Therapeutic effects of kefir grain Lactobacillus-derived extracellular vesicles in mice with 2,4,6-trinitrobenzene sulfonic acid-induced inflammatory bowel disease.

Kefir is a fermented product from yeast and lactic acid bacteria, and has been associated with various health benefits including relieving inflammatory bowel disease. Recently, it has been shown that gram-positive bacteria produce extracellular vesicles (EV). The EV could be appearing as potentially important mediators of cell to cell interaction. In this study, we explored the role of kefir grain Lactobacillus-derived EV in modulating inflammation responses via alleviating the production of inflammatory cytokines in tumor necrosis factor-α (TNF-α)-induced inflammation in Caco-2 cells and the 2,4,6-trinitrobenzene sulfonic acid-induced inflammatory bowel disease mouse model. Kefir-derived Lactobacillus EV were isolated by ultracentrifugation of the culture medium of 3 different kefir-derived strains (i.e., Lactobacillus kefir, Lactobacillus kefiranofaciens, and Lactobacillus kefirgranum). Nanoparticle tracking analysis showed that the size of isolated kefir-derived Lactobacillus EV was within 80 to 400 nm, and kefir-derived Lactobacillus EV uptake into recipient Caco-2 cells was confirmed by fluorescence labeling. Treatment of each kefir-derived Lactobacillus EV onto TNF-α-stimulated Caco-2 cells significantly reduced the level of both mRNA expression and secretion of IL-8, and Western blot analysis revealed that such an effect was related to inhibition of TNF-α signaling mediated by reducing the phosphorylation of p65, a subunit of NF-kB. Subsequent administration of kefir-derived Lactobacillus EV into inflammatory bowel disease-induced mice significantly alleviated the body weight loss and rectal bleeding, and enhanced stool consistency. Histological examination showed that kefir-derived Lactobacillus EV substantially reduced the infiltration of transmural leukocytes and loss of goblet cells within the colon, and the serum level of myeloperoxidase was significantly lower in the EV-treated group than control group. Our study demonstrates that kefir-derived Lactobacillus EV can be potentially used for developing innovative strategies for alleviating inflammatory bowel disease.

Improving the nutritional quality and shelf life of broiler meat by feeding diets supplemented with fermented pomegranate (Punica granatum L.) by-products.

1. Four experimental diets containing 0, 0.5, 1.0 and 2.0% fermented pomegranate by-products (FPB) were supplied to 320d-old broilers to evaluate the effects of FPB on growth performance, nutritional composition, fatty acid profile and oxidative stability of meat. 2. Dietary supplementation of FPB linearly increased the weight gain and feed intake of broilers with linear reduction in feed conversion ratio. 3. The crude protein, iron, magnesium, and sodium content were linearly higher, whereas cholesterol was linearly lower in the breast meat of FPB-supplemented broilers. In thigh meat, linearly lower ether extract and cholesterol with higher moisture was noted in response to increasing levels of FPB. 4. The proportion of saturated fatty acids was both linearly and quadratically lower in breast and thigh meat, whereas those of monounsaturated fatty acids of breast (linear and quadratic) and n-3 fatty acids of breast and thigh (linear) meat was higher in the FPB-supplemented broilers. The n-6/n-3 ratio of breast meat was linearly lower in response to FPB supplementation. The hypocholesterolaemic to hypercholesterolaemic ratio of thigh meat was higher in the FPB-supplemented groups. 5. The thiobarbituric acid reactive substances and pH value were lower in the breast and thigh meat of FPB-supplemented broilers. 6. Thus, additive supplementation of the diet with up to 2% FPB improved the nutritional quality, fatty acid profile and shelf life of broiler meat.

Serum cortisol level and disc displacement disorders of the temporomandibular joint.

