Effects of genomic estimated breeding value and dietary energy to protein ratio on growth performance, carcass trait, and lipogenic gene expression in Hanwoo steer.

"Genome-based precision feeding" is a concept that involves the application of customised diets to different genetic groups of cattle. We investigated the effects of the genomic estimated breeding value (gEBV) and dietary energy to protein ratio (DEP) on growth performance, carcass traits, and lipogenic gene expression in Hanwoo (Korean cattle) steers. Forty-four Hanwoo steers (BW = 636 kg, age = 26.9 months) were genotyped using the Illumina Bovine 50 K BeadChip. The gEBV was calculated using genomic best linear unbiased prediction. Animals were separated into high gEBV of marbling score or low-gMS groups based on the upper and lower 50% groupings of the reference population, respectively. Animals were assigned to one of four groups in a 2 × 2 factorial arrangement: high gMS/high DEP (0.084 MJ/g), high gMS/low DEP (0.079 MJ/g), low gMS/high DEP, and low gMS/low DEP. Steers were fed concentrate with a high or low DEP for 31 weeks. The BW tended to be higher (0.05 < P < 0.1) in the high-gMS groups compared to the low-gMS groups at 0, 4, 8, 12, and 20 weeks. The average daily gain (ADG) tended to be lower (P = 0.08) in the high-gMS group than in the low-gMS group. Final BW and measured carcass weight (CW) were positively correlated with the gEBV of carcass weight (gCW). The DEP did not affect ADG. Neither the gMS nor the DEP affected the MS and beef quality grade. The intramuscular fat (IMF) content in the longissimus thoracis (LT) tended to be higher (P = 0.08) in the high-gMS groups than in the low-gMS groups. The mRNA levels of lipogenic acetyl-CoA carboxylase and fatty acid binding protein 4 genes in the LT were higher (P < 0.05) in the high-gMS group than in the low-gMS group. Overall, the IMF content tended to be affected by the gMS, and the genetic potential (i.e., gMS) was associated with the functional activity of lipogenic gene expression. The gCW was associated with the measured BW and CW. The results demonstrated that the gMS and the gCW may be used as early prediction indexes for meat quality and growth potential of beef cattle.

Development of ruminating behavior in Holstein calves between birth and 30 days of age.

Ruminating behavior accompanies the development of the rumen and the intake of solid feed in calves. However, few studies have reported on the emergence and development of rumination. In this study, we observed ruminating behavior changes of 56 Holstein calves (body weight at birth = 40.1 ± 3.96 kg; mean ± standard deviation) from birth to 30 d of age under the feeding management of suckling calves that were only fed pelleted concentrate feed and milk. All calves were housed in individual pens equipped with infrared cameras. We explored feed intake within 30 d of age, body weight on 61 d of age, and other apparent indicators, including the age of first eating the bedding, duration of non-nutritive oral behavior at 25 and 30 d of age, total starter feed intake within 30 d of age, average daily starter feed intake within 30 d of age, and duration of ruminating behavior at 25 and 30 d of age for all calves, to further explore the effects of the age of first ruminating behavior (AFR). The AFR fitted the normal distribution and ranged from 15 to 20 d of age for 50% of the experimental population. The AFR was positively correlated with the age of first eating the bedding and duration of non-nutritive oral behavior at 30 d of age. Total starter feed intake within 30 d of age, average daily starter feed intake within 30 d of age, duration of ruminating behavior at 25 and 30 d of age, and duration of eating the bedding at 25 and 30 d of age were negatively correlated with AFR. Overall, to the best of our knowledge, this is the first study that has analyzed the correlation between AFR and other indicators. We found that earlier AFR was associated with shorter duration of non-nutritive oral behavior, longer durations of rumination and eating the bedding, and higher feed intake by 30 d of age.

[Study on bone volume harvested from the implant sites with different methods].

