BACKGROUND: Priming and boosting vaccination strategy has been widely explored for new vaccine development against tuberculosis. As an effort to identify other vaccine candidates, this study was initiated to evaluate protective efficacy of adenylate kinase (AK), nucleoside diphosphate kinase (NdK), and heat shock protein 70 (Hsp70) of Mycobacterium tuberculosis. METHOD: M. tuberculosis genes encoding AK, NdK, and Hsp70 proteins were amplified by PCR and cloned into E. coli expression vector, pQE30. Recombinant AK, NdK, and Hsp70 was purified through Ni-NTA resin. To evaluate immune responses, we performed enzyme-linked immunosorbent assay (ELISA) for IgG isotype and IFN-gamma after mice were immunized subcutaneously with recombinant proteins delivered in dimethyl dioctadecylammonium bromide (DDA). Immunized- and control groups were challenged by aerosol with M. tuberculosis. The spleens and lungs of mice were removed aseptically and cultured for CFU of M. tuberculosis. RESULT: Vaccination with recombinant proteins AK, NdK, and Hsp70 delivered in DDA elicited significant level of antibody and IFN-gamma responses to corresponding antigens but no protective immunity comparable to that achieved with Mycobacterium bovis BCG. CONCLUSION: Recombinant proteins AK, NdK, and Hsp70 do not effectively control growth of M. tuberculosis in mice when immunized with DDA as an adjuvant.
Adenylate Kinase* ; Animals ; Clone Cells ; Enzyme-Linked Immunosorbent Assay ; Heat-Shock Proteins* ; HSP70 Heat-Shock Proteins* ; Immunoglobulin G ; Lung ; Mice* ; Mycobacterium bovis ; Mycobacterium tuberculosis* ; Mycobacterium* ; Nucleoside-Diphosphate Kinase* ; Polymerase Chain Reaction ; Recombinant Proteins* ; Spleen ; Tuberculosis ; Vaccination

A novel biomass was prepared from Pichia anomala KCCM 11473, which grew well in ginseng-steaming effluent (GSE), and its physiological functionalities and enzyme activities were determined. When the strain was cultured in the GSE (pH 6.0) at 30degrees C for 48 h, 1.6 mg of biomass per ml-cultures was produced. The cell-free extract of the biomass showed high antihypertensive angiotensin I-converting enzyme inhibitory activity of 72.0% and anticholesteromia HMG-CoA reductase inhibitory activity of 46.5%. The cell-free extract also showed 13.0 U per ml and 8.5 U per ml of neutral protease activity and alkaline protease, respectively.
Acyl Coenzyme A ; Bacterial Proteins ; Biomass ; Endopeptidases ; Oxidoreductases ; Peptidyl-Dipeptidase A ; Pichia ; Sprains and Strains

Purpose: Stress hyperglycemia is common in critically ill adult patients. It is known as a predictor of increased mortality, and intensive insulin therapy has been shown to improve the prognosis in such patients. We have investigated the relationship between early stress hyperglycemia and clinical outcomes in preterm infants. Methods: In this study, 141 preterm infants with a gestational age of less than 30 weeks were enrolled. The hyperglycemic group was defined as that having maximum glucose of more than 150 mg/dL (n=61) during the first 48 h of life, and the non-hyperglycemic group was defined as that having maximum glucose of less than 150 mg/dL (n=80). Perinatal history, severity of illness using the Clinical Risk Index for Babies (CRIB) score, clinical outcomes, and mortality of the two groups were compared. Results: There was no significant difference in the gestational age between the two groups, but the birth weight (P<0.001) was significantly lower, and the CRIB score (P<0.001) was significantly higher in the hyperglycemic group. Disseminated intravascular coagulation (P<0.001) and clinically suspected sepsis (P=0.046) were more common in the hyperglycemic group. Mortality was markedly higher in the hyperglycemic group (11.3% vs. 41.0%, P<0.001). On performing a stepwise multiple logistic regression analysis, hyperglycemia (OR 3.787; 95% CI 1.324 to 10.829), the CRIB score (OR 1.252; 95% CI 1.047 to 1.496) and birth weight (OR 0.997; 95% CI 0.994 to 1.000) was independently associated with higher mortality. Conclusion: Stress hyperglycemia within the first 48 h of life is independently related to increased morbidity and mortality in preterm infants.
Adult ; Birth Weight ; Critical Illness ; Disseminated Intravascular Coagulation ; Gestational Age ; Glucose ; Humans ; Hyperglycemia ; Infant Equipment ; Infant, Newborn ; Infant, Premature ; Insulin ; Logistic Models ; Prognosis ; Sepsis

