Molecular screening of multidrug-resistance tuberculosis by a designated public health laboratory in Taiwan.

This manuscript describes our experience in early identifying MDR-TB cases in high-risk populations by setting up a single-referral molecular diagnosis laboratory in Taiwan. Taiwan Centers for Disease Control designated a single-referral laboratory to provide the GenoType MTBDRplus test for screening high-risk MDR-TB populations nationwide in 2012-2015. A total of 5,838 sputum specimens from 3,308 patients were tested within 3 days turnaround time. Compared with the conventional culture and drug susceptibility testing, the overall performance of the GenoType MTBDRplus test for detecting TB infection showed accuracy of 70.7%, sensitivity of 85.9%, specificity of 65.7%, positive predictive value of 45.5%, and negative predictive value of 93.3%. And the accuracy of detecting rifampin (RIF) resistance, isoniazid (INH) resistance, and MDR-TB (resistant to at least RIF and INH) were 96.5%, 95.2%, and 97.7%, respectively. MDR-TB contacts presented a higher rate of mutated codons 513-519, GenoType MTBDRplus banding pattern: rpoB WT3(-), and rpoB WT4(-) than the treatment failure group. The MDR-TB contact group also had a higher rate of inhA C15T mutation, banding pattern: inhA WT1(-), and inhA MUT1(+) than the recurrent group. Resistance profiles of MDR-TB isolates also varied geographically. The referral molecular diagnosis system contributed to rapid detection and initiation of appropriate therapy.

Antioxidant molecular targets of wheat bran fermented by white rot fungi and its potential modulation of antioxidative status in broiler chickens.

1. The study focused on antioxidant molecular targets of wheat bran fermented by white rot fungi (WRF) in poultry. After solid-state fermentation of wheat bran by WRF for 12 d, scanning electron microscopy found that the lignocellulose structure showed degradation and fragmentation. 2. A total of 300 1-d-old broilers were evenly divided by gender and randomly allocated into the following treatments: (1) maize-soybean meal (control group), (2) 10% of wheat bran replacing maize (10% WB group) or (3) 10% of fermented wheat bran replacing maize (10% FWB group). 3. The results indicated that the antioxidant gene expression, such as haem oxygenase-1 and glutathione-S-transferase of chicken peripheral blood mononuclear cells, of the 10% FWB group was significantly higher than that of the control group at d 35. For genes of Nicotinamide adenine dinucleotide phosphate oxygenase 1 and reactive oxygen species modulator protein 1, the expression of the 10% FWB group was lower than that of the control group at d 21 and 35. 4. In conclusion, wheat bran fermented by WRF could increase lignocellulolytic enzyme activities and the levels of active components that further regulate the expression of antioxidant molecular targets in poultry.

Probiotic Escherichia coli Nissle inhibits IL-6 and MAPK-mediated cardiac hypertrophy during STZ-induced diabetes in rats.

Escherichia coli Nissle (EcN), a probiotic bacterium protects against several disorders. Multiple reports have studied the pathways involved in cardiac hypertrophy. However, the effects of probiotic EcN against diabetes-induced cardiac hypertrophy remain to be understood. We administered five weeks old Wistar male (271±19.4 g body weight) streptozotocin-induced diabetic rats with 109 cfu of EcN via oral gavage every day for 24 days followed by subjecting the rats to echocardiography to analyse the cardiac parameters. Overexpressed interleukin (IL)-6 induced the MEK5/ERK5, JAK2/STAT3, and MAPK signalling cascades in streptozotocin-induced diabetic rats. Further, the upregulation of calcineurin, NFATc3, and p-GATA4 led to the elevation of hypertrophy markers, such as atrial and B-type natriuretic peptides. In contrast, diabetic rats supplemented with probiotic EcN exhibited significant downregulated IL-6. Moreover, the MEK5/ERK5 and JAK2/STAT3 cascades involved during eccentric hypertrophy and MAPK signalling, including phosphorylated MEK, ERK, JNK, and p-38, were significantly attenuated in diabetic rats after supplementation of EcN. Western blotting and immunofluorescence revealed the significant downregulation of NFATc3 and downstream mediators, thereby resulting in the impairment of cardiac hypertrophy. Taken together, the findings demonstrate that supplementing probiotic EcN has the potential to show cardioprotective effects by inhibiting diabetes-induced cardiomyopathies.

