Phylogenetic group distributions, virulence factors and antimicrobial resistance properties of uropathogenic Escherichia coli strains isolated from patients with urinary tract infections in South Korea.

UNLABELLED: Urinary tract infections (UTIs) are one of the most common diseases by which humans seek medical help and are caused mainly by uropathogenic Escherichia coli (UPEC). Studying the virulence and antibiotic resistance of UPEC with respect to various phylogenetic groups is of utmost importance in developing new therapeutic agents. Thus, in this study, we analysed the virulence factors, antibiotic resistance and phylogenetic groups among various UPEC isolates from children with UTIs. The phylogenetic analysis revealed that majority of the strains responsible for UTIs belonged to the phylogenetic groups B2 and D. Of the 58 E. coli isolates, 79·31% belonged to group B2, 15·51% to group D, 3·44% to group A and 1·72% to B1. Simultaneously, the number of virulence factors and antibiotic resistance exhibited were also significantly high in groups B2 and D compared to other groups. Among the isolates, 44·8% were multidrug resistant and of that 73% belonged to the phylogenetic group B2, indicating the compatibility of antibiotic resistance and certain strains carrying virulence factor genes. The antibiotic resistance profiling of UPEC strains elucidates that the antimicrobial agents such as chloramphenicol, cefoxitin, cefepime, ceftazidime might still be used in the therapy for treating UTIs. SIGNIFICANCE AND IMPACT OF THE STUDY: As the antibiotic resistance pattern of uropathogenic Escherichia coli varies depending on different geographical regions, the antibiotic resistance pattern from this study will help the physicians to effectively administer antibiotic therapy for urinary tract infections. In addition, the frequency of virulence factors and antibiotic resistance genes among various phylogenic groups could be effectively used to draw new targets for uropathogenic Escherichia coli antibiotic-independent therapies. The study emphasizes need of public awareness on multidrug resistance and for more prudent use of antimicrobials.

Psoriasis concurrent with inflammatory bowel disease.

BACKGROUND: Previous studies have indicated an association between psoriasis and inflammatory bowel disease (IBD), and the concurrence of the two diseases reportedly has higher morbidities in Caucasian populations. However, reports on the concurrence of psoriasis with IBD in the Asian population in the literature are scarce. Objective To analyse the characteristics of psoriasis concurrent with IBD and investigate the associated morbidity in the Asian population. METHODS: We retrospectively examined the medical records of 15 patients with a confirmed diagnosis of both psoriasis and IBD. Sixty age-, gender-, and ethnicity-matched patients with a confirmed diagnosis of only psoriasis were included as controls. Both cases and controls had visited the Seoul National University Hospital or Seoul National University Boramae Hospital between 1990 and 2012. The characteristics of psoriasis, presence of comorbidity and laboratory parameters were compared between the two groups. RESULTS: Compared to controls with psoriasis only, cases of psoriasis concurrent with IBD had a younger age of onset, longer duration of psoriasis and a higher Psoriasis Area Severity Index (PASI) score. A larger proportion of cases was treated with phototherapy, systemic therapy and biologics. However, all these differences above were not statistically significant. Cases of psoriasis with concurrent IBD showed higher erythrocyte sedimentation rate and C-reactive protein levels compared with the controls (both P = 0.000). Furthermore, this case group had a higher proportion of patients with psoriatic arthritis and with more than one autoimmune disease as compared with the control group (P = 0.007 and 0.005 respectively). CONCLUSION: Asian patients having psoriasis concurrent with IBD exhibited different characteristics as compared with those having psoriasis only, particularly in terms of psoriasis severity, risk of psoriatic arthritis, systemic inflammatory parameters and presence of autoimmune comorbidity. However, further studies elucidating the exact pathogenesis and including a larger number of patients are required.

Prevention of postoperative sore throat using capsicum plaster applied at the Korean hand acupuncture point.

