13 research outputs found
Assessment of individual molecular response in chronic myeloid leukemia patients with atypical BCR-ABL1 fusion transcripts. Recommendations by the EUTOS cooperative network
No full textPurpose: Approximately 1–2% of chronic myeloid leukemia (CML) patients harbor atypical BCR-ABL1 transcripts that cannot be monitored by real-time quantitative PCR (RT-qPCR) using standard methodologies. Within the European Treatment and Outcome Study (EUTOS) for CML we established and validated robust RT-qPCR methods for these patients. Methods: BCR-ABL1 transcripts were amplified and sequenced to characterize the underlying fusion. Residual disease monitoring was carried out by RT-qPCR with specific primers and probes using serial dilutions of appropriate BCR-ABL1 and GUSB plasmid DNA calibrators. Results were expressed as log reduction of the BCR-ABL1/GUSB ratio relative to the patient-specific baseline value and evaluated as an individual molecular response (IMR). Results: In total, 330 blood samples (2–34 per patient, median 8) from 33 CML patients (19 male, median age 62 years) were analyzed. Patients expressed seven different atypical BCR-ABL1 transcripts (e1a2, n = 6; e6a2, n = 1; e8a2, n = 2; e13a3, n = 4; e14a3, n = 6; e13a3/e14a3, n = 2; e19a2, n = 12). Most patients (61%) responded well to TKI therapy and achieved an IMR of at least one log reduction 3 months after diagnosis. Four patients relapsed with a significant increase of BCR-ABL1/GUSB ratios. Conclusions: Characterization of atypical BCR-ABL1 transcripts is essential for adequate patient monitoring and to avoid false-negative results. The results cannot be expressed on the International Scale (IS) and thus the common molecular milestones and guidelines for treatment are difficult to apply. We, therefore, suggest reporting IMR levels in these cases as a time-dependent log reduction of BCR-ABL1 transcript levels compared to baseline prior to therapy.</p
Laboratory recommendations for scoring deep molecular responses following treatment for chronic myeloid leukemia
No full textTreatment of chronic myeloid leukemia (CML) with tyrosine kinase inhibitors has advanced to a stage where many patients achieve very low or undetectable levels of disease. Remarkably, some of these patients remain in sustained remission when treatment is withdrawn, suggesting that they may be at least operationally cured of their disease. Accurate definition of deep molecular responses (MRs) is therefore increasingly important for optimal patient management and comparison of independent data sets. We previously published proposals for broad standardized definitions of MR at different levels of sensitivity. Here we present detailed laboratory recommendations, developed as part of the European Treatment and Outcome Study for CML (EUTOS), to enable testing laboratories to score MR in a reproducible manner for CML patients expressing the most common BCR-ABL1 variants
Raising the level of knowledge of rural population in diabetes mellitus and screening measures
No full textIMSP Centrul de Sănătate FăleștiContext
Oficiul Medicului de Familie (OMF) din s.
Musteața deservește o populație de circa 1300 de
oameni, aici activând un medic de familie, două
asistente medicale, o infirmieră. În OMF sunt create
condiții optime pentru activitate, inclusiv pentru
organizarea întrunirilor cu populația, a intervențiilor
educaționale terapeutice și de promovare a modului
sănătos de viață.
Obiectivul principal al proiectului a fost ridicarea nivelului de cunoștințe privind măsurile de
profilaxie a diabetului zaharat în populația-țintă,
persoane cu vârsta <40 de ani, din localitatea Morozeni, raionul Fălești. La data demarării proiectului
(decembrie 2018), grupul-țintă se estima la 556 de
persoane cu vârsta <40 de ani, dintre care 271 bărbați
și 285 femei. Pe parcursul realizării proiectului au
decedat 8 persoane, s-au aflat peste hotarele republicii 93 persoane, iar 62 au refuzat participarea în
activitățile proiectului.
Desfășurarea proiectului
În cadrul examinării populației din grupul-țintă
din localitatea Musteața, r. Fălești, au fost depistate
41 de persoane cu un nivel mărit al glicemiei în
sânge, dintre care 14 sunt pacienți diagnosticați
anterior și care sunt în evidență cu diabet zaharat, și
27 persoane au fost depistate primar. Anterior examinării, aceștia nu cunoșteau că au nivelul glicemiei
în sânge mărit. Nivelul de cunoștințe al persoanelor
participante privind diabetul zaharat a fost apreciat
prin completarea unui chestionar special elaborat.
