Hong In-Hee;Lee Jun-Hwa;Go Cheol-Woo;Kwak Jung-Sik;Koo Ja-Hoon
Childhood Kidney Diseases
/
v.3
no.2
/
pp.153-160
/
1999
Purpose : Present study was undertaken to find out significance of clinical presentation, initial laboratory data and renal biopsy findings on subsequent clinical course of IgA nephropathy in children. Methods : Clinical and laboratory data were analysed retrospectively from 60 children who have been admitted to the Pediatric Department of Kyungpook National University Hospital for the past 11 years and diagnosed as IgA nephropathy. Renal biopsy findings were graded according to the pathologic subclass proposed by Haas. Results : Pathologic grading according to Haas subclassification showed 10 cases in subclass I, 36 in II, 12 in IV and 2 in V and none in subclass II. Sex distribution showed male predominance (male to female ratio = 3 : 1) and mean age at onset of disease was $10.4{\pm}2.8$ years. Episodes of gross hematuria was seen in 71.7% and IgA level increased in 28.3% of children and these were not associated with pathologic grading nor clinical outcomes. With increasing subclass grading, serum protein and albumin decreased and 24 hours urinary protein excretion increased. Normalization of urinalysis (disappearance of hematuria) was seen in 14% at 1-2 years and 37.1% at 3-4 years of follow up period. In 3 cases, renal function deteriorated progressively and they belonged one each to the Haas subclass III, IV and V. Conclusion : In children with IgA nephropathy, progression to chronic renal failure appears to be quite high and pathologic grading according to Haas' subclassification seems to predict patient's outcome faily well. However, firm conclusion cannot be drawn from present study due to the small numbers of patients and short follow-up period. Therefore further multicenter study involving larger numbers of patients and longer periods of follow-up over 10 years was to be undertaken.
The crushed fruiting body of Alaskan Porodaedalea pini (Brot.) Murrill (syn. Phellinus pini) was extracted in boiling water for 4 h with the yield of 20.5% in dry mass. This hot-water extract showed significant antiviral activity by inhibiting the plaque formation in HeLa cells by coxackievirus B3 (CVB3) and also showed highest inhibitory effect against neuraminidase activity among water extracts of various mushrooms. From the water extract, the ethanol precipitate (EP) and supernatant fraction (ES) were obtained through 75% ethanol precipitation with the yield of 43.3% and 28.3% in dry mass, respectively. Whereas ES did not show any detectable level of antiviral activity, EP showed significant dose-dependent inhibition of plaque formation by CVB3 in HeLa cells with an $EC_{50}$ (50% effective concentration) of 0.45 mg/mL. The cytotoxicity on HeLa cells by EP was relatively low with the $CC_{50}$ (50% cytotoxic concentration) of 2.25 mg/mL. EP also effectively inhibited neuraminidase activity in a dose-dependent manner showing up to 75% inhibition at 1.7 mg/mL. These results suggest that the hot-water extract and its EP of P. pini fruiting body can be a candidate for the development of a potent broad-range antiviral agent against influenza virus(Flu) as well as CVB3. The major active component of EP was shown to be a heteropolysaccharide-protein complex containing glucose as the main sugar residue with mole percentage of 79.8% and other sugars like galactose (19.2%), xylose (17%), mannose (5.8%), and fucose (4.6%) and a small portion (12.7%, in mass) of protein.
