Seo, Dong Won;Park, Hee Bok;Choi, Nu Ri;Jin, Shil;Yoo, Chae Kyoung;Sultana, Hasina;Heo, Kang Nyeong;Jo, Cheorun;Lee, Jun Heon
Korean Journal of Poultry Science
/
v.42
no.1
/
pp.77-86
/
2015
Chicken is one of the major livestock, especially for supplying proteins to human. The chicken genome size is approximately one-third compared with that of the human genome and regarded as a valuable model animal for genetics and development biology. In this study, we constructed the genetic linkage map for Korean native chicken (KNC) using 131 microsatellite (MS) and 8 single nucleotide polymorphism (SNP) markers. As a result, the total map length was calculated as 2729.4 cM and the average genetic distance between markers was 19.64 cM. The marker orders and genetic distances were well matched with the consensus linkage map except for the physical order of ADL0278 and MCW0351 in GGA8. In addition, the recombination rates in marcrochromosomes were 3.7 times higher than that of microchromosomes. The average numbers of alleles, expected heterozygosity (Hexp) and polymorphic information content (PIC) values were calculated as 5.5, 0.63 and 0.58, respectively. These results will give useful information for the understanding of genetic structure and QTL studies in KNC.
Our previous studies demonstrated that relaxin in concert with estrogen promotes development of the mammary parenchyma during the last third of gestation in gilts, and the specific relaxin-binding sites were present in the mammary gland. This study was conducted to determine if relaxin-binding sites in the mammary gland were functional relaxin receptors. Three cycling cross-bred gilts were bilaterally ovariectomized on day 0 of the experiment. Beginning on day 15 and continuing through day 29 post-surgery, the gilt received an im. injection of estradiol benzoate at 12-hr intervals. Beginning on day 22 post-surgery, highly purified porcine relaxin was administered(lug/hr) into the left fourth mammary gland from the anterior end via miniature osmotic pump. Physiological saline was administered to the right fourth mammary gland. The gilt was sacrificed on day 29 post-surgery and histological characteristics of the mammary parenchyma were examined. The mammary glands treated locally with saline showed little, if any, lobulo-alveolar development, whereas the mammary glands treated with relaxin showed not only marked lobulo-alveolar development but also prominent secretions in the alveoli. The saline-treated glands were characterized by relatively dense and highly organized collagen fiber bundles. Whereas, in the relaxin-treated mammary glands, collagen fiber bundles were dispersed and loosely organized. In conclusion, relaxin-binding sites in the mammary gland are functional relaxin receptors and relaxin acts directly on the pig mammary gland to promote development of the alveoli and remodeling of the extracellular matrix.
The cDNA cloning and developmental profiles of the mRNA for A. pernyi arylphorin was determined. The complete A. pernyi arylphorin cDNA sequence comprised 2,234 bp (without the poly $A^+$ tail), including an open reading frame of 2,112 bp beginning with a methionine ATG at bp34. The A. pernyi arylphorin contained 704 amino acids which are highly enriched in aromatic amino acids, phenylalanine and tyrosine. The calculated molecular mass of the A. pernyi arylphorin from the ORF was 83,439 Da. The deduced amino acid sequence of A. pernyi arylphorin showed 78, 71, 62 and 64% identity with those of H. cecropia, M. sexta $\alpha$ subunit, M. sexta $\beta$ subunit and B. mori storage protein. In Northern blot analysis, the A. pernyi arylphorin mRNA only in the fat body of the 5th instar larvae was responsible for gene expression of the protein, and the synthetic activity of the mRNA was detected strongly in the early larvae, but not in the middle or late-stage larvae. In addition, a very weak signal in mRNA activity was detected in pupal stages, but this was considered to be inactive mRNA after reviewing the results of the labeling experiment of this protein.
Microbial lethal value and nutrient retention of sous vide processed spinach were evaluated with mathematical model prediction and experimental trial for different package sizes and pasteurization temperatures. The package size covers 500 g, 1 kg and 2 kg, while the pasteurization temperature includes 80, 90 and 97$^{\circ}C$. The basic process scheme consists of filling blanched spinach into barrier plastic film pouch, sealing under vacuum, pasteurization in hot water with over pressure and final cooling to 3$^{\circ}C$. Pasteurization condition was designed based on attainment of 6 decimal inactivation of Listeria monocytogenes at geometric center of the pouch package by heating cycle, which was determined by general method. Heat penetration property of the package and thermal destruction kinetics were combined to estimate the retention of ascorbic acid and chlorophyll. Smaller packages with shorter pasteurization time gave better nutrient retention, physical and chemical qualities. Larger package size was estimated and confirmed experimentally to give higher pasteurization value at center, lower ascorbic acid and chlorophyll contents caused by longer heat process time. Lower pasteurization temperature with longer process time was predicted to give lower pasteurization value at center and lower ascorbic acid, while chlorophyll content was affected little by the temperature. Experimental trial showed better retention of ascorbic acid and chlorophyll for smaller package and higher pasteurization temperature with shorter heating time. The beneficial effect of smaller package and higher pasteurization temperature was also observed in texture, color retention and drip production.
