Mutation-based selection and analysis of Komagataeibacter hansenii HDM1-3 for improvement in bacterial cellulose production.

AIMS: A low yield of bacterial cellulose (BC) always results from an excessive accumulation of organic acids. Screening and the selection of bacterial mutants with a low accumulation of organic acids is an efficient approach for improving BC production. METHODS AND RESULTS: In combination with the proton suicide method (medium containing NaBr-NaBrO3 ), diethyl sulphate chemical mutagenesis coupled with 60 Co-γ irradiation treatment were performed for the screening and selection of desired mutant lines with a high yield of BC. Two high-yield strains, Br-3 and Co-5, as well as a low-yield strain, Br-12, were obtained. Amplified fragment length polymorphism (AFLP) was applied to explore the differences between the mutant lines and the wild type. For the Br-12 line, three specific fragments were verified, corresponding to TonB-dependent transport (TBDT), exopolysaccharides output protein (PePr) and an unknown gene. For Co-5, two specific fragments were matched, acsD and UDP-galactose-4-epimerase. In addition, metabolic analysis for the mutant lines indicated that BC production may be limited by excessive accumulation of organic acids in the fermentation. The limitation would be resolved by the cross-talk of genes involved in BC biosynthesis. CONCLUSIONS: Reduced organic acid by-products from glucose in bypasses were found to be responsible for the high-yield BC synthesis in Komagataeibacter hansenii mutant strains. SIGNIFICANCE AND IMPACT OF THE STUDY: The metabolic process was varied by mutagenesis-induced gene disruption of the metabolic products. A new idea was provided for the targeted screening and characterization of mutants in the future.

HLA-B*13:01 and the dapsone hypersensitivity syndrome.

BACKGROUND: Dapsone is used in the treatment of infections and inflammatory diseases. The dapsone hypersensitivity syndrome, which is associated with a reported mortality of 9.9%, develops in about 0.5 to 3.6% of persons treated with the drug. Currently, no tests are available to predict the risk of the dapsone hypersensitivity syndrome. METHODS: We performed a genomewide association study involving 872 participants who had received dapsone as part of multidrug therapy for leprosy (39 participants with the dapsone hypersensitivity syndrome and 833 controls), using log-additive tests of single-nucleotide polymorphisms (SNPs) and imputed HLA molecules. For a replication analysis, we genotyped 24 SNPs in an additional 31 participants with the dapsone hypersensitivity syndrome and 1089 controls and performed next-generation sequencing for HLA-B and HLA-C typing at four-digit resolution in an independent series of 37 participants with the dapsone hypersensitivity syndrome and 201 controls. RESULTS: Genomewide association analysis showed that SNP rs2844573, located between the HLA-B and MICA loci, was significantly associated with the dapsone hypersensitivity syndrome among patients with leprosy (odds ratio, 6.18; P=3.84×10(-13)). HLA-B*13:01 was confirmed to be a risk factor for the dapsone hypersensitivity syndrome (odds ratio, 20.53; P=6.84×10(-25)). The presence of HLA-B*13:01 had a sensitivity of 85.5% and a specificity of 85.7% as a predictor of the dapsone hypersensitivity syndrome, and its absence was associated with a reduction in risk by a factor of 7 (from 1.4% to 0.2%). HLA-B*13:01 is present in about 2 to 20% of Chinese persons, 1.5% of Japanese persons, 1 to 12% of Indians, and 2 to 4% of Southeast Asians but is largely absent in Europeans and Africans. CONCLUSIONS: HLA-B*13:01 was associated with the development of the dapsone hypersensitivity syndrome among patients with leprosy. (Funded by the National Natural Science Foundation of China and others.).

IL-10 promoter SNPs and susceptibility to leprosy in ethnic groups from southwest China.