UNLABELLED: Temporomandibular disorders (TMD) are a significant public health problem, affecting approximately 5-12% of the population. OBJECTIVES: This retrospective cross-sectional study investigated the relationship between 8 AM serum cortisol levels (8ASC) and disc displacement disorders (DDD) of TMD. One hundred and forty patients with DDD were recruited. Among them, 60 patients comprised the case group of disc displacement without reduction with limited opening (DDWORWLO, age 37·7 ± 17·22), and 80 were 'other DDD' for the control group (age 36·4 ± 13·08). The independent variables included domains of demography, history, malocclusion, comorbid symptoms, comorbid TMD and 8ASC. Data were analysed with the chi-square test, logistic regression and receiver operating characteristic (ROC) curve. Results of multiple logistic regression showed that 8ASC was the only factor significantly related to DDWORWLO (P = 0·006). Receiver operating characteristic analysis of DDWORWLO and 8ASC indicated an area under the curve of 0·669, standard error of 0·049 and P value of 0·001. The adequate cut-off point of 8ASC was 12·45 (µg dL(-1) ), with sensitivity of 0·636, and specificity of 0·729. 8 AM serum cortisol level can be used as a clinical clue to differentiate DDWORWLO from other DDD.

A new allele, HLA-DQB1*05:50, identified by sequence-based typing in a Korean individual.

The new allele DQB1*05:50 showed one nucleotide difference with DQB1*05:03:01:01 at codon 39 (CGC/CAC).

The novel HLA-C*03:02:02:03 identified by cloning and genomic full-length sequencing.

The sequence of the C*03:02:02:03 differs from that of C*03:02:02:01 by one nucleotide substitution in the 5' UTR at position -57 (C>A).

HLA-B*40:02:18, a new HLA-B*40 allele.

The new allele B*40:02:18 showed one nucleotide difference with B*40:02:01 at codon 111 (CGC/CGT).

Effects of virtual reality on anxiety and pain in adult patients undergoing wound-closure procedures: A pilot randomized controlled trial.

Background: In emergency departments, suturing is a typical procedure for closing lacerated wounds but is invasive and often causes anxiety and pain. Virtual reality (VR) intervention has been reported as a relaxing measure. Objective: The study aims to examine the effects of VR intervention on anxiety, pain, physiological parameters, local anesthesia requirements and satisfaction in Chinese adult patients undergoing wound closure in emergency departments in Hong Kong. Methods: Adult patients who had lacerated wounds and were undergoing wound closure by suturing can communicate in Chinese and were hemodynamically stable were invited for this trial. Eighty patients were randomly assigned to the VR group, which received VR intervention and standard care, or to the control group, which received standard care only. The primary outcome was anxiety, and the secondary outcomes included pain, blood pressure, pulse rate, satisfactory with pain management, service satisfactory, and extra local analgesia requirement. Outcomes were conducted at baseline, during the procedure and 5 min after the procedure. Results: The VR group had a significantly greater reduction in anxiety (p < 0.001), pain (p < 0.001), systolic blood pressure (p < 0.001), diastolic blood pressure (p < 0.001), pulse rate (p = 0.003) and requested less amount of additional local anesthesia (p = 0.025). The satisfactory level with pain management (p = 0.019) and service (p = 0.002) were significantly higher in participants who received VR intervention. In addition, most participants preferred to have VR in the future, and no major adverse events associated with the use of VR were reported. Conclusion: This pilot study provides insight into the use of VR and the direction of future studies. It may effectively improve psychological and physiological outcomes in adult patients during wound-closure procedures in emergency departments.

Evaluation of probiotic strains for development of fermented Alisma canaliculatum and their effects on broiler chickens.