OBJECTIVE: To compare the volume of autogenous bone particles harvested utilizing different techniques and various implant systems during implant surgery, and to determine the advantageous method to collect autogenous bone particles. METHODS: Homogeneous epoxy resin simulated jaw bone model was enrolled. Bicon, Bego implant systems and Straumann tissue level implant systems were utilized. The two techniques were investigated. One method was low-speed drilling (50 r/min) without water irrigating, and the other one was drilling with cold water irrigating to the ideal depth, then closing the water and drilling out with low speed (50 r/min). The bone particles in the drill groove and implant beds were collected. The volumes of the bone harvested were compared between the different techniques and also among the three implant systems, then they were compared with the volume of the bone harvested by the special bone drill. The sample size of each sub-group was 10. The bone particles were weighed by electronic balance after drying. RESULTS: The harvested bone volume between the latch reamers and hand reamers of Bicon system with the first method was not significantly different. When the same size implant bed was prepared, the volume of the bone particles produced during the implant surgery with low-speed drill without water was significantly higher than that with the other method no matter Bicon [3.5 mm×10 mm hole for example (28.42±6.04) mg vs. (6.30±2.51) mg, P<0.001] or Bego system [2.8 mm×10 mm hole for example (28.95±5.39) mg vs. (4.61±3.39) mg, P<0.001] was used, and the ratio of bone volume between the first method and the second one was approximately 3.3 to 7.0 times. When using the second method to prepare the similar size implant bed, the bone volume was not significant different among Bicon, Bego and Straumann implant systems [Bicon (9.90±3.42) mg, Bego (8.70±4.09) mg, and Straumann (10.56±5.66) mg, P=0.69]. When preparing a 5 mm-diameter-10 mm-length hole with Bicon implant system and a 4.7 mm-diameter-10 mm-length with Bego implant system, the bone quantity harvested from each group was less than that harvested by special bone drill from Neo Biotech [Bicon (82.54±12.26) mg, Bego (85.07±12.64) mg vs. Neo Biotech (96.78±13.19) mg, P<0.05]. CONCLUSION: More autogenous bone can be harvested from implant beds by preparing with low-speed rolling without water than the method with water irrigation. When utilizing the same preparing method, the implant system has no impact on the volume of the bone harvested.

[Radiography study on osteotome sinus floor elevation with placed implant simultaneously with no graft augmentation].

OBJECTIVE: To investigate the change of endo-sinus bone height and bone volume in osteotome sinus floor elevation (OSFE) without bone graft but placing implants simultaneously by using cone beam computed tomography (CBCT) and three dimensional analysis, and to find the impacting factors on endo-sinus bone augmentation. METHODS: OSFE was performed in 38 edentulous patients with missing teeth at posterior maxillary region, and 44 implants were placed and referred for OSFE using no graft materials. CBCT was performed pre-surgery and 9－68 months post-surgery when the patients encountered another implant surgery. The gained bone height at mesial, distal, buccal and palatal sites around the implant in sinus were measured, volumetric measurements of the endo-sinus gained bone volume (ESGBV) in the elevated region were calculated by Mimics software. Univariate analysis and multiple linear regression were performed to investigate the impacting factors on the gained bone height and ESGBV. Marginal bone loss was recorded according to the periapical radiography after implant restoration. RESULTS: The mean residual bone height (RBH) pre-surgery was (3.41±1.23) mm, the mean protruded length (PL) into sinus of implant post-surgery was (3.41±1.28) mm, the mean endo-sinus gained bone height was (2.44±1.23) mm at distal sites, (2.88±1.20) mm at mesial sites, (2.83±1.22) mm at buccal sites and (2.96±1.16) mm at palatal sites, the mean endo-sinus gained bone height at distal sites was significantly lower than the other three sites (P < 0.05). The average endo-sinus gained bone height was (2.78±1.13) mm. The mean ESGBV was (122.15± 73.27) mm3. Univariate analysis showed the more RBH, the less bone height gained in sinus, which existed at buccal, lingual, mesial and distal sites (P < 0.001), and the more RBH, the smaller ESGBV gained (P=0.012). The ESGBV was significantly higher in the subjects whose bone generation period was more than 24 months than those whose bone generation period less than 24 months (P=0.034). The more PL, the more bone height and ESGBV gained (P=0.008). Multivariate analysis showed after adjusting factors of gender, age, smoking, width of sinus floor, thickness of sinus membrane pre-surgery, diameter and length of the implant, PL and bone generation period was positively correlated with mean endo-sinus gained bone height and ESGBV, while RBH negatively correlated with mean endo-sinus gained bone height. During the follow-up, the mean marginal bone loss was 0 (0－1.41) mm and all the implants loaded successfully. CONCLUSION: OSFE without bone graft but with placed implant simultaneously can increase endo-sinus gained bone height and ESGBV. RBH, PL and bone generation period are the significant factors impacting endo-sinus bone augmentation.