The purpose of this study was to determine whether interleukin-10 (IL-10) promoter polymorphisms are associated with leprosy or their subtypes in ethnic groups from southwest China. Genotyping using TaqMan® SNP Genotyping Master Mix and ABI 7500 real-time PCR system was performed for IL-10 T3575A, G2849A, C2763A, A1082G, C819T, and C592A in 189 healthy controls (40 ± 18 years) and 193 patients (46 ± 18 years) with leprosy [multibacillary, N = 131; paucibacillary (PB), N = 62]. The allelic frequencies of -2763C (97.9 vs 94.0%, P = 0.0074) and -1082A (92.8 vs 88.6%, P = 0.0452) in leprosy patients were significantly higher than in control subjects. The genetic frequency of -2763CC and -1082AA was not only significantly higher among leprosy patients than among control subjects [odds ratio (OR) = 3.33, 95% confidence interval (95%CI) = 1.39-7.99, P = 0.0071 and OR = 1.76, 95%CI = 1.02-3.03, P = 0.0420, respectively] but also significantly higher among PB patients than among control subjects (OR = 2.46, 95%CI = 1.22-4.96, P = 0.0115 and OR = 5.58, 95%CI = 2.06-15.12, P = 0.0007, respectively). The frequency of IL-10 haplotype 3575A/2849G/2763A/1082G/819C/592C was significantly higher among leprosy patients (OR = 5.57, 95%CI = 1.13-27.52, P = 0.0351) and PB patients (OR = 10.5, 95%CI = 1.36- 81.05, P = 0.0241) than among control subjects. IL-10 promoter -2763C/CC,-1082A/AA and haplotype 3575A/2849G/2763A/1082 G/819C/592C are associated with susceptibility to leprosy and the PB subtype in southwest China.

Studies on mode of detection of leprosy in China during the years 1981-1998.

Along with the nationwide economic reform initiated in the early 1980s and the rapid decrease of leprosy endemic after the implementation of multi-drug therapy (MDT), the leprosy programme changed from 'vertical' to 'horizontal'. An evolution in the mode of detection of leprosy cases has consequently taken place. Based on the nationwide registration of newly detected cases, the profile of patients at detection has been studied. The proportions of cases corrected significantly with calendar years in detection by dermatological clinics, contact checks, 'clue survey' and mass survey, showing a significant increase in percentage of cases detected through dermatological clinics and contact checks, and decreases through 'clue survey' and mass survey. Detection of cases through dermatological clinics and voluntary reporting have become the main modes of case-finding during 1997-1998, accounting for 37.3% and 28.6%, respectively, where contact check accounts for only 9.1%. In areas with good dermatological services, a significantly higher proportion (75.9%) of cases was detected through dermatological clinics, where voluntary reporting and 'clue survey' were the main modes of detection in endemic areas. As regards confirmation of diagnosis, the great majority of cases were confirmed by leprosy units, even though they were detected in various other situations. Only 6.5% of leprosy cases were detected and subsequently confirmed by doctors in dermatologic clinics. The present modes of detection and their relation to demographical, epidemiological, clinical factors and health services are discussed. This study emphasizes the cardinal importance of the dermatological clinics in the detection of leprosy cases in China at the present time and hence the need to strengthen the training of doctors in these clinics, while continuously encourage their involvement in leprosy control.

Leprosy in China: epidemiological trends between 1949 and 1998.

OBJECTIVE: To report the epidemiological trends of leprosy in China from 1949 to 1998. METHOD: Data for the study were obtained from the computerized database of the National System of Leprosy Surveillance. FINDINGS: A total of 474,774 leprosy patients were detected during this 50-year period. Case detection rates per 100,000 population were highest in the 1950s and 1960s, with peaks appearing in 1957-58, 1963-66, 1969-70, and 1983-84, corresponding to mass surveys or screening surveys carried out in most areas or selected areas of the country. While the duration of the disease at the time of detection fell over the period, the disability rates, which were > 50% in the early 1950s, have decreased gradually to 20.8% by 1997-98 but are still too high. More than 50% of cases were found through active methods in the periods 1955-58, 1965-66, and 1969-76, but in recent years cases are mostly detected through dermatological clinics or by voluntary reporting. The peak prevalences of the 1960s (i.e. > 2 per 10,000 population) decreased annually from the 1970s onwards. By the end of 1998 the prevalence was 0.05 per 10,000 population. CONCLUSIONS: This study shows that leprosy was well controlled in China and that the WHO goal of elimination of leprosy as a public health problem has been achieved at the national and subnational levels. However, leprosy is still unevenly distributed in the country. According to the criterion for leprosy elimination in China--defined as a prevalence of < 1 per 100,000 in county or city--there are still more than 10% of counties or cities where this target has not yet been reached. Special attention must therefore be given to achieve elimination and final eradication of leprosy in China.

Leprosy in children: a retrospective study in China, 1986-1997.

Leprosy in children forms an important link in the natural evolution and the epidemic profile of the disease. The study was undertaken on 1208 child cases detected during the years 1986-1997 in China. The number of child cases constituted 3-4 per cent of the total during this period although the incidence rates decreased dramatically. The detection rate significantly increased with age, and the average rate of 0.027 per 100,000 population was much lower than that in adults. In the pediatric cases, children in the younger age group had a higher prevalence of solitary lesions and lower prevalence of reactions and disabilities than the older group. A total of 70.01 per cent of the affected cases were infected by intra-familial contact, 35.60 per cent were detected by contact examination, and another 22.86 per cent by active surveys. The results of the study suggest that leprosy in children is not a public health problem at the present in China, but the regular screening of children in the family of patients is still one of the important ways to detect child cases and should be recommended.

