ỨNG DỤNG SẮC KÝ ÁI LỰC MIỄN DỊCH ĐỂ TINH CHẾ KHÁNG THỂ KHÁNG ĐỘC TỐ DOMOIC ACID TRONG HUYẾT THANH THỎ TRẮNG NEW ZEALAND

In order to develop an ELISA KIT for detection of domoic acid (DA) in seafood in Vietnam, DA antibody was produced by immunizing in New Zealand white rabbit using DA-DSS (disuccinimidyl suberate)-BSA (Bovine Serum Albumin) as an antigen. The DA antibody in rabbit’s serum was purified for its use as a material of ELISA KIT. In antibody/antigen purification process, immunoaffinity chromatography (IAC) is considered as the best method by both its specification and ability to enrich antibody. However, DA is a hapten, and it is impossible to apply a regular immunizing principal for making an IAC column with DA as a solid phase to catch DA antibody. This paper presents a trial using polymer sepharose EAH 4B resin (omega-aminohexyl) IAC bound with hapten DA as a solid phase for DA antibody purification from rabbit’s serum. Using DSS as a chemical linker; then, based on the reaction between a carboxyl group of DA and a carbodiimide group of the resin EAH 4B, we were successful to make the IAC column with DA-DSS as the solid phase. Recovery ratio of the made IAC column reached 91.5 ± 1.5% (n = 3) to capture the antibody against DA standard. DA antibody from white rabbit’s serum was eluted successfully using this column with 0.5 M NaCl; 0.1 M PBS and then, 0.1 M Ldy-HCl solutions (pH 2.7). The obtained antibody was qualified by Western Blot using a specific reaction with the 2nd specific antibody HRP-labeled anti-goat IgG and the confirmed result revealed that it can be used as ELISA KIT material.Nhằm phục vụ phát triển bộ KIT ELISA phát hiện nhanh độc tố vi tảo domoic acid trong sản phẩm thủy sản tại Việt Nam, kháng thể kháng độc tố này đã được sản xuất bằng liệu pháp miễn dịch trên thỏ trắng New Zealand. Với ưu điểm có khả năng tinh sạch nhanh và cô đặc kháng thể, phương pháp sắc ký ái lực được lựa chọn để tinh chế kháng thể từ huyết thanh thỏ sau miễn dịch nhằm đáp ứng yêu cầu sử dụng làm vật liệu cho KIT ELISA. Tuy nhiên, do domoic acid là một bán kháng nguyên (hapten) nên không thể áp dụng nguyên tắc miễn dịch thông thường trong chế tạo cột sắc ký ái lực miễn dịch sử dụng độc tố này làm giá bắt kháng thể. Bài báo này trình bày kết quả thử nghiệm chế tạo cột sắc ký ái lực miễn dịch sepharose omega-aminohexyl với giá bắt kháng thể là bán kháng nguyên domoic acid để tinh chế kháng thể kháng domoic acid trong huyết thanh thỏ. Sử dụng cầu nối hóa học dissuccinyl substrate để tạo khả năng bắt kháng thể của phân tử domoic acid; sau đó, trên nguyên tắc kết hợp gốc carboxyl của phân tử domoic acid với gốc carbodiimide của hạt sepharose omega-aminohexyl, chúng tôi đã chế tạo thành công cột sắc ký ái lực bắt kháng thể kháng domoic acid với hiệu suất thu hồi đạt 91,5 ± 1,5% (n = 3) khi thử nghiệm với kháng thể chuẩn. Sử dụng cột sắc ký ái lực này với dịch giải cột lần lượt là NaCl 0,5 M; PBS 0,1 M và Ldy-HCl (pH 2,7) 0,1 M; đã thu nhận được kháng thể kháng domoic acid từ huyết thanh thỏ. Western Blot với kháng thể đặc hiệu HRP-labeled anti-goat IgG cho thấy, sản phẩm kháng thể thu nhận được từ quy trình này đạt độ tinh sạch làm vật liệu cho KIT ELISA

Triterpenes and triterpene-glycoside from the leaves of Lawsonia inermis.