In a randomised, double-blind, sham-controlled study, we compared the efficacy of capsicum plaster (PAS) applied at the Korean hand acupuncture point for the prevention of postoperative sore throat in 150 patients scheduled to undergo abdominal hysterectomy. The K group had PAS applied at the K-A20 of both hands and placebo tape at both non-acupoints. The PAS was applied prior to induction of anaesthesia and removed 8 h postoperatively. The sore throat scores of Group K were significantly lower than those of other groups at 24 h following surgery (p = 0.00027). The prevalence of moderate to severe sore throat at 24 h was lower for Group K (0%) than for sham and placebo controls (16%[p = 0.038] and 19%[p = 0.032], respectively). There were no differences in the recovery room of the sore throat scores for all groups. We found that the PAS applied at the K-A20 was an alternative method for reducing postoperative sore throat.

Simultaneous removal of organic and strong nitrogen from sewage in a pilot-scale BNR process supplemented with food waste.

As the sewerage system is incomplete, sewage in Korea lacks easily biodegradable organics for nutrient removal. In this country, about 11,400 tons of food waste of high organic materials is produced daily. Therefore, the potential of food waste as an external carbon source was examined in a pilot-scale BNR (biological nutrient removal) process for a half year. It was found that as the supply of the external carbon increased, the average removal efficiencies of T-N (total nitrogen) and T-P (total phosphorus) increased from 53% and 55% to 97% and 93%, respectively. VFAs (volatile fatty acids) concentration of the external carbon source strongly affected denitrification efficiency and EBPR (enhanced biological phosphorus removal) activity. Biological phosphorus removal was increased to 93% when T-N removal efficiency increased from 78% to 97%. In this study, several kinds of PHAs (poly-hydroxyalkanoates) in cells were observed. The observed PHAs was composed of 37% 3HB (poly-3-hydroxybutyrate), 47% 3HV (poly-3-hydroxyvalerate), 9% 3HH (poly-3-hydroxyhexanoate), 5% 3HO (poly-3-hydroxyoctanoate), and 2% 3HD (poly-3-hydroxydecanoate).

Squamous cell carcinoma in vitiligo lesion after long-term PUVA therapy.

Photochemotherapy using psoralen and ultraviolet (UV)A irradiation (PUVA) is a useful treatment method for vitiligo. However, long-term PUVA therapy may cause several adverse effects including skin cancer, especially squamous cell carcinoma (SCC). We describe a case of SCC in vitiligo lesion after long-term PUVA therapy.

Ex vivo purging of leukemia cells using tumor-necrosis-factor-related apoptosis-inducing ligand in hematopoietic stem cell transplantation.

The aim of this study was to evaluate the potential of tumor-necrosis-factor-related apoptosis-inducing ligand TRAIL to eradicate leukemia cell lines, while sparing normal hematopoietic stem cells. Human Jurkat and Molt-4 cell lines were used to optimize the purging process in umbilical cord blood (UCB) mononuclear cells. The Jurkat cell line was TRAIL sensitive and TRAIL-resistant Molt-4 cell line became sensitive after being treated with TRAIL and a low dose of doxorubicin (0.1 micro M), but UCB mononuclear cells remained resistant. DR4 expression was increased when Jurkat cells were treated with TRAIL, and DR5 expression increased after exposing Molt-4 cells to TRAIL plus a low dose of doxorubicin for 24 h. The expression of DR4 and DR5 in UCB mononuclear cells was unchanged after treatment with TRAIL, a low-dose doxorubicin, or TRAIL plus a low dose of doxorubicin. In TRAIL-sensitive Jurkat cells, caspases 8, 9, 3, and 7 were activated by TRAIL treatment and activation of caspases was augmented by TRAIL plus a low dose of doxorubicin than TRAIL or a low dose of doxorubicin alone in Molt-4 cells. Experiments involving mixture of UCB mononuclear cells and Jurkat or Molt-4 cells showed a marked eradication of leukemia cells and the limiting dilution assay demonstrated an eradication rate of more than 4 logs after 24 h incubation with 100 ng/ml of TRAIL in Jurkat cells. In the case of Molt-4 cells, the eradication rate was about 3 logs when TRAIL was used in combination with a low dose of doxorubicin. No significant decrease in the number of granulocyte-macrophage colony-forming unit) (CFU-GM) colonies was detected when UCB mononuclear cells were treated with TRAIL in combination with a low dose of doxorubicin. These results suggest that TRAIL offers the possibility of being used as an ex vivo purging agent for autologous transplantation in hematologic malignancies.

