• Journal of The Korean Society of Inherited Metabolic disease
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• v.6 no.1
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• pp.15-23
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• 2006

Purpose: Glycogen storage disease type III (GSD-III), is a rare autosomal recessive disorder of glycogen metabolism. The affected enzyme is amylo-1,6-glucosidase, 4-alpha-glucanotransferase (AGL, glycogen debranching enzyme), which is responsible for the debranching of the glycogen molecule during catabolism. The disease has been demonstrated to show clinical and biochemical heterogeneity, reflecting the genotype-phenotype heterogeneity among different patients. In this study, we analyzed mutations of the AGL gene in three unrelated Korean GSD-III patients and discussed their clinical and laboratory implications. Methods: We studied three GSD-III patients and the clinical features were characterized. Sequence analysis of 35exons and part exon-intron boundaries of the AGLgene in patients were carried out by direct DNA sequencing method using genomic DNA isolated from patients' peripheral leukocytes. Results: The clinical features included hepatomegaly (in all patients), seizures (in patient 2), growth failure (in patients 1), hyperlipidemia (in patients 1 and 3), raised transaminases and creatinine kinase concentrations (in all patients) and mild EKG abnormalities (in patients 2). Liver transplantation was performed in patient 2due to progressive hepatic fibrosis. Administration of raw-corn-starch could maintain normoglycemia and improve the condition. DNA sequence analysis revealed mutations in 5 out of 6 alleles. Patient 1 was a compound heterozygote of c.1282 G>A (p.R428K) and c.1306delA (p.S603PfsX6), patient 2 with c.1510_1511insT (p.Y504LfsX10), and patient 3 with c.3416 T>C (p.L1139P) and c.l735+1 G>T (Y538_R578delfsX4) mutations. Except R428K mutation, 4 other mutations identified in3 patients were novel. Conclusion: GSD-III patients have variable phenotypic characteristics resembling GSD-Ia. The molecular defects in the AGL gene of Korean GSD-III patients were genetically heterogeneous.

• International Journal of Oral Biology
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• v.32 no.3
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• pp.85-91
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• 2007

Dlx3 is a homeodomain protein and is known to playa role in development and differentiation of many tissues. Deletion of four base pairs in DLX3 (NT3198) is causally related to tricho-dento-osseous (TDO) syndrome (OMIM # 190320), a genetic disorder manifested by taurodontism, hair abnormalities, and increased bone density in the cranium. Although the observed defects of TDO syndrome involves bone, little is known about the role of Dlx3 in bone remodeling process. In this study, we examined the effect of wild type DLX3 (wtDlx3) expression on osteoclast differentiation and compared it with that of 4-BP DEL DLX3 (TDO mtDlx3). To examine whether Dlx3 is expressed during RANKL-induced osteoclast differentiation, RAW264.7 cells were cultured in the presence of receptor activator of nuclear factor-B ligand (RANKL). Dlx3 protein level increased slightly after RANKL treatment for 1 day and peaked when the fusion of prefusion osteoclasts actively progressed. When wtDlx3 and TDO mtDlx3 were overexpressed in RAW264.7 cells, they enhanced RANKL-induced osteoclastogenesis and the expression of osteoclast differentiation marker genes such as calcitonin receptor, vitronectin receptor and cathepsin K. Since osteoclast differentiation is critically regulated by the balance between RANKL and osteoprotegerin (OPG), we examined the effect of Dlx3 overexpression on expression of RANKL and OPG in C2C12 cells in the presence of bone morphogenetic protein 2. Overexpression of wtDlx3 enhanced RANKL mRNA expression while slightly suppressed OPG expression. However, TDO mtDlx3 did not exert significant effects. This result suggests that inability of TDO mtDlx3 to regulate expression of RANKL and OPG may contribute to increased bone density in TDO syndrome patients. Taken together, it is suggested that Dlx3 playa role as a positive regulator of osteoclast differentiation via up-regulation of osteoclast differentiation-associated genes in osteoclasts, as well as via increasing the ratio of RANKL to OPG in osteoblastic cells.