Chestionarul conținea întrebări despre: cauzele îmbolnăvirii populației de diabet zaharat, factorii de risc
ce pot provoca îmbolnăvirea, semnificația regimului
alimentar, a activității fizice etc.
S-a constatat că un număr de beneficiari simt
un sentiment de rușine în expunerea cunoștințelor
cu privire la diabetul zaharat, în același timp, toți
participanții în activitățile proiectului au manifestat
interes și dorință de a cunoaște mai multe informații
despre sănătate și problema vizată. Ca urmare, echipa facilitatorilor proiectului a oferit scurte sesiuni de
educație tuturor doritorilor și a asigurat instruirea
verbală prin oferirea broșurii despre diabetul zaharat de tip 2. Primăria locală a fost receptivă și activă
în acordarea ajutorului echipei de implementare a
proiectului, mai ales la mobilizarea populației din
grupul-țintă.
Rezultatele implementării proiectului:
1. Populația, în urma procesului de comunicare
cu medicii de familie, inclusiv cu cei pregătiți în acest
domeniu, și-a îmbunătățit nivelul de cunoștințe cu
privire la problemele diabetului zaharat.
2. Au fost depistate 27 de persoane cu nivelul
glicemiei ridicat, care pe parcurs își vor afla statutul
lor și vor putea beneficia de asistența medicală
necesară.
3. Persoanele depistate cu diabet zaharat vor fi
incluse într-o listă suplimentară, care va fi prezentată
la CNAM, pentru a putea beneficia de glucometre,
teste, lanțete etc.
4. Au crescut nivelul de vigilență și nivelul de
adresabilitate a populației după asistență medicală.
5. A fost stabilită situația reală privind morbiditatea prin diabet zaharat în rândul populației
cu vârsta <40 de ani, ca urmare a examinării lor, în
perioada derulării proiectului, pentru aprecierea
glicemiei în sânge.
6. Acordarea asistenței medicale și sociale
pacienților cu diabet zaharat se va realiza prin evaluarea și monitorizarea managementului de caz.
7. A fost ridicat prestigiul lucrătorilor medicali, în primul rând al medicilor de familie, în fața
populației.
8. S-a ameliorat nivelul de asigurare a infrastructurii cu unele echipamente, dispozitive medicale,
mobilier ș.a.
9. Lucrătorii medicali care au participat la
implementarea proiectului au avut ocazia să-și
îmbunătățească aptitudinile de comunicare cu
populația despre problemele stringente de sănătate
publică.
10. Datorită implementării managementului de
caz al bolnavilor de diabet zaharat, a fost inițiată o
colaborare intersectorială cu alte servicii, cum ar fi
asistența socială, APL, ONG-uri ș.a.
Activitățile planificate pentru perioada următoare
1. Implementarea managementului de caz al
bolnavilor cu diabet zaharat împreună cu Direcția
Asistență Socială a Consiliului raional, cu Primăria
s. Musteața.
