ABSTRACT Introduction: Minimally invasive coronary artery bypass grafting (MICS CABG) offers a new paradigm in coronary revascularization. This study aims to compare the outcomes of MICS CABG with those of conventional median sternotomy CABG (MS CABG) within a growing minimally invasive cardiac surgical program in Singapore. Methods: Propensity matching produced 111 patient pairs who underwent MICS CABG or MS CABG between January 2009 and February 2020 at the National University Heart Centre, Singapore. Minimally invasive direct coronary artery bypass surgery patients were matched to single- or double-graft MS CABG patients (Group 1). Multivessel MICS CABG patients were matched to MS CABG patients with equal number of grafts (Group 2). Results: Overall, MICS CABG patients experienced shorter postoperative length of stay (P<0.071). In Group 2, procedural duration (P<0.001) was longer among MICS CABG patients, but it did not translate to adverse postoperative events. Postoperative outcomes, including 30-day mortality, reopening for bleeding, new onset atrial fibrillation as well as neurological, pulmonary, renal, and infectious complications were comparable between MICS and MS CABG groups. Conclusion: MICS CABG is a safe and effective approach for surgical revascularization of coronary artery disease and trends toward a reduction in hospital stay. Introduction Singapore Methods 11 200 202 Centre single doublegraft double graft 1. 1 . 1) 2. 2 2) Results Overall P<0.071. P0071 P P<0.071 0 071 (P<0.071) P<0.001 P0001 001 (P<0.001 events 30day day 30 mortality bleeding neurological pulmonary renal groups Conclusion 20 P007 P<0.07 07 (P<0.071 P<0.00 P000 00 (P<0.00 3 P00 P<0.0 (P<0.07 (P<0.0 P0 P<0. (P<0. P<0 (P<0 P< (P< (P

Resumo Fundamento: O antibiótico quimioterápico antraciclina doxorrubicina (DOX) pode induzir cardiotoxicidade cumulativa e levar à disfunção cardíaca. RNAs não codificantes longos (lncRNAs) podem funcionar como importantes reguladores na lesão miocárdica induzida por DOX. Objetivo: Este estudo tem como objetivo investigar o papel funcional e o mecanismo molecular do RNA antisense lncRNA OXCT1 1 (OXCT1-AS1) na lesão celular miocárdica induzida por DOX in vitro. Métodos: Cardiomiócitos humanos (AC16) foram estimulados com DOX para induzir um modelo de lesão celular miocárdica. A expressão de OXCT1-AS1, miR-874-3p e BDH1 em células AC16 foi determinada por RT-qPCR. A viabilidade das células AC16 foi medida pelo ensaio XTT. A citometria de fluxo foi empregada para avaliar a apoptose de células AC16. Western blotting foi utilizado para avaliar os níveis proteicos de marcadores relacionados à apoptose. O ensaio repórter de luciferase dupla foi conduzido para verificar a capacidade de ligação entre miR-874-3p e OXCT1-AS1 e entre miR-874-3p e BDH1. O valor de p<0,05 indicou significância estatística. Resultados: A expressão de OXCT1-AS1 foi diminuída em células AC16 tratadas com DOX. A superexpressão de OXCT1-AS1 reverteu a redução da viabilidade celular e a promoção da apoptose celular causada pela DOX. OXCT1-AS1 está ligado competitivamente ao miR-874-3p para regular positivamente o BDH1. A superexpressão de BDH1 restaurou a viabilidade das células AC16 e suprimiu a apoptose celular sob estimulação com DOX. A derrubada do BDH1 reverteu a atenuação da apoptose de células AC16 mediada por OXCT1-AS1 sob tratamento com DOX. Conclusão: LncRNA OXCT1-AS1 protege células miocárdicas humanas AC16 da apoptose induzida por DOX através do eixo miR-874-3p/BDH1. Fundamento (DOX cardíaca lncRNAs (lncRNAs Objetivo OXCT OXCT1AS1 OXCTAS AS1 AS (OXCT1-AS1 vitro Métodos AC (AC16 OXCT1AS1, AS1, miR8743p miRp miR 874 3p p BDH AC1 RTqPCR. RTqPCR RT qPCR. qPCR RT-qPCR XTT OXCT1-AS p005 0 05 p<0,0 estatística Resultados Conclusão miR8743p/BDH1. miR8743pBDH1 miRpBDH 3p/BDH1. miR-874-3p/BDH1 OXCT1AS (OXCT1-AS (AC1 87 p00 p<0, miR8743p/BDH1 pBDH miR8743pBDH 3pBDH1 3p/BDH1 miR-874-3p/BDH (AC 8 p0 p<0 miR8743p/BDH 3pBDH 3p/BDH p<

