genetic evidences to support UBXN1, UBE3B, TRIP12 and ZRANB1 as the functional candidate genes for water holding capacity of pork.Während der Fleischreifung, wenn Muskelgewebe zu Fleisch wird, so wird die Wasserbindungskapazität des Gewebes durch den Abbau myofibrillärer Proteine beeinflusst. Neben dem Calpain/Calpastatin Proteolysesystem ist die Aktivität des Ubiquitin-Systems für den Abbau von Proteinen der Intermediärfilamente sowie von Integrinen mitverantwortlich. Dies verursacht einen erhöhten Flüssigkeitsverlust der Muskelzellen. Die vorliegende Studie versucht, die Auswirkungen von vier funktionellen Kandidatengenen aus dem Ubiquitinierungsprozess, UBXN1, UBE3B, TRIP12 und ZRANB1, auf die Fleischqualität auszuwerten. Neun neue Polymorphismen wurden in der transskribierten Sequenz sowie im 3’UTR der Gene identifiziert. Sieben dieser neuen SNP wurden in insgesamt etwa 570 Tieren genotypisiert, die aus zwei Populationen stammen, Deutsche Landrasse (GL) und Pietrain × (Deutsches Edelschwein × Deutsche Landrasse) (PiF1). Die SNP der vier Kandidatengene wiesen eine signifikante Assoziation mit Indikatoren der Wasserbindungskapazität auf, nämlich der Muskelleitfähigkeit (UBXN1, UBE3B, TRIP12), dem Flüssigkeitsverlust (Drip) (UBXN1, UBE3B), dem pH-Wert (UBXN1, TRIP12 und ZRANB1) und der Rotfärbung des Fleisches (UBE3B). Die SNP der vier Kandidatengene zeigten zudem signifikante Assoziationen mit Schlachtkörpermerkmalen wie der Kotelettfläche, Lendenfettdicke und dem Fleisch-Fett Verhältnis. Darüber hinaus war die Varianz der Transkriptmenge von UBXN1, ZRANB1 and TRIP12 signifikant mit den jeweiligen Polymorphismen korreliart. Gleichzeitig war die Transkriptmenge mit Muskelleitfähigkeit, pH Wert und Flüssigkeitsverlust assoziiert. Die Transkriptmenge von UBE3B war assoziiert mit der Rotfärbung des Fleisches. Die Zusammenführung von Assoziations- und Expressionsdaten weist auf die Existenz kausaler Mutationen in den cis-regulatorischen Regionen der Kandidatengene hin, welche sich in einem unvollständigen Kopplungsungleichgewicht mit den entdeckten SNP befinden. Diese beeinflussen in erster Linie die Transkriptmenge und, als Konsequenz, Merkmale der Wasserbindungskapazität. Die vorliegende Studie demonstiert die konsistente dreiseitige Beziehung zwischen Phänotyp, Genotyp und Transkriptmenge bei allen vier Kandidatengenen. Tatsächlich befanden sich die detektierten SNP in einer Kopplungsphase mit Allelen kausaler Mutationen, welche die Transkriptmenge erhöhen und im Fall von UBXN1 und UBE3B den Flüssigkeitsverlust erhöhen, wohingegen dieser im Fall von TRIP12 and ZRANB1 verringert wurde. Darüber hinaus weisen Interaktionen zwischen den Genen des Ubiquitin-Systems und dem Ryanodin 1 Rezeptor (RYR1) auf Möglichkeiten der weitergehenden Verbesserung der Fleischqualität hin, besonders in RYR1 heterozygoten Tieren, indem der Genotyp an diesen Loci berücksichtigt wird. Die Ergebnisse dieser Studie unterstützen UBXN1, UBE3B, TRIP12 und ZRANB1 als funktionellen Kandidatengene für die Wasserbindungskapazität von Schweinefleisch

Effect of Molecular Charge Asymmetry on Even-to-odd Ratio of High-order Harmonic Generation

Recently, asymmetric molecules, such as HeH$_2^+$, CO, OCS, HCl, have been evolved much attention since its rich information in the high-order harmonic generation (HHG), whose ratio of adjacent even and odd harmonics characterizes the asymmetry of molecules. In this paper, we study the dependence of even-to-odd ratio on the asymmetric parameters, in particular, the nuclear-charge ratio, and the permanent dipole, by exploiting a simple but general model of asymmetric molecules $Z_1Z_2$ subjected to an intense laser pulse. The HHG is simulated by the numerical method of solving the time-dependent Schrödinger equation. We find out that this even-to-odd ratio strongly depends on the nuclear-charge ratio. In particular, the even-to-odd ratio reaches its maximum when the nuclear-charge ratio is about from 0.5 to 0.7. Besides, the dependence on the permanent dipole of the even-to-odd ratio has a non-trivial law. To explain, we calculate the analytical ratio of the transition dipole according to the emission of even and odd harmonics, and this ratio is well consistent with the even-to-odd ratio of the HHG

