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2.  Effects of Serial Passage on the Characteristics and Chondrogenic Differentiation of Canine Umbilical Cord Matrix Derived Mesenchymal Stem Cells 
Mesenchymal stem cells (MSCs) are often known to have a therapeutic potential in the cell-mediated repair for fatal or incurable diseases. In this study, canine umbilical cord MSCs (cUC-MSCs) were isolated from umbilical cord matrix (n = 3) and subjected to proliferative culture for 5 consecutive passages. The cells at each passage were characterized for multipotent MSC properties such as proliferation kinetics, expression patterns of MSC surface markers and self-renewal associated markers, and chondrogenic differentiation. In results, the proliferation of the cells as determined by the cumulative population doubling level was observed at its peak on passage 3 and stopped after passage 5, whereas cell doubling time dramatically increased after passage 4. Expression of MSC surface markers (CD44, CD54, CD61, CD80, CD90 and Flk-1), molecule (HMGA2) and pluripotent markers (sox2, nanog) associated with self-renewal was negatively correlated with the number of passages. However, MSC surface marker (CD105) and pluripotent marker (Oct3/4) decreased with increasing the number of subpassage. cUC-MSCs at passage 1 to 5 underwent chondrogenesis under specific culture conditions, but percentage of chondrogenic differentiation decreased with increasing the number of subpassage. Collectively, the present study suggested that sequential subpassage could affect multipotent properties of cUC-MSCs and needs to be addressed before clinical applications.
doi:10.5713/ajas.2012.12488
PMCID: PMC4093376  PMID: 25049827
Mesenchymal Stem Cell; Umbilical Cord Matrix; Canine; Multipotent; Differentiation
3.  Effects of Hydrated Potato Starch on the Quality of Low-fat Ttoekgalbi (Korean Traditional Patty) Packaged in Modified Atmosphere Conditions during Storage 
This study was carried out to investigate the effects of hydrated potato starch on the quality of low-fat ttoekgalbi (Korean traditional patty) packaged in modified atmosphere conditions during storage. The ttoekgalbi was prepared from 53.2% lean beef, 13.9% lean pork, 9.3% pork fat, and 23.6% other ingredients. Two low-fat ttoekgalbi treatments were prepared by substituting pork fat with hydrated potato starch; either by 50% fat replacement (50% FR) or 100% fat replacement (100% FR). Both 50% and 100% FR increased the moisture, crude protein, and decreased fat content, cooking loss, and hardness. For MAP studies, 200 g of ttoekgalbi were placed on the tray and filled with gas composed of 70% O2: 30% CO2 (70% O2-MAP) and 30% CO2: 70% N2 (70% N2-MAP), and were stored at 5°C for 12 d. During the storage time, both 50% and 100% FR showed higher protein deterioration, while no differences were found in CIE a*, CIE L*, lipid oxidation, and bacterial counts in comparison to control. The ttoekgalbi with 70% O2-MAP was more red, lighter in color, and showed higher TBARS values compared with 70% N2-MAP. The meat with 70% N2-MAP showed lower aerobic bacterial counts in control than those with 70% O2-MAP. The lower anaerobic bacterial counts were observed only in 50% FR and 100% FR packed with 70% N2-MAP in comparison with 70% O2-MAP. In conclusion, the fat replacement with hydrated potato starch showed no negative effects on the quality of low fat ttoekgalbi during storage and 70% N2-MAP was better than 70% O2-MAP for low-fat ttoekgalbi packaging.
doi:10.5713/ajas.2011.11280
PMCID: PMC4093120  PMID: 25049619
Low-fat Ttoekgalbi (Korean Traditional Patty); Hydrated Potato Starch; Modified Atmosphere Packaging (MAP)
4.  Farnesoid X receptor, overexpressed in pancreatic cancer with lymph node metastasis promotes cell migration and invasion 
British Journal of Cancer  2011;104(6):1027-1037.
Background:
Lymph node metastasis is one of the most important adverse prognostic factors for pancreatic cancer. The aim of this study was to identify novel lymphatic metastasis-associated markers and therapeutic targets for pancreatic cancer.
Methods:
DNA microarray study was carried out to identify genes differentially expressed between 17 pancreatic cancer tissues with lymph node metastasis and 17 pancreatic cancer tissues without lymph node metastasis. The microarray results were validated by real-time PCR. Immunohistochemistry and western blotting were used to examine the expression of farnesoid X receptor (FXR). The function of FXR was studied by small interfering RNA and treatment with FXR antagonist guggulsterone and FXR agonist GW4064.