Abstract Background: The anthracycline chemotherapeutic antibiotic doxorubicin (DOX) can induce cumulative cardiotoxicity and lead to cardiac dysfunction. Long non-coding RNAs (lncRNAs) can function as important regulators in DOX-induced myocardial injury. Objective: This study aims to investigate the functional role and molecular mechanism of lncRNA OXCT1 antisense RNA 1 (OXCT1-AS1) in DOX-induced myocardial cell injury in vitro. Methods: Human cardiomyocytes (AC16) were stimulated with DOX to induce a myocardial cell injury model. OXCT1-AS1, miR-874-3p, and BDH1 expression in AC16 cells were determined by RT-qPCR. AC16 cell viability was measured by XTT assay. Flow cytometry was employed to assess the apoptosis of AC16 cells. Western blotting was used to evaluate protein levels of apoptosis-related markers. Dual-luciferase reporter assay was conducted to verify the binding ability between miR-874-3p and OXCT1-AS1 and between miR-874-3p and BDH1. The value of p<0.05 indicated statistical significance. Results: OXCT1-AS1 expression was decreased in DOX-treated AC16 cells. Overexpression of OXCT1-AS1 reversed the reduction of cell viability and promotion of cell apoptosis caused by DOX. OXCT1-AS1 is competitively bound to miR-874-3p to upregulate BDH1. BDH1 overexpression restored AC16 cell viability and suppressed cell apoptosis under DOX stimulation. Knocking down BDH1 reversed OXCT1-AS1-mediated attenuation of AC16 cell apoptosis under DOX treatment. Conclusion: LncRNA OXCT1-AS1 protects human myocardial cells AC16 from DOX-induced apoptosis via the miR-874-3p/BDH1 axis. Background (DOX dysfunction noncoding non coding lncRNAs (lncRNAs DOXinduced induced Objective OXCT OXCT1AS1 OXCTAS AS1 AS (OXCT1-AS1 vitro Methods AC (AC16 model OXCT1AS1, AS1, miR8743p, miR8743p miRp miR 874 3p, 3p p BDH AC1 RTqPCR. RTqPCR RT qPCR. qPCR RT-qPCR apoptosisrelated related markers Dualluciferase Dual luciferase OXCT1-AS p005 0 05 p<0.0 significance Results DOXtreated treated stimulation OXCT1AS1mediated OXCTASmediated mediated treatment Conclusion miR8743p/BDH1 miR8743pBDH1 miRpBDH 3p/BDH1 miR-874-3p/BDH axis OXCT1AS (OXCT1-AS (AC1 87 p00 p<0. miR8743p/BDH pBDH miR8743pBDH 3pBDH1 3p/BDH (AC 8 p0 p<0 3pBDH p<