2. Pregătirea unei Note informative privind
rezultatele implementării proiectului, care va fi pusă
în discuție la ședința Consiliului teritorial de sănătate
publică
Assessment of individual molecular response in chronic myeloid leukemia patients with atypical BCR-ABL1 fusion transcripts: recommendations by the EUTOS cooperative network
No full textPurpose: Approximately 1–2% of chronic myeloid leukemia (CML) patients harbor atypical BCR-ABL1 transcripts that cannot be monitored by real-time quantitative PCR (RT-qPCR) using standard methodologies. Within the European Treatment and Outcome Study (EUTOS) for CML we established and validated robust RT-qPCR methods for these patients. Methods: BCR-ABL1 transcripts were amplified and sequenced to characterize the underlying fusion. Residual disease monitoring was carried out by RT-qPCR with specific primers and probes using serial dilutions of appropriate BCR-ABL1 and GUSB plasmid DNA calibrators. Results were expressed as log reduction of the BCR-ABL1/GUSB ratio relative to the patient-specific baseline value and evaluated as an individual molecular response (IMR). Results: In total, 330 blood samples (2–34 per patient, median 8) from 33 CML patients (19 male, median age 62 years) were analyzed. Patients expressed seven different atypical BCR-ABL1 transcripts (e1a2, n = 6; e6a2, n = 1; e8a2, n = 2; e13a3, n = 4; e14a3, n = 6; e13a3/e14a3, n = 2; e19a2, n = 12). Most patients (61%) responded well to TKI therapy and achieved an IMR of at least one log reduction 3 months after diagnosis. Four patients relapsed with a significant increase of BCR-ABL1/GUSB ratios. Conclusions: Characterization of atypical BCR-ABL1 transcripts is essential for adequate patient monitoring and to avoid false-negative results. The results cannot be expressed on the International Scale (IS) and thus the common molecular milestones and guidelines for treatment are difficult to apply. We, therefore, suggest reporting IMR levels in these cases as a time-dependent log reduction of BCR-ABL1 transcript levels compared to baseline prior to therapy
beta-thalassemia haplotypes in Romania in the context of genetic mixing in the Mediterranean area
No full textThe purpose of this meta-study was to investigate β-thalassemia (β-thal) mutations and their chromosomal background in order to highlight the origin and spread of thalassemia alleles in the European and Mediterranean areas. Screening of more than 100 new Romanian β-thal alleles was also conducted. The results suggest an ancient introduction of mutations at codon 39 (C > T) (HBB: c.118C > T) and IVS-I-6 (T > C) (HBB: c.92 + 6T > C) in Romania. A comparative study was performed based on restriction fragment length polymorphism (RFLP) haplotypes associated with β-thal mutations in Romania and in Mediterranean countries. Each common β-thal allele from different populations exhibits a high degree of haplotype similarity, a sign of a clear unicentric origin for the IVS-I-110 (G > A) (HBB: c.93-21G > A), IVS-I-6, IVS-II-745 (C > G) (HBB: c.316-106C > G) and codon 39 mutations (the 17a [+ - - - - + +], 13c [ - + + - - - +], 17c [ + - - - - - +] and 14a [- + + - + + + ] ancestral RFLP background, respectively), followed by recurrent recombination events. This study also showed that geographic distances played a major role in shaping the spread of the predominant β-thal alleles, whereas no genetic boundaries were detected between broad groups of populations living in the Middle East, Europe and North Africa. The analyses revealed some discrepancies concerning Morocco and Serbia, which suggest some peculiar genetic flows. Marked variations in β(A) were observed between Southeast Asia and the Mediterranean, whereas a relative genetic homogeneity was found around the Mediterranean Basin. This homogeneity is undoubtedly the result of the high level of specific historic human migrations that occurred in this area
Scanning of beta-globin gene for identification of beta-thalassemia mutation in Romanian population
No full textbeta-Thalassemia is uncommon (0.5%) in the Romanian population, but it must be considered in the differential diagnosis of hypochromic anemia. The molecular characterization of beta-thalassemia is absolutely necessary for molecular diagnosis, as well as any genetic epidemiological study in this region. Molecular analyses consist of mutation detection by molecular scanning of beta-globin gene. This gene has 3 exons and 2 introns, involved in beta-thalassemia pathogenesis. Clinical application of DNA analysis on beta-thalassemic chromosomes allowed characterization of 29 persons with different beta-thalassemia mutations among 58 patients with anemia. The experimental strategy was based on sequential PCR amplification of most of the beta-globin gene and running on denaturing gradient gel electrophoresis of amplification products. Definitive characterization of mutations in samples identified with shifted DGGE patterns was performed ARMS-PCR and/or PCR-restriction enzyme analysis methods. Eight different beta-thalassemia alleles were identified, the most common being IVS I-110 (G-A) and cd 39 (C-T). Comparison of overall frequency of mutations in the neighboring countries, shows that these results are in the frame of overall distribution of these mutations in Mediterranean area, especially in Greece and in Bulgaria. Molecular diagnosis is useful for differentiating mild from severe alleles, for genetic counseling, as well as for mutation definition in carriers, identified by hematological analysis necessary for prenatal testing and genetic counseling
Scanning of ?-globin gene for identification of ?-thalassemia mutation in Romanian population
No full textVanadium(V) Complexes with Substituted 1,5-bis(2-hydroxybenzaldehyde)carbohydrazones and Their Use As Catalyst Precursors in Oxidation of Cyclohexane
No full textSix dinuclear vanadium(V) complexes have been synthesized: NH4[(VO2)2(HLH)] (NH4[1]), NH4[(VO2)2(t‑BuLH)] (NH4[2]), NH4[(VO2)2(ClLH)]
(NH4[3]), [(VO2)(VO)(HLH)(CH3O)] (4), [(VO2)(VO)(t‑BuLH)(C2H5O)]
(5), and [(VO2)(VO)(ClLH)(CH3O)(CH3OH/H2O)] (6) (where HLH4 = 1,5-bis(2-hydroxybenzaldehyde)carbohydrazone, t‑BuLH4 = 1,5-bis(3,5-di-tert-butyl-2-hydroxybenzaldehyde)carbohydrazone, and ClLH4 = 1,5-bis(3,5-dichloro-2-hydroxybenzaldehyde)carbohydrazone).
