Chromosomal dynamics in Senna: comparative PLOP-FISH analysis of tandem repeats and flow cytometric nuclear genome size estimations.

Introduction: Tandem repeats (TRs) occur abundantly in plant genomes. They play essential roles that affect genome organization and evolution by inducing or generating chromosomal rearrangements such as duplications, deletions, inversions, and translocations. These impact gene expression and chromosome structure and even contribute to the emergence of new species. Method: We investigated the effects of TRs on speciation in Senna genus by performing a comparative analysis using fluorescence in situ hybridization (FISH) with S. tora-specific TR probes. We examined the chromosomal distribution of these TRs and compared the genome sizes of seven Senna species (estimated using flow cytometry) to better understand their evolutionary relationships. Results: Two (StoTR03_159 and StoTR04_55) of the nine studied TRs were not detected in any of the seven Senna species, whereas the remaining seven were found in all or some species with patterns that were similar to or contrasted with those of S. tora. Of these studies species, only S. angulata showed significant genome rearrangements and dysploid karyotypes resembling those of S. tora. The genome sizes varied among these species and did not positively correlate with chromosome number. Notably, S. angulata had the fewest chromosomes (2n = 22) but a relatively large genome size. Discussion: These findings reveal the dynamics of TRs and provide a cytogenetic depiction of chromosomal rearrangements during speciation in Senna. To further elucidate the dynamics of repeat sequences in Senna, future studies must include related species and extensive repeatomic studies, including those on transposable elements.

Subgenome Discrimination in Brassica and Raphanus Allopolyploids Using Microsatellites.

Intergeneric crosses between Brassica species and Raphanus sativus have produced crops with prominent shoot and root systems of Brassica and R. sativus, respectively. It is necessary to discriminate donor genomes when studying cytogenetic stability in distant crosses to identify homologous chromosome pairing, and microsatellite repeats have been used to discriminate subgenomes in allopolyploids. To identify genome-specific microsatellites, we explored the microsatellite content in three Brassica species (B. rapa, AA, B. oleracea, CC, and B. nigra, BB) and R. sativus (RR) genomes, and validated their genome specificity by fluorescence in situ hybridization. We identified three microsatellites showing A, C, and B/R genome specificity. ACBR_msat14 and ACBR_msat20 were detected in the A and C chromosomes, respectively, and ACBR_msat01 was detected in B and R genomes. However, we did not find a microsatellite that discriminated the B and R genomes. The localization of ACBR_msat20 in the 45S rDNA array in ×Brassicoraphanus 977 corroborated the association of the 45S rDNA array with genome rearrangement. Along with the rDNA and telomeric repeat probes, these microsatellites enabled the easy identification of homologous chromosomes. These data demonstrate the utility of microsatellites as probes in identifying subgenomes within closely related Brassica and Raphanus species for the analysis of genetic stability of new synthetic polyploids of these genomes.

The Change of Endotracheal Tube Cuff Pressure During Laparoscopic Surgery.

BACKGROUND: We evaluated the endotracheal tube cuff pressure (Pcuff) changes during pneumoperitoneum for laparoscopic cholecystectomy and the correlations between body mass index (BMI), pneumoperitoneum time, and Pcuff changes. METHODS: Total 60 patients undergoing laparoscopic cholecystectomy were allocated to either a study group (BMI ≥ 25 kg/m2) or a control group (BMI < 25 kg/m2). The endotracheal intubation was performed with a high-volume low-pressure cuffed oral endotracheal tube. A manometer was connected to the pilot balloon using a 3-way stopcock and the cuff was inflated. The change in Pcuff was defined as the difference between the pressure just before intra-abdominal CO2 insufflation and the pressure before CO2 desufflation. RESULTS: Pcuff increased to 5.3 ± 3.6 cmH2O in the study group and 5.7 ± 5.4 cmH2O in the control group. There was no significant difference between two groups. While BMI was not correlated with change in Pcuff (r = 0.022, p = 0.867), there was a significant correlation between change in Pcuff and the duration of pneumoperitoneum (r = 0.309, p = 0.016). CONCLUSION: The change in Pcuff was not affected by BMI and was significantly correlated with pneumoperitoneum time. We recommend regular measurement and adjustment of Pcuff during laparoscopic surgery.