Purpose: North Koreans could be at higher risk for their bone health because of previous periods of severe famine and the continuing low availability of food. This study determined the bone mineral density (BMD) status and its relationship with dietary behaviors and nutrient intake of North Korean refugees (NKR) in South Korea (SK). Methods: This cross-sectional study analyzed 110 female NKR from a NORNS cohort of a non-probability sample of adult NKR in Seoul. BMD examined by DEXA was used to divide participants into the normal group (NG) and the non-normal group (NNG) according to the WHO guideline. A self-administered questionnaire included questions on age, the socioeconomic situation in North Korea (NK) and SK, the food security in NK and SK, and the health behaviors, dietary behaviors, and food frequency questionnaire administered in SK. A one-day 24-hr recall was conducted and the results were analyzed by using CanPro. SPSS was used to analyze whether BMD and related dietary behaviors and nutrient intakes differed according to the groups. Results: NG (62.7%) was significantly younger and had a lower abdominal obesity score than NNG (p < 0.001). While 14.5% of NG reported experiencing menopause, all of NNG reported experiencing menopause. The NG more frequently consumed the dairy group of foods (9.6 times a week) than did the NNG (4.8 times a week) after the statistics were adjusted for age (p < 0.007). The NG consumed significantly more animal protein and animal calcium than did the NNG (p = 0.01, p = 0.009, respectively). Calcium intake was low with 49.3% of NG, and 78.0% of the NNG reported consuming calcium lower than the estimated average requirement. Only calcium showed an index of nutrient quality lower than one in both groups. Conclusion: These results showed that NKR women and possibly all North Korean women are at high risk for bone health and they consumed low levels of bone-related nutrients, and this should be considered for the nutrition policy for NKR and North Korea.
Liver regeneration is a result of highly coordinated proliferation of hepatocytes and non-parenchymal liver cells. At this process, induction of metallothionein (MT), which is low molecular and cysteine rich, has been reported. The present study was carried to find the ultrastructure of hepatocytes and determine the expression of MT in regenerating rat liver after partial hepatectomy. As a result, the remnant liver after PH grew fast from 1 day until 7 days. Various changes were morphologically observed. Disintegration of cell plates and liver lobule appeared shortly after PH. And hepatocytes showed the rapid proliferation, characterized by high nuclear cytoplasmic ratio, weak intercellular junctional complexes, chromatin condensation, increase of ribosomes and mitochondria, and temporary increase of lipid droplets. Finally, remodeling of the liver lobule was completed through the rearrangement of blood vessels and cell plates by 7 days after PH. On histochemistry, immunoreactivity indicating the presence of MT appeared moderately throughout the cytoplasm of control rat hepatocyte. After PH, positive reactions for MT increased at the cytoplasm and the nucleus. These results suggest that the remnant liver cells immediately entered cell proliferation and increase of MT expression after PH. It is thought that MT protein might be associated with transfer of some factors needed to cell division from the cytoplasm to the nucleus for regeneration of the liver after PH.
The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).
Background : The p53 gene codes for a DNA-binding nuclear phosphoprotein that appears to inhibit the progression of cells from the G1 to the S phase of the cell cycle. Mutations of the p53 gene are common in a wide variety of human cancers, including lung cancer. In lung cancers, point mutations of the p53 gene have been found in all histological types including approximately 45% of resected NSCLC and even more frequently in SCLC specimens. Mutant forms of the p53 protein have transforming activity and interfere with the cell-cycle regulatory function of the wild-type protein. The majority of p53 gene mutations produce proteins with altered conformation and prolonged half life; these mutant proteins accumulate in the cell nucleus and can be detected by immunohistochemical staining. But protein overexpression has been reported in the absence of mutation. p53 protein overexpression or gene mutation is reported poor prognostic factor in breast cancer, but in lung cancer, its prognostic significance is controversial. Method : We investigated the p53 abnormalities by nucleotide sequencing, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP), and immunohistochemical staining. We correlated these results with each other and survival in 75 patients with NSCLC resected with curative intent. Overexpression of the p53 protein was studied immunohistochemically in archival paraffin- embedded tumor samples using the D07(Novocastra, U.K.) antibody. Overexpression of p53 protein was defined by the nuclear staining of greater than 25% immunopositive cells in tumors. Detection of p53 gene mutation was done by PCR-SSCP and nucleotide sequencing from the exon 5-9 of p53 gene. Result: 1) Of the 75 patients, 36%(27/75) showed p53 overexpression by immunohistochemical stain. There was no survival difference between positive and negative p53 immunostaining(overall median survival of 26 months, disease free median survival of 13 months in both groups). 2) By PCR-SSCP, 27.6%(16/58) of the patients showed mobility shift. There was no significant difference in survival according to mobility shift(overall median survival of 27 in patients without mobility shift vs 20 months in patients with mobility shift, disease free median survival of 8 months vs 10 months respectively). 3) Nucleotide sequence was analysed from 29 patients, and 34.5%(10/29) had mutant p53 sequence. Patients with the presence of gene mutations showed tendency to shortened survival compared with the patients with no mutation(overall median survival of 22 vs 27 months, disease free median survival of 10 vs 20 months), but there was no statistical significance. 4) The sensitivity and specificity of immunostain based on PCR-SSCP was 67.0%, 74.0%, and that of the PCR-SSCP based on the nucleotide sequencing was 91.8%, 96.2% respectively. The concordance rate between the immunostain and PCR-SSCP was 62.5%, and the rate between the PCR-SSCP and nucleotide sequencing was 95.3%. Conclusion : In terms of detection of p53 gene mutation, PCR-SSCP was superior to immunostaining. p53 gene abnormalities either overexpression or mutation were not a significant prognostic factor in NSCLC patients resected with curative intent. However, patients with the mutated p53 gene showed the trends of early relapse.
