Safety and efficacy of fluoroscopy-guided urethral catheterization in case of failed blind or cystoscopy-assisted urethral catheterization.

This retrospective study evaluated the safety and efficacy of fluoroscopy-guided urethral catheterization in patients who failed blind or cystoscopy-assisted urethral catheterization. We utilized our institutional database between January 2011 and March 2023, and patients with failed blind or cystoscopy-assisted urethral catheterization and subsequent fluoroscopy-guided urethral catheterization were included. A 5-Fr catheter was inserted into the urethral orifice, and the retrograde urethrography (RGU) was acquired. Subsequently, the operator attempted to pass a hydrophilic guidewire to the urethra. If the guidewire and guiding catheter could be successfully passed into the bladder, but the urethral catheter failed pass due to urethral stricture, the operator determined either attempted again with a reduced catheter diameter or performed balloon dilation according to their preference. Finally, an appropriately sized urethral catheter was selected, and an endhole was created using an 18-gauge needle. The catheter was then inserted over the wire to position the tip in the bladder lumen and ballooned to secure it. We reviewed patients' medical histories, the presence of hematuria, and RGU to determine urethral abnormalities. Procedure-related data were assessed. Study enrolled a total of 179 fluoroscopy-guided urethral catheterizations from 149 patients (all males; mean age, 73.3 ± 13.3 years). A total of 225 urethral strictures were confirmed in 141 patients, while eight patients had no strictures. Urethral rupture was confirmed in 62 patients, and hematuria occurred in 34 patients after blind or cystoscopy-assisted urethral catheterization failed. Technical and clinical success rates were 100%, and procedure-related complications were observed in four patients (2.2%). The mean time from request to urethral catheter insertion was 129.7 ± 127.8 min. The mean total fluoroscopy time was 3.5 ± 2.5 min and the mean total DAP was 25.4 ± 25.1 Gy cm2. Balloon dilation was performed in 77 patients. Total procedure time was 9.2 ± 7.6 min, and the mean procedure time without balloon dilation was 7.1 ± 5.7 min. Fluoroscopy-guided urethral catheterization is a safe and efficient alternative in patients where blind or cystoscopy-assisted urethral catheterization has failed or when cystoscopy-urethral catheterization cannot be performed.

Synergistic Effect of Cold Treatment Combined with Ethyl Formate Fumigation against Drosophila suzukii (Diptera: Drosophilidae).

Drosophila suzukii is a quarantine pest that is rapidly spreading in berries. This study evaluated the synergistic effect of combination treatment with ethyl formate (EF) and cold temperature for D. suzukii control on imported grapes. A higher insecticidal effect was observed at 1 °C than at 5 °C at all developmental stages, and the pupal stage showed the strongest tolerance to cold temperature. After EF fumigation alone, eggs showed the highest tolerance at 216.67 mg·h/L (LCT99 value), and adults showed the highest susceptibility at <27.24 mg·h/L. Among the combination treatment methods, cold temperature after fumigation resulted in the best synergistic effect. The effect of this combination was significant, with 23.3% higher mortality for eggs, 22.4% for larvae, and 23.4% for pupae than observed with EF fumigation alone. Furthermore, the period of complete D. suzukii control in the 12 L desiccator was shorter in the combination treatment group at the LCT80 value than at the LCT50 value of the egg stage. EF showed a very high sorption rate (24%) after 4 h of exposure at a grape loading ratio of 15% in a 0.65 m3 fumigation chamber. As the grape loading ratio for combination treatment decreased, D. suzukii mortality increased, but when EF was administered at the LCT80 value, there was little difference in the mortalities of the eggs and larvae but not the pupae. All D. suzukii developmental stages were completely controlled within 7 days after combination treatment, and phytotoxicity was not observed in grapes. These results suggest that the combination of cold-temperature treatment and EF fumigation could be used for D. suzukii control.

Efficacy and Phytotoxicity Assessment of Successive Application of Methyl Bromide and Cold Treatment on Export Strawberry Fruits.