Effects of a Preoperative Transdermal Fentanyl Patch on Proinflammatory Cytokine and Pain Levels During the Postoperative Period: A Randomized Controlled Trial.

OBJECTIVE: The main objectives of this article were to assess the effect of preoperative transdermal fentanyl patch (TFP) on interleukin (IL)-6 and IL-8 levels and pain after laparoscopic cholecystectomy. MATERIALS AND METHODS: Patients received a TFP (25 µg/h) (patch group, n=30) or a placebo patch (control group, n=30) applied 14 hours before operation. After surgery, control group received intravenous continuous fentanyl (25 µg/h) with loading dose (25 µg). IL-6 and IL-8 levels were measured at admission and 1, 6, 12, 24, and 48 hours postoperatively. Pain score and consumption of rescue analgesic were evaluated too. RESULTS: At 24 hours postoperatively, IL-6 and IL-8 reached a peak and then decreased. The peak IL-6 levels were 21.92(±6.22) and 24.91(±6.81) pg/mL in the patch and control group. The significant differences of IL-6 between groups were shown at 6 and 12 hours postoperatively (P=0.032, 0.0001). There were no significant differences in IL-8 levels and pain score. CONCLUSIONS: Preoperative TFP attenuated the increase in IL-6 levels after surgery and provided similar analgesia to continuous fentanyl infusion. Preemptive TFP may have influence on proinflammatory reactions and pain control after surgery.

Injuries in national Olympic level judo athletes: an epidemiological study.

PURPOSE: To present an epidemiological study of injuries found among South Korea's National level Judo athletes as a foundation for future injury prevention and skill enhancement in this group. METHODS: This study is a prospective study on a 4-year injury assessment held from January 2010 to December 2013 at the training centre in South Korea for National Level athletes. Athlete's weight class, gender, injury location and injury grade (grade I=1-3 treatment days, grade II=4-7 treatment days, and grade III ≥8 treatment days) were analysed. RESULTS: There were a total of 782 injuries recorded during this period, equalling to four injuries per athlete annually. Almost half of these injuries (47%) were grade I injuries. Injury occurrence was the highest in the Lower body (44.2%). This was then followed by injuries in the upper body (29.8%), trunk (20.3%) and head and neck (5.6%). Men and women showed similar, non-significantly different trends in the proportion of body parts injured. Women experienced more grade III injuries than males (p=0.0228). Comparison between women in different weight classes also showed that heavyweights incurred more grade III injuries than lightweights (p=0.0087). Lightweights had a higher rate of injury than heavyweights in males and females, although this was statistically significant only among males (p<0.001). CONCLUSIONS: Many body regions are prone to injury in the elite judo population. Women, especially those in the heavyweight classification, were more prone to severe injuries. Lightweights experienced more injuries than heavyweights among male athletes. Specifically, further studies are needed to confirm these findings and to address the impact of rapid weight loss practices on injury risk to implement effective preventive measures.

The immunomodulatory effects of human mesenchymal stem cells on peripheral blood mononuclear cells in ALS patients.

