Background: Leprosy is a chronic infectious disease caused by Mycobacterium leprae (M. leprae), an obligate intracellular organism residing within host macrophages and Schwann cells.Currently, there is no artificial medium capable of culturing M. leprae, so it is mainly grown by inoculating animals such as nude mice and armadillos. In a laboratory environment, it is convenient to maintain and cultureM. lepraeusing athymic nude mice. Objective: The maintenance and growth ofM. lepraestrains in nude mice is important for studying leprosy. In this study, we developed an efficient purification method to collect M. lepraestrain grown in nude mice. Methods: Purification ofM. lepraestrain from nude mouse footpads was used of gentleMACS™ C Tube and DT-20 Dispersing tube without a tissue homogenizer. The isolated tissues were dissociated with gentleMACS™ C Tubes and gentleMACS Dissociators. In order to increase the bacterial recovery rate, bacilli from the remaining tissues (dermis, tendons, nerves and bones of the foot) were purified using a DT-20 Dispersing tube and IKA Tube Disperser. After the tissue dissociation, cells were separated by trypsin treatment and the contaminated bacteria were removed by sodium hydroxide (NaOH). The isolated bacteria were stained with methylene blue solution in the usual Ziehl-Neelsen method and the chromosomal DNA extracted by cracking of cell wall with bead beating was analyzed by DNA amplification by polymerase chain reaction (PCR). Results: Using gentleMACS™ C Tube and DT-20 Dispersing tube instead of a tissue homogenizer, M. lepraestrain was efficiently obtained from tissues. Trypsin was suitable for tissue dissociation, resulting in an effective cell suspension and did not affect the survival of M. lepraestrain. The procedures showed high viability and high recovery rate and allowed simultaneous purification of different bacillary strains from each nude mice without contamination by NaOH treatment. Conclusion It is a useful method for high viability rate and high recovery rate of M. lepraestrain harvested from nude mouse. The procedures were possible method for extracting several strains at the same time.

Background: Leprosy is a chronic granulomatous disease caused by Mycobacterium leprae. As leprosy has a long disease course, various dermatoses must be considered in the differential diagnosis. Objective: In Korea, due to the low incidence rate of leprosy and lack of doctors with experience of leprosy, it is difficult to make an early diagnosis. Therefore, we evaluated the importance of cooperation projects in the diagnosis of leprosy. Methods: A retrospective study was performed on 137 patients who were referred by other hospitals to the Institute for Leprosy Research, Korean Hansen Welfare Association, to make an early diagnosis of leprosy from 2009 to 2018. Results: Among 137 patients, 19 were diagnosed with leprosy, which accounted for approximately 37% of new cases of leprosy in Korea during the 10 years. By nationality, 15 were Korean, and 4 were foreigners. The most common type was lepromatous leprosy. All multibacillary patients were positive for phenolic glycolipid-1 antibody, M. leprae-specific gene polymerase chain reaction, and acid-fast bacilli stain. Among the four paucibacillary patients who showed both negative acid-fast bacilli stain and phenolic glycolipid-1, three were positive for polymerase chain reaction. Of the 55 skin biopsy specimens that were not diagnosed with leprosy, granulomatous skin disease was the most commonly diagnosed condition. Conclusion In this study, we diagnosed patients with leprosy who were referred from other hospitals due to the suspicion of leprosy, which enables early treatment of leprosy. Finally, we believe that our project has public health significance in preventing the spread of leprosy in the community.

