Comparison of the characteristics of Mycobacterium tuberculosis isolates from sputum and lung lesions in chronic tuberculosis patients.

Mycobacterium tuberculosis (Mtb) in sputum originates from lung cavities in tuberculosis (TB) patients. But drug susceptibility testing (DST) of sputum Mtb can not be conducted the same as in the lung because mutagenesis of bacilli may be happening in the lung during treatment and result in the possibility of the presence of heterogeneous drug-resistant subpopulations in the different lung lesions. This could be one of the reasons for low cure rates for multi-drug resistant (MDR)-TB. We studied the resected lungs of nine surgery patients with chronic TB. The isolates isolated from the sputum and different lung lesions of each patient were tested for phenotypic DST and genotyped using restriction fragment length polymorphism (RFLP) typing method. Genetic analysis to resistance to first and second line drugs was also performed. Five of nine patients were MDR-TB and three XDR-TB. DST results for ten anti-TB drugs were in accordance among different lung lesions in eight patients. However, only three of these eight patients showed the concordance of DST with sputum. Even though the isolates were heteroresistant, genotyping them by RFLP showed the clonal population in each individual patient. Six of eight followed-up patients achieved successful cure. In conclusion, the heteroresistance between sputum and lung lesions and a clonal population without mixed infection might provide useful information in establishing treatment regimen and surgery decision for MDR- and XDR-TB.

Continuous ohmic heating of commercially processed apple juice using five sequential electric fields results in rapid inactivation of Alicyclobacillus acidoterrestris spores.

Spores of Alicyclobacillus acidoterrestris, a spoilage bacterium, cause problems for the apple juice industry because they are resistant to thermal treatment. Here, we examined the sporicidal effect of an ohmic heating (OH) system with five sequential electric fields and compared it with that of conventional heating. Apple juice product (50kg) inoculated with A. acidoterrestris spores were subjected to OH (electric field strength=26.7V/cm; frequency=25kHz) at 85-100°C for 30-90s. The effect of conventional heating was also examined under these conditions. OH treatment at 100°C for 30s resulted in total inactivation of the inoculum, with no recovery of viable cells (initial population=4.8-4.9logCFU/ml), whereas 3.6-4.9logCFU/ml of the spores survived conventional heating. OH did not alter the quality (°Brix, color, and pH) of commercial apple juice (p>0.05). These results suggest that the OH system is superior to conventional heating for rapid sterilization (30s) of apple juice to assure microbiological quality in the absence of chemical additives.

Destruction of Bacillus cereus spores in a thick soy bean paste (doenjang) by continuous ohmic heating with five sequential electrodes.

UNLABELLED: This study selected spores from Bacillus cereus FSP-2 strain (the isolate from a commercial doenjang processing line) as the test strain which showed significantly higher thermal resistance (P < 0·05) than B. cereus reference strain (ATCC 27348). The spores in doenjang were subjected to ohmic heating (OH) at 95, 105, 115 and 125°C for 30, 60 or 90 s using a five sequential electrode system (electrical field: 26·7 V cm(-1) ; alternating current frequency: 25 kHz). OH at 105°C for 30-90 s reduced the B. cereus spore count in doenjang samples to <4 log CFU g(-1) . Since OH treatment at 115 and 125°C caused a perceivable colour change in the product (>1·5 National Bureau of Standards units), treatment at 105°C for 60 s was selected and applied on a large scale (500 kg of product). Reliable and reproducible destruction of B. cereus spores occurred; the reductions achieved (to < 4 log CFU g(-1) ) met the Korean national standards. Scanning electron microscopy revealed microstructural alterations in the spores (shrinkage and a distorted outer spore coat). OH is an effective method for destroying B. cereus spores to ensure the microbiological quality and safety of a thick, highly viscous sauce. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that an ohmic heating (OH) using a five sequential electrode system can effectively destroy highly heat-resistant Bacillus cereus spores which have been frequently found in a commercial doenjang processing line without perceivable quality change in the product. In addition, it may demonstrate high potential of the unique OH system used in this study that will further contribute to ensure microbiological quality and safety of crude sauces containing high levels of electrolyte other than doenjang as well.