In a previous study, we reported that intrathecal injection of mesenchymal stem cells (MSCs) slowed disease progression in G93A mutant superoxide dismutase1 transgenic mice. In this study, we found that intrathecal MSC administration vastly increased the infiltration of peripheral immune cells into the spinal cord of Amyotrophic lateral sclerosis (ALS) mice (G93A mutant superoxide dismutase1 transgenic). Thus, we investigated the immunomodulatory effect of MSCs on peripheral blood mononuclear cells (PBMCs) in ALS patients, focusing on regulatory T lymphocytes (Treg ; CD4(+) /CD25(high) /FoxP3(+) ) and the mRNA expression of several cytokines (IFN-γ, TNF-α, IL-17, IL-4, IL-10, IL-13, and TGF-ß). Peripheral blood samples were obtained from nine healthy controls (HC) and sixteen patients who were diagnosed with definite or probable ALS. Isolated PBMCs from the blood samples of all subjects were co-cultured with MSCs for 24 or 72 h. Based on a fluorescence-activated cell sorting analysis, we found that co-culture with MSCs increased the Treg /total T-lymphocyte ratio in the PBMCs from both groups according to the co-culture duration. Co-culture of PBMCs with MSCs for 24 h led to elevated mRNA levels of IFN-γ and IL-10 in the PBMCs from both groups. However, after co-culturing for 72 h, although the IFN-γ mRNA level had returned to the basal level in co-cultured HC PBMCs, the IFN-γ mRNA level in co-cultured ALS PBMCs remained elevated. Additionally, the levels of IL-4 and TGF-ß were markedly elevated, along with Gata3 mRNA, a Th2 transcription factor mRNA, in both HC and ALS PBMCs co-cultured for 72 h. The elevated expression of these cytokines in the co-culture supernatant was confirmed via ELISA. Furthermore, we found that the increased mRNA level of indoleamine 2,3-dioxygenase (IDO) in the co-cultured MSCs was correlated with the increase in Treg induction. These findings of Treg induction and increased anti-inflammatory cytokine expression in co-cultured ALS PBMCs provide indirect evidence that MSCs may play a role in the immunomodulation of inflammatory responses when MSC therapy is targeted to ALS patients. We propose the following mechanism for the effect of mesenchymal stem cells (MSCs) administered intrathecally in amyotrophic lateral sclerosis (ALS): MSCs increase infiltration of peripheral immune cells into CNS and skew the infiltrated immune cells toward regulatory T lymphocytes (Treg ) and Th2 lymphocytes. Treg and Th2 secret anti-inflammatory cytokines such as IL-4, IL-10, and TGF-ß. A series of immunomodulatory mechanism provides a new strategy for ALS treatment.

Newly developed glycogen synthase kinase-3 (GSK-3) inhibitors protect neuronal cells death in amyloid-beta induced cell model and in a transgenic mouse model of Alzheimer's disease.

Glycogen synthase kinase-3 (GSK-3) is emerging as a prominent therapeutic target of Alzheimer's disease (AD). A number of studies have been undertaken to develop GSK-3 inhibitors for clinical use. We report two novel GSK-3 inhibitors (C-7a and C-7b) showing good activity and pharmacokinetic (PK) profiles. IC50 of new GSK-3 inhibitors were in the range of 120-130 nM, and they effectively reduced the Aß-oligomers induced neuronal toxicity. Also, new GSK-3 inhibitors decreased the phosphorylated tau at pThr231, pSer396, pThr181, and pSer202, and inhibited the GSK-3 activity against Aß-oligomers induced neuronal cell toxicity. In B6;129-Psen1(tm1Mpm) Tg(APPSwe, tauP301L)1Lfa/Mmjax model of AD, oral administration of C-7a (20 mg/kg, 50 mg/kg) showed increased total arm entries and spontaneous alteration of Y-maze which was regarded as short-term memory. In particular, 50 mg/kg C-7a treated mice significantly decreased the level of phosphorylated tau (Ser396) in brain hippocampus. We suggest that new GSK-3 inhibitor (C-7a) is potential candidates for the treatment of AD.

Effects of valproic acid on the expression of trophic factors in human bone marrow mesenchymal stromal cells.