Acellular dermal matrix (ADM), produced by decellularization from human cadaveric skin, has been used for various biomedical applications. A manufacturing process for ADM ($SureDerm^{TM}$) using tri-n-butyl phospahate (TnBP) and deoxycholic acids as the decellularization solution has been developed. The manufacturing process for $SureDerm^{TM}$ has 70% ethanol treatment and ethylene oxide gas sterilization for inactivating infectious microorganisms. The purpose of this study was to examine the efficacy of the 70% ethanol treatment, decellularization process using 0.1% TnBP and 2% deoxycholic acids, and EO gas sterilization process in the inactivation of viruses. A variety of experimental model viruses for human pathogens, including the human immunodeficiency virus type 1 (HIV-1), bovine herpes virus (BHV), bovine viral diarrhoea virus (BVDV), hepatitis A virus (HAV), and porcine parvovirus (PPV) were all selected for this study. Enveloped viruses such as HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by 70% ethanol treatment. However HAV and PPV showed high resistance to 70% ethanol treatment with the log reduction factors of 1.85 and 1.15, respectively. HIV-1, BHV, and BVDV were effectively inactivated to undetectable levels by decellularization process. All the viruses tested were completely inactivated to undetectable levels by EO gas treatment. The cumulative log reduction factors of HIV-1, BHV, BVDV, HAV, and PPV were $\geq12.71$, $\geq18.08$, $\geq14.92$, $\geq6.57$, and $\geq7.18$, respectively. These results indicate that the production process for $SureDerm^{TM}$ has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.
The vesicle system of DPPC(dipalmitoylphosphaticylcholine)/Chol(Cholesterol) has been modified by incorporating various mole fractions of flourinated surfactant($C_8F_{17}(CH_2)_2OCO-CH_2CH(SO_3Na)COO(CH_2)_2C_8F_{17}$. Sodium bis(1H,1H,2H,2H-heptadecaflurododecyl)-2-sulfosuccinate, FS)/fluorinated fatty acid salt ($C_7F_{15}COONH_4$, ammoniumpentadecaflurooctyrate, FFS), and their physicochemical properties have been investigated in an attempt to enhance the stability of phospholipid vesicle system. The ${\zeta}$-potential measurement by use of Zetamaster sub-micron Particle Electrophoresis Analyzer (Malvern Co.) showed that a charged homogeneous DPPC/Chol/FS vesicle has been formed owing to the incorporated FFS effect on the membrane, playing a role as a cosurfactant in the bilayer between DPPC and FS components. With increase in the concentration of FFS, it was found that the particle size and also surface charge of the DPPC/Chol/FS vesicle decreased. The stability of DPPC/Chol/FS/FFS liposome was found to be enhanced significantly compared to that of DPPC/Chol/FS according to the dispersity change as a function of time. The release rate of dye molecule of Methylene Blue from the DPPC/Chol/FS/FFS vesicle was determined to be slower than that of DPPC/Chol/FS system, and it may be attributed to the increase in microviscosity of the hydrophobic region in the bilayer. The affinfinity of DPPC/Chol/FS/FFS vesicles to albumin was found to be slightly lowered compared to that of DPPC/Chol/FS. Based on these findings, it was confirmed that a more stable and homogeneous vesicle system of DPPC/Chol/FS could be prepared by addition of FFS, acting as a cosurfactant in the aggregate formation.
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