Enzyme substitution with recombinant phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is currently being explored for treatment of phenylketonuria (PKU), an autosomal recessive genetic disorder with mutations of the gene encoding phenylalanine-4-hydroxylase (EC 1.14.16.1). However, oral administration of PAL is limited because of proteolytic digestion in the gastrointestinal tract. The aim of this study was to determine the biochemical properties of PAL and delinate the susceptibility of wild-type PAL to pancreatic proteolysis by exploring several mutants, and to develop therapeutic drugs with PAL for PKU. The specific activity of PAL was assayed and its optimal pH, temperature stability, and intestinal protease susceptibility were investigated. Its $V_{max}$ values for phenylalanine and tyrosine were 1.77 and $0.47{\mu}mol$/ min/mg protein, respectively, and its $K_m$ values were $4.77{\times}10^{-4}$ and $4.37{\times}10^{-4}\;M$, respectively. PAL showed an optimal pH at 8.5, corresponding to the average pH range of the small intestine. It showed no loss of activity at $-80^{\circ}C$ for 5 months and possessed 93.4% of its activity under $4^{\circ}C$ for 4 wks. PAL was susceptible to chymotrypsin digestion and, to a lesser extent, to trypsin, elastase, carboxypeptidase A, and B. The trypsin and chymotrypsin cleaving sites were mutated to investigate protection from pancreatic digestion and the specific activities of these mutants were evaluated. The six mutants displayed low specific activities compared to the wild-type, suggesting that the primary trypsin and chymotrypsin cleaving sites may be essential for catalytic reaction. The PAL mutants could therefore be applied as a pretreatment modality without susceptibility to proteolytic attack, however, additional modification for enhancing enzymatic activity is needed to reduce the Phe levels effectively.
Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.
Liposomes having particle size from several tens to hundreds nanometers are efficient carriers for injectable drug delivery. Enhancement of liposome stability in bloodstream has been studied because of its relatively short circulation time and fast clearance from human body by reticuloendothelial system (RES) in blood vessel. In this study, new disaccharide-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) derivatives in which lactose or sucrose as the disaccharide molecule was conjugated covalently to DSPE were synthesized. Liposomes of which surface had disaccharide molecules were prepared by incorporating the disaccharide-DSPE into liposomes as one of their lipid components. Particle size of the prepared liposomes was approximately 100 nm. The liposomes of which surface were modified with the disaccharide-DSPE showed -25 mV of zeta potential value due to the presence of hydroxyl groups on their surface, while the unmodified control liposomes showed -10 mV of zeta potential value. Loading efficiency of model drug, doxorubicin, into liposomes was about 90%. Stability of the disaccharide-modified liposomes in vitro was evaluated by monitoring the amount of protein adsorption and particle size of the liposomes in serum. Disaccharide-modified liposomes were more stable in serum than unmodified control liposomes or polyethyleneglycol (PEG)-modified liposomes due to less adsorption of serum protein and hence less increase of their particle size. The liposomes of which surface was modified with disaccharide-DSPE conjugate can be used as long-circulating carriers for drugs having high toxicity or short half-life time due to their enhanced stability in blood circulatory system.
Jung Tae-Wook;Kim Sun-Lim;Moon Hyeon-Gui;Son Beom-Young;Kim Si Ju;Kim Soon Kwon
KOREAN JOURNAL OF CROP SCIENCE
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v.50
no.spc1
/
pp.161-166
/
2005
Objectives of this study were to select inbred lines which have a good eating quality and desirable segregates during inbreeding of waxy corn. The 64 inbred lines showed a large variance in their kernel shape and weight. 100-kernel weight, pericarp thickness, kernel length, kernel width, and kernel thickness ranged $11.7\~37.3g,\;11\~77{\mu}m,\;5.8\~9.6mm,\;6.5\~10.0mm$, and $4.1\~6.8mm$, respectively. The physicochemical analysis of 64 waxy corn inbred lines showed crude protein, crude fat, free sugar, and amylopectin content ranging $8.7\~15.8\%,\;2.3\~5.8\%,\;1.1\~11.0\%,\;and\;78.5\~93.8\%$, respectively. The texture property analysis of 64 inbred lines by texture analyzer showed a big difference. Gumminess, hardness, and chewiness of 64 inbred lines ranged $91\~383,\;181\~394,\;and\;73\~370$, respectively. The principal component analysis for 14 characteristics related to kernel quality showed that $73.1\%$ of the total variation could be attributed to the first five principal components. Biological meaning of the principal component was explained clearly by the correlation coefficient between principal components and characters. The first principal component appeared to correspond to small kernel and bad eating quality, The second principal component appeared to correspond to large kernel and good eating quality. The 64 inbred lines were classified into 8 groups by the cluster analysis using the first and second principal component. Among the groups, group VII and VIII included inbred lines with good eating quality that had thin pericarp thickness, low protein content, large kernel, and soft tenderness.