Inactivation of Bacillus cereus spores in a tsuyu sauce using continuous ohmic heating with five sequential elbow-type electrodes.

AIMS: The effect of ohmic heating (OH) in a pilot plant system which had a zig-zag shaped (elbow-type) ohmic heater with five sequential voltage electrodes was investigated on Bacillus cereus spores in a commercial tsuyu sauce. METHODS AND RESULTS: The electrical field was fixed at 26·7 V cm(-1) with an alternating current frequency of 25 kHz. Raw tsuyu sauce (50 l) inoculated with B. cereus spores was submitted in a 4 × 3 factorial design to the OH system and heated at 95, 105, 115, and 125°C each for 30, 60, and 90 s. Survival of B. cereus spores and colour change in the commercial tsuyu sauce were both measured before and after treatment. As the treatment temperature and time increased, the number of surviving B. cereus spores decreased. The OH treatment in a bath-type process at 105°C for more than 30 s resulted in the total inactivation of the inoculated B. cereus spores (average 5·4 log reductions to undetectable levels after treatment). The OH protocol of heating at 105°C for 60 s which ensure complete eradication of the inoculated spores without compromising product quality was chosen and investigated for its suitability for commercial application on bulk quantities of samples (500 l). Reliable and reproducible reductions in B. cereus spore counts of 4·7-5·5 log CFU ml(-1) (mean ± standard deviation = 5·1 ± 0·3 CFU ml(-1) ) were achieved by the selected protocol of the continuous OH treatment (105°C for 60 s). CONCLUSION: This study suggests that OH treatment with five sequential elbow-type electrodes has great potential as an industrial sterilizing method for liquid food contaminated with B. cereus spores. SIGNIFICANCE AND IMPACT OF THE STUDY: This procedure will enhance the microbiological quality of liquid foods while minimizing quality deterioration.

Clinical outcomes in Asian elderly kidney transplant recipients: a multicenter cohort study.

BACKGROUND: The kidney transplantation rate in elderly patients is increasing rapidly. However, the clinical outcomes of kidney transplantation in elderly patients have not yet been thoroughly evaluated. METHODS: This multicenter cohort study included adult kidney transplant recipients (KTRs) admitted to five major tertiary hospitals in Korea between 1997 and 2012. A total of 3,565 adult participants were enrolled. Patient survival, allograft survival, and biopsy-proven acute rejection (BPAR) of 242 elderly recipients (≥ 60 years) were assessed and compared with those of a younger population. RESULTS: Patients were divided into five groups according to age at time of transplantation. The proportion of elderly patients was 6.7 % (mean age, 63.1 ± 2.7 years; n = 242). The numbers of male patients (69.4%), those with diabetes mellitus history (36.3%), and those with pretransplantation ischemic heart disease history (17.7%) were significantly higher in the elderly group than in the younger age groups. Elderly patients were more likely to receive a cadaveric kidney, and overall mortality rates were significantly higher in the elderly patients (1-year survival 93.3%, 5-year survival 91.3%). However, death-censored allograft survival rate and BPAR were not affected by patient age (P = .104 and .501, respectively). Among the elderly, BPAR and female donors were independent risk factors for allograft loss. CONCLUSION: The overall survival rate of the elderly KTRs was significantly lower than that of younger KTRs. However, the death-censored allograft survival rate did not differ between groups. Kidney transplantation should not be stagnated especially in elderly patients with end-stage renal disease.

A novel transverse expression domain in the mouse cerebellum revealed by a neurofilament-associated antigen.

