• Title/Summary/Keyword: 23 kDa protein

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Effect of feed restriction on the maintenance energy requirement of broiler breeders

  • da Silva Teofilo, Guilherme Ferreira;Lizana, Rony Riveros;de Souza Camargos, Rosiane;Leme, Bruno Balbino;Morillo, Freddy Alexander Horna;Silva, Raully Lucas;Fernandes, Joao Batista Kochenborger;Sakomura, Nilva Kazue
    • Animal Bioscience
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    • v.35 no.5
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    • pp.690-697
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    • 2022
  • Objective: This study aimed to evaluate the effect of the ad libitum and restricted feeding regimen on fasting heat production (FHP) and body composition. Methods: Twelve Hubbard broilers breeders were selected with the same body weight and submitted in two feeding regimes: Restricted (T1) with feed intake of 150 g/bird/d and ad libitum (T2). The birds were randomly distributed on the treatments in two runs with three replications per treatment (per run). The birds were adapted to the feed regimens for ten days. After that, they were allocated in the open-circuit chambers and kept for three days for adaptation. On the last day, oxygen consumption (VO2) and carbon dioxide production (VCO2) were measured by 30 h under fasting. The respiratory quotient (RQ) was calculated as the VCO2/VO2 ratio, and the heat production (HP) was obtained using the Brower equation (1985). The FHP was estimated throughout the plateau of HP 12 hours after the feed deprivation. The body composition was analyzed by dual-energy X-ray absorptiometry scanning at the end of each period. Data were analyzed for one-way analysis of variance using the Minitab software. Results: The daily feed intake was 30 g higher to T2 (p<0.01) than the T1. Also, the birds of the T2 had significatively (p<0.05) more oxygen consumption (+3.1 L/kg0.75/d) and CO2 production (+2.2 L/kg0.75/d). That resulted in a higher FHP 359±14 kJ/kg0.75/d for T2 than T1 296±17.23 kJ/kg0.75/d. In contrast, the RQ was not different between treatments, with an average of 0.77 for the fasting condition. In addition, protein and fat composition were not affected by the treatment, while a tendency (p<0.1) was shown to higher bone mineral content on the T1. Conclusion: The birds under ad libitum feeding had a higher maintenance energy requirement but their body composition was not affected compared to restricted feeding.

Isolation and characterization of sigH from Corynebacterium glutamicum (Corynebacterium glutamicum의 sigH 유전자의 분리 및 기능분석)

  • Kim Tae-Hyun;Kim Hyung-Joon;Park Joon-Sung;Kim Younhee;Lee Heung-Shick
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.99-104
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    • 2005
  • Corynebacterial clones which exert regulatory effects on the expression of the glyoxylate bypass genes were isolated using a reporter plasmid carrying the enteric lacZ fused to the aceB promoter of Corynebacterium glutamicum. Some clones carried common fragments as turned out by DNA mapping technique. Subcloning analysis followed by the measurement of $\beta-galactosidase$ activity in Escherichia coli identified the region responsible for the aceB-repressing activity. Sequence analysis of the DNA fragment identified two independent ORFs of ORF1 and ORF2. Among them, ORF2 was turned out to be responsible for the aceB-repressing activity. ORF1 encoded a 23,216 Da protein composed of 206 amino acids. Sequence similarity search indicated that the ORF may encode a ECF-type $\sigma$ factor and designated sigH. To identify the function of sigH, C. glutamicum sigH mutant was constructed by gene disruption technique and the sigH mutant showed growth retardation as compared to the wild type strain. In addition, the mutant strain showed sensitivity to oxidative-stress generating agent plumbagin. This result imply that sigH is probably involved in the stress response occurring during normal cell growth.

