Certification and stability assessment of recombinant human growth hormone as a certified reference material for protein quantification.

A certified reference material (CRM) for the quantification of protein, essential to manage quality control and quality assurance in protein-related works, has been developed. Amino acid analysis with conventional acid hydrolysis and isotope dilution HPLC-MS was used to establish an SI-traceable absolute protein quantification method using recombinant human growth hormone (hGH) as a model protein. The certification method was verified by comparative studies between 1) different methods of protein quantification based on microwave-assisted hydrolysis, and 2) different labs as part of the Asian Collaboration on Reference Material project with Japan, China, and Korea. Certification, evaluation of measurement uncertainty, homogeneity testing, and stability testing were carried out, after which the candidate CRM for hGH quantification was successfully certified with excellent agreement within the certified value in the two comparative studies. Although the quantification value of hGH by amino acid analysis showed good robustness in various conditions, results of intact protein analysis showed degradation profiles in temperatures higher than 4 °C. Consequently, storage and dissemination conditions should be set in accordance with stability tests. Based on the results, this method is believed to be suitable for accurate quantification of hGH. Additionally, it can also be used as a guide to preparation of CRM, and instructions for quality management of protein work for other similar proteins.

Implementation and assessment of diverse strategies for physical activity promotion in Vietnam-A case report.

BACKGROUND: Regular physical activity (PA) has documented effects in prevention and treatment of many non-communicable diseases (NCDs). Physical inactivity is recognized as the fourth leading risk factor for premature death, worldwide. Despite these facts, physical inactivity is increasing, not only in high-income, but also in middle- and low-income countries. To address this negative trend, evidence-based methods to increase PA are needed. The purpose of this paper is to describe the implementation and assessment of 4 strategies designed to increase PA in Vietnam. METHODS: Four strategies were used: i) introduction and evaluation of an education and training program on the Swedish method of Physical Activity on Prescription (PAP) among health care professionals, ii) translation of the PAP evidence-based handbook, Physical Activity in the Prevention and Treatment of Disease (called FYSS in Swedish) into Vietnamese, iii) launch of a mass-media campaign to promote PA, and iv) advocacy to support development of PA guidelines in Vietnam. RESULTS: The evaluation indicated that the participating health care professionals had a positive attitude to PAP. However, they also reported uncertainty in prescribing PA. FYSS was translated and disseminated successfully to health care professionals. A mass-media campaign identified the beneficial effects of PA to health care professionals, journalists, policy makers, and the public. Last, the process of developing national guidelines on PA was initiated. CONCLUSION: This project led to enhanced awareness and appreciation of PA in the prevention and treatment of NCDs among health care professionals as well as initiation of national PA guidelines. Important lessons also were learned in the presentation of PAP, which will be considered when designing similar projects in the future.

Fully international system of units-traceable glycated hemoglobin quantification using two stages of isotope-dilution high-performance liquid chromatography-tandem mass spectrometry.

Glycated hemoglobin (HbA1c), defined as hemoglobin (Hb) molecules having a stable adduct of glucose on the N-terminal of the ß-chains, has been endorsed as a diagnostic tool for diabetes mellitus and a prediction indicator for the development of diabetes complications. Here we describe an accurate procedure using two stages of isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) for the quantification of HbA1c that provides full traceability to International System of Units (SI). First, synthetic peptides representing specific markers of HbA1c (G-hexa) and hemoglobin A0 (Hexa) were certified by amino acid analysis via acid hydrolysis as reference materials (RMs) for the next step. For this peptide certification, three amino acids (proline, valine, and leucine) were determined by hydrolysis with 10M hydrochloric acid at 130°C for 48h followed by ID-LC-MS/MS. Then, HbA1c content in blood was quantified with the ratio of specific proteolytic peptides from HbA1c and HbA0 via enzyme digestion using ID-LC-MS/MS with the certified peptides as RMs and isotope-labeled peptides as internal standards. Results demonstrate complete traceability to SI-units throughout this procedure. Reliability was confirmed through comparative studies with commercially available RMs for HbA1c, and other routine HbA1c diagnostic methods as well. Following full method validation, we applied this procedure to the certification of candidate hemolysate-certified RMs for HbA1c content, as well as 52 real clinical samples. All of the results showed the suitability of this method to act as a primary reference measurement procedure for HbA1c in complex biological samples.

A reference measurement procedure for amino acids in blood using isotope dilution ultra-performance liquid chromatography-tandem mass spectrometry.

We described a reference measurement procedure for amino acid (AA) quantification in blood samples based on deproteinization with 5-sulfosalicylic acid (SSA) and an isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry (LC-MS) method. The serum was deproteinized with 15% v/v SSA and the supernatant was injected directly into the LC-MS system without further processing. Compared with the use of other precipitants and water as a control, five model AAs-valine, isoleucine, leucine, tyrosine, and phenylalanine-in the SSA-treated samples showed ionization enhancement as well as stable background signals without significant ion suppression effects. Five analytes were clearly separated within 3min using gradient elution and ion-pair chromatography of water and acetonitrile containing 0.1% v/v trifluoroacetic acid. The limit of detection range of this method was 2-52fmol, and the RSDs of accuracy and precision from intra- and inter-day assays were within 2.7%. The method was applied to various blood samples including serum, whole blood and plasma, with no reasonable measurement bias revealed. The quantification accuracy of this method was then assessed using commercially available plasma certified reference material (CRM) for AA, and the results agreed well within certified values. We finally applied this method to the determination of candidate serum CRM. The optimized protocol was found to be suitable for the accurate quantification of five AAs in serum, and may satisfactorily serve as a primary method for AA measurement in various blood matrices.