Recently, spotted wing Drosophila, Drosophila suzukii, is globally prevalent and causes agricultural losses to many fruits. To export Korean strawberry, methyl bromide fumigation is required to remove D. suzukii infestations, but Korean strawberry farmers are worried about fruit damage because methyl bromide can cause phytotoxicity on fresh commodities. In this report, we assessed the efficacy and phytotoxicity of single and successive application of methyl bromide and cold treatment on an export variety of strawberry to reduce fruit damage. The currently recommended dosage of methyl bromide, 40 g/m3 for 3 h at 18 °C, was enough to control all stages of D. suzukii without phytotoxicity. A dosage of 20 g/m3 of methyl bromide treatment for 3 h, followed by 1 d of cold (0 °C) treatment, showed 100% mortality in all growth stages of D. suzukii without fruit damage. Successive application of methyl bromide and cold treatment shows potential as a method of decreasing phytotoxicity and reducing the use of methyl bromide for environmental considerations.

Fumigation Activity against Phosphine-Resistant Tribolium castaneum (Coleoptera: Tenebrionidae) Using Carbonyl Sulfide.

Phosphine resistance is occurring among stored-grain pests worldwide. This study investigated the fumigation activity of phosphine (PH3) and carbonyl sulfide (COS) against domestic strain (d-strain) Tribolium castaneum, resistance strain (r-strain) T. castaneum and Oryzaephilus surinamensis. All developmental stages of the pests were exposed to two fumigants (PH3 and COS), and the fumigation activity according to the dose and exposure time was evaluated in a 12-L desiccator and 0.5 m3 fumigation chamber. The rice sorption rate and quality following exposure to thetwofumigants were evaluated. The mortality was 2.9% in r-strain T. castaneum, 49.5% in d-strain T. castaneum and 99.2% in O. surinamensis when 2 mg/L PH3 was used in a 12-L desiccator for 4 h. However, all pest developmental stages showed 100% mortality after 24 h of exposure in the 0.5 m3 fumigation chamber, except for the r-strain T. castaneum. A mortalityof 100% was observed in all of the r-strain T. castaneum developmental stages at an exposure time of 192 h. For COS applied at 40.23 mg/L and 50 g/m3 in the 12-L desiccator and the 0.5 m3 fumigation chamber, respectively, 100% mortality was observed across all developmental stages regardless of species and strain. The sorption of COS was 10% higher than that of PH3, but there was no significant difference in rice quality compared to that in the control. Therefore, this study suggests that COS can be used for controlling T. castaneum resistant to PH3.

Susceptibility of the Cigarette Beetle Lasioderma serricorne (Fabricius) to Phosphine, Ethyl Formate and Their Combination, and the Sorption and Desorption of Fumigants on Cured Tobacco Leaves.

The susceptibility of Lasioderma serricorne to phosphine (PH3), ethyl formate (EF) and their combination (PH3 + EF) was evaluated in this study. Eggs, larvae, pupae and adults were subjected to treatment with fumigants to determine the 90% lethal concentration time (LCt90) values. Treatment with PH3 for 20 h resulted in LCt90 values of 1.15, 1.39, 14.97 and 1.78 mg h/L while treatment with EF resulted in values of 157.96, 187.75, 126.06 and 83.10 mg h/L, respectively. By contrast, the combination of PH3 + EF resulted in LCt90 values of 36.05, 44.41, 187.17 and 35.12 mg h/L after 4 h. These results show that, through treatment with PH3 + EF, control can be achieved at lower concentrations than for treatment with EF alone and at lower exposure times than for treatment with PH3 alone. The sorption rates of the fumigants on cured tobacco leaves were determined for filling ratios of 2.5%, 5.0% and 10.0% (w/v). Cured tobacco leaves were treated with either 2 mg/L PH3, 114 mg/L EF or 0.5 mg/L PH3 + 109 mg/L EF. Treatment with PH3 showed sorption rates of 0.0%, 7.1% and 14.3%. EF, however, showed higher sorption rates of 64.9%, 68.5% and 75.5%, respectively, for the indicated filling ratios. When PH3 and EF were combined, the sorption rate of PH3 was 0.0%, while the sorption rates of EF were lower (9.1%, 12.0% and 23.2%) than treatment with only EF. EF required a ventilation time of longer than 22 h to desorb from cured tobacco leaves. Therefore, PH3 + EF can effectively control L. serricorne in cured tobacco leaves, with sufficient ventilation time required after treatment for the safety of workers.

Histone acetylation together with DNA demethylation empowers higher plasticity in adipocytes to differentiate into osteoblasts.