Resumo Fundamento A hipertrofia cardíaca patológica (HC) sustentada é um fator de risco independente para aumento da incidência e mortalidade de eventos cardiovasculares. Objetivos Esta pesquisa foi projetada para desvendar o papel do RNA não codificante longo (LncRNA) CCAT2 na progressão da HC. Métodos Procedimentos de constrição aórtica transversal (TAC) foram conduzidos para construir um modelo de HC in vivo induzido por sobrecarga de pressão. O tratamento com angiotensina II (Ang II) foi utilizado para induzir células hipertróficas de cardiomiócitos de rato H9c2. Resultados Os resultados in vivo mostraram que o silenciamento de CCAT2 reduziu a área de superfície dos cardiomiócitos, aliviou a fibrose cardíaca e diminuiu os níveis de β-MHC, ANP e BNP em modelos de camundongos HC. Os resultados in vitro revelaram que o knockdown de CCAT2 reduziu a área de superfície celular e atenuou os níveis de β-MHC, ANP e BNP em células hipertróficas H9c2. Além disso, o silenciamento de CCAT2 diminuiu os níveis de β-catenina ativa, GSK-3β fosforilada e genes alvo Wnt (c-Myc, ciclinaD1 e c-Jun) em camundongos HC e células H9c2 hipertróficas. É importante ressaltar que o tratamento com o ativador da via Wnt / β-catenina LiCl reverteu a supressão do knockdown de CCAT2 na área de superfície celular H9c2 e nos níveis de MHC, ANP e BNP. Conclusões Coletivamente, o silenciamento do CCAT2 desempenha um papel protetor contra a HC através da inativação da sinalização Wnt/β-catenina, o que sugere que o CCAT2 pode se tornar um alvo terapêutico promissor para o HC. (HC cardiovasculares LncRNA (LncRNA CCAT TAC (TAC pressão Ang Hc H c βMHC, βMHC β MHC β-MHC disso βcatenina catenina ativa GSK3β GSKβ GSK 3β cMyc, cMyc Myc, Myc (c-Myc ciclinaD cJun Jun c-Jun H9c Coletivamente Wnt/βcatenina, Wntβcatenina Wnt/β catenina, Wnt/β-catenina Wnt/βcatenina Wntβ

Abstract Background Sustained pathological cardiac hypertrophy (CH) is an independent risk factor for increased incidence and mortality of cardiovascular events. Objectives This research was designed to unravel the role of long non-coding RNA (LncRNA) CCAT2 in CH progression. Methods Transverse aortic constriction (TAC) procedures were conducted to construct a pressure overload-induced in vivo CH model. Angiotensin II (Ang II) treatment was utilized to induce hypertrophic rat cardiomyocyte H9c2 cells. Results In vivo results showed that silencing of CCAT2 reduced cardiomyocyte surface area, alleviated cardiac fibrosis, and decreased β-MHC, ANP, and BNP levels in CH mouse models. In vitro results revealed that CCAT2 knockdown reduced cell surface area and attenuated β-MHC, ANP, and BNP levels in hypertrophic H9c2 cells. Besides, CCAT2 silencing decreased the levels of active β-catenin, phosphorylated-GSK-3β, and Wnt target genes (c-Myc, cyclinD1, and c-Jun) in CH mice and hypertrophic H9c2 cells. Importantly, treatment with the Wnt/β-catenin pathway activator LiCl reversed the suppression of CCAT2 knockdown on H9c2 cell surface area and MHC, ANP, and BNP levels. Conclusions Collectively, CCAT2 silencing plays a protective role against CH through inactivating the Wnt/β-catenin signaling, which suggests that CCAT2 might become a promising therapeutic target for CH. (CH events noncoding non coding LncRNA (LncRNA CCAT progression TAC (TAC overloadinduced overload induced model Ang Hc H c H9c cells fibrosis βMHC, βMHC β MHC β-MHC ANP models Besides βcatenin, βcatenin catenin, catenin β-catenin phosphorylatedGSK3β, phosphorylatedGSK3β phosphorylatedGSKβ phosphorylated GSK 3β, 3β phosphorylated-GSK-3β cMyc, cMyc Myc, Myc (c-Myc cyclinD1 cyclinD cJun Jun c-Jun Importantly Wnt/βcatenin Wntβcatenin Wnt/β Collectively signaling phosphorylatedGSK Wntβ