Immobilization of beta-fructofuranosidases from Aspergillus on methacrylamide-based polymeric beads for production of fructooligosaccharides.

beta-Fructofuranosidases from Aspergillus niger ATCC 20611 and Aspergillus japonicus TIT-KJ1 were purified and immobilized covalently onto methacrylamide-based polymeric beads. The porous, oxriane-containing support was reactive and could bind enzymes in a buffered solution at room temperature with a density up to 0.4 mg of protein g-1 of support with 100% immobilized yield. Neither the optimum temperature for the highest enzymatic activities nor the batch reaction pattern for fructooligosaccharides formation catalyzed by beta-fructofuranosidases was changed by immobilization. The amount of fructooligosaccharides produced from 50% (w/w) sucrose solution using the prepared immobilized enzymes was determined to be approximately 60% of the total sugars in the reaction mixtures. This level of fructooligosaccharides produced by the immobilized enzymes was comparable to that resulting from processes employing other immobilized biocatalysts as shown in the literature. The fraction of total fructooligosaccharides presented in the final mixture increased with the initial sucrose concentration, while fractions of glucose and fructose decreased with an increase sucrose concentration.

Antimicrobial resistance and serotype distribution of Streptococcus pneumoniae infections in Kaohsiung from 1996 through 1999.

A total of 89 isolates of Streptococcus pneumoniae were obtained from 86 patients during the period from November 1996 through September 1999 at the Kaohsiung Medical University Hospital. The purpose of this study was to determine the antimicrobial susceptibilities and the distribution of serotypes of these isolates, and to correlate these findings with the clinical characteristics of patients. Twenty-one (23.6%) isolates were obtained from patients aged below 5 years, and 38 (42.7%) from patients aged over 65 years. These 86 patients included 53 pneumonia, 13 bacteremia (including 6 with septic shock), 8 urinary tract infection, 8 soft tissue infections, 7 acute exacerbation of chronic bronchitis, 2 ophthalmic infection, and 2 cholecystitis cases. The most frequent serotypes were types 20 (10.1%), 6 (9%), 10 (9%), 11 (9%), and 23 (9%). All isolates were included in the serotypes represented in the 23-valent pneumococcal vaccine. Thirty-four (38.2%) isolates showed reduced penicillin susceptibility by the E-test. The predominant serotypes of penicillin-resistant S. pneumoniae were types 11 (17.6%), 7 (14.7%), 6 (8.8%), 8 (8.8%), and 23 (8.8%). All isolates were susceptible to vancomycin. Resistance rate to erythromycin was 49.4%, chloramphenicol, 20.2%; and trimethoprim/sulfamethoxazole, 61.8%. Multiple resistance (> or = 3 classes of antibiotics) was found in 28 (31.5%) isolates, of which the majority were serotypes 11 (14.3%), 7 (14.3%), 6 (10.7%), 8 (10.7%), and 23 (10.7%).

Calibrated nanoscale capacitance measurements using a scanning microwave microscope.

A scanning microwave microscope (SMM) for spatially resolved capacitance measurements in the attofarad-to-femtofarad regime is presented. The system is based on the combination of an atomic force microscope (AFM) and a performance network analyzer (PNA). For the determination of absolute capacitance values from PNA reflection amplitudes, a calibration sample of conductive gold pads of various sizes on a SiO(2) staircase structure was used. The thickness of the dielectric SiO(2) staircase ranged from 10 to 200 nm. The quantitative capacitance values determined from the PNA reflection amplitude were compared to control measurements using an external capacitance bridge. Depending on the area of the gold top electrode and the SiO(2) step height, the corresponding capacitance values, as measured with the SMM, ranged from 0.1 to 22 fF at a noise level of ~2 aF and a relative accuracy of 20%. The sample capacitance could be modeled to a good degree as idealized parallel plates with the SiO(2) dielectric sandwiched in between. The cantilever/sample stray capacitance was measured by lifting the tip away from the surface. By bringing the AFM tip into direct contact with the SiO(2) staircase structure, the electrical footprint of the tip was determined, resulting in an effective tip radius of ~60 nm and a tip-sample capacitance of ~20 aF at the smallest dielectric thickness.

Overproduction of D-hydantoinase and carbamoylase in a soluble form in Escherichia coli.

The production of D-hydantoinase and carbamoylase from Agrobacterium radiobacter NRRL B11291 using T7 and trc promoters, respectively, was found to cause protein aggregates in Escherichia coli. We initiated a systematic study aimed at overproducting these two proteins in a soluble form. As a result, the protein aggregate from carbamoylase overproduction could be alleviated with the aid of GroEL/GroES. In contrast, the production of a high level of D-hydantoinase in an active form can be achieved at low temperature (25 degrees C) or by the coproduction of DnaJ/DnaK. Overall, with such approaches both recombinant proteins gain more than a four-fold increase in enzyme activity. In addition, by fusion with thioredoxin, D-hydantoinase activity can be increased 25% more than the unfused counterpart in the presence of DnaJ/DnaK. These results indicate the success of our approaches to overproducing D-hydantoinase and carbamoylase in a soluble form in E. coli.