The potential of human bone marrow-mesenchymal stromal cells (hBM-MSCs) to differentiate into diverse cell types and secrete a variety of trophic factors makes them an excellent cell therapy tool for intractable diseases. However, their therapeutic efficacy has not yet been satisfied in preclinical and/or clinical trials with autologous or allogenic stem cells. To improve the efficacy of stem cell therapy, optimized conditions for stem cells need to be defined. In this study, we evaluated the effects of valproic acid (VPA), an HDAC inhibitor, in human BM-MSCs and assessed the expression of trophic factors (ANG, BDNF, ECGF1, bFGF-2, GDNF, HGF, IGF-1, PIGF, TGF-ß1, and ß-Pix) in MSCs treated with 200µg/ml VPA for 12h. Under these conditions the features of MSCs were not changed. The VPA-treated MSCs also showed an increased cell protective effect against oxidative injuries in MTT assays and improved migratory ability when examined by the Boyden chamber assay. This suggests that MSCs may be improved by treatment with an optimal VPA dose and incubation time, which may increase the efficacy of stem cell therapy.

Development of a cellular tau enzyme-linked immunosorbent assay method for screening GSK-3ß inhibitors.

Glycogen synthase kinase-3ß (GSK-3ß), a serine/threonine kinase also known as tau protein kinase I, has been implicated in the pathogenic conditions of Alzheimer's disease. Many investigators have focused on GSK-3 inhibitor as a therapeutic drug. In this study, we established a cell-based assay for the screening of novel GSK-3ß inhibitors. For this purpose, four-repeat tau cDNAs were stably expressed in human embryonic kidney 293 (HEK293) cells (HEK293-Tau). The proliferation of HEK293-Tau cells was no different from that of HEK293 cells, as measured by the bromodeoxyuridine enzyme-linked immunosorbent assay (BrdU ELISA). The concentration-dependent reduction of tau phosphorylation by GSK-3 inhibitors, LiCl, Chir98023, and SB415286, was examined by immunoblot analysis and Tau ELISA (in situ ELISA). Highly consistent data were obtained, suggesting that this novel ELISA method is highly reproducible. Using this ELISA strategy, we isolated a few candidate compounds, including compounds 114 and 149, from several hundreds of synthetic agents and demonstrated that such candidates protect nerve growth factor-differentiated PC12 cells against amyloid-ß-induced cell death. These data indicate that this Tau ELISA method in HEK293-Tau cells may be a suitable cell-based assay system to screen for GSK-3ß inhibitors.

Distribution of the Trp64Arg polymorphism in the ß 3-adrenergic receptor gene in athletes and its influence on cardiovascular function.

BACKGROUND: The Trp64Arg polymorphism in the beta3-adrenergic receptor (beta3-AR) gene was extensively studied concerning the relationship with cardiovascular (CV) diseases, but few studies addressed this issue in athletes. AIM: To examine the relationship between the Trp64Arg polymorphism in the beta3-AR gene and several CV functions such as cardiac structure and function or blood biochemical parameters in Korean male controls and athletes. METHODS: We recruited a total of 114 study subjects, including 81 male athletes (8 long distance runners, 17 soccer, 8 baseball, 10 basketball, 8 volleyball, 8 ice hockey, 8 judo, 6 taekwondo and 8 gymnastics) and 33 controls. Two dimensional echocardiography was performed in order to assess the cardiac structure and function and blood biochemical parameters were measured. Genotyping of Trp64Arg polymorphism in the beta3-AR gene was also performed by the SNaPshot method. RESULTS: The genotype and allele distribution of Trp64Arg polymorphism in the beta3-AR gene were significantly different among each sporting discipline, showing the highest Arg allele frequency in volleyball and gymnastics (p < 0.05). Also, this polymorphism was significantly associated with serum HDL-cholesterol and glucose level in athletes only (p < 0.05). CONCLUSIONS: Our data showed the significant associations between the Trp64Arg polymorphism in the beta3-AR gene and some CV parameters such as serum HDL-cholesterol and glucose levels in athletes. However, further studies of the precise mechanism behind these associations are needed.

Association of estrogen receptor-alpha gene polymorphism with pathogenesis of osteoporosis in Korean vegetarian men.