The present study was conducted to select appropriate microbial strains for the development of fermented Alisma canaliculatum A. Br. et Bouche (FAC) feed additive and to examine their effects on broilers. In experiment 1, 16 strains of Lactobacillus acidophilus, Lactobacillus plantarum, Enterococcus faecium, Bacillus subtilis, Bacillus coagulans, and Saccharomyces cerevisiae from the Korean Collection for Type Cultures (KCTC) were evaluated. The strains were tested for their acid, bile, and heat tolerance levels. Among them, L. acidophilus KCTC 3111, E. faecium KCTC 2022, B. subtilis KCTC 3239, and S. cerevisiae KCTC 7928 were selected to produce FAC. For experiment 2, 140 Ross broiler chicks were allocated to four 5-wk-long dietary groups: NC (basal diet), PC (basal diet with 0.005% oxytetracycline), AC (basal diet with 0.5% A. canaliculatum rhizome powder), and FAC (basal diet with 0.5% FAC). Final BW and total BW gain were increased (P < 0.05) upon the addition of FAC, whereas feed conversion ratio improved (P < 0.05) in the supplemented groups compared with the NC group. Crude protein content increased and crude fat decreased (P < 0.05) in the breast meat of the FAC group compared with the AC and PC groups, respectively. Proventriculus and kidney relative weights of the FAC group differed (P < 0.05) from the AC group but not from the PC group. Lower (P < 0.05) thiobarbituric acid reactive substances value of thigh meat was observed in the FAC group compared with the other groups. The FAC diet increased (P < 0.05) α-linolenic acid and linoleic acid in meat. Alteration of these fatty acids led to increased levels of polyunsaturated fatty acid, polyunsaturated fatty acid /saturated fatty acid, n-3 and n-6 fatty acids, and decreased n-6/n-3 ratio in breast or thigh meat (P < 0.05). It is concluded that FAC confers high tolerance to acid, bile and heat; and beneficially affects growth, meat composition, oxidative stability, and fatty acid composition. The FAC may be a functional feed additive in broilers.

A novel HLA-A*31 allele, A*31:34, identified by sequence-based typing.

New allele A*31:34 showed one nucleotide difference with A*31:01:02 at codon 166 (GAG/CAG).

A processed HLA-A*24:02 pseudogene found in the peripheral blood of a father and his son.

We encountered a case that exhibited a discrepancy in human leukocyte antigen-A (HLA-A) type determined by sequence-based typing (SBT) and sequence-specific primer (SSP) molecular typing. The child of this case was identified as A* 02:01 homozygote and A* 02, A* 24, respectively. The HLA-A type of his father was A* 02:01, 26:01, but low-resolution SSP also showed unexpected amplification with A* 24 primers as with the child. Serologic typing of the child and the father was A2/blank and A2/A26, respectively. Sequencing analysis of the A* 24 variant in the child and the father showed a complete deletion of all introns of the A* 24:02 allele. Though rare, this type of processed pseudogene variant can be one of the causes of discrepancies between high- and low-resolution HLA typing.

HLA-C*03:93, a novel HLA-C*03 allele identified by sequence-based typing.

The new allele C*03:93 showed one nucleotide difference with C*03:04:01 at codon 140 (GCT/ACT).

HLA-DRB1*13:99, a novel HLA-DRB1*13 allele identified by sequence-based typing.

The new allele DRB1*13:99 showed one nucleotide difference with DRB1*13:02:01 at codon 51 (ACG/AAG).

HLA-DQB1*05:06, a novel HLA-DQB1*05 allele identified by sequence-based typing.

The new allele DQB1*05:06 showed one nucleotide difference with DQB1*05:03:01 at codon 40 (TTC/TTG).

Identification of a novel HLA-B*40 null allele, HLAB*40:155N.

The new allele B*40:155N showed five nucleotide insertion between nucleotide 594 and 595 (codon 174 and 175) compared to B*40:01:01.

B*39:60, a novel HLA-B*39 allele identified by sequence-based typing.

The new allele B*39:60 showed one nucleotide difference with B*39:01:01 at codon 152 (GTG/GCG).

MICB polymorphisms and haplotypes with MICA and HLA alleles in Koreans.