Life time suicidal thoughts in an urban community in Hanoi, Vietnam.

BACKGROUND: Suicidal thought is a risk factor and a stage in the suicidal process from planning to attempting and dying by suicide. To date, studies on suicidal thought in the general population, especially in Asian communities, have been limited. METHOD: The WHO SUPRE-MISS (the multisite intervention study on suicidal behaviours) community survey questionnaire was filled in for 2,280 randomly selected residents of the DongDa district of Hanoi, Vietnam by means of face-to-face interviews. This multi-factor questionnaire includes such variables as sociodemographic information, suicidal thought and history of suicide attempts, physical health, alcohol consumption and medication. RESULTS: Prevalence rates for life time suicidal thoughts, suicide plans and suicide attempts were 8.9%, 1.1% and 0.4% respectively. Suicidal thoughts are associated with multiple characteristics, such as female gender, single/widowed/separated/divorced marital status, low income, lifestyle (use of alcohol, sedatives and pain relief medication), but not with low education or employment status. Having no religion and being a Buddhist appear to be protective factors for suicidal thought. The ratio of suicidal thoughts, suicide plans and suicide attempts on a lifetime basis is 22.3:2.8:1. CONCLUSION: In Vietnam, as in Western and other Asian countries, suicidal thoughts are associated with similar negative psychosocial risk factors, lifestyle and emotional problems, which implies that suicide preventive measure developed elsewhere can be adjusted to Vietnamese condition. Understanding the unique and common risks in a culture may assist in prediction and control.

Suicide attempts, plans, and ideation in culturally diverse sites: the WHO SUPRE-MISS community survey.

BACKGROUND: The objectives were to assess thoughts about suicide, plans to commit suicide and suicide attempts in the community, to investigate the use of health services following a suicide attempt, and to describe basic socio-cultural indices of the community. METHOD: The community survey was one component of the larger WHO multisite intervention study on suicidal behaviours (SUPRE-MISS). In each site, it aimed at randomly selecting and interviewing at least 500 subjects of the general population living in the catchment area of the emergency department where the intervention component of the study was conducted. Communities of eight SUPRE-MISS sites (in Brazil, China, Estonia, India, Iran, South Africa, Sri Lanka, and Viet Nam) participated plus two additional sites from Australia and Sweden conducting similar surveys. RESULTS: Suicide attempts (0.4-4.2%), plans (1.1-15.6%), and ideation (2.6-25.4%) varied by a factor of 10-14 across sites, but remained mostly within the ranges of previously published data. Depending on the site, the ratios between attempts, plans, and thoughts of suicide differed substantially. Medical attention following a suicide attempt varied between 22% and 88% of the attempts. CONCLUSIONS: The idea of the suicidal process as a continuous and smooth evolution from thoughts to plans and attempts of suicide needs to be further investigated as it seems to be dependent on the cultural setting. There are indications, that the burden of undetected attempted suicide is high in different cultures; an improved response from the health sector on how to identify and support these individuals is needed.

Using the Fluorescence in Situ Hybridization technique for prenatal diagnosis of Down’s syndrome

Fluorescence in Situ Hybridization (FISH) is cytogenetics - molecular technique. FISH is the rapid detection technique of chromosomal aberration by chromosome specific probes applied to interphase cells. In this study, the authors used Kit duple DNA probe for chromosome 13, 21, X, Y to prenatal diagnosis of Down’s syndrome. With 6 amniotic fluid samples, the authors obtained the results as follows: 2 samples were Down’s syndrome trisomy; 1 sample was Patau syndrome trisomy; 3 samples were normal; 1 sample male foetus died without clear causes. Cytogenetic result: 46, XX but FISH result: 46, XY; PCR (Polymerase Chain Reaction) result: TDF (Testis Determining Factor): (+). These results were confirmed by chrosome analysis.

Congenital torticolis treated by physiotherapy: report of a clinical case

An one year old girl with congenital torticolis was subjected to study. At birth, an oval neck mass was persisted at the right side, which made the head tilted. The patient was treated by massage to reduce the contraction of clavicle muscle. Infra-red was irradiated during 10 minutes in the neck and the left side lumbar regions. Exercises were practised to rotate the neck and the trunk, and to stretch the neck. Mother was trained for holding and carrying the child at home. 4 months of treatment gave good results

The relation between physical illness and suicidal thoughts in general population in DongDa district, Hanoi city

2.280 people living in DongDa district, Hanoi city were randomly selected and interviewed concerning the health status, the physical illness the emotional troubles as well as the habit of use of alcohol, sleeping medication, pain medication, and their suicidal thought in whole life. The results showed that there is an association between the suicidal thoughts and cancer, asthma. Anxiety, depression and fear were considered predictors for these thoughts, the habit for using sleeping and pain relief medications were associated statistic significantly with the suicidal thoughts but there was not an association with the use of alcohol.