TRIENNIAL GROWTH AND DEVELOPMENT SYMPOSIUM: Molecular mechanisms related to bovine intramuscular fat deposition in the longissimus muscle.

The intramuscular fat (IMF) content of the LM, also known as marbling, is particularly important in determining the price of beef in Korea, Japan, and the United States. Deposition of IMF is influenced by both genetic (e.g., breed, gender, and genotype) and nongenetic factors (e.g., castration, nutrition, stressors, animal weight, and age). Castration of bulls markedly increases deposition of IMF, resulting in improved beef quality. Here, we present a comparative gene expression approach between bulls and steers. Transcriptomic and proteomic studies have demonstrated that the combined effects of increases in lipogenesis, fatty acid uptake, and fatty acid esterification and decreased lipolysis are associated with increased IMF deposition in the LM. Several peripheral tissues (LM, adipose tissues, and the liver) are involved in lipid metabolism. Therefore, understanding the significance of the tissue network in lipid metabolism is important. Here, we demonstrate that lipid metabolism in LM tissues is crucial for IMF deposition, whereas lipid metabolism in the liver plays only a minor role. Metabolism of body fat and IMF deposition in bovine species has similarities with these processes in metabolic diseases, such as obesity in humans and rodents. Extensive studies on metabolic diseases using epigenome modification (DNA methylation, histone modification, and microRNA), microbial metagenomics, and metabolomics have been performed in humans and rodents, and new findings have been reported using these technologies. The importance of applying "omics" fields (epigenomics, metagenomics, and metabolomics) to the study of IMF deposition in cattle is described. New information on the molecular mechanisms of IMF deposition may be used to design nutritional or genetic methods to manipulate IMF deposition and to modify fatty acid composition in beef cattle. Applying nutrigenomics could maximize the expression of genetic potential of economically important traits (e.g., marbling) in animals.

Effects of Ambient Temperature on Growth Performance, Blood Metabolites, and Immune Cell Populations in Korean Cattle Steers.

Exposure to cold may affect growth performance in accordance with the metabolic and immunological activities of animals. We evaluated whether ambient temperature affects growth performance, blood metabolites, and immune cell populations in Korean cattle. Eighteen Korean cattle steers with a mean age of 10 months and a mean weight of 277 kg were used. All steers were fed a growing stage-concentrate diet at a rate of 1.5% of body weight and Timothy hay ad libitum for 8 weeks. Experimental period 1 (P1) was for four weeks from March 7 to April 3 and period 2 (P2) was four weeks from April 4 to May 1. Mean (8.7°C) and minimum (1.0°C) indoor ambient temperatures during P1 were lower (p<0.001) than those (13.0°C and 6.2°C, respectively) during P2. Daily dry matter feed intake in both the concentrate diet and forage groups was higher (p<0.001) during P2 than P1. Average daily weight gain was higher (p<0.001) during P2 (1.38 kg/d) than P1 (1.13 kg/d). Feed efficiency during P2 was higher (p = 0.015) than P1. Blood was collected three times; on March 7, April 4, and May 2. Nonesterified fatty acids (NEFA) were higher on March 7 than April 4 and May 2. Blood cortisol, glucose, and triglyceride concentrations did not differ among months. Blood CD4+, CD8+, and CD4+CD25+ T cell percentages were higher, while CD8+CD25+ T cell percentage was lower, during the colder month of March than during May, suggesting that ambient temperature affects blood T cell populations. In conclusion, colder ambient temperature decreased growth and feed efficiency in Korean cattle steers. The higher circulating NEFA concentrations observed in March compared to April suggest that lipolysis may occur at colder ambient temperatures to generate heat and maintain body temperature, resulting in lower feed efficiency in March.