Bisphosphonates inhibit bone resorption by affecting osteoclastic function and formation of osteoclasts from their precursor cells. Chondroclasts have the same origin and differentiation as osteoclasts. Thus, it is hypothesized that bisphosphonate can affect on cartilage metabolism. This study was aimed to elucidate effects of alendronate, a nitrogen containing bisphosphonate, on cartilage development in the tibial proximal and femoral distal epiphyseal plates in rats. Alendronate (1 mg/kg) was subcutaneously administered in growing rat pups for 10 days. Several parameters such as the number and size of chondroclasts, involved in cartilage resorption, size of secondary ossification center and thickness of cartilage cell layers were measured and analysed by histomorphometry. The size of the secondary ossification centers in the tibial proximal and femoral distal epiphysis was smaller in the alendronate treated group (p< 0.01). The number of osteoclasts in the both the ossification centers and chondroclasts beneath the epiphyseal plates was significantly decreased by alendronate treatment (p< 0.01). The size of chondroclasts was not significantly changed (p> 0.05). The thickness of proliferating cartilage layer was not changed, but by contrast, hypertrophied cartilage layer was increased in thickness by alendronate treatment. These findings suggest that bisphosphonates can affect cartilage cell metabolism in a chondroprotective way.
Alendronate ; Animals ; Bone Resorption ; Cartilage ; Diphosphonates ; Epiphyses ; Growth Plate* ; Knee Joint* ; Knee* ; Metabolism ; Nitrogen ; Osteoclasts ; Rats*

Bovine tuberculosis (TB) is a major zoonosis that's caused by Mycobacterium bovis (M. bovis). Being able to detect M. bovis is important to control bovine TB. We applied a molecular technique, the variable number tandem repeat (VNTR) typing method, to identify and distinguish the M. bovis isolates from Gyeonggi-do, Korea. From 2003 to 2004, 59 M. bovis clinical strains were isolated from dairy cattle in Gyeonggi-do, Korea, and these cattle had tuberculosis-like lesions. Twenty-four published MIRUVNTR markers were applied to the M. bovis isolates and ten of them showed allelic diversity. The most discriminatory locus for the M. bovis isolates in Korea was QUB 3336 (h = 0.64). QUB 26 and MIRU 31 also showed high discriminative power (h = 0.35). The allelic diversity by the combination of all VNTR loci was 0.86. Six loci (MIRU 31, ETR-A and QUB-18, -26, -3232, -3336) displayed valuable allelic diversity. Twelve genotypes were identified from the 59 M. bovis isolates that originated from 20 cattle farms that were dispersed throughout the region of Gyenggi-do. Two genotypes [designation index (d.i.) = e, g] showed the highest prevalence (20% of the total farms). For the multiple outbreaks on three farms, two successive outbreaks were caused by the same genotype at two farms. Interestingly, the third outbreak at one farm was caused by both a new genotype and a previous genotype. In conclusion, this study suggests that MIRU-VNTR typing is useful to identify and distinguish the M. bovis isolates from Gyeonggi-do, Korea.
Animals ; Cattle ; DNA Primers/genetics ; *Genetic Variation ; Genotype ; Korea/epidemiology ; Minisatellite Repeats/*genetics ; Mycobacterium bovis/*genetics ; Prevalence ; Tuberculosis, Bovine/*epidemiology/*microbiology

