• Title/Summary/Keyword: Inter-Molecular Association

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Development and evaluation of probiotic delivery systems using the rennet-induced gelation of milk proteins

  • Ha, Ho-Kyung;Hong, Ji-Young;Ayu, Istifiani Lola;Lee, Mee-Ryung;Lee, Won-Jae
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.1182-1193
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    • 2021
  • The aims of this study were to develop a milk protein-based probiotic delivery system using a modified rennet-induced gelation method and to determine how the skim milk powder concentration level and pH, which can affect the rennet-induced intra- and inter-molecular association of milk proteins, affect the physicochemical properties of the probiotic delivery systems, such as the particle size, size distribution, encapsulation efficiency, and viability of probiotics in simulated gastrointestinal tract. To prepare a milk protein-based delivery system, skim milk powder was used as a source of milk proteins with various concentration levels from 3 to 10% (w/w) and rennet was added to skim milk solutions followed by adjustment of pH from 5.4 or 6.2. Lactobacillus rhamnosus GG was used as a probiotic culture. In confocal laser scanning microscopic images, globular particles with a size ranging from 10 ㎛ to 20 ㎛ were observed, indicating that milk protein-based probiotic delivery systems were successfully created. When the skim milk powder concentration was increased from 3 to 10% (w/w), the size of the delivery system was significantly (p < 0.05) increased from 27.5 to 44.4 ㎛, while a significant (p < 0.05) increase in size from 26.3 to 34.5 ㎛ was observed as the pH was increased from 5.4 to 6.4. An increase in skim milk powder concentration level and a decrease in pH led to a significant (p < 0.05) increase in the encapsulation efficiency of probiotics. The viability of probiotics in a simulated stomach condition was increased when probiotics were encapsulated in milk protein-based delivery systems. An increase in the skim milk powder concentration and a decrease in pH resulted in an increase in the viability of probiotics in simulated stomach conditions. It was concluded that the protein content by modulating skim milk powder concentration level and pH were the key manufacturing variables affecting the physicochemical properties of milk protein-based probiotic delivery systems.

Proteomic analysis of serum proteins responsive to styrene exposure (Styrene 노출에 반응을 보이는 혈청 단백질에 대한 프로테오믹스 분석)

  • Kim, Ki-Woong;Heo, Kyung-Hwa;Won, Yong Lim;Jeong, Jin Wook;Kim, Tae Gyun;Park, Injeong
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.17 no.3
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    • pp.235-244
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    • 2007
  • By comparing the proteins from the workers exposed to styrene with the ones from controls, it may be possible to identify proteins that play a role in the occurrence and progress of occupational disease and thus to study the molecular mechanisms of occupational disease. In order to find the biomarkers for assessing the styrene effects early, before clinical symptoms develop and to understand the mechanisms of adverse health effects, we surveyed 134 employees, among whom 52 workers(30 male and 22 female) were chronically exposed to styrene in 10 glass-reinforced plastic boat manufacturing factories in Korea and 82 controls had never been occupationally exposed to hazardous chemicals including styrene. The age and drinking habits and serum biochemistry such as total protein, BUN and serum creatinine in both groups were significantly different. Exposed workers were divided into three groups according to exposure levels of styrene(G1, below 1/2 TLV; G2, 1/2 TLV to TLV; G3, above TLV). The mean concentration of airborne styrene in G1 group was $10.93{\pm}11.33ppm$, and those of urinary mandelic acid(MA) and phenylglyoxylic acid(PGA) were $0.17{\pm}0.21$ and $0.13{\pm}0.11g/g$ creatinine, respectively. The mean concentration of airborne styrene in G2 and G3 groups were $47.54{\pm}22.43$ and $65.33{\pm}33.47ppm$, respectively, and levels of urinary metabolites such as MA and PGA increased considerably as expected with the increase in exposure level of styrene. The airborne styrene concentration were significantly correlated to the urinary concentration of MA(r=0.784, p=0.000) and PGA(r=0.626, p<0.001). In the 2D electrophoresis, the concentration of five proteins including complement C3 precursor, alpha-1-antitrypsin(AAT), vitamin D binding protein precursor(DBP), alpha-1-B-glycoprotein(A1BG) and inter alpha trypsin inhibitor(ITI) heavy chain-related protein were significantly altered in workers exposed to styrene compared with controls. While expression of complement C3 precursor and AAT increased by exposure to styrene, expression of DBP, A1BG and ITI heavy chain-related protein decreased. These results suggest that the exposure of styrene might affects levels of plasma proteinase, carriers of endogenous substances and immune system. In particular, increasing of AAT with the increase in exposure level of styrene can explain the tissue damage and inflammation by the imbalance of proteinase/antiproteinase and decrease of DBP, A1BG and ITI heavy chain-related protein in workers exposed to styrene is associated with dysfunction and/or declination in immune system and signal transduction

Study for Reliability of Interpretation of the Three Phase Bone Scintigraphy in Patients with Post-traumatic Complex Regional Pain Syndrome (외상 후 복합부위통증증후군 환자에서 시행한 삼상 뼈 스캔의 판독 신뢰도에 관한 연구)