RESUMO Introdução A elevada dificuldade e o alto risco são marcas inerentes da ginástica artística. Ela exige a coordenação motora de grandes grupos musculares. Objetivo Aprimorar o treinamento do controle da coordenação corporal dos ginastas. Métodos Selecionou-se aleatoriamente 16 ginastas como voluntários de pesquisa, divididos aleatoriamente em dois grupos de mesma distribuição etária. Após o período de treinamento, implementou-se um novo protocolo de treino e analisou-se o controle da coordenação motora após essa intervenção. Resultados Não houve diferença significativa em um índice comparativo unilateral (P>٠,٠٥). Houve diferenças estatisticamente significantes no índice de equilíbrio e no índice de velocidade na direção de ponto de toque rápido nos atletas (P<0,05). Em comparação com o grupo controle, houve uma diferença significativa no índice geral de teste entre os grupos experimental (P<0,01). A coordenação motora do grupo experimental teve uma melhora significativa. Conclusão As técnicas de batida cruzada, pedalada rápida, equilíbrio, pular corda, pegada com duas mãos, drible com duas mãos, chute e outras sete técnicas avaliadas demonstraram uma grande relação com a coordenação motora dos atletas de ginástica. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

ABSTRACT Introduction The great difficulty and the high risk are inherent marks of artistic gymnastics. It demands motor coordination of large muscle groups. Objective Improve the training of body coordination control in gymnasts. Methods We randomly selected 16 gymnasts as research volunteers, randomly divided into two groups of the same age distribution. After the training period, a new training protocol was implemented, and motor coordination control was analyzed after this intervention. Results There was no significant difference in a one-sided comparative index (P>0.05). There were statistically significant differences in the balance index and the speed index in the fast touch point direction in the athletes (P<0.01). Compared with the control group, there was a significant difference in the overall test index between the experimental groups (P<0.01). The motor coordination of the experimental group had a significant improvement. Conclusion The techniques of cross-striking, fast pedaling, balance, jump rope, two-handed grip, two-handed dribbling, kicking, and seven other techniques evaluated showed a great relationship with the motor coordination of gymnastics athletes. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMEN Introducción La alta dificultad y el alto riesgo son marcas inherentes a la gimnasia artística. Exige la coordinación motriz de grandes grupos musculares. Objetivo Mejorar el entrenamiento del control de la coordinación corporal en gimnastas. Métodos Se seleccionaron al azar 16 gimnastas como voluntarios de la investigación, divididos aleatoriamente en dos grupos con la misma distribución de edad. Tras el periodo de entrenamiento, se aplicó un nuevo protocolo de entrenamiento y se analizó el control de la coordinación motora después de esta intervención. Resultados No hubo diferencias significativas en el índice comparativo unilateral (P>0,05). Hubo diferencias estadísticamente significativas en el índice de equilibrio y en el índice de velocidad en la dirección del punto de contacto rápido en los atletas (P<0,01). En comparación con el grupo de control, hubo una diferencia significativa en el índice de prueba global entre los grupos experimentales (P<0,01). La coordinación motora del grupo experimental tuvo una mejora significativa. Conclusión Las técnicas de golpe cruzado, pedaleo rápido, equilibrio, salto de cuerda, agarre con dos manos, regate con dos manos, patada y otras siete técnicas evaluadas mostraron una gran relación con la coordinación motora de los atletas de gimnasia. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

RESUMO Introdução A inovação e o desenvolvimento de movimentos complexos na ginástica aeróbica apresentam maiores exigências para a qualificação geral das atletas. Objetivo Analisar o efeito do treinamento de força especial de membro inferior no desempenho das ginastas femininas. Métodos Foram selecionadas quatro ginastas femininas de faculdades de esportes como alvos de pesquisa. O método é discutido utilizando materiais de literatura, entrevistas com especialistas, métodos experimentais e estatísticas matemáticas. Resultados Houve uma diferença significativa no índice de composição corporal das atletas antes e depois do treinamento (P<0,05). A excepcional qualidade das atletas de ginástica aeróbica feminina após o treinamento especial de força dos membros inferiores foi significativamente elevada em comparação com aquela antes do treinamento. Conclusão O método de treinamento de força utilizado neste trabalho tem um efeito significativo na melhoria da força individual dos membros inferiores das ginastas femininas. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