Computerization of leprosy records: national leprosy recording and reporting system in China.

This paper describes the national system of leprosy recording and reporting in China and the computerization of records. The system was designed for data collection at local level and data entry by optically scanned or manual mode as well as for sophisticated data analysis. The major functions include data entry, data check, sum-up, maintenance, communication, inquiry, statistics, graph and print. A total of 17 options for epidemiological and clinical data analysis are available. Through the implementation for about 10 years, the system has gained widespread acceptance. This acceptance would facilitate introduction of computer analysis to other leprosy projects and other disease control programs in China. Up to 1998, a database of more than 740,000 records covering all the leprosy patients detected since 1949 had been established by this system.

Leprosy in China: delay in the detection of cases.

In a national survey in China, 27,928 cases of leprosy detected by the health authorities between 1984 and 1998 were investigated. The delay between onset of symptoms (estimated from each case's recall) and confirmed diagnosis was < or = 2 years for 55.1% of the new patients but > 10 years for 7.0%, with a median value, overall, of 22.0 months. The median delay was longer: (1) for the multibacillary cases than the paucibacillary; (2) among farmers than among factory workers; (3) among some nationalities than among others (being longest among the Tu and shortest among the Wei); and (4) for some methods of case-detection than for others. Over the study period, the mean delay decreased with time. The delay was greatest in the areas where leprosy was endemic and/or where access to health services was poor. The later the cases were detected the more likely they were to show disability. Leprosy cases are still going undetected in China, although, over the last 14 years, case-finding has significantly improved. Age, occupation, nationality, leprosy type and detection method all appear to affect the delay.

Final report about Stage II Collaboration Project on Leprosy Rehabilitation.

OBJECTIVE: To undertake the pilot experiments of prevention of disability (POD) in 14 different geographical areas to serve as examples for future development of rehabilitation work in China and in Asia. METHODS: According to the principles and national criterion, 27 000 people affected by leprosy were selected and assessed using disability record forms at beginning and followed up regularly for observing changes of different indicators. RESULTS: A total of 197 neuritis cases were detected and treated with prednisolone out of 1 407 new or active cases. Self-care training of eyes, hands and feet were conducted for 10 500 disabled people affected by leprosy. Comprehensive therapy was given to 1 804 cases having complicated ulcers of which 1 055 cases have got their ulcers healed. Out of 706 prostheses, 613 were given to patients with satisfactory results. Surgical treatment was given to 269 cases and 251 have shown good progress. CONCLUSION: Most of patients have got benefit from the project in function or appearance which is very helpful for their going back to the society and agreed by foreign experts during the final evaluation. The experiences from the project can be implemented in the whole country.

Prediction of elimination of leprosy in leprosy endemic areas of China.

A study was carried out based upon the data from the National System for Leprosy Surveillance and using appropriate mathematical models. The results showed that of 337 counties where the national goal of basic eradication of leprosy had not been reached and in 40 counties where the WHO goal of leprosy elimination had not been achieved in 1996, the detection rates in calendar years followed exponential models with significant goodness-of-fit. In the 67 counties with downward trends of detection rates, the national goal can be met in terms of detection rate in 6% of counties before the year 2000 or 34.4% before the year 2010, or, in terms of prevalence rate in 31.3% before the year 2010. In the 11 counties with downward trends of the detection rates, the WHO target can be met in eight to ten counties within this century when the duration of disease was determined with the WHO definition. If the MB proportion among new cases increased by 10%, the target would be met one year later. However, at the same MB proportion, the change of fixed treatment schedules from PB six months and MB two years to PB nine months and MB three years will cause achievement of the goal to be postponed by two to ten years.

Studies on risk of leprosy relapses in China: relapses after treatment with dapsone monotherapy.