Low rate of advanced adenoma formation during a 5-year colonoscopy surveillance period after adequate polypectomy of non-advanced adenoma.

AIM: Patients with non-advanced adenoma (NAA) underwent surveillance colonoscopy at intervals shorter than the interval recommended by the guidelines. We aimed to assess the incidence of recurrent advanced adenoma (AA) over a 5-year period and to identify risk factors for recurrence. METHOD: Patients with and without NAA identified at baseline colonoscopy who had had at least two colonoscopy examinations during the subsequent 5 years were included in the study. Data on the patients' demographics and colorectal findings were extracted from a specially designed colonoscopy database. The primary outcome was the finding of recurrent AA formation. Multivariate analysis was used to identify factors that predict subsequent AA formation at surveillance colonoscopy. RESULTS: Among 43 155 colonoscopy procedures, 828 cases were identified with NAA (374) and without an adenoma (454). Forty-eight (51.1%) of 94 received a follow-up colonoscopy within 1 year due to an inadequate baseline colonoscopy. Patients with NAA at baseline had a low incidence of AA at an interval of 1-5 years which was not statistically different from patients without adenoma formation at the initial baseline colonoscopy (1.5% vs 2.2%, P = 0.51). The incidence of AA at follow-up colonoscopy performed at 1-3 years and 3-5 years in patients with a baseline NAA was 1.7% and 1.4% (P = 0.59). Age over 50 years and male gender were independent risk factors for AA recurrence within 5 years. CONCLUSION: Surveillance colonoscopy within 5 years is of little benefit to patients who had an adequate polypectomy of an NAA at baseline. Too frequent reexamination due to concerns about AA recurrence should be avoided.

Seasonal variation in carcass characteristics of korean cattle steers.

Climate temperature affects animal production. This study was conducted to evaluate whether climatic conditions affect beef carcass characteristics of Korean cattle steers. The monthly carcass characteristics of Korean cattle steers (n = 2,182,415) for 8 yr (2006 through 2013) were collected from the Korean Institute for Animal Products Quality Evaluation. Daily climate temperature (CT) and relative humidity (RH) data were collected from the Korean Meteorological Administration. Weather conditions in South Korea during summer were hot and humid, with a maximum temperature of 28.4°C and a maximum RH of 91.4%. The temperature-humidity index (THI), calculated based on CT and RH, ranges from 73 to 80 during summer. Winter in South Korea was cold, with a minimum temperature of -4.0°C and a wind-chill temperature of -6.2°C. Both marbling score (MS) and quality grade (QG) of Korean cattle steer carcasses were generally best (p<0.05) in autumn and worst in spring. A correlation analysis showed that MS and QG frequencies were not associated (p>0.05) with CT. Yield grade (YG) of Korean cattle steer carcasses was lowest (p<0.05) in winter (November to January) and highest in spring and summer (May to September). A correlation analysis revealed that YG frequency was strongly correlated (r≥0.71; p<0.01) with CT and THI values. The rib eye area, a positive YG parameter, was not associated with CT. Backfat thickness (BT), a negative YG factor, was highest in winter (November and December). The BT was strongly negatively correlated (r≤-0.74; p<0.01) with CTs. Therefore, the poor YG during winter is likely due in part to the high BT. In conclusion, YG in Korean cattle steer carcasses was worst in winter. QGs were not associated with winter or summer climatic conditions.

Association of DNA Methylation Levels with Tissue-specific Expression of Adipogenic and Lipogenic Genes in Longissimus dorsi Muscle of Korean Cattle.

