Background: The rapid urease test (RUT) is a major diagnostic tool for detecting Helicobacter pylori infection. This study aimed to establish an objective method for measuring the color changes in the RUT kit to improve the test’s diagnostic accuracy. Methods: A UV-visible spectrophotometer was selected as the colorimeter; experiments were conducted in three stages to objectively identify the color changes in the RUT kit. Results: First, the urea broth solution showed an identifiable color change from yellow to red as the pH increased by 0.2. The largest transmittance difference detected using the UV-visible spectrophotometer was observed at a 590-nm wavelength. Second, the commercialized RUT kit also showed a gradual color change according to the pH change detected using the UV-visible spectrophotometer. Third, 13 cases of negative RUT results with a biopsy specimen and 16 of positive RUT results were collected. The transmittance detected using the UV-visible spectrophotometer showed a clear division between the positive and negative RUT groups; the largest difference was observed at a 559-nm wavelength. The lowest transmittance in the negative RUT group was 64, while the highest in the positive RUT group was 56, at the 559-nm wavelength. The UV-visible spectrophotometry reading showed a consistency of 92.7% compared with that of manual reading. Conclusion A transmittance of 60 at a 559-nm wavelength detected using UV-visible spectrophotometer can be used as a cutoff value for interpreting RUT results; this will help develop an automatic RUT kit reader with a high accuracy.

OBJECTIVEAcne vulgaris is a chronic dermatologic problem with multiple factors involved in its pathogenesis. Alternative solutions to acne treatment were instigated by antibiotic resistance despite of its extensive use. Purified bee venom (PBV) has been proposed as a promising candidate for that purpose. The present study was designed to confirm the antibacterial effect of PBV and access the efficacy of cosmetics containing PBV in subjects with acne vulgaris.

METHODSThe skin bacterium Propionibacterium acnes was incubated with PBV at various concentrations and bacterial growth was evaluated using the colony forming unit (CFU) assay. The mechanism of PBV employed in killing P. acnes was examined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). In addition, a total of 12 subjects were randomized in a double-blind, controlled trial to receive either cosmetics containing PBV or cosmetics without PBV for two weeks. Evaluations included lesion counts and skin microorganism.

RESULTSPBV exhibited antimicrobial activity in a concentration-dependent manner, reducing the number of P. acnes CFU by approximately 6 logs at a concentration of 0.5 mg. When PBV concentration was higher than 1.0 mg, no P. acnes colonies were spotted on an agar. TEM and SEM of untreated P. acnes illustrated the normal pleomorphic structure, whereas the PBV-treated bacterium lost the integrity of surface architecture. Significant difference (P=0.027) in the grading levels based on numbers of lesion counts for inflammatory and noninflammatory was observed in favour of the PBV group compared with the control group. In terms of average decrement of skin microorganism, subjects receiving cosmetics containing PBV experienced a significant 57.5% decrease of adenosine triphosphate levels, whereas participants receiving cosmetics without PBV experienced a nonsignificant decrease of 4.7%.

CONCLUSIONThese results show that the in vitro actions of antimicrobial activity of PBV were translated in vivo. Cosmetics containing PBV provided a certain degree of efficacy in terms of lesion counts and skin microorganism concentration compared with cosmetics without PBV in subjects with acne vulgaris. PBV may be a good candidate compound for developing therapeutic drug for the treatment of acne vulgaris.

Acne Vulgaris ; drug therapy ; microbiology ; Adolescent ; Adult ; Anti-Infective Agents ; therapeutic use ; Bee Venoms ; therapeutic use ; Child ; Cosmetics ; Double-Blind Method ; Humans ; Propionibacterium acnes ; drug effects

Acne vulgaris is a chronic dermatologic problem with multiple factors involved in its pathogenesis. Alternative solutions to acne treatment were instigated by antibiotic resistance despite of its extensive use. Purified bee venom (PBV) has been proposed as a promising candidate for that purpose. The present study was designed to confirm the antibacterial effect of PBV and access the efficacy of cosmetics containing PBV in subjects with acne vulgaris.

