Towards the Anaerobic Production of Surfactin Using Bacillus subtilis.

The anaerobic growth of B. subtilis to synthesize surfactin poses an alternative strategy to conventional aerobic cultivations. In general, the strong foam formation observed during aerobic processes represents a major obstacle. Anaerobic processes have, amongst others, the distinct advantage that the total bioreactor volume can be exploited as foaming does not occur. Recent studies also reported on promising product per biomass yields. However, anaerobic growth in comparison to aerobic processes has several disadvantages. For example, the overall titers are comparably low and cultivations are time-consuming due to low growth rates. B. subtilis JABs24, a derivate of strain 168 with the ability to synthesize surfactin, was used as model strain in this study. Ammonium and nitrite were hypothesized to negatively influence anaerobic growth. Ammonium with initial concentrations up to 0.2 mol/L was shown to have no significant impact on growth, but increasing concentrations resulted in decreased surfactin titers and reduced nitrate reductase expression. Anaerobic cultivations spiked with increasing nitrite concentrations resulted in prolonged lag-phases. Indeed, growth rates were in a similar range after the lag-phase indicating that nitrite has a neglectable effect on the observed decreasing growth rates. In bioreactor cultivations, the specific growth rate decreased with increasing glucose concentrations during the time course of both batch and fed-batch processes to less than 0.05 1/h. In addition, surfactin titers, overall Y P/X and Y P/S were 53%, ∼42%, and ∼57% lower than in serum flask with 0.190 g/L, 0.344 g/g and 0.015 g/g. The Y X/S, on the contrary, was 30% lower in the serum flask with 0.044 g/g. The productivities q were similar with ∼0.005 g/(g⋅h). However, acetate strongly accumulated during cultivation and was posed as further metabolite that might negatively influence anaerobic growth. Acetate added to anaerobic cultivations in a range from 0 g/L up to 10 g/L resulted in a reduced maximum and overall growth rate µ by 44% and 30%, respectively. To conclude, acetate was identified as a promising target for future process enhancement and strain engineering. Though, the current study demonstrates that the anaerobic cultivation to synthesize surfactin represents a reasonable perspective and feasible alternative to conventional processes.

Training and financial intervention for encouraging maternal health service utilization: Results of cluster randomized trials in Shaanxi Province.

BACKGROUND: The rising maternal and child healthcare costs and the lack of training and educational resources for healthcare workers have reduced service quality in primary health centers of China. We sought to compare strategies promoting healthcare service utilization in rural western China. METHOD: A randomized community trial was carried out in Zhen'an country between 2007 and 2009. Two cross-sectional surveys were conducted to compare the outcomes of financial subsidy for pregnant women seeking antenatal care and clinical training provided to healthcare workers by difference-in-difference estimation. RESULTS: In all, 1113 women completed the questionnaires. The proportion of postnatal visits increased three times in the training group, reaching 35.7%. The number of women who received advice from their doctors regarding nutrition and warning signs necessitating immediate medical attention also improved significantly (5.8% and 8.2%, respectively). Furthermore, the percentage of women who underwent blood tests increased significantly to 19.5% in the training group. Compared to the financial group, the training group had more women who breastfed for longer than 4 months (15.8%) and provided timely complementary feeding (8.9%). CONCLUSION: The training intervention appeared to have improved prenatal care utilization. Essential obstetric training helped enhance knowledge and self-efficacy among healthcare workers.

Genome Sequence of Jannaschia aquimarina GSW-M26, a Member of the Roseobacter Clade.

The Gram-negative alphaproteobacterium Jannaschia aquimarina GSW-M26 (DSM 28248) is a member of the Roseobacter clade. The size of the draft genome is 4.1 Mb. Genome analysis revealed the presence of genes encoding a complete gene transfer agent and aerobic anoxygenic photosynthesis. The latter indicated a photoheterotrophic lifestyle.

The status of vitamin B12 and folate among Chinese women: a population-based cross-sectional study in northwest China.

