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Planetary gear transmission is subjected to base motions in many industrial applications such as wind turbines and automobiles, and its dynamic behavior will be affected by base motions. To develop a dynamic model more suitable for the planetary gear under base motions, the general modeling procedures are illustrated in detail from the perspective of energy. A rotational-translational-axial dynamic model of the planetary gear under pitching base motion is proposed and validated. Pitching base motion causes additional damping, stiffness and forced excitation, which disrupts the force symmetry of the planetary gear and results in unequal load sharing. Dynamic responses of the planetary gear are obtained by numerical integration, and spectrum analysis is conducted using the Fourier transform. Spectra of the rotational and translational vibrations of the central components show completely different characteristics because of the structure symmetry of the planetary gear. The influences rules of parameters on response spectra and load sharing conditions are derived, including the amplitude and the frequency of the pitching base motion, and the carrier rotating speed. When the base excitation frequency approaches half the translational natural frequencies, the load sharing condition of the planetary gear will deteriorate rapidly. The load sharing factor increases by the square of the base excitation amplitude. [ABSTRACT FROM AUTHOR]

With the increase of wind turbine size, gravity becomes an important non-torque excitation source. Gravity disrupts the cyclic symmetry of the planetary gear and causes unequal load-sharing. Because of the specific operation conditions, the bedplate will tilt and lead to the offset of the gear plane and vertical plane. Taking gravity, tooth separation, backside contact and bedplate tilt angle into consideration, a rotational-translational-axial dynamic model of the spur planetary gear is developed. With two different load-sharing factor models, the load-sharing characteristics of the planetary gear in horizontal axis wind turbines are numerically investigated. The effects of gravity, ring support stiffness and bedplate tilt angle on load-sharing characteristics are systematically examined. When planets move to certain positions, severe unequal load-sharing and backside contact are more likely to happen. Load-sharing characteristics change with the bedplate tilt angle and the ring support stiffness, and the variation trend is closely related to the occurrence of tooth separation and backside contact. [ABSTRACT FROM AUTHOR]

Abstract: A soil bacterium strain, capable of using p-nitrophenol (PNP) as its sole source of carbon and energy, was isolated by enrichment on minimal salt medium (MSM). On the basis of a phylogenetic analysis of 16S rRNA gene sequences the bacterium is a species of Arthrobacter, closely related to Arthrobacter ureafaciens DSM 20126. This strain has an unusually high substrate tolerance for PNP degradation in MSM. Greatest degradation of PNP was observed at 30°C and under slightly alkaline pH (pH 7–9) conditions. Effective degradation rates slowed as the concentration of PNP was increased. Addition of glucose from 0.1% to 0.5% generally enhanced the degradation of PNP at high concentration (400mg/l) although acidification as a result of glucose metabolism had a negative effect on PNP depletion. Biodegradation of PNP at high concentration was greatly accelerated by β-cyclodextrin at a concentration of 0.5%, indicating that β-cyclodextrin could be a promising addictive for effective PNP bioremediation. [Copyright &y& Elsevier]

Ochrobactrum sp. B2, a methyl parathion-degrading bacterium, was proved to be capable of using p-nitrophenol (PNP) as carbon and energy source. The effect of factors, such as temperature, pH value, and nutrition, on the growth of Ochrobactrum sp. B2 and its ability to degrade p-nitrophenol (PNP) at a higher concentration (100mgl−1) was investigated in this study. The greatest growth of B2 was observed at a temperature of 30°C and alkaline pH (pH 9–10). pH condition was proved to be a crucial factor affecting PNP degradation. Enhanced growth of B2 or PNP degradation was consistent with the increase of pH in the minimal medium, and acidic pH (6.0) did not support PNP degradation. Addition of glucose (0.05%, 0.1%) decreased the rate of PNP degradation even if increased cell growth occurred. Addition of supplemental inorganic nitrogen (ammonium chloride or ammonium sulphate) inhibited PNP degradation, whereas organic nitrogen (peptone, yeast extract, urea) accelerated degradation. [Copyright &y& Elsevier]

