Oral administration of Lactiplantibacillus plantarum NR16 isolated from Kimchi ameliorates murine allergic rhinitis.

Allergic rhinitis (AR) is a type I hypersensitivity mediated by dominant T helper 2 (Th2) response over the Th1 response after re-exposure to a specific allergen. Currently, socio-economic cost evoked by AR is quickly increasing since the prevalence of AR is gradually increasing in all ages worldwide. Several probiotic Lactobacillus strains have been described with potential immunomodulatory effects against type I hypersensitivity such as AR. Thus, the aim of the present work was to characterize basic probiotic property and immunomodulatory role of newly isolated Lactobacillus strains from Kimchi, a traditional fermented Korean food, in AR. Among the identified strains, Lactiplantibacillus plantarum NR16 revealed to be a powerful Th1 inducer since immune cells co-cultured with NR16 produced the highest quantity of interferon-γ (IFN-γ) and interleukin-12 (IL-12) but secreted a low amount of IL-4 in vitro. Therefore, NR16 was selected for the following assays conducted with mice with birch pollen-induced AR. Oral administration of NR16 reduced airway hyperresponsiveness and leukocyte infiltration in lesions of mice. In conclusion, oral administration of NR16 may mitigate symptoms of AR by inducing Th1 immune response, which might rebalance Th2/Th1 ratio by decreasing Th2 cytokine production in specific lesions of mucosa.

Oral administration of Lactobacillus plantarum CJLP133 and CJLP243 alleviates birch pollen-induced allergic rhinitis in mice.

AIMS: In this study, we evaluated the therapeutic efficacy of selected probiotics in a mouse model of birch pollen (BP)-induced allergic rhinitis. METHODS AND RESULTS: Oral administration of Lactobacillus plantarum CJLP133 and CJLP243 ameliorated the symptoms of BP-induced allergic rhinitis by reducing airway hyperresponsiveness, and both the histological scores and the number of infiltrated cells in the nasal cavities and lungs. Compared with those from vehicle-treated mice, bronchoalveolar lavage fluid and draining lymph node samples from CJLP133 and CJLP243-administrated mice showed diminished numbers of immune cells, increased secretion of a Th1-type cytokine (IFN-γ) and decreased production of Th2-type cytokines (IL-4, IL-5 and IL-13). Consistent with these results, levels of IL-4, IL-5, IL-13, serum IgE and BP-specific serum IgG1 were decreased, whereas secretion of IFN-γ and BP-specific serum IgG2a was augmented upon administration of CJLP133 and CJLP243 in mice. CONCLUSION: Oral administration of L. plantarum CJLP133 and CJLP243 alleviates symptoms of BP-induced allergic rhinitis in mice by recovering Th1/Th2 balance via enhancement of the Th1-type immune response. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus plantarum CJLP133 and CJLP243 have therapeutic effects on BP-induced allergic rhinitis in an animal model.

Effect of direct loading of phytoestrogens into the calcium phosphate scaffold on osteoporotic bone tissue regeneration.

3D porous calcium deficient hydroxyapatite (CDHA) scaffolds with phytoestrogens were fabricated for osteoporotic bone tissue regeneration through a combination of 3D printing techniques and cement chemistry as a room temperature process. Quercetin, one of the major phytoestrogens isolated from onions and apples, was directly incorporated into CDHA for local administration in place of bisphosphonates (BPs) (which are recognized as standard treatment in osteoporosis), to avoid drug side effects. The CDHA scaffolds with quercetin (QC-CDHA) showed favorable mechanical properties (compressive strength < 21 MPa) as well as pore morphology. Quercetin was steadily released with the biodegradation of CDHA scaffolds in vitro without any initial burst. The QC-CDHA scaffolds greatly influenced both osteoblast and osteoclast cell activities. The QC-CDHA scaffolds significantly increased pre-osteoblast cell (MC3T3-E1) proliferation, differentiation, and mineralization, whereas osteoclast cell (RANK treated RAW 264.7) proliferation and differentiation were dramatically suppressed. The influence of quercetin on bone tissue regeneration was superior to alendronate, which is one of the most commonly administered BPs. All results indicated that quercetin in CDHA scaffolds plays an important role in both enhancing bone formation and suppressing bone resorption. Consequently, this technology promises great potential in osteoporotic bone tissue regeneration.

Short communication: Development of a direct in vivo screening model to identify potential probiotic bacteria using Caenorhabditis elegans.

