scholarly journals Application of heteroduplex analysis for CALR mutation screening detection in patients with Ph-myeloproliferative neoplasms

2021 ◽  
Vol 16 (2) ◽  
pp. 48-55
Author(s):  
T. N. Subbotina ◽  
D. V. Kurochkin ◽  
I. E. Maslyukova ◽  
A. S. Khazieva ◽  
E. V. Vasiliev ◽  
...  
Keyword(s):  
Screening Test ◽  
Mutant Allele ◽  
Myeloproliferative Neoplasms ◽  
Mutation Screening ◽  
World Health ◽  
Heteroduplex Analysis ◽  
Pcr Product ◽  
Exon 9 ◽  
Health Organization

Background. In accordance with the World health organization clinical guidelines, the analysis of somatic mutations in the CALR gene, as well as mutations in the JAK2 and MPL genes, are included in the list of criteria for the Ph-myeloproliferative neoplasms diagnosis.More than 50 different mutation variants have been found in the CALR gene, among which the most frequent are a 52 bp deletion (c.1092_1143del), also called type 1, and a 5 bp insertion (c.1154_1155insTTGTC), also called type 2 (88 %).The remaining 12 % are other type less frequent indels or combinations thereof.It is most convenient to use sequencing methods to identify all possible variants of CALR mutations. It is also important to develop inexpensive screening test that can detect any mutations in the analyzed DNA fragment of CALR gene. This method can be heteroduplex analysis followed by electrophoresis on polyacrylamide gel (PAGE).The objective: to develop and demonstrate the feasibility of using heteroduplex analysis with separation of the PCR product by electrophoresis on non-denaturing PAGE for the CALR exon 9 mutations detection as the screening test. Materials and methods. DNA samples of 13 CALR-positive patients with different phenotypic variants of Ph-myeloproliferative neoplasms were screened by heteroduplex analysis. For the most common variants of CALR mutations (c.1092_1143del and c.1154_1155insTTGTC), a threshold determination of the mutant allele presence was analyzed.Nucleotide sequence of exon 9 fragment was determined using Sanger sequencing. Also, all 13 samples were analyzed using the pyrosequencing method to assess the allelic burden level.Results. Heteroduplex analysis revealed mutations in exon 9 of the CALR gene in all 13 patients. The threshold determinations of the method in the case of the c.1154_1155insTTGTC and c.1092_1143del analysis are 6.25 % and 3.13 % of the mutant allele presence in the patient sample, respectively.Conclusion. The proposed variant of the heteroduplex analysis with separation of the PCR product by electrophoresis on non-denaturing PAGE can be recommended for use as the preliminary screening test which is carried out before the confirming sequencing methods for the different indels (or combinations thereof) CALR mutations determine.The presence of heteroduplexes indicates the presence of a mutation, even if the mutant product is not visualized (in case of small mutations).

scholarly journals A Rapid, Sensitive and Specific Method for Quantifying Calr Mutant Allele Burden in Persons with Myeloproliferative Neoplasms

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 5210-5210
Author(s):  
Jiao Zhou ◽  
Qiumei Yao ◽  
Robert Peter Gale ◽  
Jinlan Li ◽  
Lingdi Li ◽  
...  
Keyword(s):  
Mutant Allele ◽  
Myeloproliferative Neoplasms ◽  
Mutation Screening ◽  
Heterozygous Mutation ◽  
Specific Method ◽  
Wild Type ◽  
Allele Burden ◽  
Exon 9 ◽  
Calr Mutations ◽  
Calr Mutation