• International Journal of Oral Biology
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• v.32 no.4
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• pp.119-125
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• 2007

Dlx3 is a homeodomain protein and is known to play a role in development and differentiation of many tissues. Deletion of four base pairs in DLX3 (NT3198) is causally related to tricho-dento-osseous (TDO) syndrome (OMIM #190320), a genetic disorder manifested by taurodontism, hair abnormalities, and increased bone density in the cranium. The molecular mechanisms that explain the phenotypic characteristics of TDO syndrome have not been clearly determined. In this study, we examined phenotypic characteristics of wild type DLX3(wtDlx3) and 4-BP DEL DLX3 (TDO mtDlx3) in C2C12 cells. To investigate how wtDlx3 and TDO mtDlx3 differentially regulate osteoblastic differentiation, reporter assays were performed by using luciferase reporters containing the promoters of alkaline phosphatase, bone sialoprotein or osteocalcin. Both wtDlx3 and TDO mtDlx3 enhanced significantly all the reporter activities but the effect of mtDlx3 was much weaker than that of wtDlx3. In spite of these differences in reporter activity, electrophoretic mobility shift assay showed that both wtDlx3 and TDO mtDlx3 formed similar amounts of DNA binding complexes with Dlx3 binding consensus sequence or with ALP promoter oligonucleotide bearing the Dlx3 binding core sequence. TDO mtDlx3 exhibits a longer half-life than wtDlx3 and it corresponds to PESTfind analysis result showing that potential PEST sequence was missed in carboxy terminal of TDO mtDlx3. In addition, co-immunoprecipitation demonstrated that TDO mtDlx3 binds to Msx2 more strongly than wtDlx3. Taken together, though TDO mtDlx3 acted as a weaker transcriptional activator than wtDlx3 in osteoblastic cells, there is possibility that during in vivo osteoblast differentiation TDO mtDlx3 may antagonize transcriptional repressor activity of Msx2 more effectively and for longer period than wtDlx3, resulting in enhancement of osteoblast differentiation.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.7
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• pp.1191-1201
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• 2020

Objective: The aim of this study was to evaluate the chemical-nutritional parameters, oxidative stability and volatile profile of eggs and cakes made with eggs from laying hens fed with a dietary supplementation of extruded linseed. Methods: Two thousand ISA Warren laying hens were randomly divided into two groups: a control group was fed with a standard diet while the experimental group received the same diet supplemented with 7% of extruded linseed. The trial lasted 84 days, in which three samplings of laid eggs were performed. Samples of eggs and food systems arising from eggs were then analyzed in order to obtain information about β-carotene and total flavonoid content, antioxidant activity, fatty acid profile, lipid oxidation, and volatile profile. Results: Linseed induced the increase of α-linolenic acid with consequent reduction of the ω-6/ω-3 ratio (4.3:1 in egg yolk); in addition to this, was evidenced the cholesterol reduction and the significant increase in total flavonoids and β-carotene, although no variations were detected in antioxidant capacity. Even in cooked products there was not only a direct effect of linseed in increasing α-linolenic acid, but also in inducing the reduction of cholesterol and its major oxidation product, 7-ketocholesterol. The dietary linseed integration was also shown to affect the volatile profile of baked products. Conclusion: Data confirmed that dietary supplementation with extruded linseed resulted in food products with interesting implications for human health. With regard to the volatile profile of baked products it would be necessary undertake further sensorial analysis in order to evaluate any variations on flavor and consumer acceptability.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.24 no.2
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• pp.218-229
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• 2021