Bee venom (Apis mellifera L., BV) has been used as a cosmetic ingredient for antiaging, anti-inflammatory and antibacterial functions. The aim of this study was to access the skin sensitization of BV, a Buehler test was conducted fifty healthy male Hartley guinea pigs with three groups; Group G1 (BV-sensitization group, 20 animals), group G2 (the positive control-sensitization group, 20 animals), and group G3 (the ethyl alcohol-sensitization group, 10 animals). The exposure on the left flank for induction was repeated three times at intervals of one week. Two weeks after the last induction, the challenge was performed on the right flank. No treatment-related clinical signs or body weight changes were observed during the study period. The average skin reaction evaluated by erythema and edema on the challenge sites and sensitization rate in the BV-sensitization group at 30 hours were 0.0 and 0%, respectively, which are substantially low compared with in positive control group (average skin reaction: 0.55, sensitization rate: 40%) and identical with in vehicle control group, representing a weak sensitizing potential. The average skin reaction and sensitization rate observed at 54 hours were 0.0 and 0% in the BV-sensitization group, respectively, and 0.25 and 20% in the positive control group, respectively. It was concluded that BV classified to Grade I, induced no sensitization when tested in guinea pigs and may provide a developmental basis for a cosmetic ingredient or external application for topical uses.
Animals ; Bee Venoms ; Bees ; Body Weight Changes ; Cosmetics ; Edema ; Erythema ; Guinea ; Guinea Pigs ; Humans ; Male ; Skin

Bee venom (Apis mellifera L. BV) has been used as a cosmetic ingredient for anti-ageing, anti-inflammatory and antibacterial functions. The aim of this study was to evaluate the acute toxicity after a single dermal administration of BV, BV was administered to 2 groups of Sprague-Dawley (SD) male and female rats (5 animals/group) at doses of 0 and 1,500 mg/kg body weight (BW). Mortality, clinical signs, body weight changes and gross findings were continually monitored for 15 days following the single dose. There were no unscheduled deaths in any groups during the study period. No BV related clinical signs and body weight changes were observed in any groups during the study period. There were no abnormal gross findings at necropsy on day 15 after the treatment. On the basis of the above results, it was concluded that there were no treatment-related effect on mortality, clinical signs, body weight changes and gross findings in SD rats treated with a single dermal dose of BV at dose of 1,500 mg/kg BW. Therefore, the approximate lethal dose of BV was considered to be over 1,500 mg/kg/day for both sexes of rats. BV may provide a developmental basis for a cosmetic ingredient or external application for topical uses.
Administration, Cutaneous ; Animals ; Bee Venoms ; Bees ; Body Weight ; Body Weight Changes ; Cosmetics ; Female ; Humans ; Male ; Rats

Oral administration of royal jelly (RJ) promotes wound healing in diabetic mice. Concerns have arisen regarding the efficacy of RJ on the wound healing process of normal skin cells. In this study, a wound was created by scratching normal human dermal fibroblasts, one of the major cells involved in the wound healing process. The area was promptly treated with RJ at varying concentrations of 0.1, 1.0, or 5 mg/ml for up to 48 hrs and migration was analyzed by evaluating closure of the wound margins. Furthermore, altered levels of lipids, which were recently reported to participate in the wound healing process, were analyzed by HPTLC and HPLC. Migration of fibroblasts peaked at 24 hrs after wounding. RJ treatment significantly accelerated the migration of fibroblasts in a dose-dependent manner at 8 hrs. Although RJ also accelerated the migration of fibroblasts at both 20 hrs and 24 hrs after wounding, the efficacy was less potent than at 8 hrs. Among various lipid classes within fibroblasts, the level of cholesterol was significantly decreased at 8 hrs following administration of both 0.1 ug/ml and 5 mg/ml RJ. Despite a dose-dependent increase in sphinganines, the levels of sphingosines, ceramides, and glucosylceramides were not altered with any concentration of RJ. We demonstrated that RJ enhances the migration of fibroblasts and alters the levels of various lipids involved in the wound healing process.
Administration, Oral ; Animals ; Ceramides ; Cholesterol ; Chromatography, High Pressure Liquid ; Fatty Acids ; Fibroblasts ; Glucosylceramides ; Humans ; Mice ; Skin ; Sphingosine ; Wound Healing