Major histocompatibility complex (MHC) class I chain-related gene B (MICB) is located within the human MHC class I region. The location of MICB in the MHC region may imply the presence of linkage disequilibrium with polymorphic MICA and human leukocyte antigen (HLA) loci. MICB is also polymorphic; however, MICB polymorphisms have not been investigated in Koreans. Using sequence-based typing (SBT), we estimated the allelic frequencies of MICB and haplotypes with MICA, HLA-B, and HLA-DRB1 at high resolution in a population of 139 unrelated Korean individuals. Eight MICB alleles were identified. The most frequent allele was MICB*005:02/*010 (57.2%), followed by *002 (11.5%), *004 (8.3%), *005:03 (8.3%), and *008 (6.8%). The most common two-locus haplotypes were MICB*005:02/*010-MICA*010 (19.4%), MICB*005:02/*010-DRB1*15:01 (6.5%), and MICB*005:02/*010-B*15:01 (10.4%); the most common three-locus haplotypes were B*15:01-MICA*010-MICB*005:02/*010 (5.8%) and MICA*010-MICB*005:02/*010-DRB1*04:06 (10.4%); and the most common four-locus haplotype was B*15:01-MICA*010-MICB*005:02/*010-DRB1*04:06 (5.8%). This is the first study to provide information about MICB allele frequencies and haplotypes with HLA in Koreans. These study results should help understand mechanisms of disease association between the MICB locus and neighboring loci in Koreans.

Comparison of transnasal small-caliber vs. peroral conventional esophagogastroduodenoscopy for evaluating varices in unsedated cirrhotic patients.

BACKGROUND AND STUDY AIMS: We aimed to evaluate the accuracy of transnasal small-caliber esophagogastroduodenoscopy (TNSC-EGD) compared with peroral conventional EGD (POC-EGD) for evaluating varices in unsedated patients with liver cirrhosis. The success rate, safety, endoscopist satisfaction, and patient tolerability of TNSC-EGD were also addressed. PATIENTS AND METHODS: One hundred patients with liver cirrhosis participated in this randomized crossover trial, and 84 subjects completed both procedures. Of the 84 patients, 28 had marked bleeding diathesis (platelet count ≤ 50000/mm (3) and/or prothrombin time ≥ 1.7 INR). Endoscopists and patients answered questionnaires using a 100-mm visual analog scale about, respectively, their satisfaction and their tolerance of the procedure. RESULTS: The success rate of TNSC-EGD was comparable to that of POC-EGD (96% vs. 99%). Nasal mucosal hemorrhages induced by TNSC-EGD occurred in 5 patients (6%), but were easily controlled. Compared to the POC-EGD reference test, diagnostic accuracies of TNSC-EGD for detecting esophageal varices, gastric varices, and red color signs were 98%, 98%, and 96%, respectively. Concordance rates on grading esophageal varices and gastric varices were excellent at 93% (κ = 0.85) and 96% (κ = 0.87). Endoscopist satisfaction was not significantly different between TNSC-EGD and POC-EGD, whereas patient tolerance of TNSC-EGD was significantly greater than that of POC-EGD (79.0 ± 14.4 vs. 69.5 ± 16.1; P = 0.001). CONCLUSION: TNSC-EGD without sedation was found to be feasible, safe, and accurate for evaluating esophageal varices, gastric varices, and red color signs in patients with cirrhosis - even in those with marked bleeding diathesis. Furthermore, it was significantly better tolerated by patients, without altering endoscopist satisfaction. Our findings indicate that TNSC-EGD without sedation might be viewed as a potential alternative to POC-EGD for evaluation of varices.

Artificial neural network weights of residues for the serological specificities of HLA.