Based upon the data from the Chinese National System for Leprosy Surveillance, this paper reports on the relapses in 297,343 leprosy patients [multibacillary (MB) 106,518, paucibacillary (PB) 190,825] cured by dapsone monotherapy. A total of 11,055 (MB 8675, PB 2380) patients relapsed during an accumulated follow-up period of 4,229,050 patient-years (PY), giving an overall relapse rate of 3.72 per 100 cases or 2.61 per 1000 PY, i.e., 8.14% or 5.91 per 1000 PY over an average follow-up period of 13.8 +/- 8.4 years in MB patients and 1.25% or 0.86 per 1000 PY over an average period of 14.5 +/- 8.9 years in PB patients. For either the overall relapse rate per 100 cases or per 1000 PY, the differences between MB and PB patients were statistically significant, except during 36-40 years of follow up. For both MB and PB patients, the relapse rates showed consistently significant decreases year by year, particularly in PB patients whose relapse rate per 1000 PY was 1.21 in year 10 of follow up; whereas it remained more than 10 per 1000 PY in MB patients. In view of that, the overall relapse rates in MB and PB patients cured by dapsone monotherapy were acceptably low, and most of these patients have been followed up for more than a mean incubation period of observed dapsone relapse. Along with the further extension of follow up, the risk of relapse in dapsone-cured patients will not be expected to increase. This conclusion should be considered when planning policy for the management of patients released from dapsone monotherapy.

Studies on risk of leprosy relapses in China: relapses after treatment with multidrug therapy.

Based upon the data from the Chinese National System for Leprosy Surveillance, this paper reports on the relapses in 47,276 leprosy patients cured by or released from WHO-recommended multidrug therapy (WHO/MDT). The overall relapse rate was 0.73/1000 patient-years (PY). There was a statistically significant difference in the relapse rates of WHO/MDT-MB (0.61/1000 PY) and WHO/MDT-PB (1.04/1000 PY) (chi 2 = 15.7, p < 0.01) patients. For multibacillary (MB) patients, the relapse rate in patients treated with fixed-duration MDT (0.56/1000 PY) was comparable with that in patients treated with MDT until skin-smear negativity (0.73/1000 PY) (chi 2 = 2.20, p > 0.05). Our present study suggests that fixed-duration MDT is a cost-effective regimen for the treatment of leprosy in China. The present results also show that relapse of leprosy is acceptably low and has not yet become a serious clinical or public health problem but, based upon the incubation of relapse in MDT patients, it is necessary to encourage annual follow up for at least 5 years for paucibacillary (PB) and 10 years for MB patients after being released from WHO/MDT.

An investigation of attitudes, beliefs and behaviour of leprosy patients, family members and PHC workers towards multidrug therapy in Yangzhou and Dongtai Districts of China.

To improve the operational efficiency of multidrug therapy (MDT) implementation in rural areas, an investigation into the attitudes, beliefs and behaviour of leprosy patients and their family members as well as primary health care (PHC) workers towards MDT was carried out in Yangzhou and Dongtai Districts of China. A sample of 370 leprosy patients, 594 family members and 730 PHC workers was interviewed or investigated individually using questionnaires. The results showed that: 1, the presently used MDT is acceptable to a wide range of patients although a small number of patients have various problems in their treatment; 2, the patients' habit in daily drug administration, their awareness of the risk of default and confidence in MDT have a positive influence in increasing drug compliance; and 3, the supervision and encouragement of family members to patients' treatment which is associated with their knowledge on MDT is also beneficial to patients' drug compliance. However, only half of the PHC workers had a basic knowledge of MDT and a desire to participate in MDT implementation, a finding which clearly calls for urgent attention and improvement. In order to ensure the effective implementation of MDT, there is a need to educate leprosy patients and their family members as well as PHC workers to establish the patients' correct awareness of MDT, obtain the family support and motivate the PHC workers.

An epidemiological survey of deformities and disabilities among 14,257 cases of leprosy in 11 counties.

This study was planned and conducted in Yang Zhou Prefecture, covering 11 counties that were formerly areas with a high prevalence of leprosy. Out of 14,257 leprosy patients, 8122 (56.97%) cases with deformities and disabilities were found. The disability rate is much higher in patients with MB leprosy (81.15%) than in PB leprosy (53.04%). The statistical data and the type of deformities and disabilities are presented. The influences of various host factors and disease factor which cause disability and deformity are discussed.

An epidemiological survey of deformities and disabilities among 14257 leprosy patients in 11 counties.

This study was planned and conducted in 11 counties in Yangzhou Prefecture, which had formerly had a high prevalence of leprosy. Out of 14257 leprosy patients, 8122 (56.97%) with permanent deformities and disabilities were found. The disability rate was much higher among patients with multibacillary (MB) leprosy (81.15%) than among those with paucibacillary (PB) leprosy (53.04%). The statistical data involving the patients and the types of deformities and disabilities are presented in this paper, and the influences of various host factors and disease factors are discussed.