Expression of an artificial polypeptide with a repeated tripeptide glutamyl-tryptophanyl-lysine in Saccharomyces cerevisiae.

AIMS: Artificial genes, which encode 48 or 64 repeats of a tripeptide, glutamyl-tryptophanyl-lysine have been cloned to the yeast expression vector pAM82 containing the PHO5 promoter and expressed in Saccharomyces cerevisiae AH22. METHODS AND RESULTS: When the yeast cells harbouring recombinant plasmids pALTG6-2 and pALTG4-4 were derepressed in Burkholder minimal medium (Toh-e, A., Ueda, Y., Kakimoto, S.I. and Oshima, Y. (1973) Journal of Bacteriology113, 727-738) containing low phosphate (0.03 g l-1 KH2PO4 and 1.5 g l-1 KCl), the expression was the highest after 24 h induction and the artificial polypeptides were synthesized to about 10% (pALTG6-2) and 14% (pALTG4-4) of the total cell protein. CONCLUSIONS: The artificial polypeptides produced in yeast were made to react with the rabbit antiserum against the polypeptide purified from Escherichia coli and found only in the pellet fraction of cell lysates, indicating the formation of inclusion body. Artificial polypeptide consisting of Glu-Trp-Lys may be useful as partial supplement in food and feeds. SIGNIFICANCE AND IMPACT OF THE STUDY: The production of single cell enriched with homopolymers of an essential amino acid in yeast might be an important tool of supplementing cereal diets and feed grain rations and could be used as means for improvement of the amino acid profile of single cell protein and production of pharmaceutical peptides.

Differences in sensitization rates to outdoor aeroallergens, especially citrus red mite (Panonychus citri), between urban and rural children.

BACKGROUND: A recent investigation has suggested that citrus red mite (Panonychus citri, CRM) is the most important allergen in citrus-cultivating farmers with asthma and allergic rhinitis. OBJECTIVE: A cross-sectional survey was performed to evaluate the prevalence of asthma and chronic rhinitis symptoms and sensitization to common indoor and outdoor aeroallergens, including CRM and Japanese cedar pollen, in rural and urban Korean children. METHODS: A total of 2,055 children (1,055 subjects living in rural areas with citrus farms and 1,000 controls in urban areas without citrus farms) were enrolled. They were evaluated by a questionnaire, and by skin prick tests with 13 common indoor and outdoor aeroallergens, including CRM and Japanese cedar pollen. RESULTS: The prevalence of wheezing and chronic rhinitis symptoms during the last 12 months was 8.3% and 35.7% in the rural children and 10.5% and 22.4% in the control group. The most common sensitizing allergens in order of decreasing frequency were Dermatophagoides pteronyssinus (26.6%), Dermatoplagoides farinae (22.7%), CRM (14.2%), cockroach (11.3%), and Japanese cedar pollen (9.7%) among the rural children, but the sensitization rates to CRM and Japanese cedar pollen were 1.3% and 0.2% among the control children, respectively. The prevalence of wheeze during the last 12 months was not different between rural children with sensitization to CRM or Japanese cedar pollen and those without sensitization (5.4% vs 6.1%; 6.9% vs 5.9%). However, the prevalence of chronic rhinitis during the last 12 months was higher among those with sensitization to CRM or to Japanese cedar pollen than among those without sensitization (40.8% vs 34.4%; 51.5% vs 33.5%). CONCLUSIONS: CRM is a common sensitizing allergen in rural children, and the sensitization rates to outdoor aeroallergens, especially CRM and Japanese cedar pollen, are very different between children from rural and urban areas in Korea.

Clinical and immunologic changes after allergen immunotherapy with Hop Japanese pollen.