OBJECTIVE: The purpose of this study was to investigate the association between two genetic polymorphisms (PvuII and XbaI restriction fragment length polymorphisms, RFLPs) of the estrogen receptor-alpha (ER1) gene and the quantitative ultrasound (QUS) parameters at the calcaneus. SUBJECTS AND METHODS: Two hundred and sixty-six Korean vegetarian men, mean age 50.9+/-12.0 years (range 26-80), were studied. Polymorphisms at the ER1 gene sites and the cross-sectional associates of genetic factors with calcaneal QUS parameters including broadband ultrasound attenuation (BUA) and the speed of sound (SOS) were analyzed by RFLPs using polymerase chain reaction. RESULTS: The distribution of PvuII and XbaI RFLPs in the ER1 gene was as follows: PP 11.6%, Pp 47.2%, pp 41.2%, XX 1.2%, Xx 24.4% and xx 74.4%. After adjusting for potential confounding factors such as age and body mass index, two genetic polymorphisms of the ER1 gene were independently associated with BUA, SOS and stiffness index at the calcaneus of our subjects. The QUS measurements of the subjects with the xx genotype were higher than those of the subjects with an Xx genotype, while the QUS measurements of the subjects with a Pp genotype were significantly lower than those of the subjects with PP or pp genotypes (p<0.05). CONCLUSION: The results suggest that the PvuII and XbaI RFLPs of the ER1 gene may be genetic factors that affect QUS at the calcaneus.

The 5178C/A and 16189T/C polymorphisms of mitochondrial DNA in Korean men and their associations with blood iron metabolism.

Several studies reported that there were the associations between the genetic polymorphisms in the mitochondrial DNA (mtDNA) and some blood iron markers. Thus, we tried to investigate the relationship between two genetic polymorphisms (5178C/A and 16189T/C) in the mtDNA and several blood iron markers in Korean men. A total of unrelated 131 Korean men were participated in this study. Two genetic polymorphisms in the mtDNA was determined by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) method, and hematological or biochemical assay performed by autoanalyzer. Although the 16189T/C polymorphism was not significantly associated with any iron parameters measured in this study, we found that the 5178C/A polymorphism was significantly associated with red blood cell (RBC) count and hematocrit (HCT) value in Korean men (P < 0.05). Therefore, our data suggest that the 5178C/A polymorphism in the mtDNA might be useful as a genetic marker with respect to blood iron metabolism.

The effect of training type on oxidative DNA damage and antioxidant capacity during three-dimensional space exercise.

OBJECTIVE: Orbotron training is a popular training method for fighter pilots because it replicates a high-acceleration environment with excessive G-force. The purpose of this study was to investigate the effects of 9 weeks of orbotron training on oxidative DNA damage and antioxidant capacity in humans during 3-dimensional space exercise. SUBJECTS AND METHODS: The subjects comprised 15 senior cadets from the Korea Air Force Academy who had no record of medical disorders and who participated in a regular exercise program (3 times per week). They were randomly divided into three groups consisting of 5 subjects each: a weight training group (21.97 +/- 1.12 years), a running training group (21.53 +/- 0.18 years) and an orbotron training group (21.48 +/- 0.29 years). Three-dimensional exercise tests were performed before and after training, and blood samples were taken to measure the concentration of plasma lactate, malondialdehyde (MDA), erythrocyte superoxide dismutase (SOD) activity, and leukocyte DNA damage. RESULTS: Plasma lactate concentrations decreased in all three groups when measured after training and after 30 min of recovery compared to before training (p < 0.05). The concentration of plasma MDA also decreased after training in all blood samples compared to the values obtained before training although there was no significant difference in the weight training and orbotron training groups. In contrast, the activity of erythrocyte SOD increased for all three groups compared to before training (p < 0.05). In the comet assay results, the greatest lymphocyte DNA damage was demonstrated at the end of exercise compared to the other three samples under all conditions, and these aspects were commonly observed in all three parameters of lymphocyte DNA damage (tail DNA, tail length and tail moment) (p < 0.05). CONCLUSION: It can be concluded that the three types of exercise training reduced plasma lactate concentration, improved antioxidant enzyme activity, and further protected the body against oxidative stress (lipid peroxidation and DNA damage). Although we have identified an effect of exercise training on the levels of antioxidants and oxidants, our cohort was small, so further studies are needed to evaluate the different types of exercise training.