Epigenetic factors, such as DNA methylation status, may regulate adipogenesis and lipogenesis, thus affecting intramuscular fat (IMF) deposition in longissimus dorsi muscle (LM) of beef cattle. In Korean cattle steers, the LM consists mainly of muscle tissue. However, the LM tissue also contains IMF. We compared the gene expression levels between the IMF and muscle portions of the LM after tissue separation. Real-time polymerase chain reaction analysis showed that the mRNA levels of both adipogenic peroxisome proliferator-activated receptor gamma isoform 1 (PPARG1) and lipogenic fatty acid binding protein 4 (FABP4) were higher (p<0.01) in the IMF than in the muscle portion of the LM. We determined DNA methylation levels of regulatory regions of the PPARG1 and FABP4 genes by pyrosequencing of genomic DNA. DNA methylation levels of two of three CpG sites in the PPARG1 gene promoter region were lower (p<0.05) in the IMF than in the muscle portion of the LM. DNA methylation levels of all five CpG sites from the FABP4 gene promoter region were also lower (p<0.001) in the IMF than in the muscle portion. Thus, mRNA levels of both PPARG1 and FABP4 genes were inversely correlated with DNA methylation levels in regulatory regions of CpG sites of the corresponding gene. Our findings suggest that DNA methylation status regulates tissue-specific expression of adipogenic and lipogenic genes in the IMF and muscle portions of LM tissue in Korean cattle.

Gas chromatograph-surface acoustic wave for quick real-time assessment of blood/exhaled gas ratio of propofol in humans.

BACKGROUND: Although pilot studies have reported that exhaled propofol concentrations can reflect intraoperative plasma propofol concentrations in an individual, the blood/exhaled partial pressure ratio RBE varies between patients, and the relevant factors have not yet been clearly addressed. No efficient method has been reported for the quick evaluation of RBE and its association with inter-individual variables. METHODS: We proposed a novel method that uses a surface acoustic wave (SAW) sensor combined with a fast gas chromatograph (GC) to simultaneously detect propofol concentrations in blood and exhaled gas in 28 patients who were receiving propofol i.v. A two-compartment pharmacokinetic (PK) model was established to simulate propofol concentrations in exhaled gas and blood after a bolus injection. Simulated propofol concentrations for exhaled gas and blood were used in a linear regression model to evaluate RBE. RESULTS: The fast GC-SAW system showed reliability and efficiency for simultaneous quantitative determination of propofol in blood (correlation coefficient R(2)=0.994, P<0.01) and exhaled gas (R(2)=0.991, P<0.01). The evaluation of RBE takes <50 min for a patient. The distribution of RBE in 28 patients showed inter-individual differences in RBE (median 1.27; inter-quartile range 1.07-1.59). CONCLUSIONS: Fast GC-SAW, which analyses samples in seconds, can perform both rapid monitoring of exhaled propofol concentrations and fast analysis of blood propofol concentrations. The proposed method allows early determination of the coefficient RBE in individuals. Further studies are required to quantify the distribution of RBE in a larger cohort and assess the effect of other potential factors. CLINICAL TRIAL REGISTRATION: ChiCTR-ONC-13003291.

Epigenetic modification of Nrf2 in 5-fluorouracil-resistant colon cancer cells: involvement of TET-dependent DNA demethylation.

5-Fluorouracil (5-FU) is a widely used anticancer drug for the treatment of colorectal cancer (CRC). However, resistance to 5-FU often prevents the success of chemotherapy. Nuclear factor-erythroid 2-related factor 2 (Nrf2) is a transcriptional regulator and a possible target to overcome 5-FU resistance. The present study examined epigenetic changes associated with Nrf2 induction in a human CRC cell line (SNUC5) resistant to 5-FU (SNUC5/5-FUR). Nrf2 expression, nuclear translocation, and binding to promoter were higher in SNUC5/5-FUR cells than in SNUC5 cells. The activated Nrf2 in SNUC5/5-FUR cells led to an increase in the protein expression and activity of heme oxygenase-1 (HO-1), an Nrf2-regulated gene. SNUC5/5-FUR cells produced a larger amount of reactive oxygen species (ROS) than SNUC5 cells. The siRNA- or shRNA-mediated knockdown of Nrf2 or HO-1 significantly suppressed cancer cell viability and tumor growth in vitro and in vivo, resulting in enhanced 5-FU sensitivity. Methylation-specific (MS) or real-time quantitative MS-PCR data showed hypomethylation of the Nrf2 promoter CpG islands in SNUC5/5-FUR cells compared with SNUC5 cells. Expression of the DNA demethylase ten-eleven translocation (TET) was upregulated in SNUC5/5-FUR cells. ROS generated by 5-FU upregulated TET1 expression and function, whereas antioxidant had the opposite effect. These results suggested that the mechanism underlying the acquisition of 5-FU resistance in CRC involves the upregulation of Nrf2 and HO-1 expression via epigenetic modifications of DNA demethylation.