Caenorhabditis elegans is an accepted model host to study host-bacteria interactions in the gut, in addition to being a simple model with which to study conserved aspects of biological signaling pathways in intestinal environments, because these nematode worms have similar intestinal cells to those of humans. Here, we used C. elegans to develop a new in vivo screening system for potential probiotic lactic acid bacteria (LAB). Initially, critical colonization ability of LAB strains isolated from Korean infant feces was screened in the worm intestinal tract over a period of 5 d. Furthermore, we investigated host health-promoting activities, including longevity-extending effects and immune-enhancing activities against foodborne pathogen infection. We identified 4 LAB strains that were highly persistent in the nematode gut and that significantly prolonged the longevity of C. elegans and improved the survival of C. elegans in response to infection by Staphylococcus aureus. The 4 LAB strains we identified showed resistance to acid and bile conditions, assimilated cholesterol, and were able to attach to a mucus layer. The 4 LAB isolates were identified as Lactobacillus plantarum using 16S rRNA sequencing analysis. Taken together, we developed a direct in vivo screening system using C. elegans to study host health-promoting LAB. Our system is simple, rapid, cost-effective, and reliable, and we anticipate that this system will result in the discovery of many more potential probiotic bacteria for dairy foods.

Cell-bound exopolysaccharide from probiotic bacteria induces autophagic cell death of tumour cells.

AIM: Lactic acid bacteria (LAB) are beneficial micro-organisms that have been associated with several probiotic effects in both humans and animals. Here, using proteome analysis, we investigate the antitumour effects of cell-bound exopolysaccharides (cb-EPS) isolated from Lactobacillus acidophilus 606 on colon cancer cells and explore the proteins critical for their antitumour activity. METHODS AND RESULTS: cb-EPS inhibited the proliferation of HT-29 colon cancer cells by directly affecting cell morphology and not the cell cycle. Using two-dimensional polyacrylamide gel electrophoresis coupled with matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF/MS) and immunoblot analysis, we found that cb-EPS dramatically induced Beclin-1 and GRP78, and affected Bcl-2 and Bak regulation. CONCLUSIONS: The results of this study indicate that cb-EPS are antitumourigenic against HT-29 colon cancer cells and that this activity is because of the activation of autophagic cell death promoted directly by the induction of Beclin-1 and GRP78, as well as indirectly through the induction of Bcl-2 and Bak. SIGNIFICANCE AND IMPACT OF THE STUDY: These results may contribute to understanding the novel mechanisms by which probiotic bacteria induce tumour cell death via autophagy.

Synthesis of Cu-CNT nanocomposite powder by ball milling.

In this paper, to improve the poor wettability between the Carbon nanotubes (CNTs) and metal matrix, the mechanical alloying (MA) technique by the ball milling method was employed. Also, the CNTs were pre-coated to improve the agglomeration of the CNTs with copper powder. The main aim of this work is the fabrication of Cu-CNT nanocomposite powder using the MA by ball milling. The as-received dendritic copper powders change their shape sequentially to flaky, disk-typed particles and finally, to globular type particles. Also, the tendency of globular formation was prominent in the milled Cu-Cu coated MWNTs (multi-walled carbon nanotubes) powder and the grain size of the sintered Cu coated MWNTs specimen decreased more than that of the pure copper specimen. Finally, the homogeneous Cu-CNT nanocomposite intermetallic particles having fine grains was produced by the MA method of ball milling.

Molecular characterization of hepatitis A virus isolated from acute gastroenteritis patients in the Seoul region of Korea.

Hepatitis A virus (HAV) is a major public health problem throughout the world. As a result of declining HAV endemic in Korea, an increasing number of children and adolescents have become susceptible to HAV infection. HAV is related with sanitation conditions of the environment and is transmitted via the fecal-oral route, either through person-to-person contact or by contaminated water and food. The present study has been carried out to determine the phylogenetic analysis and circulating patterns of HAV strains detected from hospitalized patients with acute gastroenteritis (AGE) in the Seoul region of Korea. In total, 2,782 stool specimens from hospitalized patients with AGE collected in October 2006 to September 2007 in Seoul were tested for HAV. A pair comparison of the nucleic acid sequence of a 159-bp base region at the putative VP1/2A junction of 85 Seoul isolates revealed that the most common HAV strain circulating in the region during 2006-2007 was subgenotype IA. HAV phylogenetic studies can provide important information on the genetic characteristics of HAV from AGE patients who may subsequently become the source of infection in Korea.