BACKGROUND AND OBJECTIVE: Hop Japanese (Hop J) pollen has been reported as one of the major causative pollen allergens in the autumn season. There have been no published data regarding the clinical and immunologic effects of Hop J pollen immunotherapy in sensitized patients. In this study, we evaluated clinical and immunologic effects of Hop J immunotherapy. PATIENTS AND METHODS: Pollens were collected in our area, and "Depo-Hop J" was prepared in the laboratory of Allergopharma (Reinbek, Germany). Fifteen asthmatic patients who had Hop J immunotherapy for > 1 year were enrolled. Their clinical parameters, such as asthma symptom scores, were monitored. Skin reactivity to Hop J and degree of airway hyperresponsiveness to methacholine were measured before and 1 year after the immunotherapy. Sera were collected before the immunotherapy, at the end of initial therapy, and 1 year after the therapy. Serum total IgE levels were compared by radioimmunoassay. Serum-specific IgE, IgG1, and IgG4 levels to Hop J were compared by ELISA. To evaluate the changes of cellular mechanisms, soluble CD30 (sCD30), soluble interleukin (IL)-2 receptor (sIL-2R), soluble CD23 (sCD23), and IL-10 levels were measured by ELISA. RESULTS: Specific IgG1 and IgG4 levels began to increase at the end of the initial therapy (P < 0.05) with significant decreases in symptom scores (P < 0.05), whereas total and specific IgE levels showed variable responses during the immunotherapy with no statistical significance (P > 0.05). Serum sIL-2R and sCD30 levels decreased significantly (P < 0.05) 1 year after immunotherapy. No significant changes were noted in sCD23, IL-10, skin reactivity to Hop J, or airway responsiveness to methacholine (P > 0.05). CONCLUSIONS: We are certain that Hop J allergen immunotherapy, if carried out properly according to suitable indications, can favorably influence asthma. Thus, an increase in specific IgG4 and IgG1 antibodies and reduction of a possible Th2 lymphocyte marker (sCD30) may be associated with symptomatic improvements.

Occupational asthma and rhinitis caused by multiple herbal agents in a pharmacist.

BACKGROUND: Herb agents have been widely used for centuries in the Orient and they have been cultivated throughout Asia. There have been a few cases of occupational allergy caused by herb materials. We report a case of occupational asthma and rhinitis caused by six herb materials in a pharmacist working at a pharmacy. OBJECTIVE: We sought the role of immediate hypersensitivity in herbal agent-induced asthma in a pharmacist. METHODS AND RESULTS: The patient had strong positive responses on skin prick test to extracts of six herb materials: Chunkung (Cnidii rhizoma), Banha (Pinellia ternata), Sanyak (Dioscorea radix), Kangwhal (Ostericum koreanum), Danggui (Angelica radix), and Kunkang (Zingiberis rhizoma). Bronchoprovocation tests showed an early asthmatic response to Danggui extract. Serum specific IgE antibodies to Chunkung, Banha, and Sanyak were detected by ELISA with no specific IgE bindings to Kangwhal, Danggui, and Kunkang extracts. Twelve percent sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and IgE immunoblotting revealed one IgE binding component (60 kD) within Chunkung extract, two (10, 25 kD) in Banha, and four (33, 34, 65, 98 kD) in Sanyak. Basophil histamine release test revealed that Danggui extract could release a greater amount of histamine from basophils in the patient than in a healthy control. CONCLUSIONS: Chunkung, Banha, and Sanyak may induce IgE-mediated bronchoconstriction in an exposed worker, and Danggui can cause bronchoconstriction by direct histamine-releasing effect from mast cells in a sensitized patient.

Are there any links between Hop Japanese pollen and other weed pollens or food allergens on skin prick tests?

Recent investigations suggest that the importance of Hop Japanese pollen, which has been known as one of the major causative weed pollens, is increasing in this country. There have been few data dealing with the allergenic relationship between Hop J pollen and other food or inhalant allergens. Among 2909 patients who visited the Allergy Clinic of Ajou University Hospital, Suwon, Korea, 471 patients sensitized to Hop J, mugwort, or ragweed pollens on skin-prick test were enrolled. Positive rates to common inhalant or food allergens and their allergenic relationships with other pollens or food allergens were analyzed based upon skin-prick test results. The positive rates to sunflower, fat hen, nettle, grass (Bermuda, Orchard) and tree (alder, birch, and poplar) pollen were significantly higher in those sensitized to Hop J pollen than in those of negative responders (p < 0.05, respectively). No significant associations were noted with ragweed or mugwort pollen (p > 0.05, respectively). In regard to food-related allergens, an association was noted between Hop (Humulus lupulus) or celery allergens in those sensitized to Hop J pollen (p < 0.05, respectively). Hop J pollen may have possible links with celery, Hop, and sunflower pollens on skin-prick test. Further in vitro investigations will be needed to evaluate the possibility of cross-reacting components between them.