Results:
Farnesoid X receptor overexpression in pancreatic cancer tissues with lymph node metastasis is associated with poor patient survival. Small interfering RNA-mediated downregulation of FXR and guggulsterone-mediated FXR inhibition resulted in a marked reduction in cell migration and invasion. In addition, downregulation of FXR reduced NF-κB activation and conditioned medium from FXR siRNA-transfected cells showed reduced VEGF levels. Moreover, GW4064-mediated FXR activation increased cell migration and invasion.
Conclusions:
These findings indicated that FXR overexpression plays an important role in lymphatic metastasis of pancreatic cancer and that downregulation of FXR is an effective approach for inhibition of pancreatic tumour progression.
doi:10.1038/bjc.2011.37
PMCID: PMC3065277  PMID: 21364590
pancreatic cancer; lymph node metastasis; FXR; DNA microarray; siRNA
5.  Novel WDR72 Mutation and Cytoplasmic Localization 
Journal of Dental Research  2010;89(12):1378-1382.
The proven candidate genes for amelogenesis imperfecta (AI) are AMELX, ENAM, MMP20, KLK4, FAM83H, and WDR72. We performed mutation analyses on seven families with hypomaturation AI. A novel WDR72 dinucleotide deletion mutation (g.57,426_57,427delAT; c.1467_ 1468delAT; p.V491fsX497) was identified in both alleles of probands from Mexico and Turkey. Haplotype analyses showed that the mutations arose independently in the two families. The disease perfectly segregated with the genotype. Only persons with both copies of the mutant allele were affected. Their hypomineralized enamel suffered attrition and orange-brown staining following eruption. Expression of WDR72 fused to green fluorescent protein showed a cytoplasmic localization exclusively and was absent from the nucleus. We conclude that WDR72 is a cytoplasmic protein that is critical for dental enamel formation.
doi:10.1177/0022034510382117
PMCID: PMC3144073  PMID: 20938048
protein localization; enamel maturation; enamel; tooth; Amelogenesis imperfect
6.  MMP20 Hemopexin Domain Mutation in Amelogenesis Imperfecta 
Journal of Dental Research  2010;89(1):46-50.
Proteolytic enzymes serve important functions during dental enamel formation, and mutations in the kallikrein 4 (KLK4) and enamelysin (MMP20) genes cause autosomal-recessive amelogenesis imperfecta (ARAI). So far, only 1 KLK4 and 3 MMP20 mutations have been reported in ARAI kindreds. To determine whether ARAI in a family with a hypomaturation-type enamel defect is caused by mutations in the genes encoding enamel proteolytic enzymes, we performed mutational analysis on candidate genes. Mutational and haplotype analyses revealed an ARAI-causing point mutation (c.910G>A, p.A304T) in exon 6 of MMP20 that results in a single amino acid substitution in the hemopexin domain. Western blot analysis showed decreased expression of the mutant protein, but zymogram analysis demonstrated that this mutant was a functional protein. The proband and an affected brother were homozygous for the mutation, and both unaffected parents were carriers. The enamel of newly erupted teeth had normal thickness, but was chalky white and became darker with age.
doi:10.1177/0022034509352844
PMCID: PMC3318044  PMID: 19966041
MMP20; hemopexin; enamel; tooth; amelogenesis imperfecta
7.  Relation between cognitive dysfunction and reduced vital capacity in amyotrophic lateral sclerosis 
Background
Many patients with amyotrophic lateral sclerosis (ALS) with cognitive impairment have fronto‐temporal dysfunction. Whereas in some patients with ALS the fronto‐temporal dysfunction is undoubtedly due to the degenerative process associated with the disease, in others dysfunction cannot be accounted for by an irreversible degenerative process alone, as it also appears to involve a reversible process. We hypothesised that reduced vital capacity can be a key contributor to the fronto‐temporal dysfunction observed in patients with ALS.
Objective
To investigate the association between fronto‐temporal dysfunction and reduced vital capacity in ALS.
Methods
16 consecutive patients who conformed to a diagnosis of definite or probable ALS (El escorial criteria) were grouped by vital capacity, and their clinical characteristics and cognitive functions, including disease duration, attention, executive function and memory, were measured.
Results
Patients with a reduced vital capacity performed significantly poorer in memory retention (p = 0.028), retrieval efficacy (p = 0.003), spoken verbal fluency (p = 0.03) and spoken verbal fluency indexes (p = 0.016) than those with a normal vital capacity.