  • Park, Jung-Mi;Kim, Seon-Jung;Chung, Seung-Hyun;Lee, Yong-Taek
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.1
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    • pp.44-51
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    • 2008
  • Purpose: We performed this study to evaluate reliability on interpretation of three phase bone scintigraphy (TPBS) in patients with post-traumatic complex regional pain syndrome (PT-CRPS). Methods: Based on International Association for the Study of Pain guideline in 1994, 34 patients with PT-CRPS were selected for this study. Two nuclear medicine physicians evaluated identical TPBS according to the uptake pattern, extent and intensity of the lesion, and their agreements (kappa values) were analysed. The final diagnosis based on arbitrary criteria of each physician were compared with those obtained by the criteria for PT-CRPS established in this study, which are hyperactivity on all phases (criteria 1), hyperactivity of whole joints on delayed phase (criteria 2), and hyperactivity of either whole or FDGal joints on delayed phase (criteria 3). Results: Intra-observer agreements were good for uptake pattern, intensity, and extent on TPBS. Inter-observer agreements were also good, except extent on blood pool phase (0.55). The inter-observer agreements on final diagnosis improved when criteria 1-3 were applied (0.77-0.88), compared to when physician's own criteria were used (0.63). Those also improved from 0.29 to 0.47-0.82 for acute stage, and from 0.37 to 1.0 for chronic stage. The sensitivities of chronic stage were relatively lower to those of acute stage. Conclusions: Inter-observer's variations in diagnosis of the patients with PT-CRPS using TPBS were observed. These results were attributed to different criteria set by observers. In order to improve agreement on interpretation of TPBS, common positive criteria should be established, especially considering uptake pattern and clinical stages.

Association of A/T Rich Microsatellites with Responses to Artificial Selection for Larval Developmental Duration in the Silkworm Bombyx mori

  • Pradeep, Appukuttan Nair Retnabhavan;Awasthi, Arvind Kumar;Urs, Raje Siddaraje
    • Molecules and Cells
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    • v.25 no.4
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    • pp.467-478
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    • 2008
  • Simple sequence repeats (SSRs) and interSSR (ISSR) marker systems were used in this study to reveal genetic changes induced by artificial selection for short/long larval duration in the tropical strain Nistari of the silkworm Bombyx mori. Artificial selection separated longer larval duration (LLD) ($29.428{\pm}0.723days$) and shorter larval duration (SLD) ($22.573{\pm}0.839days$) lines from a base, inbred population of Nistari (larval span of $23.143{\pm}0.35days$). SSR polymorphism was observed between the LLD and SLD lines at one microsatellite locus, Bmsat106 ($CA_7$) and at two loci of 1074 bp and 823 bp generated with the ISSR primer UBC873. Each of these loci was present only in the LLD line. The loci segregated in the third generation of selection and were fixed in opposite directions. In the $F_2$ generation of the $LLD{\times}SLD$ lines, the alleles of Bmsat106 and $UBC873_{1074bp}$ segregated in a 1:1 ratio and the loci were present only in the LLD individuals. $UBC873_{823bp}$ was homozygous. Single factor ANOVA showed a significant association between the segregating loci and longer larval duration. Together, the two alleles contributed to an 18% increase in larval duration. The nucleotide sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci had 67% A/T content and consisted of direct, reverse, complementary and palindromic repeats. The repeats appeared to be "nested" (59%) in larger repeats or as clustered elements adjacent to other repeats. Of 203 microsatellites identified, dinucleotides (67.8%) predominated and were rich in A/T and T/A motifs. The sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci showed similarity (E = 0.0) to contigs located in Scaffold 010774 and Scaffold 000139, respectively, of the B. mori genome. BLASTN analysis of the $UBC873_{1074bp}$ sequence showed significant homology of (nt.) 45-122 with upstream region of three exons from Bombyx. The complete sequence of this locus showed ~49% nucleotide conservation with transposon 412 of Drosophila melanogaster and the Ikirara insertions of Anopheles gambiae. The A + T richness and lack of coding potential of these small loci, and their absence in the SLD line, reflect the active process of genetic change associated with the switch to short larval duration as an adaptation to the tropics.

Monitoring of genetically close Tsaiya duck populations using novel microsatellite markers with high polymorphism

  • Lai, Fang-Yu;Chang, Yi-Ying;Chen, Yi-Chen;Lin, En-Chung;Liu, Hsiu-Chou;Huang, Jeng-Fang;Ding, Shih-Torng;Wang, Pei-Hwa
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.888-901
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    • 2020
  • Objective: A set of microsatellite markers with high polymorphism from Tsaiya duck were used for the genetic monitoring and genetic structure analysis of Brown and White Tsaiya duck populations in Taiwan. Methods: The synthetic short tandem repeated probes were used to isolate new microsatellite markers from the genomic DNA of Tsaiya ducks. Eight populations, a total of 566 samples, sourced from Ilan Branch, Livestock Research Institute were genotyped through novel and known markers. The population genetic variables were calculated using optional programs in order to describe and monitor the genetic variability and the genetic structures of these Tsaiya duck populations. Results: In total 24 primer pairs, including 17 novel microsatellite loci from this study and seven previously known loci, were constructed for the detection of genetic variations in duck populations. The average values for the allele number, the effective number of alleles, the observed heterozygosity, the expected heterozygosity, and the polymorphism information content were 11.29, 5.370, 0.591, 0.746, and 0.708, respectively. The results of analysis of molecular variance and principal component analysis indicated a contracting Brown Tsaiya duck cluster and a spreading White Tsaiya duck cluster. The Brown Tsaiya ducks and the White Tsaiya ducks with Pekin ducks were just split to six clusters and three clusters when K was set equal to 6 and 3 in the Bayesian cluster analysis. The individual phylogenetic tree revealed eight taxa, and each individual was assigned to its own population. Conclusion: According to our study, the 24 novel microsatellite markers exhibited a high capacity to analyze relationships of inter- and intra-population in those populations with a relatively limited degree of genetic diversity. We suggest that duck farms in Taiwan could use the new (novel) microsatellite set to monitor the genetic characteristics and structures of their Tsaiya duck populations at various intervals in order to ensure quality breeding and conservation strategies.