Gracilaria bursa-pastoris (Gmelin) Silva extract attenuates ultraviolet B radiation-induced oxidative stress in human keratinocytes.

The purpose of this study was to assess the protective effects of an ethanol extract derived from the red alga Gracilaria bursa-pastoris (Gmelin) Silva (GBE) on ultraviolet B (UVB)-irradiated human HaCaT keratinocytes. GBE exhibited scavenging activity against intracellular reactive oxygen species that were induced by either hydrogen peroxide or UVB radiation. In addition, both the superoxide anion and the hydroxyl radical were scavenged by GBE in cell-free systems. GBE absorbed light in the UVB range (280-320 nm) of the electromagnetic spectrum and lessened the extent of UVB-induced oxidative damage to cellular lipids, proteins, and DNA. Finally, GBE-treated keratinocytes showed a reduction in UVB-induced apoptosis, as exemplified by fewer apoptotic bodies. These results suggest that GBE exerts cytoprotective actions against UVB-stimulated oxidative stress by scavenging ROS and absorbing UVB rays, thereby attenuating injury to cellular constituents and preventing cell death.

A clinical prediction model and its application for bleeding in chronic liver failure patients with esophageal varices.

OBJECTIVES: We aimed to explore the model for end-stage liver disease (MELD), Child-Turcotte-Pugh (CTP) score, endotoxin, bleeding score and dynamic changes of D-dimer in chronic liver failure patients with portal hypertension and esophageal varices, and explored their potential contact with bleeding in short-term prognosis. PATIENTS AND METHODS: Chronic liver failure patients with esophageal varices were divided into 2 groups: bleeding group (Group A, n=50) and non-bleeding group (Group B, n=50). MELD, CTP score, endotoxin and plasma D-dimer was compared at different time point. The receiver operating characteristic curve (ROC) was drawn. The predictive model based on their cut off value in esophageal varices bleeding patients was evaluated. RESULTS: Infection and endotoxin levels were related with bleeding and death in chronic liver failure patients with esophageal varices, and affect the patient's coagulation and fibrinolysis activity. The criteria of predictive model for predicting hemorrhage of esophageal varices in patients with chronic liver failure is: MELD ≥ 26; bleeding grading score ≥ 10; and/or plasma D-dimer > 700 ug/L. Plasma D-dimer, MELD score and death rate showed significant differences between two groups. Patients with chronic liver failure occurred bleeding and eventually dead have persistent anomaly plasma D-dimer level. In our model-group patients, the D-dimer and CTP score has statistical difference between surviving and death patients (p < 0.05). CONCLUSIONS: This model could predict the prognosis of bleeding in chronic liver failure patients with esophageal varices. And has the short-term prognostic value for clinical application.

Volatile anesthetic isoflurane inhibits LTP induction of hippocampal CA1 neurons through α4ß2 nAChR subtype-mediated mechanisms.