PURPOSE: To report our experience in the diagnosis and treatment of a localized corneal amyloidosis secondary to trichiasis. METHODS: Case 1. A 55-year-old woman visited our clinic due to discomfort of her right eye. Thirty years previously, she received a lower lid blepharoplasty due to lower lid entropion. Biomicroscopy revealed some trichiasis and a 3 mm, grayish-white nodule at the center of the cornea. Case 2. A 30-year-old woman visited our clinic due to chronic irritation of both eyes. Ten and 3 years previously, she received a lower lid blepharoplasty. Biomicroscopy revealed some trichiasis of the right lower lid and a grayish-white, patch-like lesion at the inferior cornea of both eyes. Lamellar keratectomy and amniotic membrane transplantation with careful electrolysis were performed. RESULTS: Hematoxylin and eosin stain revealed a pink, amorphous, hyaline material under the epithelium, and Congo-red stain showed birefringence of the whitish lesion. Electron microscopy revealed multidirectional, fibrillar arrangement. Secondary, localized amyloidosis of the cornea was diagnosed without any systemic involvement. No clues to the origin of the amyloid were found by immunohistochemical staining. By the sixth month after operation, the patients showed favorable vision and no evidence of recurrence. CONCLUSIONS: In the identification of a corneal mass, a secondary, localized amyloidosis should be considered, which can be managed successfully.
Adult ; Amnion ; Amyloid ; Amyloidosis* ; Birefringence ; Blepharoplasty ; Cornea ; Diagnosis ; Electrolysis ; Entropion ; Eosine Yellowish-(YS) ; Epithelium ; Female ; Hematoxylin ; Humans ; Hyalin ; Microscopy, Electron ; Middle Aged ; Recurrence ; Trichiasis*

BACKGROUND: Isoniazid (INH, H) is a key drug of the standard first-line regimen for the treatment of tuberculosis (TB), yet some reports have suggested that treatment efficacy was maintained even though INH was omitted from the treatment regimen. METHODS: One hundred forty C57BL/6 mice were infected with the H37Rv strain of M. tuberculosis with using a Glas-Col aerosol generation device, and this resulted in depositing about 100 bacilli in the lung. Four weeks after infection, anti-TB treatment was initiated with varying regimens for 4-8 weeks; Group 1: no treatment (control), Group 2 (4HREZ): 4 weeks of INH, rifampicin (R), pyrazinamide (Z) and ethambutol (E), Group 3: 1HREZ/3REZ, Group 4: 4REZ, Group 5: 4HREZ/4HRE, Group 6: 1HREZ/3REZ/4RE, and Group 7: 4REZ/4RE. The lungs and spleens were harvested at several time points until 28 weeks after infection, and the colony-forming unit (CFU) counts were determined. RESULTS: The CFU counts increased steadily after infection in the control group. In the 4-week treatment groups (Group 2-4), even though the culture was negative at treatment completion, the bacilli grew again at the 12-week and 20-week time points after completion of treatment. In the 8-week treatment groups (Groups 5-7), the bacilli did not grow in the lung at 4 weeks after treatment initiation and thereafter. In the spleens of Group 7 in which INH was omitted from the treatment regimen, the culture was negative at 4-weeks after treatment initiation and thereafter. However, in Groups 5 and 6 in which INH was taken continuously or intermittently, the bacilli grew in the spleen at some time points after completion of treatment. CONCLUSION: TThe exclusion of INH from the standard first-line regimen did not affect the treatment outcome in a murine model of TB in the early stage of disease. Further studies using a murine model of chronic TB are necessary to clarify the role of INH in the standard first-line regimen for treating TB.
Animals ; Ethambutol ; Isoniazid ; Lung ; Mice ; Pyrazinamide ; Rifampin ; Spleen ; Sprains and Strains ; Stem Cells ; Treatment Outcome ; Tuberculosis