Conclusion
The fronto‐temporal dysfunction in ALS might be attributable to potentially reversible secondary effects associated with reduced vital capacity, as well as to the primary degenerative process.
doi:10.1136/jnnp.2006.111195
PMCID: PMC2095584  PMID: 17557798
8.  Phase II study and biomarker analysis of cetuximab combined with modified FOLFOX6 in advanced gastric cancer 
British Journal of Cancer  2009;100(2):298-304.
This prospective study was conducted with the Korean Cancer Study Group to evaluate the efficacy and safety of cetuximab combined with modified FOLFOX6 (mFOLFOX6) as first-line treatment in recurrent or metastatic gastric cancer and to identify potential predictive biomarkers. Patients received cetuximab 400 mg m−2 at week 1 and 250 mg m−2 weekly thereafter until disease progression. Oxaliplatin (100 mg m−2) and leucovorin (100 mg m−2) were administered as a 2-h infusion followed by a 46-h continuous infusion of 5-fluorouracil (2400 mg m−2) every 2 weeks for a maximum of 12 cycles. Biomarkers potentially associated with efficacy were analysed. Among 38 evaluable patients, confirmed response rate (RR) was 50.0% (95% CI 34.1–65.9). Median time-to-progression (TTP) was 5.5 months (95% CI 4.5–6.5) and overall survival (OS) 9.9 months. Eleven patients having tumour EGFR expression by immunohistochemistry with low serum EGF and TGF-α levels showed a 100% RR compared to 37.0% in the remaining 27 patients (P<0.001). Moreover, ligand level increased when disease progressed in seven out of eight patients with EGFR expression and low baseline ligand level. No patient exhibited EGFR amplification or K-ras mutations. Gastric cancer patients with EGFR expression and low ligand levels had better outcomes with cetuximab/mFOLFOX6 treatment.
doi:10.1038/sj.bjc.6604861
PMCID: PMC2634707  PMID: 19127259
Cetuximab; chemotherapy; epidermal growth factor; epidermal growth factor receptor; gastric cancer; transforming growth factor-α
9.  Evidence of thalamic disinhibition in patients with hemichorea: semiquantitative analysis using SPECT 
Kim, J | Lee, K | Lee, K | Kim, Y | Kim, B | Chung, Y | Chung, S
Objectives: Hemichorea sometimes occurs after lesions that selectively involve the caudate nucleus, putamen, and globus pallidus. Some reports have hypothesised that the loss of subthalamic nucleus control on the internal segment of the globus pallidus, followed by the disinhibition of the thalamus may contribute to chorea. However, the pathophysiology is poorly understood. Therefore, clinicoradiological localisation was evaluated and a comparison of the haemodynamic status of the basal ganglia and thalamus was made.
Methods: Six patients presenting with acute onset of hemichorea were assessed. Neuroimaging studies, including MRI and SPECT examinations in addition to detailed biochemical tests, were performed. A semiquantitative analysis was performed by comparing the ratio of blood flow between patients and normal controls. In addition, the ratio of perfusion asymmetry was calculated as the ratio between each area contralateral to the chorea and that homolateral to the chorea. The comparison was made with a two sample t test.
Results: The causes of hemichorea found consisted of four cases of acute stroke, one non-ketotic hyperglycaemia, and one systemic lupus erythematosus. Brain MRI indicated lesion sites in the contralateral putamen, globus pallidus, caudate nucleus, and subthalamic nucleus. A significant decrease in the ratio of blood flow in the basal ganglia contralateral to the chorea and a significant increase in the thalamus was found when comparing the perfusion asymmetries, which were calculated as the ratio of cerebral blood flow (CBF) for each region to that in the homolateral occipital area (p<0.05).
Conclusion: An alteration in CBF in both the contralateral thalamus and basal ganglia reflect the loss of pallidal inhibitory input from the pallidum to the thalamus. This change in CBF may be one of epiphenomena, which implicates an occurrence of hemichorea in humans.
doi:10.1136/jnnp.72.3.329
PMCID: PMC1737786  PMID: 11861689
10.  Emergence of vaccine-induced escape mutant of hepatitis B virus with multiple surface gene mutations in a Korean child. 
Journal of Korean Medical Science  2001;16(3):359-362.