Identification of the novel HLA-G*010112 allele by sequencing genomic DNA of Korean individuals.

Human leukocyte antigen (HLA)-G is an immune modulator that inhibits the functions of cytotoxic T lymphocytes and natural killer cells. Immune modulation of HLA-G plays an important role in the prevention of fetal rejection. We found a novel substitution at nucleotide position 195 (G-T), codon 41 (Ala), in exon 2 of HLA-G. This substitution does not change the amino acid (GCG-->GCT).

Identification of a novel HLA-G allele, HLA-G*010113, in a Korean individual.

In the present study, a novel human leucocyte antigen-G allele, G*010113, was identified via direct sequencing from donor gDNA in a Korean population. This allele differs from G*01010201 by a single nucleotide substitution, C to T, in codon 169 of exon 3 but did not result in any amino acid change.

Identification of novel HLA-G subtype, HLA-G*0109, by sequencing in the Korean population.

We identified the novel human leukocyte antigen-G*0109 allele in the Korean population by direct sequencing. This allele has a single-nucleotide substitution at nucleotide residue 547 in codon 159 that results in an amino acid change from tyrosine to histidine.

Identification of stathmin 1 expression induced by Epstein-Barr virus in human B lymphocytes.

INTRODUCTION: The Epstein-Barr virus transforms resting B cells into proliferating lymphoblastoid cells, the origin of cell lines. METHOD AND RESULTS: Our cDNA microarray analyses led to the identification of 232 up-regulated and 112 down-regulated genes with more than a 3-fold difference in lymphoblastoid cell lines compared to resting B cells. The functional classification of these genes exhibited the distinct expression signature for cell proliferation, cell cycle and an immune response. Among them, we verified the differential expression of several oncogenes such as stathmin 1 (STMN1), RAB27A, RAB9A, BACH1 and BACH2 using quantitative real-time reverse transcriptase-polymerase chain reactions or Western blot analysis. Expression of STMN1 (which is involved in regulation of the microtubule filament system, cell growth and S-phase of cell cycle) was increased in lymphoblastoid cell line as well as in 7-day post-Epstein-Barr virus infection B cells, compared to resting B cells. CONCLUSION: Thus, this study suggests that Epstein-Barr virus infection induces STMN1 expression, which play a role in cell cycle progression and proliferation in the human B lymphocyte.

Hepatocellular carcinoma with metastasis to the spleen in a Holstein cow.

Hepatocellular carcinoma (HCC) with metastasis to the spleen in a Holstein cow was studied by histopathologic and immunohistochemical methods. The tumor was characterized by a pseudoglandular (acinar) pattern with an associated fibrous stroma. Individual cells often had a "hepatoid" appearance but were interspersed with scattered cells exhibiting a clear, periodic acid-Schiff (PAS)-positive cytoplasm and small eccentric nuclei. This pattern was present in nodules found in both liver and spleen. Moreover, hepatoid tumor cells were positive for alpha-fetoprotein. Immunohistochemical studies suggest that myofibroblasts were responsible for the production of fibrous septa surrounding the pseudoglandular structures of bovine HCC. In summary, our histologic and immunohistochemical findings support a diagnosis of primary HCC with splenic metastasis. Furthermore, the associated stromal response appears to be of a myofibroblast origin. The primary etiology of bovine HCC and the significance of the intralesional, PAS-positive clear cells remain undetermined.

The intracellular pathway of the acetylcholine-induced contraction in cat detrusor muscle cells.