Kang, Hwan Ku;Kim, Chan Ho;Bang, Han Tae;Kim, Ji Hyuk;Kim, Min Ji;Kim, Dong Woon;Na, Jae Cheon;Hwangbo, Jong;Yang, Young Rok;Choi, Hee Cheol;Moon, Hong Kil
Korean Journal of Poultry Science
/
v.41
no.4
/
pp.227-233
/
2014
This study investigated the effects of dietary supplementation with the effective microorganism (EM) on the growth performance, blood parameter, small intestinal microflora, and noxious gas emission of broilers. A total 720 1-d old ROSS 308 was randomly assigned to 6 dietary treatment groups: control, virginiamycin (6 mg/kg), 0.1% PB 0.1% EM, 0.5% EM, and 1.0% EM. Each treatment was fed to 4 replicates of 30 birds per diet for d 35. Two-phase feeding program with a starter diet from 0 to 3 wk, and a finisher diet from 4 to 5 wk was used in the experiment. Within each phase, a diet was formulated to meet or exceed NRC requirements of broilers for macro- and micronutrients. The diet and water were available ad libitum. Result indicated that during overall periods of the experiment, final weight, body weight gain, and feed intake were not different among dietary treatments. Feed conversion ratio was less (P<0.05) for EM treatments than control, antibiotics, and PB. Total cholesterol (TC), triglyceride (TRG), glucose (GLU), total protein (TP), calcium (CA), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were not different among dietary treatments. White blood cell (WBC), hemoglobin (Hb), heterophils (HE), lymphocyte (LY), monocytes (MO), and eosinophils (EO) were not different among dietary treatments. HE:LY was less (P<0.05) for EM0.5 treatments than control, antibiotics, and PB. Lactobacillus was greater (P<0.05) for EM treatments than control and antibiotics. E. coli and Salmonella were not different among dietary treatments. $NH_3$ and $CO_2$ wereless (P<0.05) for EM treatments than control. These results indicated that EM treatments were effective feed conversion ratio, noxious gas emission and micro flora population on the cecum in broilers.
A series of rearing experiments were conducted to determine the growth rates and feed conversion efficiencies of tilapia in accordance with body size or age in nearly total closed system glass aquariums ($270\;\ell$ each in water volume) and concrete tanks ($4000\;\ell$) from April 10 to October 16, 1987. The fish used for the experiments was a Japanese strain of Oreochromis niloticus, and the size of the fish ranged from 7 g to more than 1,000 g in body weight. The starting stocking rates for each experimental lot were 10 to 20 kg in the glass aquarium ($3.7{\%}$ to $7.4{\%}$ of water volume) and 200 kg in the concrete tank ($5{\%}$ of water volume). A single experimental rearing term was 14 days with slight variations on occasions. Water temperature was designed to be kept at $26^{\circ}C$ but slight fluctuations were inevitable. Dissolved oxygen level was designed to be maintained at around $3\;mg/\ell$, but it also showed some variations. The ammonia level in the glass aquarium section once reached up to $18\;mg/\ell$, but generally remained at around $4\;mg/\ell$, and in the concrete tank section it was maintained at around $1\;mg/ell$. The feed was composed of mainly soybean meal with a small amount of fish meal as the protein source, and the crude protein content was about $32{\%}$. Mean daily growth rate was $3.5{\%}$ of body weight with 0.9 in food conversion ratio in the glass aquarium when the mean weight of fish was around 10 g with gradually reduced performances as the fish grew bigger. When the mean weight was 800 g, mean daily growth rate was $0.5{\%}$ with about 1.5 in food coversion for fish in the glass aquarium, and $0.8{\%}$ and 1.6 for fish in the concrete tank, respectively. According to the mean growth rate obtained from this experiment, it was calculated that the fish reared in the concrete tank require 223 days from 50 g to reach 1,000 g which is the ideal size for market in Korea, at the conditions provided as above, and 302 days from 10 g fingerlings to 800 g fish in the glass aquarium conditions of the closed recirculating water system.