The S protein of hepatitis B virus is the principal component of virus envelope and the primary target of anti-HBs response. Mutants or variants that escape neutralization by anti-HBs have been selected during immunoprophylaxis of HBV after birth and liver transplantation. We investigated a case of a Korean child who was vaccinated at birth against hepatitis B and also given hepatitis B immunoglobulin, but nevertheless later became infected with the virus. Hepatitis B virus-specific deoxyribonucleic acid covering the region of genome encoding the predominant "a" determinant of hepatitis surface antigen was amplified using polymerase chain reaction, and the nucleotide sequence was determined. We present for the first time in Korea the independent emergence of an escape mutant with substitution of arginine for glycine at amino acid 145 and proline for glutamate at amino acid 120 in "a" determinant after immunization.
PMCID: PMC3054753  PMID: 11410701
11.  Anorectal dysfunction in systemic sclerosis. 
Journal of Korean Medical Science  1996;11(3):244-249.
This study was aimed to evaluate the anorectal dysfunction in systemic sclerosis (SSc) and propose the clinical significance of anorectal manometry in patients with SSc. Seven patients with SSc were evaluated with manometry for anorectal function and an additional 11 normal subjects were collected as a control group. The study group underwent esophageal manometry as well and the correlation between the degree of anorectal and esophageal dysfunction was evaluated. Patients showed a lower tolerance for balloon distention of the rectum than controls (minimal sensory volume and urgency volume, P < 0.05). The resting and squeezing pressure of the anal sphincter and the functional length of the anal canal showed no significant difference in these two groups. Rectoanal inhibitory reflex was absent in one (14%) and diminished in two (29%) of seven patients with SSc. SSc patients also showed abnormal esophageal manometry findings, notably decreased LES pressure and body amplitude of distal 2/3 esophagus. The comparison between manometric profiles of anorectum and esophagus showed no significant correlation by statistical analysis. In conclusion, our data could suggest that anorectal function may be impaired in patients with SSc which could reflect the involvement of the anorectum by the disease, and that anorectal manometric studies can be useful to detect such dysfunction in patients with SSc, even before clinical symptoms.
PMCID: PMC3054055  PMID: 8843007
12.  Study on the Korean adult cranial capacity. 
Journal of Korean Medical Science  1995;10(4):239-242.
Cranial capacity was measured in Korean adult skulls. The cavity was filled with rice seeds and the volume of the seeds were measured in a graduated cylinder. The results were 1470 +/- 107 (mean +/- standard deviation) in male and 1317 +/- 117 cc in female skulls. These values were in good accordance with those previously reported. In addition, regression formulae were obtained with the product of the length, breadth, and height of the skull as an independent parameter and the measured capacity as a dependent one. With known external measurements, the expected cranial capacity was as follows: when using baso-bregmatic height, male: capacity = 307.5 + 333 x 10(-6) x (length.breadth.baso-bregmatic height) female: capacity = -12.0 + 435 x 10(-6) x (length.breadth.baso-bregmatic height) and, when using auriculo-bregmatic height, male: capacity = 214.6 + 429 x 10(-6) x (length.breadth.auriculo-bregmatic height) female: capacity = 131.6 + 461 x 10(-6) x (length.breadth.auriculo-bregmatic height).
PMCID: PMC3054067  PMID: 8593202
13.  A validation study of four single locus probes (MS1, MS31, MS43 and g3) in a Korean population--further evaluation for paternity testing. 
Journal of Korean Medical Science  1995;10(4):243-249.
Hypervariable tandem repetitive regions in human DNA are proving to be increasingly useful for genetic analysis in humans. We chose four single locus probes (SLP; MS1, MS43, MS8 and g3) for a validation test among Koreans. The specimens were from 216 unrelated individuals and 33 paternity inclusion families. Extracted DNA from EDTA blood was restricted by Hinfl and electrophoresed in 0.7% agarose gel, transferred and hybridized with chemiluminescent probes. Heterozygosity was over 90% by all of the probes. Total numbers of unassignable mutant bands from 33 paternity inclusion cases were 5, and the highest mutation rate was determined in probe MS1(0.045). The probability of having the same DNA band between two unrelated individuals was 5.7 x 10(-10) when four SLPs were used at the same time. The data presented here on allele frequencies and mutation rates provide preliminary data supporting the validity of these probes in paternity analysis and forensic investigators in the Korean population.
PMCID: PMC3054068  PMID: 8593203
14.  Lymphomatoid granulomatosis with isolated involvement of the brain--case report. 
Lymphomatoid granulomatosis usually presents as a primary lung affliction with secondary metastatic spread to the central nervous system(CNS), and its initial manifestation purely as a CNS disease is rare. A 57-year-old man with histologically proven lymphomatoid granulomatosis of the brain as the sole manifestation of the disease is presented.