Bone regeneration has been a challenge for both researchers and clinicians. In the field of tissue engineering, much effort has been made to identify cell sources including stem cells. The present study aimed to induce trans-differentiation from adipocytes to osteoblasts using epigenetic modifiers; 5-aza-dC and/or trichostatin-A (TSA). 3 T3-L1 preadipocytes were treated with TSA (100 nM) and then with Wnt3a (50 ng/ml). Microscopic observation showed trans-differentiated cell morphology. Methylation-specific PCR and immunoblotting were performed to analyze the DNA methylation and histone acetylation patterns. The gene expression was determined by real-time PCR. Based on these in vitro experiments, in vivo mouse experiments supplemented the possibility of trans-differentiation by epigenetic modification. TSA induced the acetylation of lysine9 on histone H3, and a sequential Wnt3a treatment stimulated the expression of bone marker genes in adipocytes, suppressing adipogenesis and stimulating osteogenesis. Furthermore, TSA induced DNA hypomethylation, and a combined treatment with TSA and 5-aza-dC showed a synergistic effect in epigenetic modifications. The number of adipocytes and DNA methylation patterns of old (15 months) and young (6 weeks) mice were significantly different, and TSA and sequential Wnt3a treatments increased bone formation in the old mice. Collectively, our results confirmed cell trans-differentiation via epigenetic modifications and osteogenic signaling from adipocytes to osteoblasts for the bone regeneration in vitro and in vivo, and indicated that histone acetylation could induce DNA hypomethylation, enhancing the chance of trans-differentiation.

Comparison of the Efficacy and Phytotoxicity of Phosphine and Ethyl Formate for Controlling Pseudococcus longispinus (Hemiptera: Pseudococcidae) and Pseudococcus orchidicola on Imported Foliage Nursery Plants.

The fumigation activity of phosphine (PH3) and ethyl formate (EF) and their phytotoxicity to 13 imported foliage nursery plant species were evaluated. The lethal concentration and time (LCT99) values of the PH3 indicated that the susceptibility of the nymphs (3.95 and <0.45 mg·h/liter, respectively) was higher than that of the adults (5.29 and 3.66 mg·h/liter, respectively) of two mealybugs [Pseudococcus longispinus (Targioni-Tozzetti) and P. orchidicola Takahashi]. The highest concentration reduction rate of PH3 and EF on the 13 foliage nursery plants in the 12-liter desiccator was 41.5% for Heteropanax fragrans and 71.7% for Schefflera arboricola, respectively, which indicates that PH3 has a lower sorption rate than EF. The phytotoxicities of PH3-treated foliage nursery plants did not significantly differ from those of the nontreated plants, but EF caused phytotoxicity in 11 foliage nursery plants a week after treatment. When the exposure time of PH3 increased to 24 h, the adults and nymphs of both mealybug species showed 100% mortality in the 0.5 m3 fumigation chamber. In the 10 m3 fumigation container used in the field, there was 100% mortality of both mealybugs after treatment with 2 g/m3 PH3 for 24 h at 16°C. These results indicate that EF is not a suitable mealybug fumigant due to its high sorption and phytotoxicity to foliage nursery plants, despite fumigation activity against the two species. However, PH3 seems to be suitable for mealybug fumigation in foliage nursery plants and can be used as a substitute for methyl bromide.

Efficacy and Phytotoxicity of Phosphine as Fumigants for Frankliniella occidentalis (Thysanoptera: Thripidae) on Asparagus.

The insecticidal activity of phosphine (PH3) and ethyl formate (EF) toward Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) and their phytotoxicity to asparagus were evaluated. Both the PH3 and EF fumigants showed higher lethal concentration and time (LCT) values at lower temperatures. The LCT99 values of PH3 and EF at 5°C in a 12 liters desiccator for 4 h showed the following ranking: eggs (64.69 mg·h/liter for PH3 and EF indicating phytotoxicity to asparagus), nymphs (5.54 and 17.48 mg·h/liter, respectively), and adults (3.83 and 14.67 mg·h/liter, respectively). The adsorption of PH3 was approximately 11% at 2°C and 13% at 5°C, whereas the adsorption of EF increased sharply to 88% at 2°C and 97% at 5°C. The hatching rate of F. occidentalis eggs was approximately 95% at all locations (top, middle, and bottom) in the presence of 4 mg/liter PH3 at 5°C in a 0.65-m3 fumigation chamber for 24 h. However, extension of the treatment to 48 h resulted in 100% inhibition of egg hatching. The atmospheric level of PH3 decreased below the threshold limit value after 80 min, and phytotoxicity was not observed. The results revealed that EF is highly absorbed by asparagus and is not suitable as a fumigant, but PH3 is a suitable alternative to the fumigant methyl bromide for the control of western flower thrips in asparagus.