The mammalian cerebellum is composed of a highly reproducible array of transverse zones, each of which is subdivided into parasagittal stripes. By using a combination of Purkinje cell antigenic markers and afferent tracing, four transverse zones have been identified: the anterior zone (AZ: approximately lobules I-V), the central zone (CZ: approximately lobules VI-VII), the posterior zone (PZ: approximately lobules VIII-dorsal IX) and the nodular zone (NZ: approximately ventral lobule IX+lobule X). Neurofilament-associated antigen (NAA) is an epitope recognized by a monoclonal antibody, which is expressed strongly in association with neurofilaments. During perinatal cerebellar development, anti-NAA immunocytochemistry reveals novel features of cerebellar organization. In particular, the CZ is reproducibly subdivided into anterior and posterior components. Between embryonic day 17 and postnatal day 7 NAA immunoreactivity is expressed selectively by a parallel fiber bundle that is restricted to lobule VII, thereby distinguishing the CZ anterior (lobules VIa, b) from the CZ posterior (lobule VII). The novel restriction boundary at lobule VII/VIII, which is also reflected in the morphology of the external granular layer and aligns with a gap in the developing Purkinje cell layer, precedes the morphological appearance of the posterior superior fissure between lobules VIb and VII. In addition, afferent axons to the CZ terminate in an array of parasagittal stripes that is probably a specific climbing fiber projection. Thus, the transverse zone architecture of the mouse cerebellum is more complex than had previously been appreciated.

Training two-layered feedforward networks with variable projection method.

The variable projection (VP) method for separable nonlinear least squares (SNLLS) is presented and incorporated into the Levenberg-Marquardt optimization algorithm for training two-layered feedforward neural networks. It is shown that the Jacobian of variable projected networks can be computed by simple modification of the backpropagation algorithm. The suggested algorithm is efficient compared to conventional techniques such as conventional Levenberg-Marquardt algorithm (LMA), hybrid gradient algorithm (HGA), and extreme learning machine (ELM).

The absence of phosphorylated tyrosine hydroxylase expression in the purkinje cells of the ataxic mutant pogo mouse.

The pogo mouse is a new ataxic autosomal recessive mutant that arose in Korean wild mice (KJR/Mskist). Its ataxic phenotype includes difficulty in maintaining a normal posture and the inability to walk in a straight line. Several studies have reported that tyrosine hydroxylase (TH) is persistently ectopically expressed in particular subsets of Purkinje cells in a parasagittal banding pattern in several ataxic mutant mice, e.g. tottering alleles and pogo mice. In this present study, we examined the expression of an enzymatically active form of TH and phosphorylated TH at Ser(40) (phospho-TH) by using immunohistochemistry and double immunofluorescence in the cerebellum of pogo mice. TH immunostaining appeared in some Purkinje cells in pogo, but in only a few of Purkinje cells of their heterozygous littermate controls. In all groups of mice, no phospho-TH immunoreactive Purkinje cells were observed in the cerebellum, although subsets of TH immunoreactive Purkinje cells were found in adjacent sections. This study suggests that TH expression in the Purkinje cells of pogo abnormally increases without activation of this enzyme by phosphorylation. This may mean that TH in the Purkinje cells of these mutants does not catalyse the conversion of tyrosine to l-DOPA, and is not related to catecholamine synthesis.

Corticotropin-releasing factor immunoreactivity increases in the cerebellar climbing fibers in the novel ataxic mutant mouse, pogo.

The ataxic pogo mouse (pogo/pogo) is a novel neurological mutant, which was derived as an inbred strain (KJR/MsKist) from a Korean wild mouse. The pathological manifestations include a difficulty in maintaining a normal posture, the failure of inter-limb coordination and an inability to walk straight. In this study, we examined the distribution of corticotropin-releasing factor (CRF) immunoreactive cerebellar climbing fibres and their projections to tyrosine hydroxylase (TH) immunoreactive Purkinje cells in the cerebellum of the pogo mutant mouse using immunohistochemistry. In the pogo/pogo mouse, a subset of climbing fibres was stained more intensely for CRF than in the control. Moreover, ataxic pogo mouse, neurons of the inferior olivary nucleus projecting climbing fibres were also more intensely stained for CRF than in the control. In the pogo/pogo mouse, TH immunoreactivity was located in the Purkinje cells, whereas no TH expression was found in the control. Double immunostaining for CRF and TH in the pogo/pogo cerebellum revealed that the distribution of TH-immunoreactive Purkinje cells corresponded to terminal fields of CRF-immunoreactive climbing fibres but not to the CRF-immunoreactive mossy fibres. Therefore, we suggest that an increase of CRF level may alter the function of targeted Purkinje cells and that it is related to the ataxic phenotype in the pogo mutant mouse.