ABSTRACT Introduction The innovation and development of complex movements in aerobic gymnastics present higher demands for the athletes’ overall qualifications. Objective Analyze the effect of special lower limb strength training on the performance of female gymnasts. Methods Four female gymnasts from sports colleges were selected as research targets. The method is discussed using literature materials, expert interviews, experimental methods, and mathematical statistics. Results There was a significant difference in the athletes’ body composition index before and after training (P<0.05). The exceptional quality of female aerobic gymnastics athletes after the special lower limb strength training was significantly elevated compared with that before the training. Conclusion The strength training method used in this paper significantly improves the individual strength of the lower limbs of female gymnasts. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMEN Introducción La innovación y el desarrollo de movimientos complejos en la gimnasia aeróbica presentan mayores exigencias para la cualificación general de los atletas. Objetivo Analizar el efecto del entrenamiento especial de la fuerza de los miembros inferiores en el rendimiento de las gimnastas. Métodos Se seleccionaron cuatro gimnastas de escuelas deportivas como objetivos de la investigación. El método se discute utilizando material bibliográfico, entrevistas a expertos, métodos experimentales y estadísticas matemáticas. Resultados Hubo una diferencia significativa en el índice de composición corporal de los atletas antes y después del entrenamiento (P<0,05). La calidad excepcional de las atletas de gimnasia aeróbica tras el entrenamiento especial de fuerza de las extremidades inferiores fue significativamente mayor que antes del entrenamiento. Conclusión El método de entrenamiento de fuerza utilizado en este trabajo tiene un efecto significativo en la mejora de la fuerza individual de las extremidades inferiores de las gimnastas. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

Abstract Background: Diabetic kidney disease (DKD) is a serious microvascular complication of diabetes that affects both type 1 and type 2 diabetes patients at a high incidence rate. Naja Naja atra venom (NNAV) has been shown to have protective effects and improved renal function in diabetic rats. However, its mechanism of action is still unclear. This study aims to unravel the effectiveness and mechanisms of NNAV on DKD. Methods: We conducted in vitro experiments in which Human Kidney-2 (HK-2) cells were stimulated with high glucose, and exposed to varying concentrations of NNAV. Cell morphology, as well as α-SMA, TGF-β1, and E-cadherin levels, were analyzed using immunofluorescence and western blot. In vivo experiments involved a diabetic rat model, where varying concentrations of cobra α-neurotoxin (CTX) were administrated via gastric treatment. We observed and noted pathomorphological changes, measured biochemical and oxidative stress indices, and used western blot to assess podocin and nephrin levels. Results: High glucose levels can induce a decrease in E-cadherin expression and an increase in α-SMA and transforming growth factor-β1 (TGF-β1) expression in HK-2 cells. NNAV can inhibit the transdifferentiation of HK-2 cells to myofibroblast (MyoF) in a high glucose environment and reduce the expression of TGF-β1. Cobra α-neurotoxin (CTX) can reduce urine protein in diabetes model rats at an early stage, which is dose-independent and has a time application range. CTX can regulate the expression of nephrin and podocin. Conclusion: The present study indicates that CTX and NNAV attenuate STZ and high glucose-induced DKD. Its mechanisms of action are associated with inhibiting oxidative stress and TEMT. The study suggests that NNAV and CTX might be a potential therapeutic drug for treating DKD. Background DKD (DKD rate (NNAV However unclear Methods Kidney2 Kidney Kidney- HK2 HK (HK-2 morphology αSMA, αSMA α SMA, SMA TGFβ1, TGFβ1 TGFβ TGF β1, β1 β TGF-β1 Ecadherin E cadherin αneurotoxin neurotoxin (CTX treatment changes indices Results factorβ1 factorβ factor factor-β (TGF-β1 HK- MyoF (MyoF TGFβ1. β1. stage doseindependent dose independent range Conclusion glucoseinduced induced TEMT (HK- TGF-β (TGF-β (HK