Purpose: Serological tests of tissue transglutaminase (TTG) and deamidated gliadin (DGP) antibodies for celiac disease diagnosis show conflicting correlation with histology in young children and in type 1 diabetes mellitus (T1DM). Tests' ability to predict histology and cutoff values based on age and T1DM was evaluated. Methods: A retrospective study of children who had celiac serological tests between 6/1/2002 and 12/31/2014 at a pediatric hospital. Results: TTG IgA displayed similar results in predicting histology between <4.0 and ≥4.0 years age groups with sensitivity 98% and 93%, and specificity 88% and 86%, respectively. In children <4.0 years old, sensitivity for DGP antibodies was 100% and specificity 94%; in ≥4.0 years age groups, sensitivity was 60%, 88% for DGP IgA and IgG and specificity 95%, 96%, respectively. TTG IgA had low specificity in patients with T1DM compared with non-T1DM, 42% vs. 91%. Positive TTG IgA with normal histology was associated with higher T1DM prevalence at 36% compared with negative tests at 4%. Finally, the TTG IgA cutoff value was higher in T1DM at 36 vs. 16.3 units in non-T1DM. DGP IgG cutoff showed similar values between age groups; TTG IgA and DGP IgA cutoffs were lower in <4.0 years at 8.3 and 11.9 units than ≥4.0 years at 23.4 and 19.9, respectively. Conclusion: TTG IgA is sufficient for the <4.0 years age group and DGP antibodies had no advantage over TTG IgA in older children. The cutoff value to determine a positive TTG IgA should be higher for children with T1DM.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1513-1513
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• 2001

Present Food Legislation compels dairy industry to carry out analyses in order to guarantee the food safety and quality of products. Furthermore, in many cases industry pays milk according to bacteriological or/and nutritional quality. In order to do these analyses, several expensive instruments are needed (Milkoscan, Fossomatic, Bactoscan). NIRS technology Provides a unique instrument to deal with all analytical requirements. It offers as main advantages its speed and, specially, its versatility, since not only allows determine all the parameters required in milk analysis, but also allows analyse other dairy products, like cheese or whey. The objective of this study is to develop NIRS calibration equations to predict several quality parameters in goat milk, cheese and whey. Three sets of 123 milk samples, 190 cheese samples and 109 whey samples, have been analysed in a FOSS NIR Systems 6500 I spectrophotometer equipped with a spinning module. Milk and whey were analysed by folded transmission, using circular cells with gold surface and pathlength of 0.1 m, while intact cheese was analysed by reflectance using standard circular cells. NIRS calibrations were obtained for the prediction of chemical composition in goat milk, for fat (r$^2$=0.92; SECV=0.20%), total solids (r$^2$=0.95: SECV=0.22%), protein (r$^2$=0.94; SECV=0.07%), casein (r$^2$=0.93; SECV=0.07%) and lactose (r$^2$=0.89; SECV=0.05%). Moreover, equations have been performed to determine somatic cells (r$^2$=0.81; SECV=276.89%) and total bacteria (r$^2$=0.58; SECV=499.32%) counts in goat milk. In the case of cheese, calibrations were obtained for the prediction of fat (r$^2$=0.92; SECV=0.57), total solids (r$^2$=0.80; SECV=0.92%) and protein (r$^2$=0.70; SECV=0.63%). In whey, fat (r$^2$=0.66; SECV=0.08%), total solids (r$^2$=0.67; SECV=0.19%) and protein (r$^2$=0.76; SECV=0.07%) NIRS equations were obtained. These results proved the viability of NIRS technology to predict chemical and microbiological parameters and somatic cells count in goat milk, as well as chemical composition of goat cheese and whey.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.10
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• pp.4567-4575
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• 2016

A number of studies have evidenced the effect of modifiable lifestyle factors such as diet, breastfeeding and nutritional status on breast cancer risk. However, none have addressed the missing data problem in nutritional epidemiologic research in South America. Missing data is a frequent problem in breast cancer studies and epidemiological settings in general. Estimates of effect obtained from these studies may be biased, if no appropriate method for handling missing data is applied. We performed Multiple Imputation for missing values on covariates in a breast cancer case-control study of $C{\acute{o}}rdoba$ (Argentina) to optimize risk estimates. Data was obtained from a breast cancer case control study from 2008 to 2015 (318 cases, 526 controls). Complete case analysis and multiple imputation using chained equations were the methods applied to estimate the effects of a Traditional dietary pattern and other recognized factors associated with breast cancer. Physical activity and socioeconomic status were imputed. Logistic regression models were performed. When complete case analysis was performed only 31% of women were considered. Although a positive association of Traditional dietary pattern and breast cancer was observed from both approaches (complete case analysis OR=1.3, 95%CI=1.0-1.7; multiple imputation OR=1.4, 95%CI=1.2-1.7), effects of other covariates, like BMI and breastfeeding, were only identified when multiple imputation was considered. A Traditional dietary pattern, BMI and breastfeeding are associated with the occurrence of breast cancer in this Argentinean population when multiple imputation is appropriately performed. Multiple Imputation is suggested in Latin America's epidemiologic studies to optimize effect estimates in the future.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.23 no.6
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• pp.558-566
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• 2020