OBJECTIVE: To assess the status of the vitamin B12 and folate of Chinese women living in northwest China. METHODS: A population-based cross-sectional study was conducted in 2008 among Chinese women aged 10-49 years living in Shaanxi province of northwest China. A stratified multistage random sampling method was adopted to obtain a sample of 1170 women. The women were interviewed for collection of their background information and their plasma vitamin B12 and folate were measured with the immunoassay method. The status of both vitamins was evaluated and the prevalence of deficiency was estimated. RESULTS: The median value of the women was 214.5 pg/mL for vitamin B12 and 4.6 ng/mL for folate. The urban women had a significantly higher vitamin B12 (254.1 vs. 195.9 pg/mL) but lower folate (4.4 vs. 4.7 ng/mL) than rural women. Total prevalence of deficiency was 45.5% (95% CI: 42.6% ∼ 48.4%) for vitamin B12 and 14.7% (95% CI: 12.6% ∼ 16.8%) for folate. About 36% of women presented vitamin B12 deficiency alone, 5.2% belonged to folate deficiency alone and 9.5% was combined deficiency in both vitamins. More than 25% of the women were in marginal vitamin B12 status (200-299 pg/mL) and 60% in marginal status of folate (3-6 ng/mL). About 75.2% of rural women with folate deficiency were deficient in vitamin B12 and 46% for urban women. Quantile regression model found decreasing coefficient of folate status across 73 different quantiles of vitamin B12, which indicated that the women with folate deficiency had lower vitamin B12 significantly compared with those with no deficiency. CONCLUSIONS: The deficiency of vitamin B12 and folate is still prevalent among the Chinese women in northwest China. Vitamin B12 deficiency could be more serious and the improvement of poor vitamin B12 status should be invoked when practicing the supplementation of folate against the neural tube defects in northwest China.

Equity in use of maternal health services in Western Rural China: a survey from Shaanxi province.

BACKGROUND: The 20th century was marked by a significant improvement in worldwide human health and access to healthcare. However, these improvements were not completely or uniformly distributed among, or even within, nations. This study was designed to assess the use of maternal health services by pregnant women in China, with a focus on the inequity related to family income level. METHODS: Two population-based cross-sectional surveys were carried out in the Zhenan and Lantian counties in March 2007 and from December 2008 to March 2009. A total of 2562 women completed the questionnaires, including 948 who were pregnant in 2006 and 1614 from 2008-2009. The concentration index (CI) was calculated and used to analyze the parameters of maternal health care in the two counties surveyed. RESULTS: The responses in both 2006 and 2008-2009 indicated a bias towards higher (rich) economic statuses for the use of maternal and child health services. The CI of 'delivery at health facility' was 0.0206 (95% confidence interval between 0.0114 and 0.0299) for 2006 and 0.0053 (95% confidence interval between 0.0015 and 0.0091) for 2008, which represented a statistically significant inequity for women of lower (poor) economic statuses. Similar CI was observed in 'receiving antenatal care within 12 weeks' for 2006 (CI2006 = 0.0956, 95% confidence interval between 0.0516 and 0.1396). The CIs of 'postnatal visit' and 'postnatal visit >3-times' was positive (except for 2006), indicating that the poor used postnatal care less than the non-poor. In 2008, poor women had C-sections more often than non-poor women (CI2008 = -0.0629, 95% confidence interval between-0.1165 and -0.0093), but such a difference was not observed in 2006. CONCLUSIONS: In 2006 and 2008, the use of maternal health services in western rural China was significantly unequal between pregnant women of poor and non-poor economic statuses. Financial support that enables poorer pregnant women to use health services will be beneficial. Utilization of maternal healthcare services can be improved if out-of-pocket expenses can be minimized.

Utilisation, contents and costs of prenatal care under a rural health insurance (New Co-operative Medical System) in rural China: lessons from implementation.