Abstract Background: CALR mutations were recently identified in a substantial proportion of persons with essential thrombocythemia (ET) and with primary myelofibrosis (PMF) without JAK2V617F. Consequently rapid, sensitive and specific methods to detect and quantify these mutations are needed. Methods: We studied samples from 1088 persons with myeloproliferative neoplasms (MPNs) including 421 JAK2V617F negative subjects with ET, PMF, polycythemia vera (PV), chronic myeloid leukemia (CML) and hyper-eosinophilic syndrome (HES). Detection of CALR exon 9 mutations was done by PCR amplification followed by fragment length analysis and direct sequencing. Dilution assays were used to determine CALR mutant allele burden. Results: We detected CALR mutations in blood and bone marrow samples from 152 subjects with ET and with PMF but not in samples from normal or persons with PV, CML or HES. CALR mutant peaks were distinct from wild-type peaks and dilution experiments indicated a sensitivity level of 0.5-5% for a CALR mutant allele in a wild-type background. Diverse types of mutations were detected including deletions, insertions and complex indels. All mutations were confirmed by direct sequencing. We also used dilution experiments to quantify mutant allele burden. We were able to reproducibly detect mutant allele levels as low 5% (0.5-5%) in a wild-type background. Conclusions: PCR amplification followed by fragment length analysis is a rapid, sensitive and specific method for screening persons with MPNs for CALR mutations, especially those with ET and PV with JAK2V617F and for estimating mutant allele burden. Figure 1. Standard curve for the detection of mutant allele burden. Figure 1. Standard curve for the detection of mutant allele burden. Figure 2. Titration analyses of sensitivity of CALR mutation screening by sequencing and fragment analyses. Figure 2. Titration analyses of sensitivity of CALR mutation screening by sequencing and fragment analyses. Figure 3. Figure 3 Sequencing traces show heterozygous mutation of CALR. Gene scan electropherogram from PCR method and partial sequence of CALR exon 9 from sequencing method (numbering according to GenBank access number: NC_000019.9). A-P: Detected a wild type and 15 CALR mutation types by sequencing and fragment analysis methods. A: wild type, B-I: deletions, H-L: insertions, M-P: complex indels. Figure 3. Figure 3 Sequencing traces show heterozygous mutation of CALR. Gene scan electropherogram from PCR method and partial sequence of CALR exon 9 from sequencing method (numbering according to GenBank access number: NC_000019.9). A-P: Detected a wild type and 15 CALR mutation types by sequencing and fragment analysis methods. A: wild type, B-I: deletions, H-L: insertions, M-P: complex indels. Disclosures No relevant conflicts of interest to declare.

scholarly journals A review of current knowledge of myeloproliferative disorders in the horse

2021 ◽  
Vol 63 (1) ◽  
Author(s):  
Katy Satué ◽  
Juan Carlos Gardon ◽  
Ana Muñoz
Keyword(s):  
Bone Marrow ◽  
Myeloid Leukemia ◽  
Current Knowledge ◽  
Myeloproliferative Neoplasms ◽  
Myeloproliferative Disorders ◽  
World Health ◽  
Chronic Granulocytic Leukemia ◽  
Current State ◽  
Multiple Cell ◽  
Health Organization

AbstractMyeloid disorders are conditions being characterized by abnormal proliferation and development of myeloid lineage including granulocytes (neutrophils, eosinophils and basophils), monocytes, erythroids, and megakaryocytes precursor cells. Myeloid leukemia, based on clinical presentation and proliferative rate of neoplastic cells, is divided into acute (AML) and myeloproliferative neoplasms (MPN). The most commonly myeloid leukemia reported in horses are AML-M4 (myelomonocytic) and AML-M5 (monocytic). Isolated cases of AML-M6B (acute erythroid leukemia), and chronic granulocytic leukemia have also been reported. Additionally, bone marrow disorders with dysplastic alterations and ineffective hematopoiesis affecting single or multiple cell lineages or myelodysplastic diseases (MDS), have also been reported in horses. MDSs have increased myeloblasts numbers in blood or bone marrow, although less than 20%, which is the minimum level required for diagnosis of AML. This review performed a detailed description of the current state of knowlegde of the myeloproliferative disorders in horses following the criteria established by the World Health Organization.

scholarly journals Study of Well Waters from High-Level Natural Radiation Areas in Northern Vietnam

2021 ◽  
Vol 18 (2) ◽  
pp. 469
Author(s):  
Van-Hao Duong ◽  
Thanh-Duong Nguyen ◽  
Miklos Hegedus ◽  
Erika Kocsis ◽  
Tibor Kovacs
Keyword(s):  
Natural Radionuclide ◽  
World Health ◽  
Natural Radiation ◽  
Lifetime Cancer Risk ◽  
Northern Vietnam ◽  
Health Organization ◽  
High Level ◽  
Radiological Hazard Indices ◽  
Well Waters