BACKGROUND AND PURPOSE: Volatile anesthetic isoflurane contributes to postoperative cognitive dysfunction and inhibition of long-term potentiation (LTP), a synaptic model of learning and memory, but the mechanisms are uncertain. Central neuronal α4ß2 subtype nicotinic acetylcholine receptors (nAChRs) are involved in the induction of LTP in the hippocampus. Isoflurane inhibits α4ß2 nAChRs at concentrations lower than those used for anesthesia. Therefore, we hypothesized that isoflurane-inhibited LTP induction of hippocampal CA1 neurons via α4ß2 nAChRs subtype inhibition. METHODS: Transverse hippocampal slices (400µm thick) were obtained from male rats (6-8 weeks old). Population spikes were evoked using extracellular electrodes by electrical stimulation of the Schaffer collateral-commissural pathway of rat hippocampal slices. LTP was induced using high frequency stimulation (HFS; 100Hz, 1s). Clinically relevant concentrations (0.125-0.5mM) of isoflurane with or without nicotine (nAChRs agonist), mecamylamine (nAChRs antagonist), 3-[2(S)-2-azetidinylmethoxy] pyridine (A85380) and epibatidine (α4ß2 nAChRs agonist), dihydro ß erythroidine (DHßE) (α4ß2 nAChRs antagonist) were added to the perfusion solution 20min before HFS to test their effects on LTP by HFS respectively. RESULTS: A brief HFS induced stable LTP in rat hippocampal slices, but LTP was significantly inhibited in the presence of isoflurane at concentrations of 0.125-0.5mM. The inhibitive effect of isoflurane on LTP was not only reversible and could be prevented by nAChRs agonist nicotine and α4ß2 nAChRs agonist A85380 and epibatidine, but also mimicked and potentiated by nAChRs antagonist mecamylamine and α4ß2 nAChRs antagonist DHßE. CONCLUSIONS: Inhibition of α4ß2 nAChRs subtype of hippocampus participates in isoflurane-mediated LTP inhibition.

Characterization of Cellulolytic and Xylanolytic Enzymes of Bacillus licheniformis JK7 Isolated from the Rumen of a Native Korean Goat.

A facultative bacterium producing cellulolytic and hemicellulolytic enzymes was isolated from the rumen of a native Korean goat. The bacterium was identified as a Bacillus licheniformis on the basis of biochemical and morphological characteristics and 16S rDNA sequences, and has been designated Bacillus licheniformis JK7. Endoglucanase activities were higher than those of ß-glucosidase and xylanase at all temperatures. Xylanase had the lowest activity among the three enzymes examined. The optimum temperature for the enzymes of Bacillus licheniformis JK7 was 70°C for endoglucanase (0.75 U/ml) and 50°C for ß-glucosidase and xylanase (0.63 U/ml, 0.44 U/ml, respectively). All three enzymes were stable at a temperature range of 20 to 50°C. At 50°C, endoglucanse, ß-glucosidase, and xylanase had 90.29, 94.80, and 88.69% residual activity, respectively. The optimal pH for the three enzymes was 5.0, at which their activity was 1.46, 1.10, and 1.08 U/ml, respectively. The activity of all three enzymes was stable in the pH range of 3.0 to 6.0. Endoglucanase activity was increased 113% by K(+), while K(+), Zn(+), and tween 20 enhanced ß-glucosidase activity. Xylanase showed considerable activity even in presence of selected chemical additives, with the exception of Mn(2+) and Cu(2+). The broad range of optimum temperatures (20 to 40°C) and the stability under acidic pH (4 to 6) suggest that the cellulolytic enzymes of Bacillus licheniformis JK7 may be good candidates for use in the biofuel industry.

Arg555Gln mutation of TGFBI gene in geographical-type Reis-Bücklers corneal dystrophy in a Chinese family.

OBJECTIVE: Mutations of the transforming growth factor ß-induced (TGFBI) gene were studied in a Chinese family with Reis-Bücklers corneal dystrophy (RBCD). METHODS: Six family members with RBCD and six unaffected family members were investigated. The pedigree showed a typical dominant inheritance pattern. Genomic DNA was extracted from peripheral leucocytes from all study participants. Exons 4, 12 and 14 of the TGFBI gene were analysed using polymerase chain reaction, and standard automated sequencing was performed. Corneal tissue sampled from the proband during phototherapeutic keratectomy was examined using transmission electron microscopy (TEM). RESULTS: A typical geographical pattern of fine opacities in Bowman's layer of the cornea was seen in all six patients on slit-lamp examination. An Arg555Gln (R555Q) mutation of the TGFBI gene was identified in all six patients but was absent in all unaffected family members. TEM revealed rod-shaped bodies in Bowman's layer of the cornea. CONCLUSIONS: In this Chinese family an R555Q mutation of the TGFBI gene was associated with RBCD. As the RBCD phenotype is usually associated with an R124L mutation, this novel genotype-phenotype correlation may prompt further investigation of Bowman's layer corneal dystrophy.