Human angiogenin presents neuroprotective and migration effects in neuroblastoma cells.

Human angiogenin (ANG) has been highlighted as an angiogenic factor which supports primary and metastatic tumor growth. Recent genetic studies have shown that ANG is presented as a susceptibility gene for amyotrophic lateral sclerosis (ALS) and ALS-frontotemporal dementia (ALS-FTD). They found several missense mutations, including K40I, which present the weakest functional activity in ANG variants. In this study, we investigate whether human wild type ANG (wANG) and its variant K40I (mANG) maintain their divergent functional capacities in neuronal cells. To evaluate this, SH-SY5Y neuroblastoma cells were transfected with wANG and mANG DNA and identified both wild and mutant ANG are localized to nuclei and have no effects on proliferation. We have shown that human wANG prevented cell death under H(2)O(2)-induced oxidative stress in both SH-SY5Y and NSC-34 cells, tested by MTT assay. These effects were more enhanced in motor neuron cell NSC-34. wANG also played a role in cell migration, while mANG decreased these functional activities. Immunoblot analysis revealed that the intracellular signaling of ERK1/2 (at Thr183/Tyr185) was increased following transfection of the wANG gene, and significantly decreased by mANG in neuronal cells. These findings suggest that human ANG plays a critical role in cell protection and migration following alterations in ERK1/2 signaling in SH-SY5Y cells. This may provide the possible relationship between mutations in hANG and other neurodegenerative diseases as well as ALS.

Association between an AluI polymorphism in the calcitonin receptor gene and quantitative ultrasound parameters in Korean men.

OBJECTIVE: The purpose of this study was to investigate the association between an AluI RFLP of the calcitonin receptor (CTR) gene and quantitative ultrasound (QUS) parameters in Korean men, and the interaction with nutrition as a lifestyle factor. MATERIALS AND METHODS: Broadband ultrasound attenuation, speed of sound and stiffness index of the calcaneus were measured using an ultrasound bone densitometer in 201 Korean men (mean age +/- SD: 51.6 +/- 11.7 years). The PCR-RFLP method was used to analyze an AluI polymorphism in the CTR gene. RESULTS: In all subjects, the distribution of CC, CT and TT genotypes occurred with frequencies of 87.1, 12.4 and 0.5%, respectively. When stratified by omnivore and vegetarian groups, there was a significant association between an AluI polymorphism in the CTR gene and QUS parameters such as speed of sound and stiffness index in only vegetarian subjects. CONCLUSION: Our data suggest that the AluI polymorphism of the CTR gene can be useful as a genetic marker in the interindividual susceptibility of QUS parameters by the interaction with nutritional status as a lifestyle factor.

Genetic variation in the renin-angiotensin system and response to endurance training.

OBJECTIVE: To investigate the relationship between genetic variation in the renin-angiotensin system and the effect of 12-week endurance training in Korean women. MATERIALS AND METHODS: Seventeen women who participated in an endurance training program for 12 weeks were genotyped for the angiotensinogen M235T polymorphism, angiotensin II type 1 receptor A1166C polymorphism, angiotensin-converting enzyme (ACE) T-3892C polymorphism, and angiotensin II type 2 receptor C3123A polymorphism. The following clinical parameters were measured before and after the endurance training program: blood pressure, body composition, ventilatory response, total cholesterol, triglyceride, and glucose. RESULTS: Of the genetic markers investigated, the frequency of the T allele for the ACE T-3892C polymorphism was significantly associated with the response in body mass index and VO(2max) after 12 weeks of endurance training (p< 0.05). None of the other polymorphisms were significantly associated with the effect of training. CONCLUSION: The significant association between ACE T-3892C and the change in body mass index and VO(2max) in Korean women are attributed to training, suggesting that this genetic variation is a useful genetic marker for clarifying the interindividual response to endurance training.

Genomic fingerprinting of antituberculosis agents-resistant Lactobacillus ruminus SPM0211 using the Microbial Uniprimer kit.