Pneumatosis intestinalis (PI) following severe traumatic brain injury.

PRIMARY OBJECTIVE: To report a case of pneumatosis intestinalis complicated with sorbitol containing medicines. RESEARCH DESIGN: Case report. METHODS AND PROCEDURES: Clinical course and literature review. EXPERIMENTAL INTERVENTIONS: Chart review. MAIN OUTCOMES AND RESULTS: Large amount of sorbital may cause a reversible gastrointestinal complication, pneumatosis intestinalis. CONCLUSIONS: In the assessment of enteral abnormalities, all components of medications, active and 'inactive', must be evaluated as possible aetiological factors of pneumatosis intestinalis.

Complex regional pain syndrome (type I) after electrical injury: a case report of treatment with continuous epidural block.

A 26-year-old man presented with severe complex regional pain syndrome type I of the affected limb after a work-related electrical injury. He suffered causalgia-like pain with no electrodiagnostic evidence of nerve injury. Early steroid and analgesic regimens did not adequately relieve these symptoms. His symptoms were temporarily relieved several times with stellate ganglion blocks. The patient underwent a cervical epidural block with a local anesthetic as well as a narcotic agonist over a 4-day period, which resulted in prompt, remarkable pain relief. Vocational rehabilitation was instituted as the pain subsided.

Bladder volume measurement with electrical impedance analysis in spinal cord-injured patients.

The purpose of the study contained herein was to determine the usefulness of electrical impedance for measurement of bladder volume in spinal cord-injured patients, with an assessment of the relationship between electrical impedance and bladder volume exclusively. The study was performed during urodynamic studies to match the exact bladder volume. Thirteen patients with complete spinal cord injuries were recruited. We used silver-silver chloride compound electrodes composed of one pair of current and amplitude electrodes to minimize the influence of superficial skin impedance. Each compound electrode was attached on the lower abdomen bilaterally after skin cleansing. Constant current (60 kHz-1.0 mA), converted from 9 V of direct current, was applied, and corresponding electrical impedance (omega) was measured at "pre" (before urodynamic empty bladder), "full" (with a urodynamic filled bladder), and "post" (after urodynamic empty bladder) status. Electrical impedance at the full status was definitely lower than that at the pre and post statuses in all subjects, with a statistically significant difference (P < 0. 001). The correlation between electrical impedance and bladder volume was negative (r = -0.7988), and the fact of how much the variation in electrical impedance could be explained by variation in bladder volume was estimated (r2 = 0.6381). From these findings, we have determined that the electrical impedance analysis technique can be an alternative measure of bladder volume indirectly.

Outcome of chemotherapy in 107 patients with pulmonary tuberculosis resistant to isoniazid and rifampin.

SETTING: National Masan Tuberculosis Hospital, Korea. OBJECTIVE: Treatment for multidrug-resistant tuberculosis (MDR-TB) is considered to be clinically important, but there are few reports on this topic. We therefore retrospectively evaluated the outcomes of chemotherapy only for pulmonary MDR-TB. DESIGN: We reviewed the clinical courses of 107 patients with pulmonary disease due to Mycobacterium tuberculosis resistant to rifampin and isoniazid who were under follow-up between March 1996 and June 1996 after hospitalization between January 1993 and January 1996. We performed a retrospective cohort study for all the patients' records. Their regimens were selected individually and preferably included four medications that they had not been given previously and to which the strain was fully susceptible. RESULTS: The 107 patients (mean age 38.3 years) had previously received a mean of five drugs, and were shedding bacilli that were resistant to a mean of four drugs. Of 63 patients with sufficient follow-up data, 52 (82.5%) responded to chemotherapy (as indicated by negative sputum cultures for at least three consecutive months); 11 (17.5%) had no response, as shown by continually positive cultures. In a univariate analysis, an unfavorable response was significantly associated with resistance to a greater number of drugs before the current courses of treatment (relative risk 21.5; 95% confidence interval 1.2-3.0; P < 0.05). The mean period of follow-up was 17 months. There was no subsequent relapse among the patients with responses, and there were no tuberculosis-related deaths. CONCLUSION: In this study, multidrug-resistant pulmonary tuberculosis responded relatively well to carefully selected regimens.