PMCID: PMC3049706  PMID: 1777130
15.  Recovery of inspiratory intercostal muscle activity following high cervical hemisection 
Anatomical and neurophysiological evidence indicates that thoracic interneurons can serve a commissural function and activate contralateral motoneurons. Accordingly, we hypothesized that respiratory-related intercostal (IC) muscle electromyogram (EMG) activity would be only modestly impaired by a unilateral cervical spinal cord injury. Inspiratory tidal volume (VT) was recorded using pneumotachography and EMG activity was recorded bilaterally from the 1st to 2nd intercostal space in anesthetized, spontaneously breathing rats. Studies were conducted at 1–3 days, 2 wks or 8 wks following C2 spinal cord hemisection (C2HS). Data were collected during baseline breathing and a brief respiratory challenge (7% CO2). A substantial reduction in inspiratory intercostal EMG bursting ipsilateral to the lesion was observed at 1–3 days post-C2HS. However, a time-dependent return of activity occurred such that by 2 wks post-injury inspiratory intercostal EMG bursts ipsilateral to the lesion were similar to age-matched, uninjured controls. The increases in ipsilateral intercostal EMG activity occurred in parallel with increases in VT following the injury (R = 0.55; P < 0.001). We conclude that plasticity occurring within a “crossed-intercostal” circuitry enables a robust, spontaneous recovery of ipsilateral intercostal activity following C2HS in rats.
doi:10.1016/j.resp.2012.06.006
PMCID: PMC4288928  PMID: 22705013
Spinal cord injury; Intercostal; Plasticity
17.  Effects of Dietary Fermented Chlorella vulgaris (CBT®) on Growth Performance, Relative Organ Weights, Cecal Microflora, Tibia Bone Characteristics, and Meat Qualities in Pekin Ducks 
Fermented Chlorella vulgaris was examined for its effects on growth performance, cecal microflora, tibia bone strength, and meat qualities in commercial Pekin ducks. A total of three hundred, day-old male Pekin ducks were divided into three groups with five replicates (n = 20 ducklings per replicate) and offered diets supplemented with commercial fermented C. vulgaris (CBT®) at the level of 0, 1,000 or 2,000 mg/kg, respectively for 6 wks. The final body weight was linearly (p = 0.001) increased as the addition of fermented C. vulgaris into diets increased. Similarly, dietary C. vulgaris linearly increased body weight gain (p = 0.001) and feed intake (p = 0.001) especially at the later days of the feeding trial. However, there was no C. vulgaris effect on feed efficiency. Relative weights of liver were significantly lowered by dietary fermented C. vulgaris (linear effect at p = 0.044). Dietary fermented C. vulgaris did not affect total microbes, lactic acid bacteria, and coliforms in cecal contents. Finally, meat quality parameters such as meat color (i.e., yellowness), shear force, pH, or water holding capacity were altered by adding fermented C. vulgaris into the diet. In our knowledge, this is the first report to show that dietary fermented C. vulgaris enhanced meat qualities of duck meats. In conclusion, our study indicates that dietary fermented C. vulgaris exerted benefits on productivity and can be employed as a novel, nutrition-based strategy to produce value-added duck meats.
doi:10.5713/ajas.14.0473
PMCID: PMC4283193  PMID: 25557680
Fermented Chlorella vulgaris; Meat Quality; Performance; Pekin Duck
18.  THE ROLE OF ANDROGEN RECEPTOR IN TRANSCRIPTIONAL MODULATION OF CANNABINOID RECEPTOR TYPE 1 GENE IN RAT TRIGEMINAL GANGLIA 
Neuroscience  2013;254:10.1016/j.neuroscience.2013.09.014.