Aflatoxin B 1 (AFB 1), a naturally-occurring mycotoxin, can cause severe toxicological and carcinogenic effects in livestock and humans. Given that the chicken is one of the most important food-producing animals, knowledge regarding AFB 1 metabolism and enzymes responsible for AFB 1 transformation in the chicken has important implications for chicken production and food safety. Previously, we have successfully expressed chicken CYP1A5 and CYP3A37 monooxygenases in E. coli , and reconstituted them into a functional CYP system consisting of CYP1A5 or CYP3A37, CPR and cytochrome b 5. In this study, we aimed to investigate the roles of CYP1A5 and CYP3A37 in the bioconversion of AFB 1 to AFM 1. Our results showed that chicken CYP1A5 was able to hydroxylate AFB 1 to AFM 1. The formation of AFM 1 followed the typical Michaelis–Menten kinetics. The kinetics parameters of V max and K m were determined as 0.83 ± 0.039 nmol/min/nmol P450 and 26.9 ± 4.52 μM respectively. Docking simulations further revealed that AFB 1 adopts a "side-on" conformation in chicken CYP1A5, facilitating the hydroxylation of the C9a atom and the production of AFM 1. On the other hand, AFB 1 assumes a "face-on" conformation in chicken CYP3A37, leading to the displacement of the C9a atom from the heme iron and explaining the lack of AFM 1 hydroxylation activity. The results demonstrate that chicken CYP1A5 possesses efficient hydroxylase activity towards AFB 1 to form AFM 1. [Display omitted] • E. coli expressed chicken CYP1A5 is able to hydroxylate aflatoxin B 1 to aflatoxin M 1. • The formation of aflatoxin M 1 followed typical Michaelis–Menten kinetics. • Chicken CYP1A5 is an efficient enzyme catalyzing aflatoxin B 1 hydroxylation. • AFB 1 adopts a "side-on" conformation in chicken CYP1A5, facilitating the hydroxylation of the C9a atom. [ABSTRACT FROM AUTHOR]

Abstract: A complete cDNA encoding the NADPH–cytochrome P450 reductase (haCPR) and its genomic sequence from the cotton bollworm Helicoverpa armigera were cloned and sequenced. The open reading frame of haCPR codes for a protein of 687 amino acid residues with a calculated molecular mass of 77.4kDa. The haCPR gene spans over 11kb and its coding region is interrupted by 11 introns. haCPR is ubiquitously expressed in various tissues and at various stages of development. Escherichia coli produced haCPR enzyme exhibited catalytic activity for NADPH-dependent reduction of cytochrome c, following Michaelis–Menten kinetics. The functionality of CPR was further demonstrated by its capacity to support cytochrome P450 (e.g. haCYP9A14 and chicken CYP1A5) mediated O-dealkylation activity of alkoxyresorufins. The flavoprotein-specific inhibitor (diphenyleneiodonium chloride, DPI) showed a potent inhibition to haCPR activity (IC50 =1.69μM). Inhibitory effect of secondary metabolites in the host plants (tannic acid, quercetin and gossypol) on CPR activity (with an IC50 value ranged from 15 to 90μM) was also observed. [Copyright &y& Elsevier]

Insect cuticular hydrocarbons (CHCs) have dual functions as physical barrier and chemical signals. The last step of CHC biosynthesis is known to be catalyzed by cytochrome P450 CYP4G in a number of insects. Until recently, studies on CYP4Gs in the context of functional evolution are rare. In this study, we analyzed sequence similarity and temporal-spatial expression patterns of the five CYP4G genes in the cotton bollworm Helicoverpa armigera , an important agricultural pest and also typical representative of lepidopteran insects. Moreover, the CRISPR/Cas9-induced knockout was used to clarify the roles of the five CYP4Gs in CHC biosynthesis. Temporal-spatial expression patterns revealed that CYP4G8 was highly expressed at all developmental stages and in most tissues examined. Larvae with CYP4G8 knocked out could not produce methyl-branched CHCs and failed to pupate, while larvae with the other four CYP4G genes knocked out (4G1-type-KO) showed no significant changes in their CHC profiles, weight gain and survival. Comparative transcriptomics revealed that knocking out CYP4G8 affected the global gene expression in larvae, especially down-regulated the expression of genes in the fatty acid biosynthetic pathway, while no significant change in 4G1-type-KO transcriptome was observed. These findings indicate that the five members of the CYP4G subfamily have undergone functional divergence: CYP4G8 maintains the essential function in CHC biosynthesis, while the function of the other four CYP4G genes remains unclear. Intriguingly, CYP4G8 has evolved to be a P450 enzyme responsible for the synthesis of larval methyl-branched hydrocarbons. The observation that CYP4G8 knockout is lethal strongly suggest that CYP4G8 may serve as a candidate target for the development of insecticidal agents for the control of cotton bollworms. [Display omitted] • CRISPR/Cas9 was used to characterize the function of five CYP4G genes from Helicoverpa armigera. • CYP4G paralogs diverge in spatio-temporal expression and function. • CYP4G8, rather than the other four CYP4Gs, is the major player in cuticular hydrocarbon biosynthesis. • CYP4G8 knockout is lethal. [ABSTRACT FROM AUTHOR]

Abstract: Quinoxaline derivatives (quinoxalines) comprise a class of drugs that have been widely used as animal antimicrobial agents and feed additives. Although the metabolism of quinoxaline drugs has been mostly studied using chicken liver microsomes, the biochemical mechanism of biotransformation of these chemicals in the chicken has yet to be characterized. In this study, using bacteria produced enzymes, we demonstrated that both CYP1A4 and CYP1A5 participate in the oxidative metabolism of quinoxalines. For CYP1A5, three hydroxylated metabolites of quinocetone were generated. In addition, CYP1A5 is able to hydroxylate carbadox. For CYP1A4, only one hydroxylated product of quinocetone on the phenyl ring was identified. Neither CYP1A5 nor CYP1A4 showed hydroxylation activity towards mequindox and cyadox. Our results suggest that CYP1A4 and CYP1A5 have different and somewhat overlapping substrate specificity in quinoxaline metabolism, and CYP1A5 represents a crucial enzyme in hydroxylation of both quinocetone and carbadox. [Copyright &y& Elsevier]