Effects of multiagent chemotherapy and independent risk factors in the treatment of high-risk GTT--25 years experiences of KRI-TRD.

UNLABELLED: A retrospective and comparative study of high-risk gestational trophoblastic tumor (GTT) treated with different chemoregimen from 1971 to 1995 was performed and to find most effective chemotherapy regimen and independent risk factors. Three hundred seven patients in scoring over 8 points in WHO classification were categorized into high-risk group among 802 GTT cases received chemotherapy in the 2,418 GTD patients registered at KRI-TRD (Korean Research Institute for Gestational Trophoblastic Disease), Catholic University Medical College in Korea. Study groups of multiagent combination chemotherapy in 227 patients of the high-risk GTT were divided such as 49 cases of combination chemotherapy with MTX + folinic acid and Act-D, 40 cases of MAC regimen, 42 cases of CHAMOCA regimen, and 96 cases of EMA/CO. Initial tumor response according to hCG titer decrease was found in good response (log fall) 69.8%, of EMA /CO regimen group. On the other hand, good response was shown in only 24.5% of MTX + ACT-D, 32.5% of MAC regimen, and 52.4%, of CHAMOCA regimen respectively. Remission rate of EMA/CO regimen was 90.6% (87/96) and courses of chemotherapy until remission was 8.5+/-2.2. However, remission rate of other regimens of MTX + Act-D, MAC, and CHAMOCA were 63.3%, (31/49) 67.5% (27/40) and 76.2% (32/45) respectively, with 10.0+/-4.0, 10.7+/-4.3, 9.1+/-3.9 chemotherapy courses respectively until remission. Therefore, EMA/CO regimen groups were found to have low drug toxicity, early remission and a low failure rate. In the study of independent risk factors in the 165 cases of high-risk gestational trophoblastic tumor patients received EMA/CO regimen, stepwise Coxs proportional hazard's regression of prognostic factors using multivariate analysis revealed tumor age, number of metastatic organs, metastatic site and inadequate previous chemotherapy. According to the performance of fitted logistic regression model, the prediction rate of death and survival was 80.5%. CONCLUSIONS: The most effective chemotherapy to high-risk GTT was EMA/CO regimen than other regimens. The following factors showed poor prognosis; 1) Tumor age is over 12 month, 2) more than 2 organs had metastatic lesion, 3) inadequate previous therapy that includes unplanned operation and inadequate previous chemotherapy.

Effects of multiagent chemotherapy and independent risk factors in the treatment of high-risk GTT -25 years experiences of KRI-TRD.