Background: One of the Sequelae of facial nerve paralysis in leprosy patients is severe eye and lip deformity. Eye deformities are most common, followed by unilateral lip deviation. When there is bilateral failal nerve paralysis, the lower lip drooping may occur. The lower lip drooping makes mouth closure incomplete, so the fluid or saliva may flow out when eating. And the teeth may be exposed or even the gum may be visible, which causes serious cosmetic and functional impairments. Objective: It is to evaluate the masseter muscle sling (MMS) method in the treatment of the recurrent lower lip drooping in patients with Hansen's disease. Methods: In the case of masseter muscle sling, there is a traditional method of detaching, separating and raising anterior masseter muscle in half from the mandibular jaw line and fixing it to the lip muscle. However, the authors observed a 3cm gap between the anterior border of masseter muscle and the lateral edge of orbicularis oris muscle. Under the local anesthesia the nasolabial line was incised and 3-4 places were pulled up between both musle edges and plicated with a 2/0 PDS??threads the gap-space. Results: During 5 years (2016-2020), there were 17 recurrent lower lip palsy patients. The mean age was 71.3 years and the ratio of male and female was 9:8. Of 17 patents, 12 received temporal muscle transfer (TMT) attached with tensor fascia lata (TFL), 2 received temporal muscle transfer attached with Alloderm, and 3 received masseter muscle sling alone. The postoperative results of corrective masseter muscle sling were satisfactory in all 17 patients and there were no serious complication and recurrence during follow-up. Conclusion Masseter muscle sling is effective method in correction of recurrent lower lip drooping in patient with leprosy who treated previously with temporal muscle transfer attached with tensor fascia lata or temporal muscle transfer attached with Alloderm. In addition, masseter muscle sling may be considered as the first-line treatment in the lower lip paralysis.

Background: The prevalence of leprosy has almost reached the goal of World Health Organization. Accordingly, attention to leprosy and the number of skilled doctors who can diagnosis the leprosy upon clinical manifestations were decreased which caused the delayed diagnosis of leprosy. Objective: Serum C-reactive protein, interferon gamma-induced protein 10, anti-phenolic-glycolipid-1 IgM are biomarkers associated with multi-bacillary leprosy. Therefore, we investigated whether the use of multiple biomarkers has diagnostic value. Methods: The samples were collected from August 2012 to December 2019 at our institute. Through various statistic methods, multiple biomarkers and equation were compared in multi-bacillary patients, pauci-bacillary patients, contacts and different skin disease. Results: Samples were collected from 21patients (17 multi-bacillary, 4 pauci-bacillary) and 17 from non-patients (7 from contacts, 10 from other skin disease). Serum level of C-reactive protein, interferon gamma-induced protein 10, anti-phenolic-glycolipid-1 IgM and equation showed statistical significance compared from patients to non-patients. Comparing multi-bacillary patients to pauci-bacillary patients, only anti-phenolic-glycolipid-1 IgM showed statistical significance. Each area under receiver operating characteristic curve of those biomarkers and equation was ≥0.70 and p≤0.05. The results of Equation using 3 indicators showed higher area under receiver operating characteristic curve and lower P value compared when each indicator was used. Also sensitivity and specificity of them presented meaningful data. Conclusion We confirmed that using integrated analysis of three biomarkers is more significant than the single indicator in the diagnosis of leprosy. In addition, the use of multiple biomarkers is thought to contribute to the management of leprosy in the community.

Background: Apolipoprotein E protein is a family of apolipoproteins that play an important role in lipid metabolism. Apolipoprotein E genotype and single-nucleotide polymorphisms of genes are known to induce neurodegenerative diseases and cardiovascular diseases. Recently, it has been reported that apolipoprotein E is related to Hansen's disease by regulating genes related to lipid metabolism. Objective: In this study, we investigated that relationship of apolipoprotein E genotype with leprosy type and single-nucleotide polymorphism in korean persons affected by leprosy. Methods: The Apolipoprotein E genotype was confirmed using the polymerase chain reaction from korean persons affected by leprosy. Real-time polymerase chain reaction using probes confirmed the rs405509 and rs439401 of Apolipoprotein E. The identified genotype was cross-analyzed with each single-nucleotide polymorphism results and leprosy type by chi-squared test. Results: The distribution of apolipoprotein E genotypes was E3/E3 76.7%, E3/E4 17.4%, E2/E2 2.3%, E2/E3 2.3%, E2/E4 1.2% were in order. The frequencies of the apolipoprotein E allele were identified in the order of ε2 4.1%, ε3 86.6%, and ε4 9.3%. By type of leprosy, ε2 allele increased 2.2 times in multibacillary than pauacibacillary(4.8% vs 2.2%), and ε4 allele decreased 2.8 times in multibacillary than pauacibacillary(6.3% vs 17.4%). ApoE genotype was cross-analyzed with rs405509·rs439401, and there was a statistically significant association. Conclusion Apolipoprotein E genotype is associated with leprosy types in korean persons affected by leprosy. The research for apolipoprotein E gene is thought to be helpful in basic research on the pathogenesis of leprosy.who underwent surgery.

