Postharvest processing decreases the richness of bacterial taxa in the phyllosphere of broccoli.

AIMS: Diverse bacterial communities residing on the surfaces of fresh vegetables are important for food quality and safety; however, knowledge of the phyllosphere microbiota on fresh vegetables and of how it changes during postharvest stage is poorly understood. METHODS AND RESULTS: We used culturing to quantify bacterial abundance and 16S rRNA 454 pyrosequencing to analyse the bacterial community composition on broccoli florets collected from farms (preharvest) and retail stores (postharvest). The bacterial community compositions of the preharvest and postharvest broccoli were significantly different. The number of non-Escherichia coli coliform bacteria (Hafnia sp. and Rahnella sp.) was higher in the postharvest broccoli than in the preharvest broccoli. Minor bacterial taxa at the phylum and genus levels had markedly disappeared in the postharvest broccoli, resulting in low bacterial species richness in the postharvest broccoli. CONCLUSIONS: The dominant bacterial taxa persist and prevail in the phyllosphere of broccoli during the postharvest stage. SIGNIFICANCE AND IMPACT OF THE STUDY: A popular crop of leafy vegetables, broccoli, is of great agricultural and nutritional importance. This study provides a detailed description of changes in the bacterial community of broccoli in harvest and storage. This study presents novel data on the impact of postharvest conditions on the bacterial populations on broccoli florets.

Robotic thyroid surgery using bilateral axillo-breast approach: personal initial experience over two years.

AIM: Although endoscopic thyroid surgery is gaining wide acceptance, however, manual endoscopic operation also has shown several limitations. The advent of robotic surgical systems, such as the da Vinci surgical system (Intuitive Surgical, Mountain View, CA, USA), is expected to make it possible to overcome some limitations of manual endoscopic operation. Herein we report a single surgeon (H.Y.K.)'s initial two-year experience of new robotic thyroid operations using the bilateral axillo-breast approach (BABA), the approach which has definite advantages and recently has been widely used for the traditional endoscopic thyroid surgery. METHODS: Between July 2008 and July 2010, 93 patients underwent robotic thyroid surgery using the BABA, with the da Vinci-S surgical system, at the Korea University Anam Hospital, Seoul, Korea. The data on the patients' clinicopathological characteristics, operation types, operation times, surgical results, postoperative hospital stays and complications were collected in a prospective manner, and later evaluated. RESULTS: Seventy-two total thyroidectomies with or without central neck dissections mostly for the papillary carcinomas, twenty lobectomies with or without central neck dissections for the minute ­ smaller than 0.5 cm in their maximal diameter ­ papillary carcinomas, follicular neoplasms and benign tumors, and a bilateral subtotal lobectomy for the multinodular goiter were performed robotically. There was no conversion of robotic procedure to traditional endoscopic or open procedure. The mean total operation time was 288.5±48.0 minutes. The mean number of retrieved lymph nodes by the central neck dissection was 5.1±1.97 (range, 0-12). The mean hospital stay of the patients was 2.8±1.2 days. And the mean postoperative 3rd month serum thyroglobulin level in patients undergone total thyroidectomy was 0.3±0.14 ng/mL (range, 0.08-1.95). Three (3.2%) patients suffered from transient hoarseness postoperatively, but all of them recovered in three months. Transient hypocalcemias were observed in 17 out of 72 (23.6%) patients who had undergone total thyroidectomy, but none of them left permanent. No other complication, such as bleeding, infection, neither fluid collection, was observed. CONCLUSION: Our initial surgical results of robotic thyroid surgery using BABA demonstrate the feasibility and safety of the procedure in the treatment of benign tumors and early differentiated carcinomas.

Molecular analysis of the GJB2, GJB6 and SLC26A4 genes in Korean deafness patients.