Abstract: The chicken (Gallus gallus) is one of the most economically important domestic animals and also an avian model species. Chickens have two CYP1A genes (CYP1A4 and CYP1A5) which are orthologous to mammalian CYP1A1 and CYP1A2. Although the importance of chicken CYP1As in metabolism of endogenous compounds and xenobiotics is well recognized, their enzymatic properties, substrate preference and inhibitor selectivity remain poorly understood. In this study, functional enzymes of chicken CYP1A4 and CYP1A5 were successfully produced in Escherichia coli (E. coli). The substrate preference and inhibitor specificity of the two chicken CYP1As were compared. Kinetic results showed that the enzymatic parameters (Km, Vmax, Vmax/Km) for ethoxyresorufin O-deethylase (EROD) and benzyloxyresorufin O-debenzylase (BROD) differed between CYP1A4 and CYP1A5, while no significant difference was observed for methoxyresorufin O-demethylase (MROD). Lower Km of CYP1A4 for BROD suggests that CYP1A4 has a greater binding affinity to benzyloxyresorufin than either ethoxyresorufin or methoxyresorufin. The highest Vmax/Km ratio was seen in BROD activity for CYP1A4 and in MROD for CYP1A5 respectively. These results indicate that substrate preference of chicken CYP1As is more notably distinguished by BROD activity and CYP1A5 prefers shorter alkoxyresorufins resembling its mammalian ortholog CYP1A2. Differential patterns of MROD inhibition were observed between CYP1As and among the five CYP inhibitors (α-naphthoflavone, furafylline, piperonyl butoxide, erythromycin and ketoconazole). α-Naphthoflavone was determined to be a potent MROD inhibitor of both CYP1A4 and CYP1A5. In contrast, no or only a trace inhibitory effect (<15%) was observed by erythromycin at a concentration of 500μM. Stronger inhibition of MROD activity was found in CYP1A5 than CYP1A4 by relatively small molecules α-naphthoflavone, piperonyl butoxide and furafylline. AROD kinetics and inhibition profiles between chicken CYP1A4 and CYP1A5 demonstrate that the two paralogous members of the CYP1A subfamily have distinct enzymatic properties, reflecting differences in the active site geometry between CYP1A4 and CYP1A5. These findings suggest that CYP1A4 and CYP1A5 play partially overlapping but distinctly different physiological and toxicological roles in the chicken. [Copyright &y& Elsevier]

Abstract: Cytochrome P450 monooxygenases have been well known to be responsible for the synthesis of endogenous compounds and the metabolism of exogenous compounds in almost all living organisms, which require NADPH-cytochrome P450 oxidoreductase (POR) as an electron donor to function. In this study, a 2031bp open reading frame of POR gene was cloned from 35-day-old Roman hen liver, encoding an enzyme of 676 amino acids. Sequence analysis showed that chicken POR shares high homology with other vertebrates PORs and possesses the conserved binding domains of FAD, FMN, and NADPH. The genomic sequences of POR genes from chicken and other four vertebrates have highly conserved exon/intron organization structure. By fusion with bacterial signal peptide, chicken POR gene was functionally expressed in E. coli membrane and showed activities in reduction of cytochrome c and oxidation of NADPH. The Km values for cytochrome c and NADPH were 21.9±2.3μM and 2.4±0.3μM respectively. A Ping-Pong mechanism was proposed for chicken POR. [ABSTRACT FROM AUTHOR]

Abstract: Four microbial communities capable of degrading acetochlor, designated A, D, E, and J, were obtained from acetochlor-contaminated soil and sludge. Acetochlor at an initial concentration of 55mg/L was completely degraded by the four mixed cultures after 4 days. At 80mg/L acetochlor, more than 99% degradation was observed with D, 84% with A and E, and 88% with J after 9 days. There are primary eight strains of bacteria in community A, three in community D, E, and J, respectively. No single isolate was able to degrade acetochlor efficiently. The acetochlor biodegradation products were identified by gas chromatography–mass spectrometry. The probable degradative pathways of acetochlor involved dechlorination, hydroxylation, deethoxymethylation, cyclization, carboxylation, and decarboxylation. Propachlor, alachlor, and metolachlor, which are also the main components of the chloroacetanilide herbicide, could be degraded by the four mixed cultures to some degree. Given the high degradation rates observed here, the four mixed cultures obtained may be useful in the degradation processes of acetochlor. [Copyright &y& Elsevier]