A Lactobacillus isolate was collected from the feces of a healthy Korean individual and named as Lactobacillus ruminus SPM0211. It was further characterized by subjecting it to an antibiotic resistance test and genetic analysis. In the antibiotic resistance test, all tested Lactobacillus spp. were classified as "high resistance" for multiple antibiotics, such as isoniazid, ethambutol, cycloserine, and vancomycin. L. ruminus SPM0211 was classified as "high resistance" for streptomycin also, while the other tested Lactobacillus spp. were classified as low resistance. This suggests that the antimicrobial spectra may be a good indicator in the discrimination of this strain among the tested Lactobacillus spp. In a polymerase chain reaction-random amplified polymorphic DNA (PCR-RAPD) analysis using the Microbial Uniprimer kit, L. ruminus SPM0211, and L. suebicus were clustered as a group with a 74.3% similarity level, suggesting that these two species are genetically related. Thus, our data suggest that the PCR-RADP method using the Microbial Uniprimer kit may be valuable in discriminating L. ruminus SPM0211 from other Lactobacillus spp.

Identification of Lactobacillus ruminus SPM0211 isolated from healthy Koreans and its antimicrobial activity against some pathogens.

The intestinal microbiota are important to the host with regard to resistance they impart against bacterial infections and their involvement in mediating metabolic functions. Lactic acid producing bacteria such as Lactobacillus play an important physiological role in these matters. The aim of the present study was to isolate Lactobacillus sp. that inhibits enteric pathogens. Initially, 17 isolates from healthy Koreans were collected on Lactobacillus selective medium. Resistance of the isolates to antibiotics including rifampicin, streptomycin, clindamycin and vancomycin was measured. One of the isolate was identified as Lactobacillus ruminus on the basis of bacterial cell morphology, cultural characteristic and biochemical characteristics, 16S rRNA sequence analysis and PCR-RAPD. Antimicrobial activity of the bacterium against Vancomycin Intermediate Resistant Staphylococcus aureus (VISA) and Vancomycin-Resistant Enterococci (VRE) was measured. About 10(4) cells of VISA or VRE were mixed with 1, 5, and 9 mL of L. ruminus SPM 0211 and the final volume was adjusted to 10 mL with brain heart infusion (BHI) broth. The cell suspension was incubated for 3, 6, 9, and 24 h, serially diluted and then plated on BHI agar plates. As numbers of L. ruminus SPM 0211 were increased, viable cell count of VISA and VRE decreased. The strongest antimicrobial activity of SPM 0211 was observed after 9 h incubation in any mixture, almost completely inhibiting the growth of these two bacteria. The results suggest that the freshly isolated L. ruminus SPM 0211 may be used as a pro-biotic microbe that prevents the colonization of enteric pathogens and can thereby promote good gastrointestinal health.

Safety assessment of potential lactic acid bacteria Bifidobacterium longum SPM1205 isolated from healthy Koreans.

The safety assessment of Bifidobacterium longum SPM1205 isolated from healthy Koreans and this strain's inhibitory effects on fecal harmful enzymes of intestinal microflora were investigated. The overall safety of this strain was investigated during a feeding trial. Groups of SD rats were orally administered a test strain or commercial reference strain B. longum 1 x 10(9) CFU/kg body weight/day for four weeks. Throughout this time, their feed intake, water intake and live body weight were monitored. Fecal samples were periodically collected to test harmful enzyme activities of intestinal microflora. At the end of the four-week observation period, samples of blood, liver, spleen, kidney, and gut tissues were collected to determine for hematological parameters and histological differences. The results obtained in this experiment demonstrated that four weeks of consumption of this Bifidobacterium strain had no adverse effects on rat's general health status, blood biochemical parameters or histology. Therefore, it is likely to be safe for human use. Fecal harmful enzymes such as beta-glucosidase, beta-glucuronidase, tryptophanase and urease, were effectively inhibited during the administration of the B. longum SPM1205. These results suggested that this B. longum SPM 1205 could be used for humans as a probiotic strain.