Morphological comparison of small nerve fibres in gastric mucosa in non-diabetic and Type 2 diabetic subjects.

AIM: To determine changes in small nerve fibres in gastric mucosa in patients with Type 2 diabetes by morphological observation. METHODS: In twenty-five non-diabetic and 21 Type 2 diabetic participants, gastric mucosal biopsy under endoscopy was performed. Innervation in gastric mucosa was detected using immunohistochemical staining. Anti-protein gene product (PGP) 9.5 positive nerves underwent morphological observation and quantitative analysis. RESULTS: Small nerve fibres in gastric mucosa were shortened in the diabetic subjects. The ratio of gastric mucosal protrusions maintaining nerve fibres between gastric pits to total observed protrusions was lower in patients with Type 2 diabetes compared with the non-diabetic subjects (ratio of innervated protrusion/total protrusion: 0.49 +/- 0.12 vs. 0.89 +/- 0.06, P < 0.05). CONCLUSIONS: This study sets the scene for further research to investigate the relationship between gastric mucosal nerves and autonomic neuropathy or diabetic peripheral neuropathy.

Orientational distributions and nematic order of rodlike magnetic nanoparticles in dispersions.

Using small-angle neutron scattering (SANS), we have investigated the orientational order of iron nanoparticles dispersed in cyclohexanone. The particles have rodlike shape and size distributions with an average length of 200 nm and an average diameter of 25 nm. SANS shows an anisotropy, which is a measure of orientational order, in magnetic dispersions with a volume fraction of 3.2% and 3.9% iron particles in shear flow and/or magnetic field. The scattering anisotropy can be fitted by a model assuming an Onsager distribution of the orientation of the particles in shear flow. The orientational distribution of particles oriented by a magnetic field can be described by a different model assuming the Maier-Saupe orientational distribution for uniaxial ferromagnetic particles. The orientational distribution parameter m for the Maier-Saupe distribution or alpha for the Onsager distribution and the orientational order parameter S have been determined at shear rates gamma[over ] of to 0-4000 s(-1) and in magnetic fields of 0-18 mT. The S values indicate that the particles start to orient either in a shear flow of 100 s(-1) or in a magnetic field of 6 mT. Applying only shear results in an orientational order, with the dispersion returning to the disordered state when the shear rate is decreased to zero. In sharp contrast, application of magnetic fields greater than 6 mT results in orientational order in the field-increasing cycle, and two-thirds of the orientational order remains when the field is decreased to zero. This shows that the order in a magnetic field is different from the order in a shear flow, the action of magnetizing the particles along a certain direction is irreversible, and the orientational order parameter exhibits hysteresis.

Shear- and magnetic-field-induced ordering in magnetic nanoparticle dispersion from small-angle neutron scattering.

Small-angle neutron scattering experiments have been performed to investigate orientational ordering of a dispersion of rod-shaped ferromagnetic nanoparticles under the influence of shear flow and static magnetic field. In this experiment, the flow and flow gradient directions are perpendicular to the direction of the applied magnetic field. The scattering intensity is isotropic in zero-shear-rate or zero-applied-field conditions, indicating that the particles are randomly oriented. Anisotropic scattering is observed both in a shear flow and in a static magnetic field, showing that both flow and field induce orientational order in the dispersion. The anisotropy increases with the increase of field and with the increase of shear rate. Three states of order have been observed with the application of both shear flow and magnetic field. At low shear rates, the particles are aligned in the field direction. When increasing shear rate is applied, the particles revert to random orientations at a characteristic shear rate that depends on the strength of the applied magnetic field. Above the characteristic shear rate, the particles align along the flow direction. The experimental results agree qualitatively with the predictions of a mean field model.