Tooth movement is the result of bone metabolism in the periodontium, where various cytokines take important roles. Interleukin-6(II-6) and nitrous oxide (NO) were reported to be secreted from osteoblasts in the process of bone resorption. The mechanism of the process has not been clearly understood, but the activation of mitogen-activated protein kinase (MAPK) was known to be an important process in the release of the inflammatory cytotines in macrophages. In this regard, to prove the role of MAPK in the release of IL-6 and NO in MC3T3E-1 osteoblasts, Northern blot analysis, Western blot analysis and immune complex kinase assay were used. As a result, the treatment of MC3T3E-1 osteoblast cultures with combined $interferon-\gamma(IFN-\gamma)$, lipopolysaccharide (LPS) and tumor necrosis $factor-\alpha(TNF-\alpha)$ induces expressions of inducible nitric oxide synthase (iNOS) and IL-6, resulting in sustained releases of large amounts of NO and IL-6. However, $IFN-\gamma,\;LPS,\;and\;TNF-\alpha$ individually induce a non-detectable or small amount of NO and IL-6 in MC3T3E-1 osteoblasts. The role of MAPK activation in the early intracellular signal transduction involved in iNOS and IL-6 transcription in the combined agents-stimulated osteoblasts has been investigated. The p38 MAPK pathway is specifically involved in the combined agents-induced NO and IL-6 release, since NO and IL-6 release in the presence of a specific inhibitor of p38 MAPK, 4-(4-fluorophenyl)-2-(4-metylsulfinylphenyl)-5-(4-metylsulfinylphenyl)-5-(4-pyridyl)imidazole) (SB203580), were significantly diminished. In contrast, PD98059, a specific inhibitor of MEK1, had no effect on NO and IL-6 release. Northern blot analysis showed that the p3a MAPK pathway controlled the iNOS and IL-6 transcription level. These data suggest that p38 MAPK play an important role in the secretion of NO and IL-6 in $LPS/IFN{\gamma}-or\;TNF-\gamma-treated\;MC3T3E-1$ osteoblasts.
Rice is one of the most important crop in the world, in particular for food resources. With its small genome size of 383 Mb, the Oryza sativa is a model plant for genome research. Indeed, it's grain provides human with a source of carbohydrates and proteins. Rice grain has relatively low protein contents (around 8%) compared to other legume seeds (around 40%). Osborne classified seed proteins into water soluble albumin, salt soluble globulin, alcohol soluble prolamin and acidic/alkaline solution soluble glutelin. Glutelin and prolamin are the major storage proteins in rice. For the gene expression study of seed storage proteins, we analyzed 33,192 EST clones at immature stages in a rice cultivar (Oryza sativa L. cv. 'Ilpum'). Based on the expression analysis, we cloned 11 glutelin genes and figured out the 8 genes are located on Chromosome 2. The expression of glutelin genes appears to be about 28.2% of total level in immature seeds. Interestingly, glu-04 is duplicated as inverted sequences on the same chromosomes as far 4.5 kb. Our results indicate that glutelin genes, evolutionarily, were replicated on the chromosome and thus expressed as specific manners. In a whole protein composition analysis, glu05 (type B7) contains the highest lysin contents (4.51%) among the 11 rice glutelin genes. It will be an interesting future work to increase lysin contents by the gene overexpressor strategy with the aim of improved diet nutritionally fortified.
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