From the leaves of  Lawsonia inermis (syn. L. alba), two triterpenes augustic acid (1) and 1b,2a,3a,19a-tetrahydroxy-12-ursen-28-oic acid (2), and a triterpene-glycoside suavissimoside R1 (3) were isolated by using various chromatoghraphies. Their structures were characterized on the basis of the spectroscopic data (1D-NMR, HSQC, HMBC, ESI-MS) in comparison with the literature. This is the first report of 1 - 3 from Lawsonia species. Keywords: Lawsonia inermis, Lythraceae, Triterpene

Low perception of malaria risk among the Ra-glai ethnic minority in south-central Vietnam: implications for forest malaria control

BACKGROUND: Despite Vietnam's success in reducing malaria mortality and morbidity over the last decade, malaria persists in the forested and mountainous areas of the central and southern provinces, where more than 50% of the clinical cases and 90% of severe cases and malaria deaths occur. METHODS: Between July 2005 and September 2006, a multi-method study, triangulating a malariometric cross-sectional survey and qualitative data from focused ethnography, was carried out among the Ra-glai ethnic minority in the hilly forested areas of south-central Vietnam. RESULTS: Despite the relatively high malaria burden among the Ra-glai and their general awareness that mosquitoes can transmit an unspecific kind of fever (84.2%), the use of bed nets, distributed free of charge by the national malaria control programme, remains low at the farmers' forest fields where the malaria risk is the highest. However, to meet work requirements during the labour intensive malaria transmission and rainy season, Ra-glai farmers combine living in government supported villages along the road with a second home or shelter at their slash and burn fields located in the forest. Bed net use was 84.6% in the villages but only 52.9% at the forest fields; 20.6% of the respondents slept unprotected in both places. Such low use may be explained by the low perception of the risk for malaria, decreasing the perceived need to sleep protected. Several reasons may account for this: (1) only 15.6% acknowledged the higher risk of contracting malaria in the forest than in the village; (2) perceived mosquito biting times only partially coincided with Anopheles dirus ss and Anopheles minimus A true biting times; (3) the disease locally identified as 'malaria' was hardly perceived as having an impact on forest farmers' daily lives as they were unaware of the specific kind of fevers from which they had suffered even after being diagnosed with malaria at the health centre (20.9%). CONCLUSIONS: The progressive confinement of malaria to minority groups and settings in the Greater Mekong sub-region implies that further success in malaria control will be linked to research into these specific socio-cultural contexts. Findings highlight the need for context sensitive malaria control policies; not only to reduce the local malaria burden but also to minimize the risk of malaria spreading to other areas where transmission has virtually ceased

Malaria in central Vietnam: analysis of risk factors by multivariate analysis and classification tree models

BACKGROUND: In Central Vietnam, forest malaria remains difficult to control due to the complex interactions between human, vector and environmental factors. METHODS: Prior to a community-based intervention to assess the efficacy of long-lasting insecticidal hammocks, a complete census (18,646 individuals) and a baseline cross-sectional survey for determining malaria prevalence and related risk factors were carried out. Multivariate analysis using survey logistic regression was combined to a classification tree model (CART) to better define the relative importance and inter-relations between the different risk factors. RESULTS: The study population was mostly from the Ra-glai ethnic group (88%), with both low education and socio-economic status and engaged mainly in forest activities (58%). The multivariate analysis confirmed forest activity, bed net use, ethnicity, age and education as risk factors for malaria infections, but could not handle multiple interactions. The CART analysis showed that the most important risk factor for malaria was the wealth category, the wealthiest group being much less infected (8.9%) than the lower and medium wealth category (16.6%). In the former, forest activity and bed net use were the most determinant risk factors for malaria, while in the lower and medium wealth category, insecticide treated nets were most important, although the latter were less protective among Ra-glai people. CONCLUSION: The combination of CART and multivariate analysis constitute a novel analytical approach, providing an accurate and dynamic picture of the main risk factors for malaria infection. Results show that the control of forest malaria remains an extremely complex task that has to address poverty-related risk factors such as education, ethnicity and housing conditions

Long-Lasting Insecticidal Hammocks for controlling forest malaria: a community-based trial in a rural area of central Vietnam.

BACKGROUND: In Vietnam, malaria remains a problem in some remote areas located along its international borders and in the central highlands, partly due to the bionomics of the local vector, mainly found in forested areas and less vulnerable to standard control measures. Long Lasting Insecticidal Hammocks (LLIH), a tailored and user-friendly tool for forest workers, may further contribute in reducing the malaria burden. Their effectiveness was tested in a large community-based intervention trial carried out in Ninh Thuan province in Central Vietnam. METHODS AND FINDINGS: Thirty villages (population 18,646) were assembled in 20 clusters (1,000 individuals per cluster) that were randomly allocated to either the intervention or control group (no LLIH) after stratification according to the pre-intervention P. falciparum antibody prevalence ( or =30%). LLIH were distributed to the intervention group in December 2004. For the following 2 years, the incidence of clinical malaria and the prevalence of infection were determined by passive case detection at community level and by bi-annual malariometric surveys. A 2-fold larger effect on malaria incidence in the intervention as compared to the control group was observed. Similarly, malaria prevalence decreased more substantially in the intervention (1.6-fold greater reduction) than in the control group. Both for incidence and prevalence, a stronger and earlier effect of the intervention was observed in the high endemicity stratum. The number of malaria cases and infections averted by the intervention overall was estimated at 10.5 per 1,000 persons and 5.6/100 individuals, respectively, for the last half of 2006. In the high endemicity stratum, the impact was much higher, i.e. 29/1000 malaria cases and 15.7 infections/100 individuals averted. CONCLUSIONS: LLIH reduced malaria incidence and prevalence in this remote and forested area of Central Vietnam. As the targets of the newly-launched Global Malaria Action Plan include the 75% reduction of the global malaria cases by 2015 and eventually the elimination/eradication of malaria in the long term, LLIH may represent an additional tool for reaching such objectives, particularly in high endemicity areas where standard control tools have a modest impact, such as in remote and forested areas of Southeast Asia and possibly South America. TRIAL REGISTRATION: ClinicalTrials.gov NCT00853281