We have previously shown that anti-hyperalgesic effects of cannabinoid agonists under inflammatory condition are much greater in male than female, and that inflammatory cytokines upregulate cannabinoid receptor type 1 (CB1) expression in male, but not female, trigeminal ganglia (TG) in a testosterone-dependent manner. In this study, we investigated the mechanisms underlying the testosterone-mediated regulation of peripheral CB1 expression. We hypothesized that testosterone upregulates CB1 through transcriptional modulation by androgen receptor (AR). Interleukin-1 beta (IL-1β), a proinflammatory cytokine, upregulated CB1 mRNA expression in TG of male rats. The cytokine-induced upregulation was prevented by the pre-treatment with flutamide, a specific antagonist for AR, but not by ICI 182,780, a specific antagonist for estrogen receptor, suggesting that the effects of testosterone are not mediated by estradiol, a testosterone metabolite. The expression levels of AR and IL-1β receptors were comparable between male and female TG, suggesting that the male specific IL-1β effects on CB1 upregulation occurs downstream to these receptors. The chromatin immunoprecipitation assay showed AR binding to the CB1 promoter in the rat TG. Furthermore, luciferase reporter assay revealed that AR activated the CB1 gene in response to testosterone or dihydrotestosterone treatment. These experiments provided compelling evidence that testosterone regulates CB1 gene transcription in TG through AR following cytokine stimulation. These results should provide mechanistic bases for understanding cytokine–hormone–neuron interactions in peripheral cannabinoid systems, and have important clinical implications for pain patients in whom testosterone level is naturally low, gradually declining or pharmacologically compromised.
doi:10.1016/j.neuroscience.2013.09.014
PMCID: PMC3870904  PMID: 24055403
testosterone; inflammation; cytokines; sensory neurons; rat
19.  Lack of positive effect of intravitreal bevacizumab in central serous chorioretinopathy: meta-analysis and review 
Eye  2013;27(12):1339-1346.
Purpose
To review and evaluate the effects of intravitreal bevacizumab injection (IVB) in centralserous chorioretinopathy (CSC) by meta-analysis.
Patients and methods
Clinical controlled studies that evaluated the effect of IVB in CSC were identified through systematic searches of Embase, PubMed, and the Cochrane Central Register of Controlled Trials. Data on the best-corrected visual acuity (BCVA) in logMAR and central macular thickness (CMT) in μm at baseline and 6 months after IVB were extracted and compared with those treated by simple observation.
Results
Four clinical controlled studies were included in the meta-analysis. The IVB injection group achieved better BCVA at a follow-up of 6 months. However, the analysis showed that there were no significant differences of BCVA at 6 months after injection between IVB group and the observation group (−0.02 logMAR, 95% CI −0.14 to 0.11, P=0.80). The analysis of the reduction in CMT revealed that the difference between groups was not statistically significant (−8.37 μm, 95% CI −97.26 to 80.52, P=0.85). No report assessed severe complications or side effects of IVB in patients with CSC.
Conclusions
Meta-analysis failed to verify the positive effect of IVB in CSC based on the epidemiological literature published to date.
doi:10.1038/eye.2013.236
PMCID: PMC3869506  PMID: 24202051
central serous chorioretinopathy; intravitreal bevacizumab injection; meta-analysis
20.  Evaluation of Dietary Multiple Enzyme Preparation (Natuzyme) in Laying Hens 
The current experiment was designed to evaluate the efficacy of adding the multi-enzyme mixture (Natuzyme) into layers’ diets with different levels of energy and available phosphorus in relation to laying performance, egg qualities, blood cholesterol level, microflora and intestinal viscosity. Two hundred and fifty 43-wk-old Hy-Line commercial layers were divided into five groups with five replicates per group (10 birds per replicate) and fed one of five experimental diets. A corn and soybean meal-based control diet was formulated and used as a control diet. Two experimental control diets were formulated to reduce energy and crude protein contents (rE) or energy, crude protein and phosphorus contents (rEP). In addition, Natuzyme was added into either rE (rE-Natu500) or rEP (rEP-Natu500) diet to reach a concentration of 500 mg per kg of diet. The experiment lasted 8 weeks. There were no significant differences in feed intake, egg production, egg weight, egg qualities such as eggshell color or Haugh unit, total cholesterol, relative organ weights and cecal microflora profiles between any dietary treatments. Natu500 supplementation into the rE diet, but not rEP diet significantly increased egg mass and eggshell qualities such as strength and thickness, but it decreased cecal ammonia concentration and intestinal viscosity in laying hens. In conclusion, the present study shows that adding multiple enzyme preparation could improve performance of laying hens fed energy and protein restricted diets.
doi:10.5713/ajas.2014.14294
PMCID: PMC4213687  PMID: 25358369
Multiple Enzyme Preparation; Egg Qualities; Performance; Intestinal Viscosity
21.  Impaired Transcriptional Response of the Murine Heart To Cigarette Smoke in the Setting of High Fat Diet and Obesity 
Chemical research in toxicology  2013;26(7):1034-1042.