A Combination Treatment Using Ethyl Formate and Phosphine to Control Planococcus citri (Hemiptera: Pseudococcidae) on Pineapples.

Citrus mealybug, Planococcus citri (Risso), is a known quarantine pest that is difficult to control with phosphine (PH3) or low concentrations of ethyl formate (EF), particularly at low temperatures. Methyl bromide (MB) is a fumigant used for quarantine and preshipment (QPS) that can eradicate target pests with short fumigation periods. However, MB, which is an ozone-depleting substance, is scheduled to be phased out in South Korea over the next decade. There is no ideal alternative fumigant to replace MB for QPS of perishable commodities. A laboratory study was conducted to compare the individual effects of EF and PH3 individually, and the effects of EF mixed with PH3 as an MB alternative for the control of P. citri adults, nymphs, and eggs. In comparison to treatments with EF and PH3 individually, EF mixed with PH3 resulted in high toxicity to all stages of P. citri. The eggs were more tolerant than the nymphs and adults. A mixed treatment of EF and PH3 achieved complete control of eggs infesting pineapples at concentrations of 25.1/1.0 (EF/PH3) mg/liter at 8 °C for 4 h of exposures. This new combined EF/PH3 fumigation technology could offer shorter exposure times and less damage to perishable commodities at low temperatures, and could potentially be extended to controlling other quarantine pests as a replacement treatment for fruit and vegetables in which methyl bromide is currently being used.

Postharvest Disease Control of Colletotrichum gloeosporioides and Penicillium expansum on Stored Apples by Gamma Irradiation Combined with Fumigation.

To study the control of postharvest decay caused by Colletotrichum gloeosporioides and Penicillium expansum, gamma irradiation alone or in combination with fumigation was evaluated to extend the shelf life of apples in South Korea. An irradiation dose of 2.0 kGy resulted in the maximum inhibition of C. gloeosporioides and P. expansum spore germination. The gamma irradiation dose required to reduce the spore germination by 90% was 0.22 and 0.35 kGy for C. gloeosporioides and P. expansum, respectively. Microscopic observations revealed that when the fungal spores were treated with gamma irradiation (4.0 kGy), conidial germination was stopped completely resulting in no germ tube formation in C. gloeosporioides. Treatment with the eco-friendly fumigant ethanedinitrile had a greater antifungal activity against C. gloeosporioides and P. expansum in comparison with the non-treated control under in vitro conditions. The in vitro antifungal effects of the gamma irradiation and fumigation treatments allowed us to further study the effects of the combined treatments to control postharvest decay on stored apples. Interestingly, when apples were treated with gamma irradiation in combined with fumigation, disease inhibition increased more at lower (< 0.4 kGy) than at higher doses of irradiation, suggesting that combined treatments reduced the necessary irradiation dose in phytosanitary irradiation processing under storage conditions.

Postharvest Control of Botrytis cinerea and Monilinia fructigena in Apples by Gamma Irradiation Combined with Fumigation.

To extend the shelf life of apples in South Korea, we evaluated the effect of gamma irradiation alone or gamma irradiation combined with fumigation on the control of postharvest decay caused by Botrytis cinerea and Monilinia fructigena. An irradiation dose of 1.0 kGy caused the maximal inhibition of B. cinerea and M. fructigena spore germination. The gamma irradiation dose required to reduce the spore germination by 90% was 0.76 and 0.78 kGy for B. cinerea and M. fructigena, respectively. Inhibition of conidial germination of both fungal pathogens occurred at a greater level at the doses of 0.2 to 1.0 kGy compared with the nontreated control; 0.2 kGy caused 90.5 and 73.9% inhibition of B. cinerea and M. fructigena, respectively. Treatment in vitro with the ecofriendly fumigant ethanedinitrile had a greater effect compared with the nontreated control. The in vitro antifungal effects of the gamma irradiation and fumigation treatments allowed us to further study the effects of the combined treatments. Interestingly, when irradiation was combined with fumigation, the percentage of disease inhibition increased more at lower (<0.4 kGy) than at higher doses of irradiation, suggesting that the combined treatments reduced the necessary irradiation dose in phytosanitary irradiation processing under storage conditions.