PRELIMINARY STUDY ON CHEMICAL MECHANISM OF DECREASE OF TETRODOTOXIN CONTENT IN OVARIES OF A PUFFER FISH Lagocephalus inermis (Temminck and Schlegel, 1850) FERMENTED WITH RICE BRAN

Tetrodotoxins in salted and fermented ovaries with rice bran of a puffer fish Lagocephalus inermis (Temminck Schlegel, 1850) during five months were determined by high-performance liquid chromatography with fluorometric detector (HPLC- FLD). The analysed results showed that the original ovaries contained only tetrodotoxin with content of 7.59 µg/g fresh weight. 4,9-Anhydrotetrodotoxin - an isomer which is less toxic than tetrodotoxin has been found in all ovary samples since the second month. Tetrodotoxin in the ovaries disappeared in the 4th month of the experiment. In addition, 4,9-anhydrotetrodotoxin level in ovaries increased during fermentation. The present study contributes to clarifying the chemical mechanism of decreasing tetrodotoxin content in fermented ovary of puffer fish.Tetrodotoxins in salted and fermented ovaries with rice bran of a puffer fish Lagocephalus inermis (Temminck and Schlegel, 1850) during five months were determined by high-performance liquid chromatography with fluorometric detector (HPLC- FLD). The analysed results showed that the original ovaries contained only Tetrodotoxin with content of 7.59 µg/g fresh weight. 4,9-Anhydrotetrodotoxin- an isomer which is less toxic than Tetrodotoxin has been found in all ovary samples since the second month. Tetrodotoxin in the ovaries disappeared in the 4th month of the experiment. In addition, 4,9-Anhydrotetrodotoxin level in ovaries increased during fermentation. The present study contributes to clarify the chemical mechanism of decreasing Tetrodotoxin content in fermented ovary of puffer fish

MỘT SỐ ĐẶC TÍNH CỦA CANXI HYDROXYAPATIT CHIẾT XUẤT TỪ XƯƠNG CÁ NGỪ VẰN Katsuwonus pelamis

This paper is concerned with certain properties of calcium hydroxyapatite from skipjack tuna bone (Katsuwonus pelamis) which are by-products of fish export industry. Hydroxyapatite Ca10(PO4)6(OH)2 and β-tricalcium phosphate Ca3(PO4)2, the high-value compounds, have been successfully extracted from skipjack tuna bones. The bones were heated at different temperatures of 600oC, 900oC, 1200oC. While at 600oC hydroxyapatites were obtained with Ca/P ratio of 1.658, comparable to the value of 1.67 found in human bone; the hydroxyapatite crystals of average size of 0.25 µm were formed with the same size distribution. In case of heated bone samples at 900°C and 1200°C, the calcium formed were biphasic calcium phosphate composed of hydroxyapatite and β-tricalcium phosphate; the Ca/P ratio was between 1.660–1.665; the calcium crystals of more than 1 µm were highly porous and connected to each other in priority orientation of tube direction.Bài báo này trình bày kết quả nghiên cứu về một số đặc tính của canxi hydroxyapatit chiết xuất từ xương cá ngừ Katsuwonus pelamis, một sản phẩm phụ từ ngành xuất khẩu thịt cá ngừ. Các hợp chất có giá trị là hydroxyapatit Ca10(PO4)6(OH)2 và β-tricanxi phosphat β-TCP Ca3(PO4)2 đã được chiết xuất thành công từ xương cá ngừ vằn. Xương cá ngừ được nung ở các nhiệt độ khác nhau 600, 900 và 1.200oC. Dạng canxi thu được khi nung mẫu ở 600oC là hydroxyapatit với tỉ lệ Ca/P 1,658 gần với tỉ lệ Ca/P 1,67 trong xương người; các tinh thể hydroxyapatit có kích thước trung bình 0,25 µm và sự phân bố kích thước tương đối đồng đều. Đối với mẫu xương được xử lý ở 900 và 1.200oC, canxi thu được là hỗn hợp hai pha hydroxyapatit Ca10(PO4)6(OH)2 và β-tricanxi phosphat Ca3(PO4)2; tỉ lệ Ca/P từ 1,660–1,665, các tinh thể canxi có độ xốp cao và liên kết với nhau theo hướng ưu tiên kiểu hình ống với kích thước lớn hơn 1 µm