1. The present study was aimed to investigate intracellular pathways involved in acetylcholine (ACh)-induced contraction in cat detrusor muscle cells 2. Contraction was expressed as per cent shortening of length of individually isolated smooth muscle cells obtained by enzymatic digestion. Dispersed intact and permeabilized cells were prepared for the treatment of drugs and antibody to enzymes, respectively. Using Western blot, we confirmed the presence of related proteins. 3. The maximal contraction to ACh was generated at 10(-11) M. This response was preferentially antagonized by M3 muscarinic receptor antagonist rho-fluoro-hexahydrosiladifenidol (rhoF-HSD) but not by the M1 antagonist pirenzepine and the M2 muscarinic receptor antagonist methoctramine. We identified G-proteins (Gq/11), (Gs), (G0), (Gi1), (Gi2) and (Gi3) in the bladder detrusor muscle. ACh-induced contraction was selectively inhibited by (Gq/11) antibody but not to other G subunit. 4. The phosphatidylinositol-specific phospholipase C (PI-PLC) inhibitor neomycin reduced ACh-induced contraction. However, the inhibitors of the phospholipase D, the phospholipase A2 and protein kinase C did not attenuate the ACh-induced contraction. ACh-induced contraction was inhibited by antibody to PLC-beta1 but not PLC-beta3 and PLC-gamma. Thapsigargin or strontium, which depletes or blocks intracellular calcium release, inhibited ACh-induced contraction. Inositol 1,4,5-triphosphate IP3 receptor inhibitor heparin reduced ACh-induced contraction. 5. These results suggest that in cat detrusor muscle contraction induced by ACh is mediated via M3 muscarinic receptor-dependent activation of Gq/11 and PLC-beta1 and IP3-dependent Ca(2+) release.

Regulation of ribosome synthesis in Escherichia coli: effects of temperature and dilution rate changes.

The effect of temperature on the synthesis of ribosome in Escherichia coli K-12 was investigated. In continuous fermentation, the total and functioning ribosome contents decreased with increasing temperature, while the non-functioning ribosome content remained unchanged. Cells contained higher amounts of functioning ribosome at lower temperatures to compensate for the decrease in translational activity. A transient study was performed to investigate the dynamic response of the cell to changes in the dilution rate. In response to the dilution rate shift-up, the cell mass decreased until the cells produced a sufficient amount of ribosomes to support the new higher growth rate. However, the response to the dilution rate shift-down resulted in an immediate increase in cell mass. This may be due to the fact that the cell already contains enough ribosomes to support a lower growth rate corresponding to the new low dilution rate. Based on the experimental results, a mathematical model was developed to describe the cell growth at transient as well as steady states. The total ribosome content was included as a variable because it affects the growth rate of the cell.

Aucubin: potential antidote for alpha-amanitin poisoning.

Aucubin, an iridoid glucoside isolated from Aucuba japonica (Cornaceae), exhibited significant protective activity against alpha-amanitin intoxication in mice. When a single dose of aucubin was administered intraperitoneally, a 50% survival rate was obtained even when the treatment was withheld for 12 hr after alpha-amanitin administration. A possible mechanism of protective activity is partly due to a competitive effect of aucubin on alpha-amanitin inhibition of liver RNA biosynthesis.

Protective effect of Aucuba japonica against carbon tetrachloride-induced liver damage in rats.

Pretreatment of rats with ethanol extract from leaves of Aucuba japonica (600 mg/kg/day, po) for two days protected against CCl4-induced depression in plasma disappearance and biliary excretion of injected sulfobromophthalein (BSP) determined 24 hr after the CCl4 challenge (0.5 ml/kg, ip). Percent recovery of BSP in bile in 60 min for control, CCl4, extract + CCl4 treated rats was 66.8 +/- 1.9, 56.2 +/- 1.4, and 68.9 +/- 2.2, respectively. Pretreatment of the extract also protected CCl4-induced increased serum glutamic-pyruvic transaminase activity and liver necrosis as demonstrated by histological evaluations. However, pretreatment of the extract did not modify the intensity of CCl4-induced lipid peroxidation process or cytochrome P-450 destruction. The results suggest that ethanol extract of Aucuba japonica protects CCl4 hepatotoxicity at a site in the chain events leading to necrosis but not the activation step of CCl4 to X CCl3 and X C1 free radicals.

Protective activities of aucubin against carbon tetrachloride-induced liver damage in mice.

An iridoid glucoside, aucubin was isolated from Aucuba japonica leaves and its protective activities against CCl4-induced hepatotoxicity were evaluated by measuring the duration of hypnosis induced by hexobarbital after CCl4 challenge (0.2 ml/kg/day, po) and the levels of serum glutamic-oxalacetic (GOT) and serum glutamic-pyruvic transaminase (GPT). The duration of hypnosis for the saline control group, the CCl4 alone treated group and the aucubin plus CCl4 treated group was 24.8 +/- 8.5, 60.5 +/- 9.5 and 28.0 +/- 3.2 min, respectively. Treatment of mice with aucubin also effectively protected against CCl4-induced increased serum GOT and GPT activities. It was found that aucubin inhibited hepatic RNA and protein syntheses in vivo. Such inhibitory effects of aucubin might be responsible for protective activities against CCl4-induced liver damage.