BACKGROUND: The treatment success rates in patients with tuberculosis are known to be lower in the private sector compared to the public sector. To improve treatment outcomes in the private sector we developed a public-private collaboration model for strengthening health education and case holding activities with public health nursing in the private sector. METHODS: We performed a prospective cohort study in new smear positive pulmonary tuberculosis patients treated at private hospitals, selected by non-randomization, with an intervention consisting of health education and case holding activities by specially trained public health nurses (intervention group) results were compared with cases treated without the intervention (conventional group). Physicians were asked to treat both groups routinely. The treatment outcomes of patients under treatment by the National Tuberculosis Programme were also analyzed for comparison. RESULTS: There were 172 cases each in the intervention and conventional groups. The mean age was 48.9+/-19.0 and 48.2+/-19.7 in the respective groups (p=0.66). The PHN interacted with the cases in the intervention group by initial face to face interview and telephone calls an average of 7.1+/-9.2 times during the initial six months. The intervention group showed a significantly higher treatment success rate, 91.6%, (Rate Ratio [RR]; 1.23, 95% Confidence Interval [CI]; 1.12~1.36), lower default, 3.6%, (RR; 0.31, 95% CI; 0.13~0.75) and transfer-out rate, 3.0%, (RR; 0.32, 95% CI; 0.12~0.86) than the conventional group where they were: 75.0%, 11.6%, 9.3%, respectively. The success rate was even higher than the rate (80.5%) of 1,027 cases treated in health centers (RR; 1.11, 95% CI; 1.05~1.17). Of the completed cases in the intervention group, 82.2% regarded the role of the public health nurse as very helpful. CONCLUSION: The treatment success rate, of tuberculosis patients in the private sector, was significantly improved by an intervention using a public-private collaboration model.
Cohort Studies ; Cooperative Behavior ; Health Education ; Hospitals, Private ; Humans ; Private Sector ; Prospective Studies ; Public Health Nursing ; Public Sector ; Telephone ; Tuberculosis ; Tuberculosis, Pulmonary

BACKGROUND: Ethambutol(EMB) is one of the first-line drugs included in short-course anti-tuberculosis therapy. The point mutations in embB gene have been speculated to be associated EMB resistance. However, detection of embB mutations at these positions have been observed in both EMB-susceptible isolates; thus, it remains controversial whether these mutations are associated with EMB resistance METHODS: The 36 M. tuberculosis isolates were selected from clinical isolates which tested susceptible to EMB and resistant to at least one drug. DNA extracted from the isolates was analyzed by amplifying embB gene. The PCR products were purified and directly sequenced. We reviewed the history of past drug susceptibility test results. RESULTS: Out of 36 EMB-susceptible strains, 3 strains (8.3%) had a mutation in codon 306 or 406 of the embB gene. These three strains had at least isoniazid resistance. They grew at 1.0 mcg/ml of EMB in Lowenstein-Jensen media. The patients of the strains were continuously smear-positive for over 3 years despite taking TB therapy. One strain had been EMB-resistant in past drug susceptibility tests. CONCLUSION: EMB-susceptible strains containing embB mutation may be caused by decreased viability in vitro test not by itself.
Codon ; DNA ; Drug Resistance ; Ethambutol ; Humans ; Isoniazid ; Korea ; Mycobacterium tuberculosis* ; Mycobacterium* ; Point Mutation ; Polymerase Chain Reaction ; Tuberculosis

BACKGROUND: Fluoroquinolone drugs are an important anti-tuberculous agent for the treatment of multi-drug resistant tuberculosis. However, many drugs belonging to the fluoroquinolones have different cross resistance to each other. METHODS: Sixty-three ofloxacin (OFX) resistant and 10 pan-susceptible M. tuberculosis isolates were selected, and compared for their cross resistance using a proportion method on Lowenstein-Jensen media, containing ofloxacin (OFX), ciprofloxacin (CIP), levofloxacin (LVX), moxifloxacin (MXF), gatifloxacin (GAT) and sparfloxacin (SPX), at concentrations ranging from 0.5 to 3microgram/ml. DNA extracted from the isolates was directly sequenced after amplifying from the gyrA and gyrB genes. RESULTS: The 63 OFX resistant M. tuberculosis isolates showed complete cross resistance to CIP, but only 90.5, 44.4, 36.5 and 46.0% to LVX, MXF, GAT, and to SPX, respectively. Fifty-one of the isolates (81.0%) had point mutations in codons 88, 90, 91 and 94 in gyrA, which are known to be correlated with OFX resistance. The Gly88Ala, Ala90Valand Asp94Ala mutations in gyrA showed a tendency to be susceptible to MXF, GAT and SPX. Only 4 isolates had mutations in the gyrB gene, which did not affect the OFX resistance. CONCLUSION: About 60% of the OFX resistant M. tuberculosis isolates were susceptible to GAT, SPX and MXF. These fluoroquinolones may be useful in the treatment of TB patients showing OFX resistance.
Ciprofloxacin ; Codon ; DNA ; Fluoroquinolones ; Genotype ; Humans ; Levofloxacin ; Mycobacterium tuberculosis* ; Mycobacterium* ; Ofloxacin ; Point Mutation ; Tuberculosis ; Tuberculosis, Multidrug-Resistant

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