Purpose: Progressive familial intrahepatic cholestasis (PFIC) is a rare genetic autosomal recessive disease caused by mutations in ATP8B1, ABCB11 or ABCB4. Mutational analysis of these genes is a reliable approach to identify the disorder. Methods: We collected and analyzed relevant data related to clinical diagnosis, biological investigation, and molecular determination in nine children carrying these gene mutations, who were from unrelated families in South China. Results: Of the nine patients (five males, four females) with PFIC, one case of PFIC1, four cases of PFIC2, and four cases of PFIC3 were diagnosed. Except in patient no. 8, jaundice and severe pruritus were the major clinical signs in all forms. γ-glutamyl transpeptidase was low in patients with PFIC1/PFIC2, and remained mildly elevated in patients with PFIC3. We identified 15 different mutations, including nine novel mutations (p.R470HfsX8, p.Q794X and p.I1170T of ABCB11 gene mutations, p.G319R, p.A1047P, p.G1074R, p.T830NfsX11, p.A1047PfsX8 and p.N1048TfsX of ABCB4 gene mutations) and six known mutations (p.G446R and p.F529del of ATP8B1 gene mutations, p.A588V, p.G1004D and p.R1057X of ABCB11 gene mutations, p.P479L of ABCB4 gene mutations). The results showed that compared with other regions, these three types of PFIC genes had different mutational spectrum in China. Conclusion: The study expands the genotypic spectrum of PFIC. We identified nine novel mutations of PFIC and our findings could help in the diagnosis and treatment of this disease.

• Horticultural Science & Technology
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• v.33 no.5
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• pp.722-729
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• 2015

Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria ${\times}$ ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An $F_2$ population derived from a cross between PM-resistance 'Seolhyang' and PM-susceptibility 'Akihime' was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.

• Proceedings of the Korean Society of Near Infrared Spectroscopy Conference
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• 2001.06a
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• pp.1615-1615
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• 2001

During the past ten years, Near Infrared Spectroscopy has been successfully applied to the analysis of a great variety of agriculture products. Previous works (Morra et al., 1991; Salgo et al., 1998) have shown the potential of this technology for soil analysis, estimating different parameters just with one single scan. The main advantages of NIR applications in soils are the speed of response, allowing the increase of the number of samples analysed to define a particular soil, and the instantaneous elaboration of recommendations for fertilization and soil amendment. Another advantage is to avoid the use of chemical reagents at all, being an environmentally safe technique. In this paper, we have studied a set of 129 soil samples selected from representative glasshouse soils from Southern Spain. The samples were dried, milled, and sieved to pass a 2 mm sieve and then analysed for organic carbon, total nitrogen, inorganic nitrogen (nitrate ammonium), hygroscopic humidity, pH and electrical conductivity in the 1:1 extract. NIR spectra of all samples were obtained in reflectance mode using a Foss NIR Systems 6500 spectrophotometer equipped with a spinning module. Calibration equations were developed for seven analytical parameters (ph, Total nitrogen, organic nitrogen, organic carbon, C/N ratio and Electric Conductivity). Preliminary results show good correlation coefficients and standard errors of cross validation in equations obtained for Organic Carbon, Organic Nitrogen, Total Nitrogen and C/N ratio. Calibrations for nitrates and nitrites, ammonia and electric conductivity were not acceptable. Calibration obtained for pH had an acceptable SECV, but the determination coefficient was found very poor probably due to the reduced range in reference values. Since the estimation of Organic Carbon and C/N ratio are acceptable NIIRS could be used as a fast method to assess the necessity of organic amendments in soils from Mediterranean regions where the low level of organic matter in soils constitutes an important agronomic problem. Furthermore, the possibility of a single and fast estimation of Total Nitrogen (tedious determination by modifications of the Kjeldahl procedure) could provide and interesting data to use in the estimation of nitrogen fertilizer rates by means of nitrogen balances.