Multiplexed Cassegrain Reflector Antenna for Simultaneous Generation of Three Orbital Angular Momentum (OAM) Modes.

A multiplexed Cassegrain reflector antenna with a 2 × 2 open-ended rectangular waveguide (OERW) matrix feed and an orbital angular momentum (OAM) mode mux is proposed for the simultaneous generation of three OAM modes (l = 0, ±1). The OAM mode mux (OMM) was designed using sequential combinations of quadrature hybrids, crossovers, and phase shifters to multiplex and demultiplex three OAM modes at the same time. The 2 × 2 OERW matrix feed and the OMM were separately measured and their performances were verified according to proposed theories. A near-field antenna measurement for a multiplexed Cassegrain reflector antenna was conducted to obtain the far-field magnitude and phase patterns around polar elevation angle θ and azimuthal angle ϕ, thus confirming that our antenna can produce three OAM modes simultaneously. We also measured the communication link characteristics of two identical multiplexed antennas. The measurement results show that the channel isolation of three OAM modes is more than 12.7 [dB] and 17 [dB] for fixed and compensated receiver positions, respectively, indicating that the proposed antenna system can be used for independent communication links with the same frequency and polarisation.

Pin1-mediated Modification Prolongs the Nuclear Retention of ß-Catenin in Wnt3a-induced Osteoblast Differentiation.

The canonical Wnt signaling pathway, in which ß-catenin nuclear localization is a crucial step, plays an important role in osteoblast differentiation. Pin1, a prolyl isomerase, is also known as a key enzyme in osteogenesis. However, the role of Pin1 in canonical Wnt signal-induced osteoblast differentiation is poorly understood. We found that Pin1 deficiency caused osteopenia and reduction of ß-catenin in bone lining cells. Similarly, Pin1 knockdown or treatment with Pin1 inhibitors strongly decreased the nuclear ß-catenin level, TOP flash activity, and expression of bone marker genes induced by canonical Wnt activation and vice versa in Pin1 overexpression. Pin1 interacts directly with and isomerizes ß-catenin in the nucleus. The isomerized ß-catenin could not bind to nuclear adenomatous polyposis coli, which drives ß-catenin out of the nucleus for proteasomal degradation, which consequently increases the retention of ß-catenin in the nucleus and might explain the decrease of ß-catenin ubiquitination. These results indicate that Pin1 could be a critical target to modulate ß-catenin-mediated osteogenesis.

Effect of alumina nanofiller on properties of heat-treated glass composite sealants.

Alkali/alkaline-earth borosilicate glass-alumina composites containing 10 vol% Al2O3 were prepared for use as solid oxide fuel cell sealants. The effect of heat treatment and Al2O3, addition on the viscosities and electrical conductivities was investigated to improve cyclic sealing performance. Upon a 48-h heat treatment, the viscosity of the glass-alumina composites at 750 degrees C was approximately four orders of magnitude higher than that of the base glass owing to the crystallization of the glass in the presence of Al2O3. Heat treatment increased the electrical conductivities of both the base glass and the glass-alumina composites. The electrical conductivities of glass-alumina composites in the range from 400 degrees C to 550 degrees C were three times higher than those of the base glass regardless of heat treatment. This increase in the conductivities and viscosities by heat treatment was attributed to the devitrification and structural densification of the sealing glass and the partial dissolution of the Al2O3 filler in alkali/alkaline-earth borosilicate sealing glass.

Bacillus anthracis capsule activates caspase-1 and induces interleukin-1beta release from differentiated THP-1 and human monocyte-derived dendritic cells.