BACKGROUND: In China, the New Co-operative Medical System (NCMS), a rural health insurance system, has expanded nationwide since 2003. This study aims to describe prenatal care use, content and costs of care in one county where prenatal care is included in the NCMS and two counties where it is not. It also explores the perceptions of stakeholders of the prenatal care benefit package in order to understand the strengths and weaknesses of the approach in the context of rural China and to draw lessons from early implementation. METHODS: This study is based on the data from a cross-sectional survey and a qualitative investigation conducted in 2009. A survey recruited women giving birth in 2008, including 544 women in RC County (which covered prenatal care) and 619, and 1071 in other two counties (which did not). The qualitative investigation in RC included focus group discussions with women giving birth before or after 2007, individual interviews with local policy makers and health managers, NCMS managers and obstetric doctors in township hospitals. RESULTS: There were no significant differences in prenatal care use between RC County (which covered prenatal care) and other two counties (which did not): over 70% of women started prenatal visits early and over 60% had five or more visits. In the three counties: a small proportion of women received the number of haemoglobin and urine tests recommended by the national guideline; 90% of women received more ultrasound tests than recommended; and the out-of-pocket expenditure for prenatal care consumed a high proportion of women's annual income in the low income group. In RC: only 20% of NCMS members claimed the reimbursement; the qualitative study found that the reimbursement for prenatal care was not well understood by women and had little influence on women's decisions to make prenatal visits; and several women indicated that doctors suggested them taking more expensive tests. CONCLUSIONS: Whether or not prenatal care was included in the NCMS, prenatal care use was high, but the contents of care were not provided following the national guideline and more expensive tests were recommended by doctors. Costs were substantial for the poor.

Utilization of delivery care among rural women in China: does the health insurance make a difference? a cross-sectional study.

BACKGROUND: Since 2003, the New Cooperative Medical Scheme (NCMS) has been implemented throughout rural China, usually covering delivery services in its benefit package. The objective of this study was to compare the difference of utilization of delivery services, expenditures, and local women's perceived affordability between women with and without reimbursement from NCMS. METHODS: A cross-sectional survey was carried out in two rural counties in Shaanxi province, China, during December 2008-March 2009. Women giving birth from April 2008 to March 2009 were interviewed by a structured questionnaire to collect information on utilization of delivery services. Multivariable analyses were used to compare the differences in outcomes between women with and without reimbursement from NCMS. RESULTS: Of the total 1613 women interviewed, 747(46.3%) got reimbursement to cover their expenditure on delivery care (NCMS group) and 866(53.7%) paid delivery services entirely out of their own pocket (Non-NCMS group). Compared with the Non-NCMS group, the NCMS group had significantly more women who delivered at hospital. The rate of Caesarean section (CS), proportion of women seeking higher level services, and length of hospitalization were similar between the two groups. The total hospital costs for delivery services in the NCMS group was significantly smaller and after being reimbursed, the out-of-pocket payment in the NCMS group was less than a half of that in the Non-NCMS group. Fewer women in the NCMS group than in the Non-NCMS group considered their payment for delivery services expensive. CONCLUSIONS: There was no evidence of overuse delivery services among the women reimbursed by NCMS. Total hospital costs and women's costs for delivery services were found lower in the NCMS group, subsequently alleviation on women's perceived financial affordability.

[The introduction of breastfeeding in children under age of three in the counties of western China in 2005].

OBJECTIVE: The study was to describe the breastfeeding status of children under the age of three in counties of western China and to provide evidence to the government for decision-making on intervention. METHODS: A cross-sectional study with probability-proportional-to-size (PPS) sampling method was used. The information on breastfeeding was obtained through memory of the mothers. Fourteen thousand and seventy-seven children were studied. Data on breastfed status in counties of western China was compared with those of the children from the survey of the counties of western China in 2001. RESULTS: The breastfeeding rate of children under 3 years old in western China was 96.5%. However, the overall breastfed rate of children under 6 months were only 33.4%, with rates of 11.4% and 22.0% on exclusively and predominantly breastfed groups respectively. Timely first-suckling rate was 43.5% with the continued breastfeeding rate (1 year) as 64.9%, but the continued breastfeeding rate (2 year) was only 9.7%. Reasons causing mothers to wean would include according to her own intention and to be able to attend the physical labor while exclusive breastfeeding under 6 months was for the growth and development of children, which might reduce the two-week prevalence of diarrhea. Major risks of exclusive breastfeeding of children under 6 months were seen as: level of education of the mothers, economic depression of the counties and mother's nationality (if as minority). CONCLUSION: Most of the children were ever or being breastfed at the time of interview with timely first-suckling took place earlier than in 2001. However continued breastfeeding did not last long. During these five years, the exclusive breastfeeding rate had been at low level, especially at the economic depression and the minority area.

[Assessment on nutritional status among children under 3 years using anthropometry in 40 counties of western China].