Background: Glucose-6-phosphate dehydrogenase deficiency is the most common enzyme deficiency disease. Drugs that cause deficiency-related hemolytic responses include dapsone, which is taken as a therapeutic drug for Hansen's disease. Due to the hemolytic adverse effects, the deficiency test should be performed before the dapsone administration, but it is not conducted on the basis of low incidence in Korea. Thus, the monitoring of deficiency in patients with Hansen's disease was needed and the study was undertaken. Objective: The aims of this study were to determine the prevalence of deficiency and gene mutations from persons affected by Hansen's disease in Korea. Methods: Hemanalysis and activity test by venipuncture were performed against the 339 persons affected by Hansen's disease. All subjects were tested for detection of mutations by the polymerase chain reaction. Results: The prevalence of anemia was 45.5% and the prevalence of deficiency was 1.2% from persons affected by Hansen's disease. 2 gene mutations, Mahidol and Mediterranean, were detected in foreign participants (Myanmar and Nepal). Conclusion In this study, we detected that the prevalence of deficiency and gene mutation was low, thus inferred that anemia was relatively less relevant to deficiency in persons affected by Hansen's disease. However, the number of participant in our study is small, and many studies reported that deficiency test should be performed before the dapsone administration, therefore, we suggest that continuous monitoring of patients with Hansen's disease is necessary.

Background: One of the Sequelae of facial nerve paralysis in leprosy patients is severe eye and lip deformity. Eye deformities are most common, followed by unilateral lip deviation. When there is bilateral failal nerve paralysis, the lower lip drooping may occur. The lower lip drooping makes mouth closure incomplete, so the fluid or saliva may flow out when eating. And the teeth may be exposed or even the gum may be visible, which causes serious cosmetic and functional impairments. Objective: It is to evaluate the masseter muscle sling (MMS) method in the treatment of the recurrent lower lip drooping in patients with Hansen's disease. Methods: In the case of masseter muscle sling, there is a traditional method of detaching, separating and raising anterior masseter muscle in half from the mandibular jaw line and fixing it to the lip muscle. However, the authors observed a 3cm gap between the anterior border of masseter muscle and the lateral edge of orbicularis oris muscle. Under the local anesthesia the nasolabial line was incised and 3-4 places were pulled up between both musle edges and plicated with a 2/0 PDS??threads the gap-space. Results: During 5 years (2016-2020), there were 17 recurrent lower lip palsy patients. The mean age was 71.3 years and the ratio of male and female was 9:8. Of 17 patents, 12 received temporal muscle transfer (TMT) attached with tensor fascia lata (TFL), 2 received temporal muscle transfer attached with Alloderm, and 3 received masseter muscle sling alone. The postoperative results of corrective masseter muscle sling were satisfactory in all 17 patients and there were no serious complication and recurrence during follow-up. Conclusion Masseter muscle sling is effective method in correction of recurrent lower lip drooping in patient with leprosy who treated previously with temporal muscle transfer attached with tensor fascia lata or temporal muscle transfer attached with Alloderm. In addition, masseter muscle sling may be considered as the first-line treatment in the lower lip paralysis.