A comparison for donor-acceptor interactions between E(PH3)2 and NHEMe ligands (E = C to Pb) of W(CO)5 complexes using energy decomposition analysis method with natural orbitals for chemical valence theory

Quantum chemical calculations at the BP86/TZ2P+ level of theory are performed for a comparison of density functional theory (DFT) between tetrylones [(CO)5W-{E(PH3)2}] (W5-EP2) and tetrylenes [(CO)5W-{NHEMe}] (W5-NHEMe) when E = C to Pb. The EDA-NOCV results suggest that the W-E bond dissociation energies (BDEs) in tetrylone complexes increase from the lighter to the heavier homologues. The W-E bond dissociation energies (BDEs) trend in W5-EP2 comes from the increase in (CO)5W←E(PH3)2 donation and strong electrostatic attraction, and that the ligands E(PH3)2 (EP2) are strong s-donors and very weak π-donors. The W-E BDEs trend in tetrylene complexes W5-NHEMe is opposite to that of the W5-EP2 complexes which decrease from the lighter to the heavier homologues. The NHEMe ligands are strong s-donors and weak π-acceptors. NOCV pairs were used in a description of the chemical bond between the W(CO)5 fragment and the ligands in the transition-metal complexes and the results indicated that the NOCV pairs lead to very valuable description of the bonding situation of the fragment-ligand bond in complexes. Keywords. Density functional theory; Bond dissociation energies (BDEs); Energy decomposition analysis (EDA); Natural Orbitals for Chemical Valence (NOCV)

Structural Variations and Chemical Bonding in Platinum Complexes of group 14 heavier Tetrylene Homologues (Germylene – Plumbylene)

We computationally investigated the structures of Pt(II) complexes containing the heavier homologues of germylene, stannylene, and plumbylene (called heavier tetrylenes) [PtCl2-{NHEMe}] (Pt-NHE) with E = Ge – Pb using density functional theory (DFT) calculations at the BP86 level with the various basis sets def2-SVP, def2-TZVPP, and TZ2P+. The bonding situation of complexes was analyzed with charge and energy decomposition analysis methods. The results of bonding analysis showed that NHEMe ligands exhibit donor-acceptor bonds with the s lone pair electrons of heavier NHEMe donated into the vacant orbital of the metal fragment and the Pt-E bonds have PtCl2←NHEMe strong s-donation. The divalent heavier tetrylenes(II) have some degree of role as the divalent heavier tetrylones(0) character with the ligand can retain the two lone pairs at E atom. Current efforts in experimental laboratories are directed towards the synthesis of tetrylenes Pt(II) complexes from natural products, so the results in this study will provide properties and chemical bonding of complexes investigated for experimental researches

A quantum chemical computation insight into the donor-acceptor bond interaction of silver complexes with tetrylene

We computationally investigate the nature of chemical bonding from linear to bent structures of N-heterocyclic carbene-analogues of silver complexes (called tetrylene) AgCl-NHEMe (Ag-NHE) with E = C – Pb using quantum chemical calculations at the BP86 level with the various basis sets def2-SVP, def2-TZVPP, and TZ2P+. The geometry calculations find that the equilibrium structures of Ag-NHE system show major differences in the bonded orientation of NHPb ligand in Ag-NHPb compared with NHE ligands the slighter homologues Ag-NHE (E = C - Sn). The bond dissociation energy results show that the Ag-carbene bond in Ag-NHC is a strong bond and decreases from the slighter to the heavier homologues. The EDA-NOCV results indicate that the ligand NHE in complexes is strong s-donors and very weak π donor. The NOCV pairs of the bonding show small π-back donation from the Ag to the NHEMe ligands. Keywords. N-heterocyclic tetrylene, bond dissociation energy, quantum chemical calculations, bonding analysis

Urinary catecholamine excretion, cardiovascular variability, and outcomes in tetanus

Severe tetanus is characterized by muscle spasm and cardiovascular system disturbance. The pathophysiology of muscle spasm is relatively well understood and involves inhibition of central inhibitory synapses by tetanus toxin. That of cardiovascular disturbance is less clear, but is believed to relate to disinhibition of the autonomic nervous system. The clinical syndrome of autonomic nervous system dysfunction (ANSD) seen in severe tetanus is characterized principally by changes in heart rate and blood pressure which have been linked to increased circulating catecholamines. Previous studies have described varying relationships between catecholamines and signs of ANSD in tetanus, but are limited by confounders and assays used. In this study, we aimed to perform detailed characterization of the relationship between catecholamines (adrenaline and noradrenaline), cardiovascular parameters (heart rate and blood pressure) and clinical outcomes (ANSD, mechanical ventilation required, and length of intensive care unit stay) in adults with tetanus, as well as examine whether intrathecal antitoxin administration affected subsequent catecholamine excretion. Noradrenaline and adrenaline were measured by ELISA from 24-h urine collections taken on day 5 of hospitalization in 272 patients enrolled in a 2 × 2 factorial-blinded randomized controlled trial in a Vietnamese hospital. Catecholamine results measured from 263 patients were available for analysis. After adjustment for potential confounders (i.e., age, sex, intervention treatment, and medications), there were indications of non-linear relationships between urinary catecholamines and heart rate. Adrenaline and noradrenaline were associated with subsequent development of ANSD, and length of ICU stay