The determination of natural radionuclide concentrations plays an important role for assuring public health and in the estimation of the radiological hazards. This is especially true for high level radiation areas. In this study, 226Ra, 228Ra and 238U concentrations were measured in well waters surrounding eight of the high-level natural radiation areas in northern Vietnam. The 226Ra, 228Ra and 238U activity concentrations vary from <1.2 × 10−3–2.7 (0.46), <2.6 × 10−3–0.43 (0.07) and <38 × 10−3–5.32 Bq/L (0.50 of median), respectively. 226Ra and 238U isotopes in most areas are in equilibrium, except for the DT-Thai Nguyen area. The calculated radiological hazard indices are generally higher than WHO (World Health Organization) recommendations. Average annual effective dose and excess lifetime cancer risk values due to drinking well water range from to 130 to 540 μSv/year and 7.4 × 10−6 to 3.1 × 10−5, respectively.

scholarly journals HCV Detection, Discrimination, and Genotyping Technologies

Sensors ◽  
2018 ◽  
Vol 18 (10) ◽  
pp. 3423 ◽  
Author(s):  
Shrikant Warkad ◽  
Satish Nimse ◽  
Keum-Soo Song ◽  
Taisun Kim
Keyword(s):  
Hepatitis C ◽  
Hcv Infection ◽  
World Health ◽  
Clinical Samples ◽  
Infected People ◽  
Advantages And Disadvantages ◽  
Highly Sensitive ◽  
The World ◽  
Health Organization

According to the World Health Organization (WHO), 71 million people were living with Hepatitis C virus (HCV) infection worldwide in 2015. Each year, about 399,000 HCV-infected people succumb to cirrhosis, hepatocellular carcinoma, and liver failure. Therefore, screening of HCV infection with simple, rapid, but highly sensitive and specific methods can help to curb the global burden on HCV healthcare. Apart from the determination of viral load/viral clearance, the identification of specific HCV genotype is also critical for successful treatment of hepatitis C. This critical review focuses on the technologies used for the detection, discrimination, and genotyping of HCV in clinical samples. This article also focuses on advantages and disadvantages of the reported methods used for HCV detection, quantification, and genotyping.

scholarly journals LEVELS OF PERSONAL MATURITY AMONG THE STUDENTS OF HUMANITARIAN UNIVERSITY

2020 ◽  
Author(s):  
Valentyn Savchenko ◽  
Нalyna Kharchenko ◽  
Olga Buriak ◽  
Iryna Omeri ◽  
Jevgenija Nevedomsjka ◽  
...  
Keyword(s):  
University Students ◽  
Religious Beliefs ◽  
World Health ◽  
Religious Traditions ◽  
Internal Development ◽  
Inner World ◽  
Health Organization ◽  
Senior Students ◽  
Junior Students

Introduction. Maturity of personality, their inner world plays a substantial role in health development. For a reason, following the definition by the World Health Organization spirituality is one of human health components. Still, the level of personal maturity of an adult does not always comply with their age and can be lower. Aim is to determine the levels of personal maturity (internal development) of the humanitarian university students and to identify the peculiarities of their distribution depending on gender, duration of studying, specialty, confession of faith, and attitude to religion. Material and methods. 207 students of Borys Grinchenko Kyiv University, average age of the surveyed 19.46 ± 2.12. There were 105 males (50.7%), and 102 females (49.3%). For determination of the levels of personal maturity the questionnaire by Jose Stevens was implemented. There were 5 levels specificated: «baby», «junior», «teen», «young man/woman», and «adult». Results. Among the humanitarian university students of the specialties «physical therapy, ergotherapy» and «physical education and sports», in 62.8% (130/207) cases high levels of personal maturity were identified, in 32.4% (67/207) cases there were low levels. Among one fifth of the students (19.8%; 41/207) the combinations of several levels of personal maturity were identified. It was indicative of the youth’s failure to having an integral idea of their inner world and spirituality. The female students were more internally developed than the male ones, which was indicative of statistically substantial predominance of high levels of personal maturity among them — 66.7%; 68/102 (among the males — 48.6%; 51/105). Ideas of personal maturity among the senior students (the 4th and 5th years of studies) were more accomplished than the ones of the junior students (the 1st and 2nd years of studies). Following religious traditions both by the holders of religious beliefs and not, led to development of higher levels of personal maturity compared to those students not following religious traditions. Conclusions. One third of the humanitarian university students have low level of personal maturity (internal development). According to the level of personal maturity, females prevail over males. The students following religious traditions (the holders of religious beliefs and not) are more internally developed than those not following such traditions.