Cyanide detoxification efficiency of injection and soak of hydroxocobalamin, sodium nitrite and sodium thiosulfate for sea water ornamental fish

The Oceanographic Museum offers interesting exhibits of several marine lives for tourist sightseeing and entertainment. These sea water ornamental fish are all caught in the wild. However, its health can be affected by cyanide poisoning during human fishing. Depending on the level of cyanide poisoning, fish can die after one and two weeks that caused economic damages for the museum. The present study is concerned with results of cyanide detoxification by using direct injection into cinnamon clownfish or soak of hydroxocobalamin, sodium nitrite and sodium thiosulfate with the aim of improving the health, survival and life time for fish, contributing to increasing economic efficiency for the Oceanographic Museum

Rapid decrease of malaria morbidity following the introduction of community-based monitoring in a rural area of central Vietnam

<p>Abstract</p> <p>Background</p> <p>Despite a successful control programme, malaria has not completely disappeared in Vietnam; it remains endemic in remote areas of central Vietnam, where standard control activities seem to be less effective. The evolution of malaria prevalence and incidence over two and half years in a rural area of central Vietnam, after the introduction of community-based monitoring of malaria cases, is presented.</p> <p>Methods</p> <p>After a complete census, six cross-sectional surveys and passive detection of malaria cases (by village and commune health workers using rapid diagnostic tests) were carried out between March 2004 and December 2006 in Ninh-Thuan province, in a population of about 10,000 individuals. The prevalence of malaria infection and the incidence of clinical cases were estimated.</p> <p>Results</p> <p>Malaria prevalence significantly decreased from 13.6% (281/2,068) in December 2004 to 4.0% (80/2,019) in December 2006. <it>Plasmodium falciparum </it>and <it>Plasmodium vivax </it>were the most common infections with few <it>Plasmodium malariae </it>mono-infections and some mixed infections. During the study period, malaria incidence decreased by more than 50%, from 25.7/1,000 population at risk in the second half of 2004 to 12.3/1,000 in the second half of 2006. The incidence showed seasonal variations, with a yearly peak between June and December, except in 2006 when the peak observed in the previous years did not occur.</p> <p>Conclusion</p> <p>Over a 2.5-year follow-up period, malaria prevalence and incidence decreased by more than 70% and 50%, respectively. Possibly, this could be attributed to the setting up of a passive case detection system based on village health workers, indicating that a major impact on the malaria burden can be obtained whenever prompt diagnosis and adequate treatment are available.</p

Social Determinants of Long Lasting Insecticidal Hammock-Use Among the Ra-Glai Ethnic Minority in Vietnam: Implications for Forest Malaria Control

BACKGROUND: Long-lasting insecticidal hammocks (LLIHs) are being evaluated as an additional malaria prevention tool in settings where standard control strategies have a limited impact. This is the case among the Ra-glai ethnic minority communities of Ninh Thuan, one of the forested and mountainous provinces of Central Vietnam where malaria morbidity persist due to the sylvatic nature of the main malaria vector An. dirus and the dependence of the population on the forest for subsistence - as is the case for many impoverished ethnic minorities in Southeast Asia. METHODS: A social science study was carried out ancillary to a community-based cluster randomized trial on the effectiveness of LLIHs to control forest malaria. The social science research strategy consisted of a mixed methods study triangulating qualitative data from focused ethnography and quantitative data collected during a malariometric cross-sectional survey on a random sample of 2,045 study participants. RESULTS: To meet work requirements during the labor intensive malaria transmission and rainy season, Ra-glai slash and burn farmers combine living in government supported villages along the road with a second home at their fields located in the forest. LLIH use was evaluated in both locations. During daytime, LLIH use at village level was reported by 69.3% of all respondents, and in forest fields this was 73.2%. In the evening, 54.1% used the LLIHs in the villages, while at the fields this was 20.7%. At night, LLIH use was minimal, regardless of the location (village 4.4%; forest 6.4%). DISCUSSION: Despite the free distribution of insecticide-treated nets (ITNs) and LLIHs, around half the local population remains largely unprotected when sleeping in their forest plot huts. In order to tackle forest malaria more effectively, control policies should explicitly target forest fields where ethnic minority farmers are more vulnerable to malaria