OBJECTIVES: Mutations in the GJB2, GJB6 and SLC26A4 genes are a frequent cause of hearing loss in a number of populations. However, little is known about the genetic causes of hearing loss in the Korean population. METHODS: We sequenced the GJB2 and GJB6 genes to examine the role of mutations in these genes in 22 hearing loss patients. We also sequenced the SLC26A4 gene in seven patients with inner ear malformations, including enlarged vestibular aqueduct (EVA) revealed by computer tomography. RESULTS: Coding sequence mutations in GJB2 were identified in 13.6% of the patients screened. Two different mutations, 235delC and T86R were found in three unrelated patients. The 235delC was the most prevalent mutation with an allele frequency of 6.9% in our patient group. No mutations, including 342-kb deletion, were found in GJB6 gene. Three different variants of SLC26A4 were identified in the EVA patients, including one novel mutation. Four EVA patients carried two mutant alleles of SLC26A4, and at least one allele in all patients was the H723R mutation, which accounted for 75% of all mutant alleles. CONCLUSIONS: Our results suggest that GJB2 and SLC26A4 mutations together make up a major cause of congenital hearing loss in the Korean population. Further studies may be able to identify other common variants that account for a significant fraction of hearing loss in the Korean population.

Considerable interindividual variation in the pharmacokinetics of tolperisone HCl.

OBJECTIVE: The aim of this study was to determine the pharmacokinetic profiles of oral tolperisone hydrochloride in healthy volunteers. METHODS: After the oral administration of tolperisone hydrochloride, the plasma concentrations of tolperisone were measured in 15 healthy male Korean volunteers. The tolperisone concentration was determined using high-performance liquid chromatography with a C18 reverse-phase column. RESULTS: Very large interindividual differences in the AUC and Cmax were detected after oral tolperisone HCl. The AUC0-infinity, varied from 125.9-1,241.3 ng/ml x h, and the Cmax varied from 64.2 and 784.9 ng/ml. The tmax of tolperisone was 0.90 +/- 0.31 h and the mean half-life was 1.00 +/- 0.28 h. CONCLUSION: These results suggest that the pharmacological effect of oral tolperisone HCl varies between individuals, and the oral tolperisone HCl dose might need to be individualized.

Correlation between the in vitro ATP-based chemosensitivity assay and HER2/neu expression in women with breast cancer.

Several in vitro chemosensitivity tests have been developed to predict the chemotherapeutic response of tumours prior to initiation of individualized treatment for breast cancer. This study investigated whether the in vitro chemosensitivity response of cell lines derived from breast cancer patients was affected by HER2/neu expression. We cultured breast cancer cell lines from 50 patients and the adenosine triphosphatebased chemotherapy response assay (ATPCRA) was performed with 5-fluorouracil, gemcitabine, docetaxel, doxorubicin, methotrexate, vinorelbine and paclitaxel. 5-fluorouracil combined a high median cell death rate (32.4%) with the narrowest range of cytotoxic effects (7.3-65.7%). In addition, gemcitabine showed significantly greater activity in HER2/neupositive patients. In contrast, docetaxel was significantly less effective in HER2/neu-positive patients. No significant correlation was found between the other agents and HER2/neu expression. The use of the ATP-CRA test for metastatic tissue from patients with recurrent disease might be a useful approach to determine the most effective chemotherapy regimen.

Low loss high mesa optical waveguides based on InGaAsP/InP heterostructures.

Low loss high mesa optical waveguides were fabricated on InGaAsP/InP heterostructures by utilizing inductively-coupled-plasma reactive ion etching (ICP-RIE) and electron beam lithography technique. The fabrication process was optimized by measuring sidewall roughness of deep-etched waveguides. Atomic force microscope loaded with carbon nanotude was used to obtain three-dimensional image of the etched sidewall of waveguides. The obtained statistical information such as rms roughness and correlation length was used to theoretically calculate scattering loss of waveguides. Several waveguides with different number of sharp bends and the length were fabricated and their propagation losses were measured by modified Fabry-Perot method. The measured propagation losses were compared with theoretically calculated losses.