scholarly journals HEAVY METAL CONTENT IN SOME SEAFOODS COLLECTED FROM MUARA ANGKE FISH AUCTION PLACE, JAKARTA.

1987 ◽  
Vol 26 ◽  
pp. 51-58
Author(s):  
Horas Hutagalung ◽  
Suwirma Syamsu
Keyword(s):  
Heavy Metals ◽  
Heavy Metal ◽  
Metal Content ◽  
Heavy Metal Content ◽  
World Health ◽  
Human Consumption ◽  
Zn Content ◽  
Standard Set ◽  
Health Organization

Determination of  heavy  metal  content  (Hg, Pb, Cd, Cu, Cr, Zn) in some seafoods collected from  Muara  Angke  Fish  Auction  Place  were carried out during February to July 1979. The results showed  that Zn  content  in seafoods a nalyzed  was always much higher than the other heavy metals. The concentration levels of  the  six  heavy  metals observed were low. It is almost of the same level as the metal concentration of  other  seafoods  observed  in various parts of the world oceans, but still lower  than the maximum standard set  for human  consumption  as  established  by   Tasmania, Australia and  World Health Organization. The  mean  per capita  consumption of  fish  in Jakarta is only  about 165 g  per week, while  the  Provisional Tolerated Weekly Intake for Hg, set by WHO/FAO at 300 ug level, would be attained by consuming 1500 g Rastrelliger sp., or 698 gPenaeus setiferus, or 545 g Sepia sp., and or 1250 g Pecten sp.  Therefore,  at   present the  level of metal content in fishes sold at TPI  Muara  Angke  can be  considered  as  not  dangerous  to human health as long as these quantities are not surpassed.

Haematopathology of classic myeloproliferative neoplasms

2020 ◽  
pp. 45-62
Author(s):  
Hans Michael Kvasnicka ◽  
Jürgen Thiele
Keyword(s):  
Continuous Improvement ◽  
Who Classification ◽  
Myeloproliferative Neoplasms ◽  
Primary Myelofibrosis ◽  
World Health ◽  
Polycythaemia Vera ◽  
The World ◽  
Health Organization ◽  
Diagnostic Importance

The classification of the World Health Organization (WHO) continues to advocate the diagnostic importance of bone marrow (BM) morphology in the diagnostic workup of myeloproliferative neoplasms (MPN). In this regard, distinctive histological BM patterns characterize specific subtypes of MPN and are the key to a meaningful clinical and molecular-defined risk stratification of patients. In this regard, the morphological denominator includes a characteristic megakaryocytic proliferation along with variable changes in the granulopoiesis and erythropoiesis. Importantly, diagnosis of MPN requires absence of relevant dysgranulopoiesis or dyserythropoiesis. In terms of clinical practice, the concept of precursor stages provides the possibility of an early intervention by appropriate therapeutic regimens that might prevent fatal complications like thrombosis and haemorrhage, especially in early stages of polycythaemia vera or in primary myelofibrosis. However, the WHO classification is not aimed to capture all biological true cases of MPN or guarantee a complete diagnostic specificity and thus might be in need of continuous improvement following clinical experience.