In vitro function of S rnases in Lycopersicon peruvianum.

S RNases are products of the S locus that are expressed in the stylar tissue of Lycopersicon peruvianum with the gametophytic self-incompatibility (SI) system. Two S RNases (S12 and Sa) with RNase activity from the S12Sa genotype of L. peruvianum were purified using gel filtration and cation-exchange chromatography. The molecular masses of the two RNases, S12 and Sa, were 21 and 23.1 kDa, respectively. The specific activities of S12 and Sa for torula yeast rRNA as a substrate were 8,500 and 6,000 units/ml, respectively. Of various reagents tested for RNase activities, ZnSO4 and CuSO4 were found to remarkably reduce its activity. The growth of S12Sa pollen was inhibited when it was cultured in a pollen germination medium that contained the purified S12 RNase. The result suggested that the S RNase was either a probable inhibitor of pollen growth or controlled pollen growth. Additionally, 512Sa pollens germinated well in vitro in a germination medium that contained S12 RNase in the presence of ZnSO4 and CuSO4. Our finding suggests that the treatment of S RNase with its inhibitor destroys the SI ability on an in vitro self-pollen growth test.

Selenite negatively regulates caspase-3 through a redox mechanism.

Selenium, an essential biological trace element, exerts its modulatory effects in a variety of cellular events including cell survival and death. In our study we observed that selenite protects HEK293 cells from cell death induced by ultraviolet B radiation (UVB). Exposure of HEK293 cells to UVB radiation resulted in the activation of caspase-3-like protease activity, and pretreatment of the cells with z-DEVD-fmk (N-benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethylketone), a caspase-3 inhibitor, prevented UVB-induced cell death. Interestingly, enzymatic activity of caspase-3-like protease in cell lysates of UVB-exposed cells was repressed in vitro by the presence of selenite. Selenite also inhibited the in vitro activity of purified recombinant caspase-3 in cleaving Ac-DEVD-pNA (N-acetyl-Asp-Glu-Asp-p-nitroanilide) or ICAD(L) (inhibitor of a caspase-activated deoxyribonuclease) and in the induction of DNA fragmentation. The inhibitory action of selenite on a recombinant active caspase-3 could be reversed by sulfhydryl reducing agents, such as dithiothreitol and beta-mercaptoethanol. Furthermore, pretreatment of cells with selenite suppressed the stimulation of the caspase-3-like protease activity in UVB-exposed cells, whereas dithiothreitol and beta-mercaptoethanol reversed this suppression of the enzymatic activity. Taken together, our data suggest that selenite inhibits caspase-3-like protease activity through a redox mechanism and that inhibition of caspase-3-like protease activity may be the mechanism by which selenite exerts its protective effect against UVB-induced cell death.

Allium sativum potentiates suicide gene therapy for murine transitional cell carcinoma.

This study evaluated the synergistic effect of Allium sativum (AS) with suicide gene therapy for transitional cell carcinoma (TCC) of the bladder. Subcutaneous TCCs were established in syngeneic C3H/He mice with 1 x 10(5) MBT-2 cells. AS liquid extract was injected at the site of tumor transplantation on Day 1 for three weeks (Experiment I) and into the established tumors weekly for five weeks (Experiment II) in combination with or without gene therapy using a replication-defective adenoviral vector containing a herpes simplex virus thymidine kinase (HSV-TK) gene under the transcriptional control of Rous sarcoma virus (RSV) promoter (Ad-RSV-TK, 5 x 10(8) plaque-forming units) plus ganciclovir (20 mg/kg/day i.p.). AS demonstrated a statistically significant reduction in incidence of TCC (cumulative dose 25 mg of AS). Combination AS-suicide gene therapy significantly inhibited the tumor growth compared with the controls, which was evidenced by apoptosis on histomorphological and immunohistochemical studies. These results suggest that AS had a definite antitumor effect in inhibiting tumorigenesis and growth of TCC in a murine model. AS treatment combined with suicide gene therapy had significant additive antitumor effects on TCC and may provide a novel and effective treatment modality for TCC of the bladder.