ABSTRACT This study evaluated the changes in rumen microbiome during the process of age development of farming Guizhou White goats from Southwest China. We conducted high-throughput 16S rRNA gene sequencing to investigate the diversity, structure, and composition of goat rumen microbiota (RM) of 21 goats of different age groups (1, 6, and 12 months). We found that volatile fatty acids (i.e., acetate, propionate, and butyrate) fermented by microbes were found to increase significantly in the six- and one-month-old goats. Results of the genera abundance analysis showed that abundance of eight and seven taxa decreased in six- and one-month-old goats, respectively, compared with that in 12-month-old goats. Additionally, differences in six taxa in six-month-old goats and in one taxon in one-month-old goats were found. In addition, specific gut microbiome was found, which was significantly correlated with rumen fermentation parameters in Guizhou White goats. These results revealed the signature microbiota in RM during various developmental stages in goats raised in Southwest China and can also provide a guiding tool for evaluating rumen health of ruminants worldwide.

ABSTRACT In this study, the numerical simulations are used to simulate the discharge process of a 10-cm dual-stage 4-grid (DS4G) radiofrequency (RF) ion thruster, and the effects of RF coil turns, and electrical parameters on the key plasma parameters inside the discharge chamber are investigated. Thus, the optimal design parameters of the thruster are obtained. The RF plasma source has high inductive coupling discharge efficiency when the number of coil turns is six and the operational frequency is 2 MHz. The simulation results show that increasing the number of coil turns can effectively increase the plasma density inside the discharge chamber, and thus improve the efficiency of the inductive coupling discharge, and the operational frequency affects the characteristics of the inductively coupled discharge by influencing the depth of the plasma skinning layer. An experimental thruster demonstrated the 10-cm DS4G RF ion thruster can achieve super high specific impulse. The correctness of the simulation model is verified by the experiment results of the thruster, which will be useful for the optimal design of radio ion thruster or ion sources.

RESUMEN Los recientes avances en Microbiología y la aplicación de la Biología Molecular, han logrado un mejoramiento, complementario para las técnicas clásicas de identificación bacteriana, también esto, ha permitido el descubrimiento o reagrupamiento de géneros y especies bacterianas, las cuales no habían logrado ser identificadas correctamente, por métodos bioquímicos y fenotípicos, ya que estos, presentan algunas limitaciones como falsos positivos o baja especificidad. Dentro de las herramientas moleculares empleadas para la identificación del género Aeromonas, se han empleado la secuenciación a partir de gen 16S de rRNA, genes de mantenimiento como rpoD o gyrB, los cuales poseen mayor poder de discriminación que el gen 16S de rRNA, la técnica de RFLP-PCR y más recientemente se ha incorporado el uso de la técnica de Espectrofotometría de Masas por Matriz Asistida por laser Desorción-Ionización de Tiempo de Vuelo (MALDI-TOF MS), la cual es capaz de identificar especies de Aeromonas de forma precisa y rápidamente. Lo cual es de gran importancia, debido al incremento de evidencias que indican que estas especies se encuentran ampliamente distribuidas en el ambiente y que pueden causar una variedad de infecciones en animales y en el humano.

ABSTRACT Advances in Microbiology and applications of Molecular Biology, has been achieved an improvement, complementary to the classical techniques of bacterial identification, this also allowed the discovery or regrouping of bacterial genera and species, which had failed to be identified correctly, by biochemical and phenotypic methods, since these have some limitations such as false positives or low specificity. Among the molecular tools used to identify the Aeromonas genus, we have used sequencing from 16S rRNA gene, maintenance genes such as rpoD or gyrB, which have a greater discrimination power than the 16S rRNA gene, the technique of RFLP-PCR and more recently the technique of Mass Spectrometry has been increased by Laser Assisted Matrix Desorption-Ionization of Flight Time (MALDI-TOF MS) which is able to identify species of Aeromonas quickly and accurately. This is important because of the increasing evidence that these species are widely distributed in the environment and can cause a variety of infections in animals and humans.