Background: The prevalence of leprosy has almost reached the goal of World Health Organization. Accordingly, attention to leprosy and the number of skilled doctors who can diagnosis the leprosy upon clinical manifestations were decreased which caused the delayed diagnosis of leprosy. Objective: Serum C-reactive protein, interferon gamma-induced protein 10, anti-phenolic-glycolipid-1 IgM are biomarkers associated with multi-bacillary leprosy. Therefore, we investigated whether the use of multiple biomarkers has diagnostic value. Methods: The samples were collected from August 2012 to December 2019 at our institute. Through various statistic methods, multiple biomarkers and equation were compared in multi-bacillary patients, pauci-bacillary patients, contacts and different skin disease. Results: Samples were collected from 21patients (17 multi-bacillary, 4 pauci-bacillary) and 17 from non-patients (7 from contacts, 10 from other skin disease). Serum level of C-reactive protein, interferon gamma-induced protein 10, anti-phenolic-glycolipid-1 IgM and equation showed statistical significance compared from patients to non-patients. Comparing multi-bacillary patients to pauci-bacillary patients, only anti-phenolic-glycolipid-1 IgM showed statistical significance. Each area under receiver operating characteristic curve of those biomarkers and equation was ≥0.70 and p≤0.05. The results of Equation using 3 indicators showed higher area under receiver operating characteristic curve and lower P value compared when each indicator was used. Also sensitivity and specificity of them presented meaningful data. Conclusion We confirmed that using integrated analysis of three biomarkers is more significant than the single indicator in the diagnosis of leprosy. In addition, the use of multiple biomarkers is thought to contribute to the management of leprosy in the community.

Background: Apolipoprotein E protein is a family of apolipoproteins that play an important role in lipid metabolism. Apolipoprotein E genotype and single-nucleotide polymorphisms of genes are known to induce neurodegenerative diseases and cardiovascular diseases. Recently, it has been reported that apolipoprotein E is related to Hansen's disease by regulating genes related to lipid metabolism. Objective: In this study, we investigated that relationship of apolipoprotein E genotype with leprosy type and single-nucleotide polymorphism in korean persons affected by leprosy. Methods: The Apolipoprotein E genotype was confirmed using the polymerase chain reaction from korean persons affected by leprosy. Real-time polymerase chain reaction using probes confirmed the rs405509 and rs439401 of Apolipoprotein E. The identified genotype was cross-analyzed with each single-nucleotide polymorphism results and leprosy type by chi-squared test. Results: The distribution of apolipoprotein E genotypes was E3/E3 76.7%, E3/E4 17.4%, E2/E2 2.3%, E2/E3 2.3%, E2/E4 1.2% were in order. The frequencies of the apolipoprotein E allele were identified in the order of ε2 4.1%, ε3 86.6%, and ε4 9.3%. By type of leprosy, ε2 allele increased 2.2 times in multibacillary than pauacibacillary(4.8% vs 2.2%), and ε4 allele decreased 2.8 times in multibacillary than pauacibacillary(6.3% vs 17.4%). ApoE genotype was cross-analyzed with rs405509·rs439401, and there was a statistically significant association. Conclusion Apolipoprotein E genotype is associated with leprosy types in korean persons affected by leprosy. The research for apolipoprotein E gene is thought to be helpful in basic research on the pathogenesis of leprosy.who underwent surgery.

Background: Glucose-6-phosphate dehydrogenase deficiency is the most common enzyme deficiency disease. Drugs that cause deficiency-related hemolytic responses include dapsone, which is taken as a therapeutic drug for Hansen's disease. Due to the hemolytic adverse effects, the deficiency test should be performed before the dapsone administration, but it is not conducted on the basis of low incidence in Korea. Thus, the monitoring of deficiency in patients with Hansen's disease was needed and the study was undertaken. Objective: The aims of this study were to determine the prevalence of deficiency and gene mutations from persons affected by Hansen's disease in Korea. Methods: Hemanalysis and activity test by venipuncture were performed against the 339 persons affected by Hansen's disease. All subjects were tested for detection of mutations by the polymerase chain reaction. Results: The prevalence of anemia was 45.5% and the prevalence of deficiency was 1.2% from persons affected by Hansen's disease. 2 gene mutations, Mahidol and Mediterranean, were detected in foreign participants (Myanmar and Nepal). Conclusion In this study, we detected that the prevalence of deficiency and gene mutation was low, thus inferred that anemia was relatively less relevant to deficiency in persons affected by Hansen's disease. However, the number of participant in our study is small, and many studies reported that deficiency test should be performed before the dapsone administration, therefore, we suggest that continuous monitoring of patients with Hansen's disease is necessary.