Safety and efficacy of fluoxetine on functional outcome after acute stroke (AFFINITY): a randomised, double-blind, placebo-controlled trial

Background Trials of fluoxetine for recovery after stroke report conflicting results. The Assessment oF FluoxetINe In sTroke recoverY (AFFINITY) trial aimed to show if daily oral fluoxetine for 6 months after stroke improves functional outcome in an ethnically diverse population. Methods AFFINITY was a randomised, parallel-group, double-blind, placebo-controlled trial done in 43 hospital stroke units in Australia (n=29), New Zealand (four), and Vietnam (ten). Eligible patients were adults (aged ≥18 years) with a clinical diagnosis of acute stroke in the previous 2–15 days, brain imaging consistent with ischaemic or haemorrhagic stroke, and a persisting neurological deficit that produced a modified Rankin Scale (mRS) score of 1 or more. Patients were randomly assigned 1:1 via a web-based system using a minimisation algorithm to once daily, oral fluoxetine 20 mg capsules or matching placebo for 6 months. Patients, carers, investigators, and outcome assessors were masked to the treatment allocation. The primary outcome was functional status, measured by the mRS, at 6 months. The primary analysis was an ordinal logistic regression of the mRS at 6 months, adjusted for minimisation variables. Primary and safety analyses were done according to the patient's treatment allocation. The trial is registered with the Australian New Zealand Clinical Trials Registry, ACTRN12611000774921. Findings Between Jan 11, 2013, and June 30, 2019, 1280 patients were recruited in Australia (n=532), New Zealand (n=42), and Vietnam (n=706), of whom 642 were randomly assigned to fluoxetine and 638 were randomly assigned to placebo. Mean duration of trial treatment was 167 days (SD 48·1). At 6 months, mRS data were available in 624 (97%) patients in the fluoxetine group and 632 (99%) in the placebo group. The distribution of mRS categories was similar in the fluoxetine and placebo groups (adjusted common odds ratio 0·94, 95% CI 0·76–1·15; p=0·53). Compared with patients in the placebo group, patients in the fluoxetine group had more falls (20 [3%] vs seven [1%]; p=0·018), bone fractures (19 [3%] vs six [1%]; p=0·014), and epileptic seizures (ten [2%] vs two [<1%]; p=0·038) at 6 months. Interpretation Oral fluoxetine 20 mg daily for 6 months after acute stroke did not improve functional outcome and increased the risk of falls, bone fractures, and epileptic seizures. These results do not support the use of fluoxetine to improve functional outcome after stroke

A theoretical study of structure, bonding and property of platinum(II)-8-hydroxyquinolines complexes with carbene and heavier homologues

In this work, a theoretical study for platinum(II)-8-hydroxyquinoline-tetrylene complexes [{PtCl–C9H6NO}–NHEPh] (Pt–EPh) is carried out for the first time by using the density functional theory (DFT). Quantum chemical calculations with DFT and charge methods at the BP86 level with basic sets SVP and TZVPP have been perfomed to get insight into the structures and property of Pt–EPh. The optimization of equilibrium geometries of the ligands EPh in Pt–EPh, bonded in the distorted end-on way to the Pt fragment is studied, in which the bending angle slightly decreases from carbene Pt–CPh to germylene Pt–GePh. Quantum chemical parameters such as EHOMO, ELUMO, the energy gap (ELUMO – EHOMO), electronegativity, global hardness, and global softness in the neutral molecules have been calculated and discussed. Bond dissociation energies decrease from the slighter to the heavier homologues. The hybridization of atoms E has large p characters, while the hybridization of atom Pt has a greater d character. Thus, the Pt–E bond possesses not only NHEPh→{PtCl–C9H6NO} strong -donation but also a significant contribution of π-donation NHEPh→{PtCl–C9H6NO}, and a weak π-backdonation metal-ligand NHEPh←{PtCl-C9H6NO} in complexes Pt-EPh is also considered

Antioxidant activity of some natural essential oils in Vietnam: Comparison between QSAR simulation and experimental study

The antioxidant activity of essential oils from leaves of Piper betle L. (T) and Cleistocalyx operculatus L. (V) and aerial parts of Ageratum conyzoides L. (H), indigenously grown in Thua Thien Hue province, Vietnam, is investigated. The quantitative structure–activity relationship (QSAR) model comprising 4-hydroxy-chromene-2H-one and its 26 derivatives is used to predict the radical scavenging activity of T, V, and H. The radical scavenging activity of the oils is experimentally determined with DPPH (1,1-diphenyl-2-picrylhydrazyl) via IC50 values. The experimental IC50 values are in good agreement with those obtained from the QSAR model. The IC50 value of Piper betle L. is 3.71 mg/mL, comparable to that of the strong antioxidant ascorbic acid (3.03 mg /mL)