Smoking and obesity are each well-established risk factors for cardiovascular heart disease, which together impose earlier onset and greater severity of disease. To identify early signaling events in the response of the heart to cigarette smoke exposure within the setting of obesity, we exposed normal weight and high fat diet-induced obese (DIO) C57BL/6 mice to repeated inhaled doses of mainstream (MS) or sidestream (SS) cigarette smoke administered over a two week period, monitoring effects on both cardiac and pulmonary transcriptomes. MS smoke (250 µg wet total particulate matter (WTPM)/L, 5h/day) exposures elicited robust cellular and molecular inflammatory responses in the lung with 1466 differentially expressed pulmonary genes (p<0.01) in normal weight animals, and a muchattenuated response (463 genes) in the hearts of the same animals. In contrast, exposures to SS smoke (85 µg WTPM/L) with an equivalent CO concentration as that of MS smoke (~250 CO ppm), induced a weak pulmonary response (328 genes), but an extensive cardiac response (1590 genes). SS smoke, and to a lesser extent MS smoke preferentially elicited hypoxia- and stress-responsive genes as well as genes predicting early changes of vascular smooth muscle and endothelium, precursors of cardiovascular disease. The most sensitive smoke-induced cardiac transcriptional changes of normal weight mice were largely absent in DIO mice after smoke exposure, while genes involved in fatty acid utilization were unaffected. At the same time, smoke exposure suppressed multiple proteome maintenance genes induced in the hearts of DIO mice. Together these results underscore the sensitivity of the heart to SS smoke and reveal adaptive responses in healthy individuals that are absent in the setting of high fat diet and obesity.
doi:10.1021/tx400078b
PMCID: PMC4234196  PMID: 23786483
heart transcriptome; mainstream cigarette smoke; sidestream cigarette smoke; hypoxia; peroxisomal proliferator-activated receptor-α; high fat diet; obesity
22.  Histomorphometric comparison after fixation with formaldehyde or glyoxal 
Formaldehyde has long been the fixative of choice for histological examination of tissue. The use of alternatives to formaldehyde has grown, however, owing to the serious hazards associated with its use. Companies have striven to maintain the morphological characteristics of formaldehyde-fixed tissue when developing alternatives. Glyoxal-based fixatives now are among the most popular formaldehyde alternatives. Although there are many studies that compare staining quality and immunoreactivity, there have been no studies that quantify possible structural differences. Histomorphometric analysis commonly is used to evaluate diseased tissue. We compared fixation with formaldehyde and glyoxal with regard to the histomorphological properties of plantar foot tissue using a combination of stereological methods and quantitative morphology. We measured skin thickness, interdigitation index, elastic septa thickness, and adipocyte area and diameter. No significant differences were observed between formaldehyde and glyoxal fixation for any feature measured. The glyoxal-based fixative used therefore is a suitable fixative for structural evaluation of plantar soft tissue. Measurements obtained from the glyoxal-fixed tissue can be combined with data obtained from formalin-fixed for analysis.
doi:10.3109/10520295.2010.520275
PMCID: PMC4227596  PMID: 20854226
formaldehyde; glyoxal; heel pad; plantar soft tissue; stereology; structural properties
23.  Protected Organic Acid Blends as an Alternative to Antibiotics in Finishing Pigs 
A total of 120 finishing pigs ([Yorkshire×Landrace]×Duroc) with an average body weight (BW) of 49.72 ±1.72 kg were used in 12-wk trial to evaluate the effects of protected organic acids on growth performance, nutrient digestibility, fecal micro flora, meat quality and fecal gas emission. Pigs were randomly allotted to one of three dietary treatments (10 replication pens with 4 pigs per pen) in a randomly complete block design based on their initial BW. Each dietary treatment consisted of: Control (CON/basal diet), OA1 (basal diet+0.1% organic acids) and OA2 (basal diet+0.2% organic acids). Dietary treatment with protected organic acid blends linearly improved (p<0.001) average daily gain during 0 to 6 week, 6 to 12 week as well as overall with the increase in their inclusion level in the diet. The dry matter, N, and energy digestibility was higher (linear effect, p<0.001) with the increase in the dose of protected organic acid blends during 12 week. During week 6, a decrease (linear effect, p = 0.01) in fecal ammonia contents was observed with the increase in the level of protected organic acid blends on d 3 and d 5 of fermentation. Moreover, acetic acid emission decreased linearly (p = 0.02) on d7 of fermentation with the increase in the level of protected organic acid blends. During 12 weeks, linear decrease (p<0.001) in fecal ammonia on d 3 and d 5 and acetic acid content on d 5 of fermentation was observed with the increase in the level of protected organic acid blends. Supplementation of protected organic acid blends linearly increased the longissimus muscle area with the increasing concentration of organic acids. Moreover, color of meat increased (linear effect, quadratic effect, p<0.001, p<0.002 respectively) and firmness of meat showed quadratic effect (p = 0.003) with the inclusion of increasing level of protected organic acid in the diet. During the 6 week, increment in the level of protected organic acid blends decreased (linear effect, p = 0.01) Escherichia coli (E. coli) counts and increased (linear effect, p = 0.004) Lactobacillus counts. During 12-wk of experimental trial, feces from pigs fed diet supplemented with organic acid blends showed linear reduction (p<0.001) of E. coli counts and the tendency of linear increase (p = 0.06) in Lactobacillus count with the increase in the level of organic acid blends. In conclusion, 0.2% protected organic acids blends positively affected growth performance, nutrient digestibility, fecal gas emission and meat quality in finishing pigs without any adverse effects on blood parameters.