Pharmacokinetics and pharmacodynamic effects of a new antibody glycoprotein IIb/IIIa inhibitor (YM337) in healthy subjects.

BACKGROUND: This study describes the first administration of YM337, the Fab fragment of a humanized monoclonal antibody against the fibrinogen GP IIb/IIIa receptor, in healthy male humans. METHODS AND RESULTS: Platelet aggregation (20 micromol/L ADP), platelet adhesion, fibrinogen binding, bleeding time, and YM337 concentrations in plasma were studied in substudy 1 after single boluses of 0.025, 0. 05, 0.1, 0.2, and 0.4 mg/kg YM337 and in substudy 2 after a bolus (0. 35 mg/kg) plus 6 hours of infusion at different dose levels of YM337 (0.5, 0.75, 1.0, 1.5 microg. kg(-1) x min(-1)), with abciximab as reference drug (n=5 or 6 subjects per group). After the 0.2-mg/kg and 0.4-mg/kg boluses, fibrinogen binding was reduced by >80% and bleeding time was prolonged to approximately 60 minutes. Bolus followed by infusion of 1.0 and 1.5 microg x kg(-1) x min(-1) YM337 maintained inhibition of platelet aggregation >80%. Aggregation and bleeding time returned to normal within 24 hours. A bolus of 0.25 mg/kg of abciximab followed by an infusion of 0.125 microg x kg(-1) x min(-1) showed effects similar to those observed with the 0.5- and 0. 75-microg x kg(-1) x min(-1) infusion of YM337. In 53 subjects exposed to YM337, 1 case of transient thrombocytopenia and 3 minor bleeding events occurred. No human anti-chimeric antibodies were detected 2 weeks and 2 months after administration. CONCLUSIONS: YM337 effectively inhibits IIb/IIIa-mediated platelet aggregation and adhesion in humans. The results of this phase 1 study will give rise to further clinical evaluation of YM337.

Expression of MMP-8 and MMP-13 genes in the periodontal ligament during tooth movement in rats.

Periodontal ligament tissue is remodeled on both the tension and compression sides of moving teeth during orthodontic tooth movement. The present study was designed to clarify the hypothesis that the expression of MMP-8 and MMP-13 mRNA is promoted during the remodeling of periodontal ligament tissue in orthodontic tooth movement. We used the in situ hybridization method and semi-quantitative reverse-transcription/polymerase chain-reaction analysis to elucidate the gene expression of MMP-8 and MMP-13 mRNA. Expression of MMP-8 and MMP-13 mRNA transiently increased on both the compression and tension sides during active tooth movement in vivo. The gene expression of MMP-8 and MMP-13 was induced by tension, while compression indirectly promoted the gene expression of MMP-8 and MMP-13 through soluble factors in vitro. Thus, we concluded that the expression of MMP-8 and MMP-13 is differentially regulated by tension and compression, and plays an important role in the remodeling of the periodontal ligament.

T-lymphocyte subsets in patients with AJCC stage III gastric cancer during postoperative adjuvant chemotherapy. American Joint Committee on Cancer.

BACKGROUND AND AIMS: Advanced neoplastic diseases alter the immune response in cancer patients. The aim of this study was to evaluate the changes of T-lymphocyte subsets during postoperative adjuvant chemotherapy, and the relationship between T-lymphocyte subsets and tumor recurrence in AJCC stage III gastric cancers. MATERIAL AND METHODS: Analysis of T-lymphocyte subsets was performed in 39 patients with stage III gastric adenocarcinoma who had undergone a curative gastric resection and postoperative chemotherapy. CirculatingT-lymphocyte subsets were measured on venous blood by using flow cytometry and monoclonal antibodies on preoperative day 1, and postoperative months 1, 3, and 6. RESULTS: The 5-year disease-free survival rates of patients with stage 3a and 3b gastric cancer were 57.1% and 33.3%, respectively (p = 0.06). Values of CD3+ and CD4+ T-cells, and CD4+/CD8+ ratios were consistently lower in the recurrence group throughout the observation period. CD4+ T-cell counts were significantly lower in the recurrence group on preoperative day 1, and postoperative months 1 and 6. However, most values of the T-lymphocyte subsets showed no statistically significant difference when comparing the stage 3a and 3b disease patient groups. CONCLUSIONS: The results of this study suggest that immunosuppression associated with CD3+ and CD4+ T-cell depression is a risk factor for postoperative recurrence in patients with stage III gastric cancer.