ABSTRACT Odanacatib (ODN) is a selective inhibitor of cathepsin K. The cysteine protease cathepsin K has been implicated in cardiac hypertrophy. Resistine is an adipokine which is identified to promote cardiac hypertrophy. Here, we hypothesize that ODN mitigates resistin-induced myocyte hypertrophy. Cell surface area and protein synthesis were measured after treatment with resistin and ODN in H9c2 cells. The expression of cardiomyocyte hypertrophy marker BNP and β-MHC was detected by RT-qPCR. The expression and phosphorylation of AMPK and LKB1 were analyzed with Western blot. Resistin could significantly increase cardiomyocyte cell surface area, protein synthesis, and embryonic gene BNP and β-MHC expression, inhibit phosphorylation of AMPK and LKB1. ODN could significantly reverse the effects of resistin. Collectively, our data suggest that ODN can inhibit cardiomyocyte hypertrophy induced by resistin and the underlying mechanism may be involved in LKB1/AMPK pathway.

Background: 3-Ketosteroid-∆¹-dehydrogenase (KSDD), a flavoprotein enzyme, catalyzes the bioconversion of 4-androstene-3,17-dione (AD) to androst-1,4-diene-3,17-dione (ADD). To date, there has been no report about characterization of KSDD from Mycobacterium neoaurum strains, which were usually employed to produce AD or ADD by fermentation. Results: In this work, Corynebacterium crenatum was chosen asa new host for heterologous expression of KSDD from M. neoaurum JC-12 after codon optimization of the KSDD gene. SDS-PAGE and western blotting results indicated that the recombinant C. crenatum harboring the optimized ksdd (ksdd n) gene showed significantly improved ability to express KSDD. The expression level of KSDD was about 1.6-fold increased C. crenatum after codon optimization. After purification of the protein, we first characterized KSDD from M. neoaurum JC-12, and the results showed that the optimum temperature and pH for KSDD activity were 30°C and pH 7.0, respectively. The Km and Vmax values of purified KSDD were 8.91 µM and 6.43 mM/min. In this work, C. crenatum as a novel whole-cell catalyst was also employed and validated for bioconversion of AD to ADD. The highest transformation rate of AD to ADD by recombinant C. crenatum was about 83.87% after 10 h reaction time, which was more efficient than M. neoaurum JC-12 (only 3.56% at 10 h). Conclusions: In this work, basing on the codon optimization, overexpression, purification and characterization of KSDD, we constructed a novel system, the recombinant C. crenatum SYPA 5-5 expressing KSDD, to accumulate ADDfromADefficiently. This work provided new insights into strengthening sterol catabolism by overexpressing the key enzyme KSDD, for efficient ADD production.

We investigated inheritance and carried out gene fine mapping of aroma in crosses between the aromatic elite hybrid rice Oryza sativa indica variety Chuanxiang-29B (Ch-29B) and the non-aromatic rice O. sativa indica variety R2 and O. sativa japonica Lemont (Le). The F1 grains and leaves were non-aromatic while the F2 non-aroma to aroma segregation pattern was 3:1. The F3 segregation ratio was consistent with the expected 1:2:1 for a single recessive aroma gene in Ch-29B. Linkage analysis between simple sequence repeat (SSR) markers and the aroma locus for the aromatic F2 plants mapped the Ch-29B aroma gene to a chromosome 8 region flanked by SSR markers RM23120 at 0.52 cM and RM3459 at 1.23 cM, a replicate F2 population confirming these results. Three bacterial artificial chromosome (BAC) clones cover chromosome 8 markers RM23120 and RM3459. Our molecular mapping data from the two populations indicated that the aroma locus occurs in a 142.85 kb interval on BAC clones AP005301 or AP005537, implying that it might be the same gene reported by Bradbury et al (2005a; Plant Biotec J. 3:363-370). The flanking markers Aro7, RM23120 and RM3459 identified by us could greatly accelerate the efficiency and precision of aromatic rice breeding programs.