doi:10.5713/ajas.2014.14356
PMCID: PMC4213705  PMID: 25358320
Digestibility; Finishing Pigs; Growth Performance; Micro Flora; Protected Organic Acids
24.  Carcass Characteristics and Meat Quality of Korean Native Ducks and Commercial Meat-type Ducks Raised under Same Feeding and Rearing Conditions 
This study was conducted to compare carcass characteristics and physico-chemical meat quality in two different genotype ducks raised under identical feeding and rearing conditions. A total of ninety 1-d-old Korean native ducks (KND, n = 45) and commercial meat-type ducks (Grimaud, n = 45) were fed same experimental diets during 56 d and 42 d, respectively to obtain similar slaughter weights. The experimental diet for starter period contained 20% crude protein (CP) and 2,900 kcal nitrogen corrected true metabolizable energy (TMEn)/kg of diet and that for grower period contained 17% CP and 3,050 TMEn/kg of diet. Average daily gain and feed efficiency of KND were inferior to those of commercial meat-type ducks (p<0.05). Carcass weight was not different between two genetically different ducks, but carcass yield of KND was significantly higher (p<0.05) than that of commercial meat-type ducks. There were no significant differences in cooking loss and pH of breast meat between two genetically different ducks, but water holding capacity of KND was significantly higher than that of commercial meat-type ducks. The linoleic acid and total polyunsaturated fatty acid of breast meat from KND were significantly higher (p<0.05) than the corresponding part from commercial meat-type ducks. Significant differences were detected in water holding capacity and the content of linoleic acid and polyunsaturated fatty acid, which were significantly higher in KND, whereas growth performance tended to be superior in commercial ducks. At the market weight, the meat from KND was judged to have better qualities with regard to higher water holding capacity and greater content of polyunsaturated fatty acid compare with meat from commercial meat-type duck.
doi:10.5713/ajas.2014.14191
PMCID: PMC4213710  PMID: 25358325
Carcass Characteristic; Meat Quality; Identical Feeding Condition; Commercial Meat-type Duck; Korean Native Duck
25.  Neuritin can normalize neural deficits of Alzheimer's disease 
Cell Death & Disease  2014;5(11):e1523-.
Reductions in hippocampal neurite complexity and synaptic plasticity are believed to contribute to the progressive impairment in episodic memory and the mild cognitive decline that occur particularly in the early stages of Alzheimer's disease (AD). Despite the functional and therapeutic importance for patients with AD, intervention to rescue or normalize dendritic elaboration and synaptic plasticity is scarcely provided. Here we show that overexpression of neuritin, an activity-dependent protein, promoted neurite outgrowth and maturation of synapses in parallel with enhanced basal synaptic transmission in cultured hippocampal neurons. Importantly, exogenous application of recombinant neuritin fully restored dendritic complexity as well as spine density in hippocampal neurons prepared from Tg2576 mice, whereas it did not affect neurite branching of neurons from their wild-type littermates. We also showed that soluble recombinant neuritin, when chronically infused into the brains of Tg2576 mice, normalized synaptic plasticity in acute hippocampal slices, leading to intact long-term potentiation. By revealing the protective actions of soluble neuritin against AD-related neural defects, we provide a potential therapeutic approach for patients with AD.
doi:10.1038/cddis.2014.478
PMCID: PMC4260736  PMID: 25393479

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