Effect and putative mechanism of action of ginseng on the formation of glycated hemoglobin in vitro.

As a non-enzymatic, covalent binding between glucose and protein, glycation is a well-known cause of various forms of diabetic complications. This study was undertaken to determine the effect and mechanism of action of ginseng on the formation of glycated protein. A solution containing hemoglobin and glucose was incubated for 5 days by adding ginseng extract or glutathione. The quantitative measurement of glycated hemoglobin was determined using the ion capture component set. The anti-oxidative effect of ginseng and quercetin was determined through the TBA method. The amount of glycated hemoglobin (%GHb) significantly increased with the addition of glucose (27.8 mM) compared to the non-addition group. Nonetheless, this significantly decreased when ginseng extract (2 g/dl) was added and further dropped when glutathione (50 mM) was added. The amount of hemoglobin A1c (%HbA1c), a sub-fraction of glycated hemoglobin, was lower than that of glycated hemoglobin, but it showed a similar tendency. Compared to a non-addition group, the optical density of the organic layer that was separated through the addition of chloroform in oxidized linoleic acid significantly and dose-dependently decreased when ginseng extract or quercetin was added. These results suggest that the inhibitory effect of ginseng on the formation of glycated hemoglobin could be attributed to the anti-oxidative activity of ginseng.

Delay in diagnosis of pulmonary tuberculosis at a primary health clinic in Vitoria, Brazil.

SETTING: Primary health clinics in Vitoria, Espirito Santo, Brazil. OBJECTIVE: To identify risk factors associated with patient and health care delays among patients seeking care at primary health clinics. METHODS: A prospective study among tuberculosis (TB) patients diagnosed in Vitoria between 1 January 2003 and 30 December 2007. A questionnaire ascertained the date of onset and duration of TB symptoms and medical records were reviewed. Between-group distributions of delay were compared and multivariate logistic regression was performed. RESULTS: Of 304 patients, 296 (97%) reported at least one TB symptom presenting for the first time to a qualified health service; 244 (80%) reported cough > 3 weeks. Median health care delay was 30 days (range 5-68), and median total delay was 110 days (range 26-784). Multivariate analysis revealed any cough (OR(adj) 7.35, 95%CI 2.40-22.5) and weight at TB diagnosis < 60 kg (OR(adj) 5.92, 95%CI 1.83-19.1) to be associated with patient delay of ≥ 30 days. Factors increasing risk of prolonged delay (≥ 90 days) were age ≥ 30 years (OR(adj) 1.93, 95%CI 1.09-3.43) and chest pain (OR(adj) 2.42, 95%CI 1.29-4.53). CONCLUSION: Improving health care workers' education regarding TB symptoms and implementing active case finding in targeted populations may reduce delays.

GLUT-4 degradation rate: reduction in rat adipocytes in fasting and streptozotocin-induced diabetes.

With the use of [3H]leucine pulse-chase and immunoprecipitation methods, we measured the rate of GLUT-4 degradation in rat adipocytes in the steady state at 37 degrees C. We also studied the relationship of the reduced GLUT-4 levels observed in fasted and streptozotocin (STZ)-induced diabetic rats on degradation. GLUT-4 degradation was a simple, first-order decay process. The decay was describable by a single, first-order rate constant (k). A k value of 0.061/h was estimated in control rat adipocytes. In the adipocytes of fasted and STZ-induced diabetic rats, cellular GLUT-4 contents were reduced to 36 and 43% of the control, respectively. The rates of GLUT-4 degradation were also reduced significantly, with kappa values of 0.038 and 0.041/h, respectively. These changes were reversible; the decreased values returned to control values when GLUT-4 contents were normalized by refeeding and insulin injection. These findings demonstrate the presence of a posttranslational mechanism in rat adipocytes that reduces the GLUT-4 degradation rate constant when the cellular GLUT-4 level is reduced by a pretranslational defect.