The poly-gamma-d-glutamic acid (PGA) capsule is one of the major virulence factors of Bacillus anthracis, which causes a highly lethal infection. The antiphagocytic PGA capsule disguises the bacilli from immune surveillance and allows unimpeded growth of bacilli in the host. Recently, efforts have been made to include PGA as a component of anthrax vaccine; however, the innate immune response of PGA itself has been poorly investigated. In this study, we characterized the innate immune response elicited by PGA in the human monocytic cell line THP-1, which was differentiated into macrophages with phorbol 12-myristate 13-acetate (PMA) and human monocyte-derived dendritic cells (hMoDCs). PGA capsules were isolated from the culture supernatant of either the pXO1-cured strain of B. anthracis H9401 or B. licheniformis ATCC 9945a. PGA treatment of differentiated THP-1 cells and hMoDCs led to the specific extracellular release of interleukin-1beta (IL-1beta) in a dose-dependent manner. Evaluation of IL-1beta processing by Western blotting revealed that cleaved IL-1beta increased in THP-1 cells and hMoDCs after PGA treatment. Enhanced processing of IL-1beta directly correlated with increased activation of its upstream regulator, caspase-1, also known as IL-1beta-converting enzyme (ICE). The extracellular release of IL-1beta in response to PGA was ICE dependent, since the administration of an ICE inhibitor prior to PGA treatment blocked induction of IL-1beta. These results demonstrate that B. anthracis PGA elicits IL-1beta production through activation of ICE in PMA-differentiated THP-1 cells and hMoDCs, suggesting the potential for PGA as a therapeutic target for anthrax.

Poly-gamma-d-glutamic acid and protective antigen conjugate vaccines induce functional antibodies against the protective antigen and capsule of Bacillus anthracis in guinea-pigs and rabbits.

Anthrax is a lethal infectious disease caused by the spore-forming Bacillus anthracis. The two major virulence factors of B. anthracis are exotoxin and the poly-gamma-d-glutamic acid (PGA) capsule. The three components of the exotoxin, protective antigen (PA), lethal factor and edema factor act in a binary combination, which results in massive edema and organ failure in the progress of anthrax disease. The antiphagocytic PGA capsule disguises the bacilli from immune surveillance and allows unimpeded growth of bacilli in the host. Because PA can elicit a protective immune response, it has been a target of the anthrax vaccine. In addition to PA, efforts have been made to include PGA as a component of the anthrax vaccine. In this study, we report that PA-PGA conjugates induce expressions of anti-PA, anti-PGA and toxin-neutralizing antibodies in guinea-pigs and completely protect guinea-pigs against a 50 x LD(50) challenge with fully virulent B. anthracis spores. Polyclonal rabbit antisera produced against either PA or ovalbumin conjugated to a PGA-15mer offer a partial passive protection to guinea-pigs against B. anthracis infection, indicating that anti-PGA antibodies play a protective role. Our results demonstrate that PA-PGA conjugate vaccines are effective in the guinea-pig model, in addition to the previously reported mouse model.

Bacillus anthracis lethal toxin attenuates lipoteichoic acid-induced maturation and activation of dendritic cells through a unique mechanism.

Lethal toxin (LT), produced by the gram-positive bacterium Bacillus anthracis, was identified as a major etiologic agent causing anthrax due to its strong immunotoxicity. Gram-positive bacteria express lipoteichoic acid (LTA), which is considered as a counterpart to lipopolysaccharide (LPS) of gram-negative bacteria, but differs from LPS in the structure and function. Since dendritic cells (DCs) are essential for the appropriate initiation of immune response, we investigated the effect of LT on LTA-induced DC maturation using immature DCs prepared by differentiation of C57BL/6 mouse bone marrow cells. When immature DCs were matured with LTA in the presence of LT, the expression of representative markers for DC maturation such as CD80, CD83, and CD86 together with MHC class I and II molecules was inhibited. LT ameliorated the attenuation of endocytic capacity during DC maturation by LTA while such effect was not observed in LPS-matured DCs. Furthermore, exposure to LT resulted in a decrease in the expression of pro-inflammatory cytokines including IL-6, TNF-alpha, and IL-12p40 in LTA-stimulated DCs as in LPS-stimulated DCs. Interestingly, LT showed a minimal change in LTA-induced IL-1beta expression while LT highly enhanced the LPS-induced IL-1beta expression. Those inhibitory effects might be associated with LT interference of LTA-signaling pathways mediated through mitogen-activated protein kinases (MAPKs) since LT suppressed phosphorylation of MAPK, which was induced by LTA. Meanwhile, no change was observed in the expression of putative anthrax toxin receptors, TEM8 and CMG2, or Toll-like receptor 2. These results suggest that LT suppresses the maturation and activation of DCs stimulated with LTA, similar to the suppression in the LPS-stimulated DCs, but via a distinct mechanism.