New human leucocyte antigen (HLA) alleles are assigned largely based on their sequence homologies due to lack of information on the serological reactivities. Artificial neural networks (ANNs) have been tested as a possible tool for helping to predict the serology of new alleles in the absence of serological information. However, an ANN analysis per se imparts no information regarding which residues are important in determining serological specificity. To address this issue, we extracted ANN weights of HLA residues. The ANN was trained using 139 HLA-A, 302 HLA-B and 136 HLA-DRB1 alleles, for which serological specificities were assigned in the 2004 Nomenclature Report. When the trained ANN was evaluated using alleles that were contained in the HLA Dictionary 2008 but had not been employed in the training set, the accuracy was 91% (29/32) for HLA-A, 91% (40/44) for HLA-B and 90% (9/10) for HLA-DR. Finally, ANN residue weights were extracted by summing the weights of directly connected ANN nodes. When we assessed the significance of the ANN residue weights by comparing our data with the results of epitope studies conducted by El-Awar and colleagues, we found that the ANN weights tended to be high at the epitopes described by El-Awar et al. Furthermore, the ANN weights extracted in this work could be used to explain ambiguous characteristics of serological specificities to some extent. Our data are thus considered to support the results of the epitope studies conducted by El-Awar and advance our understanding of the ambiguous serological specificities of some alleles.

Osteoconductivity of Porous Titanium Structure on Implants in Osteoporosis.

In compromised bone conditions such as osteoporosis, developments of the implant surface are necessary to secure the stability of implants. This study investigated the effect of the surface porous titanium structure (PS) on the osseointegration of implants in osteoporotic bone. Bilateral ovariectomy (OVX) was performed in 4 female beagle dogs to induce osteoporosis for 32 wk. Success of induction was based on the evaluation of bone mineral density by Hounsfield units (HU) in computed tomography images. Posterior teeth in both mandibles were extracted 1 wk after OVX, and a total of 30 implants (15 implants in each group) were placed after 32 wk of osteoporosis induction. The control group implant underwent resorbable blast media (RBM) surface treatment, whereas the test group underwent RBM surface treatment in the coronal two-thirds and a PS added to the apical 3-mm portion. HU values in the mandibular trabecular bone, lumbar, and femoral head significantly decreased 32 wk after OVX, confirming osteoporotic condition after induction. Resonance frequency analysis and removal torque test showed comparable values between the 2 groups at 4 wk after implant placement. The surface topography of the implant after removal showed hard tissue integration at the PS in the test group. Bone-to-implant contact length was greater in the apical portion of the test group, although statistical significance was not found between the groups. Interthread bone area in the apical portion of the test group showed a significant increase compared to the control group (control: 0.059 ± 0.041 mm2, test: 0.121 ± 0.060 mm2, P = 0.028) with the histological feature of bone ingrowth at the PS. The findings of the study demonstrated that the surface PS could improve osteoconductivity in the osteoporotic trabecular bone by bone ingrowth at the pore space, thereby enhancing the osseointegration and stability of the implants.

Anthropological analysis of Koreans using HLA class II diversity among East Asians.

Human leukocyte antigens (HLAs) are useful markers for anthropological investigations because the allele and haplotype distributions at these loci vary widely among ethnic groups. HLA frequencies in Koreans, however, have not previously been analyzed on a phylogenetic basis. We determined the allele frequencies of four HLA class II (HLA-DRB1, -DQA1, -DQB1, and -DPB1) loci in 149 unrelated Korean individuals using a sequence-based typing method. A total of 29 HLA-DRB1, 17 HLA-DQA1, 16 HLA-DQB1, and 15 HLA-DPB1 alleles were identified. The most common allele at each locus was DRB1*0901, DQA1*0102, DQB1*0301, and DPB1*0501, respectively. Four-locus allelic association analysis showed the existence of 25 DRB1-DQA1-DQB1-DPB1 haplotypes with a frequency greater than 0.010. A dataset comprising ethnicity-specific information from published literature and the dbMHC database, as well as the allele frequencies determined in this study, was subjected to phylogenetic analysis. The analysis showed that Koreans are most closely related to Japanese and Han Chinese from Shandong province. Correspondence analyses showed that the current Korean population is located in a position intermediate between the northern and southern East Asian groups, supporting the theory of a bi- and/or multidirectional route of migration of early Korean settlers. This report can be used for anthropological studies, and may also be of use in the International Hematopoietic Stem Cell Sharing program.