Participation in the World Health Organization

2020 ◽  
Vol 17 (1) ◽  
pp. 261-290 ◽  
Author(s):  
Steven A. Solomon ◽  
Claudia Nannini
Keyword(s):  
Public Health ◽  
Global Health ◽  
World Health Organization ◽  
Regional Level ◽  
World Health ◽  
Global Public Health ◽  
The World ◽  
Health Organization ◽  
Collaborative Efforts

Participation in the World Health Organization (WHO) is a multifaceted matter and should be understood as not only referring to the governance of WHO, but also to its scientific and technical work as well as its collaborative efforts towards advancing global public health more generally. The article is concerned, in particular, with the legal and political framework surrounding attendance and participation of states and various entities in the governing bodies of the Organization, at the global and regional level. It shows that participation in the governance of WHO is still today a domain reserved to the determination of its Member States. At the same time, solutions have been found and continued efforts are necessary to take into account geopolitical considerations and to ensure a meaningful and inclusive participation of all relevant actors in global health discussions.

Determination of Heavy Metals in Damascus Drinking Water Using Total Reflection X-Ray Fluorescence

1999 ◽  
Vol 34 (2) ◽  
pp. 305-316 ◽  
Author(s):  
E.H. Bakraji ◽  
J. Karajo
Keyword(s):  
Trace Elements ◽  
Drinking Water ◽  
Total Reflection ◽  
World Health ◽  
X Ray ◽  
Quality Guidelines ◽  
The World ◽  
The Mean ◽  
Health Organization

Abstract Total reflection X-ray fluorescence spectrometry and chemical preconcentration have been applied for multi-elemental analysis of Damascus drinking water. Water was taken directly from taps of several city sectors and analyzed for the following trace elements: Ti, V, Cr, Fe, Co, Ni, Cu, Zn, Se and Pb. The detection limits were found to be in the range of 0.1 to 0.4 µg/L. The mean levels of trace elements in the Damascus drinking water were below the World Health Organization drinking water quality guidelines.

scholarly journals Comparison of Western Immunobloting to an Enzyme-Linked Immunosorbent Assay for the Determination of Anti-Bordetella pertussis Antibodies

2011 ◽  
Vol 18 (4) ◽  
pp. 615-620 ◽  
Author(s):  
Stephen D. Merrigan ◽  
Ryan J. Welch ◽  
Christine M. Litwin
Keyword(s):  
Immunosorbent Assay ◽  
Bordetella Pertussis ◽  
World Health ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recent Infection ◽  
Content Type ◽  
Igm Elisa ◽  
Iga Antibodies ◽  
Health Organization

ABSTRACTDuringBordetella pertussisinfection, it has been established that an increase of anti-pertussis toxin (PT) and anti-filamentous hemagglutinin (FHA) antibodies occurs. Immunoblots from two manufacturers using FHA and PT antigens were compared with an enzyme-linked immunosorbent assay (ELISA) that used both FHA and PT. One manufacturer used two concentrations of PT bands for the IgG immunoblot, calibrated to the World Health Organization standard for PT in international units (IU/ml), 100 IU/ml (PT-100) and 8 IU/ml (PT). The second immunoblot kit measured antibodies to a single calibrated PT band. Both kits measured IgA antibodies, and one additionally measured IgM antibodies. Two of 41 (5%) ELISA IgM positives were confirmed positive by IgM immunoblotting, suggesting poor specificity of the IgM ELISA. The agreements of the IgG and IgA immunoblots with the ELISA ranged from 72.5% to 85.3%, with only 38 to 51% of IgA positives confirmed by immunoblotting and only 61 to 68% of IgG positives confirmed by immunoblotting. The two immunoblots correlated well with each other, with 91.7% and 94.3% agreement for IgG and IgA, respectively. When the FHA band was used with the PT band as the criterion for positivity, significant differences existed in specificity compared to the ELISA (IgG, 84.1% versus 33.3%; IgA, 82.4% versus 71.0%). When the positive IgA immunoblots (evidence of natural recent infection) were compared to the positive PT-100 IgG immunoblots (evidence of recent infection or vaccination), the PT-100 blot showed a 71% sensitivity in detecting natural recent infection.B. pertussisimmunoblots, alone or in combination with ELISAs, can aid in the diagnosis ofB. pertussisinfection.

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