OBJECTIVE: To assess nutritional status among children younger than 3 years in 40 counties of western China, using anthropometry. METHODS: With cross-sectional study and random sampling, a survey with a sample size of 7252 was conducted in 40 counties of western China on children under 3 years old. Height and weight were measured and nutrition status was evaluated with WHO/NCHS reference using Z scores. RESULTS: SD of height for age (HA) and weight for age (WA) were 1.10 - 1.30 and 1.06 - 1.40, respectively and SD of WAZ for infants varied. The distribution of HA Z and WAZ shifted downward to 1 unit and 0.4 unit for distribution of weight for height (WHZ). The prevalence of stunting, underweight and wasting were 23.0%, 22.6% and 7.5% respectively. The prevalence rates were low among those yourger than 3 months. Stunting and underweight increased sharply after 6 months and reached 20% - 30%. Significant correlation was found between underweight and stunting or wasting with coefficients of 0.815 and 0.636, respectively. No correlation was found between stunting and wasting. CONCLUSION: The nutrition status adversely affected children as a whole but not only for individuals. There was a higher prevalence of malnutrition mainly on low height to children. Weight for age must be interpreted cautiously because of the inability of this indicator in distinguishing stunting and wasting just by itself. Assessment by anthropometry called for accurate information on age and measurement of height and weight.

Transcriptional activation function of hepatitis B virus Pre S1 protein in yeast.

OBJECTIVE: To explore the feasibility of cloning of the hepatocyte receptor interacting with the Pre S1 protein of HBV by two-hybrid system. METHODS: Yeast expression plasmids encoding fusion proteins of full length or portions of Pre S1 of HBV and DNA binding domain of yeast protein GAL4 were constructed and used to transform yeast reporter strain SFY526. Reporter gene product beta-galactosidase activity was assayed as a measure of transcriptional activation in yeast. Mammalian expression plasmid encoding fusion proteins of full length Pre S1 and DNA binding domain of GAL4 was constructed and used to cotransfect hepatoma cell line Huh-7 together with CAT reporter plasmid. Cell extracts were assayed for CAT activity by thin-layer chromatography. RESULTS: The fusion proteins of full length Pre S1 protein and GAL4 DNA binding domain presented transcriptional activation function in yeast. The transcription activating sequence was localized to the 21 to 47 amino acids of Pre S1 protein. Fusion proteins of full length Pre S1 and GAL4 DNA binding domain did not show transcriptional activation function in mammalian cells. CONCLUSIONS: The transcription activating sequence of HBV Pre S1 protein in yeast overlaps the hepatocyte receptor binding site. The transcriptional activation function of HBV Pre S1 protein in yeast may prevent researchers from using yeast two-hybrid system to clone HBV receptor interacting with Pre S1 protein. However, the Pre S1 protein does not show transcriptional activation function in mammalian cells. Mammalian two-hybrid system may be a practical method to clone the HBV hepatocyte receptor interacting with Pre S1 protein.

[Transcriptional activation function of hepatitis B virus Pre S1 protein in yeast].

BACKGROUND: To explore the feasibility of cloning of the hepatocyte receptor interacting with the Pre S1 protein of HBV by two hybrid system. METHODS: Yeast expression plasmids encoding fusion proteins of full length or portions of Pre S1 of HBV and DNA binding domain of yeast protein GAL4 were constructed and used to transform yeast reporter strain SFY526. Reporter gene product ?galactosidase activity was assayed as a measure of transcription activation in yeast. Mammalian expression plasmid encoding fusion proteins of full length Pre S1 and DNA binding domain of GAL4 was constructed and used to cotransfect hepatoma cell line Huh?7 together with CAT reporter plasmid. Cell extracts were assayed for CAT activity by thin?layer chromatography. RESULTS: The fusion proteins of full length Pre S1 protein and GAL4 DNA binding domain present transcriptional activation function in yeast. The transcription activating sequence is localized to the 21 to 47 amino acids of Pre S1 protein Fusion proteins of full length Pre S 1 and GAL 4 DNA binding domain do not show transcriptional activation function in mammalian cells. CONCLUSION: The transcriptional activating sequence of HBVPre S1 protein in yeast overlaps the hepatocyte receptor binding site. The transcriptional activation function of HBV Pre S1 protein in yeast may prevent researchers?from using yeast two hybrid system to clone HBV receptor interacting with Pre S1 protein. However, the Pre S1 protein does not show transcriptional activation function in mammalian cells. Mammalian two?hybrid system may be a practical method to clone the HBV hepatocyte receptor interacting with Pre S1 protein.