Simple sequence repeat (SSR)-based gene diversity in Burkholderia pseudomallei and Burkholderia mallei.

Pathogens Burkholderia pseudomallei (Bp) and Burkholderia mallei (Bm) contain a large number (> 12,000) of Simple Sequence Repeats (SSRs). To study the extent to which these features have contributed to the diversification of genes, we have conducted comparative studies with nineteen genomes of these bacteria. We found 210 genes with characteristic types of SSR variations. SSRs with nonamer repeat units were the most abundant, followed by hexamers and trimers. Amino acids with smaller and nonpolar R-groups are preferred to be encoded by the variant SSRs, perhaps due to their minimal impacts to protein functionality. A majority of these genes appears to code for surface or secreted proteins that may directly interact with the host factors during pathogenesis or other environmental factors. There also are others that encode diverse functions in the cytoplasm, and this protein variability may reflect an extensive involvement of phase variation in survival and adaptation of these pathogens.

National surveillance of antifungal susceptibility of Candida species in South Korean hospitals.

We investigated the species distribution and antifungal susceptibility of Candida isolates from tertiary and non-tertiary hospitals in South Korea from 2002-2004. Of the 612 Candida isolates that were collected, Candida albicans, C. parapsilosis, C. tropicalis, and C. glabrata occurred most frequently, accounting for 97.3% and 96.8% of the isolates in tertiary and non-tertiary hospitals, respectively. C. albicans was the most common isolate, but the incidence of non-C. albicansCandida species was higher than that of C. albicans in tertiary hospitals. The Candida species had much lower MIC(90) to voriconazole (tertiary hospitals: 0.5 microg/ml, non-tertiary hospitals: 0.25 microg/ml) than to fluconazole (tertiary hospitals: 8 microg/ml, non-tertiary hospitals: 4 microg/ml). The MIC(90) of Candida isolates to 5-flucytosine in non-tertiary hospitals was two times higher than that observed in tertiary facilities. The C. glabrata isolates showed a tendency toward strong resistance to fluconazole, but C. parapsilosis isolates were susceptible to all of the evaluated antifungal agents. Voriconazole showed strong in vitro activity against Candida species, especially C. krusei, which is resistant to fluconazole and 5-flucytosine. To our knowledge, this is the first report of Candida antifungal susceptibility that includes non-tertiary hospitals in South Korea.

Construction of a Vibrio cholerae prototype vaccine strain O395-N1-E1 which accumulates cell-associated cholera toxin B subunit.

Because of its production and use in Vietnam, the most widely used oral cholera vaccine consists of heat- or formalin-killed Vibrio cholerae whole cells (WC). An earlier version of this type of vaccine called whole cell-recombinant B subunit vaccine (BS-WC) produced in Sweden also contained the B subunit of cholera toxin (CTB). Both WC and BS-WC vaccines produced moderate levels of protection in field trials designed to evaluate their cholera efficacy. V. cholerae cells in these vaccines induce antibacterial immunity, and CTB contributes to the vaccine's efficacy presumably by stimulating production of anti-toxin neutralizing antibody. Although more effective than the WC vaccine, the BS-WC vaccine has not been adopted for manufacture by developing world countries primarily because the CTB component is difficult to manufacture and include in the vaccine in the doses needed to induce significant immune responses. We reasoned this was a technical problem that might be solved by engineering strains of V. cholerae that express cell-associated CTB that would co-purify with the bacterial cell fraction during the manufacture of WC vaccine. Here we report that construction of a V. cholerae O1 classical strain, O395-N1-E1, that has been engineered to accumulate CTB in the periplasmic fraction by disrupting the epsE gene of type II secretion pathway. O395-N1-E1 induces anti-CTB IgG and vibriocidal antibodies in mice immunized with two doses of formalin killed whole cells. Intraperitoneal immunization of mice with O395-N1-E1 induced a significantly higher anti-CTB antibody response compared to that of the parental strain, O395-N1. Our results suggest that this prototype cholera vaccine candidate strain may assist in preparing improved and inexpensive oral BS-WC cholera vaccine without the need to purify CTB separately.