The structures of NH4[1] and 4–6 have been determined by X-ray diffraction
(XRD) analysis. In all complexes, the triply deprotonated ligand accommodates
two V ions, using two different binding sites ONN and ONO separated
by a diazine unit −N–N–. In two pockets of NH4[1], two identical VO2+ entities are present, whereas, in those of 4–6, two different VO2+ and VO3+ are bound. The highest oxidation state of V ions was corroborated
by X-ray data, indicating the presence of alkoxido ligand bound to
VO3+ in 4–6, charge density
measurements on 4, magnetic susceptibility, NMR spectroscopy,
spectroelectrochemistry, and density functional theory (DFT) calculations.
All four complexes characterized by XRD form dimeric associates in
the solid state, which, however, do not remain intact in solution.
Compounds NH4[1], NH4[2], and 4–6 were applied as alternative
selective homogeneous catalysts for the industrially significant oxidation
of cyclohexane to cyclohexanol and cyclohexanone. The peroxidative
(with tert-butyl hydroperoxide, TBHP) oxidation of
cyclohexane was performed under solvent-free and additive-free conditions
and under low-power microwave (MW) irradiation. Cyclohexanol and cyclohexanone
were the only products obtained (high selectivity), after 1.5 h of
MW irradiation. Theoretical calculations suggest a key mechanistic
role played by the carbohydrazone ligand, which can undergo reduction,
instead of the metal itself, to form an active reduced form of the
catalyst
A certified plasmid reference material for the standardisation of BCR-ABL1 mRNA quantification by real time quantitative PCR
No full textSerial quantification of BCR–ABL1 mRNA is an important therapeutic indicator in chronic myeloid leukaemia, but there is a substantial variation in results reported by different laboratories. To improve comparability, an internationally accepted plasmid certified reference material (CRM) was developed according to ISO Guide 34:2009. Fragments of BCR–ABL1 (e14a2 mRNA fusion), BCR and GUSB transcripts were amplified and cloned into pUC18 to yield plasmid pIRMM0099. Six different linearised plasmid solutions were produced with the following copy number concentrations, assigned by digital PCR, and expanded uncertainties: 1.08±0.13 × 106, 1.08±0.11 × 105, 1.03±0.10 × 104, 1.02±0.09 × 103, 1.04±0.10 × 102 and 10.0±1.5 copies/?l. The certification of the material for the number of specific DNA fragments per plasmid, copy number concentration of the plasmid solutions and the assessment of inter-unit heterogeneity and stability were performed according to ISO Guide 35:2006. Two suitability studies performed by 63 BCR–ABL1 testing laboratories demonstrated that this set of 6 plasmid CRMs can help to standardise a number of measured transcripts of e14a2 BCR–ABL1 and three control genes (ABL1, BCR and GUSB). The set of six plasmid CRMs is distributed worldwide by the Institute for Reference Materials and Measurements (Belgium) and its authorised distributors (https://ec.europa.eu/jrc/en/reference-materials/catalogue/; CRM code ERM-AD623a-f)