A retrospective and comparative study of high-risk gestational trophoblastic tumor (GTT) treated with different chemoregimen from 1971 to 1995 was performed and to find most effective chemotherapy regimen and independent risk factors. Three hundred seven patients in scoring over 8 points in WHO classification were categorized into high-risk group among 802 GTT cases received chemotherapy in the 2,418 GTD patients registered at KRI-TRD (Korean Research Institute for Gestational Trophoblastic Disease), Catholic University Medical College in Korea. Study groups of multiagent combination chemotherapy in 227 patients of the high-risk GTT were divided such as 49 cases of combination chemotherapy with MTX + folinic acid and Act-D, 40 cases of MAC regimen, 42 cases of CHAMOCA regimen, and 96 cases of EMA/CO. Initial tumor response according to hCG titer decrease was found in good response (log fall) 69.8%, of EMA /CO regimen group. On the other hand, good response was shown in only 24.5% of MTX + ACT-D, 32.5% of MAC regimen, and 52.4%, of CHAMOCA regimen respectively. Remission rate of EMA/CO regimen was 90.6% (87/96) and courses of chemotherapy until remission was 8.5 ± 2.2. However, remission rate of other regimens of MTX + Act-D, MAC, and CHAMOCA were 63.3%, (31/49) 67.5% (27/40) and 76.2% (32/45) respectively, with 10.0 ± 4.0, 10.7 ± 4.3, 9.1 ± 3.9 chemotherapy courses respectively until remission. Therefore, EMA/CO regimen groups were found to have low drug toxicity, early remission and a low failure rate. In the study of independent risk factors in the 165 cases of high-risk gestational trophoblastic tumor patients received EMA/CO regimen, stepwise Coxs proportional hazard's regression of prognostic factors using multivariate analysis revealed tumor age, number of metastatic organs, metastatic site and inadequate previous chemotherapy. According to the performance of fitted logistic regression model, the prediction rate of death and survival was 80.5%. CONCLUSIONS: The most effective chemotherapy to high-risk GTT was EMA/CO regimen than other regimens. The following factors showed poor prognosis; 1) Tumor age is over 12 month, 2) more than 2 organs had metastatic lesion, 3) inadequate previous therapy that includes unplanned operation and inadequate previous chemotherapy.

Bioelectrical impedance changes in regional extracellular fluid alterations.

The purpose of this study was to determine whether changes in bioelectrical impedance (BI) can reveal regional extracellular fluid (ECF) alterations. ECF changes were induced by holding various arm positions for ten minutes, and they were evaluated through the measurement of BI in eight normal adult subjects. A low frequency current (100 Hz, 0.50 mA) was applied using an electromyography machine through two current electrodes, and the voltage signals were recorded by two amplitude electrodes. The corresponding BI was calculated by Ohm's law, resistance = voltage/current (R = V/I). There was no significant difference between BI and time interval, but between BI and arm position (two-way ANOVA with replicate, p = 0.954, p < 0.001). BI has a negative correlation with wrist circumference. These results show that this experimental method can be used as reflection of ECF changes and that both position and wrist circumference are major factors for BI in the upper extremity.

Sympathetic skin response recorded by 4 channel recording system.

The main purpose of this article is to determine a method of supporting the sympathetic skin response (SSR) as a sensitive clinical test. Using a non-invasive technique the SSRs are simultaneously recorded by 4 channel EMG machine. Thirty adults (10 women and 20 men, aged 19 to 46 years), normal and healthy, participated in this experiment. Not only did the latencies recorded on both palms respond faster than those on both soles, but the amplitudes measured on the palms were also higher. From these observations, one is bound to conclude that the SSR is not a segmental response but a long systemic response. More than two channel EMG recordings are desirable to see whether or not there is a lesion in any part of the SSR's pathway. Comparing the SSRs made both on the palms and soles simultaneously is recommendable in order to increase the its sensitivity.

[Effect of in vivo administration of Tetrahymena pyriformis on the in vitro toxoplasmacidal activity of mouse peritoneal macrophages].

Tetrahymena pyriformis is a free-living ciliate protozoan in the freshwater system. Experiments were carried out to determine whether intraperitoneal administration of T. pyriformis (GL strain) to mice activates macrophages to be able to kill Toxoplasma gondii tachyzoites in vitro. Mice were also injected intraperitoneally with several synthetic activators; dimethyldioctadecylammonium bromide (DDA), dextran sulfate, complete Freund's adjuvant (CFA) as well as Toxoplasma and Tetrahymena lysates in order to activate mouse peritoneal macrophages. One week after the administration of activators, peritoneal cells were harvested and the adherent macrophages were challenged with Toxoplasma tachyzoites. Macrophage monolayers were then fixed with absolute methanol after washing, and stained with Giemsa solution. The percentage of the adherent cells infected and total number of organisms per 100 macrophages were calculated to make toxoplasmacidal activity of macrophages according to the cultivation time. Peritoneal macrophages from mice administered with Tetrahymena exhibited significant protection against target parasites as compared with those treated with synthetic activators. Among non-biological synthetic activators, DDA was evaluated as an excellent activator.