Differentiation of Human Embryonic Stem Cells into Germ Cell and Culture Condition for Single Embryonic Stem Cells Dissociated by Enzyme (인간 배아줄기세포의 생식세포로의 분화 및 효소에 의해 분리된 단일줄기세포 배양조건)

  • Chi, Hee-Jun;Choi, Soon-Young;Chung, Da-Yeon
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.13-23
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    • 2010
  • Objective: The present study was carried out to induce differentiation of human embryonic stem cells (hESCs) into germ cells and to establish a culture condition for single hESCs dissociated by enzyme. Methods: Embryonic body (EB) was formed by hanging drop culture for 3 days from hESCs colony. The EBs were cultured in the medium supplemented with retionic acid (RA) or/and bone morphogenetic protein-4 (BMP4) for 14 days to differentiate into germ cells. Germ cell specific markers, c-kit and VASA were used for immunohistochemistry of EB. Human ESCs colonies were dissociated into single cells by Collagenase, Tryple and Accutase, and then colony formation rate of the single cells was examined. Rho-associated kinase inhibitor (ROCK inhibitor, Y27632) was added into the culture medium of single cells to reduce the apoptotic damage during the dissociation. Results: Single cells dissociated with Tryple or Accutase showed higher colony formation rates compared to the cells dissociated with Collagenase. Seeding of $5{\times}10^3$ cells/well (4 well dish) was efficient to obtain high colony formation rate compared to other concentrations of seeding cell. Addition of Y27632 significantly increased the colony formation rate of the single cells dissociated by Tryple. Immunohistochemistry of EB with c-kit and VASA markers showed a weak fluorescence signals compared to the signals from the testicular tissue. Conclusion: Dissociation with Tryple was useful to obtain healthy single cells and addition of Y27632 was beneficial for survival and colony formation of the single cells. Unlike other studies, we just observed a dim fluorescence staining of the germ cell markers, probably caused by the short-term culture for the differentiation of EB compared to other studies.

Composition and Physicochemical Properties of Unripe Korean Peaches According to Cultivars (국내산 복숭아 유과의 품종별 성분 분석 및 품질특성)

  • Kim, Da-Mi;Kim, Kyung-Hee;Choi, In-Ja;Yook, Hong-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.2
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    • pp.221-226
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    • 2012
  • For the investigation of a possibility as a useful functional material, 6 cultivars (Takinosawa Gold, Kawanakawase Hakuto, Madoka, Yumefuji, Nagasawa Hakuho, Hong Bak) of Prunus persica L. Batsch were studied at unripe stage to determine the physicochemical properties and chemical compositions. The cultivars were picked in late May, and all samples were analyzed for external properties, physicochemical properties, pH, Brix value, Hunter's color value, hardness, vitamin C, and reducing sugar. The size of the fruit from all six cultivars was compared, and it was determined that cultivars, fruit from Madoka was the largest, while that from Yumefuji was the smallest. Comparing fresh weight, the fruit from Yumefuji was lowest in moisture contents (89.13~89.96%), and that from Nagasawa Hakuho had significantly higher crude protein (1.02~1.62%). The contents of crude lipids (0.18~0.23%) and carbohydrates (8.00~9.35%) were not significantly different between cultivars and Madoka included higher crude ash contents (0.32~0.69%) than other cultivars. The pH of 6 cultivars from unripe peaches were significantly higher from Kawanakawase Hakuto, and the Brix value was also highest from Kawanakawase Hakuto, followed by Yumefuji, Madoka, Nagasawa Hakuho, Takinosawa Gold, and Hong Bak. In chromaticity, the L value, the indicator of brightness, was significantly higher in fruit from Nagasawa Hakuho. The a value, the indicator of redness, was the highest with Hong Bak and overall lower than -5. The b value, the indicator of yellowness, was the highest in fruit from Madoka and ranged from 16.51 to 18.33. In physical characteristics, the hardness of the unripe peaches was the highest in fruit from Hong Bak, and overall, white peaches have a higher hardness value than yellow peaches. The vitamin C content of the fruit didn't show any significant differences between cultivars, and the reducing sugar showed a higher percent than 6.34% in fruit from all cultivars. These results suggest that unripe peaches were commensurate with the development of natural pigment and as a functional foods.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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