The detection of circulating breast cancer cells in peripheral blood by reverse transcriptase-polymerase chain reaction.

Some circulating cancer cells in the blood play a central role in the metastatic process and may have a major influence on patient progress. Their numbers can be very small and techniques for their detection need to be both sensitive and specific. Polymerase chain reaction (PCR) has been successfully used to detect small numbers of tumor cells in cancer. We used a reverse transcriptase-polymerase chain reaction (RT-PCR) to detect circulating breast cancer cells in venous blood samples before operations and assessed cytokeratin-19 (CK-19) and cytokeratin-20 (CK-20) as target mRNA markers in the blood of healthy donors (n=6) and breast cancer patients (n=30) with American Joint Committee on Cancer stages 0 to IIIa. CK-19 mRNA was expressed in all blood samples of healthy donors and patients. But CK-20 was the only mRNA marker not detected in the blood from healthy donors. Seven of 30 (23%) venous blood isolates of breast cancer patients yielded a CK-20 mRNA with positive results. There was no correlating CK-20 mRNA expression with stage and axillary lymph node status. In conclusion, CK-19 showed no diagnostic value as a mRNA marker in the detection of circulating cancer cells by RT-PCR assay because this was expressed in the blood of healthy donors. CK-20 mRNA was an useful marker to detect circulating cancer cells in breast cancers.

Geobacillus toebii sp. nov., a novel thermophilic bacterium isolated from hay compost.

A thermophilic, spore-forming rod isolated from hay compost in Korea was subjected to a taxonomic study. The micro-organism, designated strain SK-1(T), was identified as being aerobic, Gram-positive, motile and rod-shaped. Growth of the isolate was observed at 45-70 degrees C (optimum 60 degrees C) and pH 6.0-9.0 (optimum pH 7.5). The G+C content of the genomic DNA was 43.9 mol%. Chemotaxonomic characteristics of the isolate included the presence of mesodiaminopimelic acid in the cell wall and iso-C15:0 and iso-C17:0 as the major cellular fatty acids. The predominant isoprenoid quinone was MK-7. The chemotaxonomic characteristics of strain SK-1(T) were the same as those of the genus Geobacillus. Phylogenetic analysis based on 16S rDNA sequences showed that strain SK-1(T) is most closely related to Geobacillus thermoglucosidasius. However, the phenotypic properties of strain SK-1(T) were clearly different from those of G. thermoglucosidasius. The level of DNA-DNA relatedness between strain SK-1(T) and the type strain of G. thermoglucosidasius was 27%. On the basis of the phenotypic traits and molecular systematic data, strain SK-1(T) represents a novel species within the genus Geobacillus, for which the name Geobacillus toebii sp. nov. is proposed. The type strain is strain SK-1(T) (= KCTC 0306BP(T) - DSM 14590(T)).

Macroamylasemia in a patient with acute appendicitis: a case report.

Macroamylasemia is a condition of persistent, elevated serum amylase activity with no apparent clinical symptoms of a pancreatic disorder. In Korea, however, no such case has been reported to date. We report a case of a 17-year-old female diagnosed with macroamylasemia and acute appendicitis. One day earlier, she developed epigastric and right lower quadrant abdominal pain. She was characterized by high level of serum amylase, but normal lipase. Amylase isoenzyme analysis demonstrated increased fraction of salivary type and follow-up amylase level was persistently increased. Immunofixation disclosed the macroamylase binding with an immunoglobulin, consisting of IgA and kappa chain. The patient was treated by appendectomy, and the abdominal pain subsided.