Oilseed rape allergy presented as occupational asthma in the grain industry.

BACKGROUND: There have been several reports on respiratory allergic symptoms induced by pollen of oilseed rape. To the best of our knowledge, this is the first report dealing with oilseed rape dust mainly composed of seeds, as an occupational allergen in the grain industry. In this paper, we present a case of occupational asthma caused by oilseed rape dust from the Animal Feed Industry, which proved to be induced by an IgE-mediated reaction. METHODS AND RESULTS: The patient displayed positive responses to Dermatophagoides farinae as well as oilseed rape dust extract. The bronchoprovocation test showed an early asthmatic response to oilseed rape dust extract. Serum specific IgE antibody to oilseed rape antigen was detected by enzyme-linked immunosorbent assay (ELISA). ELISA inhibition test showed significant inhibitions with addition of oilseed rape antigen. In order to further identify the allergenic components of extract, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis were performed. Fourteen IgE-binding components ranging from 10 to 160kDa were detected within the oilseed rape extract. CONCLUSION: These results suggest that the inhalation of oilseed rape dust, not pollen, can cause IgE mediated bronchoconstriction in an exposed worker of the grain industry.

Analysis of induced sputum for studying allergen-specific IgE antibodies in airway secretion from asthmatic patients.

BACKGROUND: Allergen-specific IgE antibodies have been considered to play an important role in the pathogenesis of atopic asthma. However, studies on allergen-specific IgE antibodies in airway secretion from asthmatic patients are very rare compared with those in serum. OBJECTIVES: The present study was undertaken to determine whether induced sputum might provide a useful method for analysing allergen-specific IgE antibodies in airway secretions from asthmatic patients. METHODS: Specific IgE antibodies to house dust mite (HDM) antigen were measured in induced sputum from 10 HDM-sensitive asthmatic patients and 12 non-allergic controls by enzyme-linked immunosorbent assay. HDM-specific IgE was regarded as positive when the absorbance value was higher than mean + 2SD of controls. Their antigen-binding characteristics were determined by immunoblot analysis. RESULTS: HDM-specific IgE was positive in induced sputum from seven of 10 HDM-sensitive asthmatics. The IgE binding to HDM antigen could be inhibited by fluid phase HDM antigen in a dose-dependent manner, not by mugwort antigen. Treatment of induced sputum with dithiothreitol decreased the antigen-specific bindings, and increased the non-specific bindings on the measurement of HDM-specific IgE. These effects were significant in a concentration of dithiothreitol greater than 0.05%. Immunoblot analysis revealed that HDM-specific IgE antibodies in induced sputum recognized the HDM antigens with molecular weights of 42, 34, 32, 25 and 14 kDa. These antigen binding characteristics were similar to those in serum. CONCLUSION: We conclude that analysis of induced sputum is a useful non-invasive method for studying allergen-specific IgE antibodies in airway secretion from asthmatic patients.

Protein surface recognition by synthetic agents: design and structural requirements of a family of artificial receptors that bind to cytochrome c.

The design, synthesis, and evaluation of a novel series of receptors for protein surface recognition are described. The design of these agents is based around the attachment of four constrained peptide loops onto a central calix[4]arene scaffold. This arrangement mimics the role of the hypervariable loops in antibody combining regions and defines a large surface area for binding to a complementary region of the exterior of a target protein. Using affinity and gel filtration chromatographies we show that one particular receptor binds strongly to the surface of cytochrome c. The site of binding is presumably close to the heme edge region, which contains several charged lysine residues. This is supported by the observation that the receptor inhibits the reduction of Fe(III) cytochrome c to its Fe(II) form. We also show that binding is strongly dependent on the nature of the substituents on the lower rim of the calixarene. The nmr and computational studies suggest that this effect may be due to conformational differences among the differently substituted receptors.