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Dettagli della rivista
Formato
Rivista
eISSN
2284-5623
Pubblicato per la prima volta
08 Aug 2013
Periodo di pubblicazione
4 volte all'anno
Lingue
Inglese

Cerca

Volume 28 (2020): Edizione 3 (July 2020)

Dettagli della rivista
Formato
Rivista
eISSN
2284-5623
Pubblicato per la prima volta
08 Aug 2013
Periodo di pubblicazione
4 volte all'anno
Lingue
Inglese

Cerca

10 Articoli
Accesso libero

Metabolomic biomarkers of polycystic ovary syndrome related-obesity: a review of the literature

Pubblicato online: 27 Jul 2020
Pagine: 241 - 255

Astratto

Abstract

Background and objectives: Polycystic ovary syndrome (PCOS) displays a phenotype-dependent cardio-metabolic risk. By performing a systematic search of the literature, we aimed to summarize metabolomic signatures associated with obesity in PCOS women.

Data sources and study eligibility criteria: We conducted a comprehensive search including: Embase, PubMed, and Web of Science until 31st of May 2019. We used the terms: metabolomics and polycystic ovary syndrome. We excluded the following papers: animal studies, studies that included only lean PCOS women, reviews, meta-analyses, results of interventional studies, those that did not apply metabolomic techniques.

Results: The lipid signature in obese women with PCOS showed increased levels of free fatty acids (carnitine, adipic acid, linoleic acid, oleic acid) and lower levels of lysophosphatidylcholines and glycerolphosphocholine compared with non-obese PCOS women. Regarding carbohydrate metabolism, a decrease in citric and lactic acid levels characterized obese PCOS women. Decreased lactic acid in obese PCOS women suggests augmented insulin stimulated glucose muscle use in lean, but not in obese women. Considering amino acid metabolomic markers, valine, glycine, serine, threonine, isoleucine and lysine were higher in obese PCOS women. Patients with visceral obesity presented a diminished uptake of essential amino acids, BCAA, leucine and serine in the skeletal muscle. α-ketoglutarate was significantly higher in obese women with PCOS in comparison with lean women with PCOS, distinguishing these 2 subgroups of PCOS with high ‘predictive accuracy’.

Limitations: Overall, a small number of studies have focused on the impact of obesity on the metabolic fingerprints of PCOS women. There is need for properly controlled, high-quality studies.

Conclusions: There is compelling evidence of significant alterations in carbohydrate, lipid, and amino acid metabolism in women with PCOS and obesity. Metabolomics may identify new metabolic pathways involved in PCOS and improve our understanding of the complex relation between PCOS and obesity in order to personalize PCOS therapy.

Parole chiave

  • untargeted metabolomics
  • polycystic ovary syndrome
  • obesity
  • biomarkers
  • targeted metabolomics
Accesso libero

High frequency of BRCA recurrent mutations in a consecutive series of unselected ovarian cancer patients

Pubblicato online: 27 Jul 2020
Pagine: 257 - 266

Astratto

Abstract

Hereditary predisposition to breast and ovarian cancer (HBOC) is diagnosed by molecular analysis of deleterious mutations in BRCA genes, allowing oncogenetic follow-up of patients and of their families. BRCA testing addresses only to HBOC families, using restrictive inclusion criteria based on familial history of cancer and age at diagnosis. Sporadic ovarian cancer has high incidence and mortality in Romania, with low median age of diagnosis and possibly a higher magnitude of hereditary contribution comparing to othe populations. However, sporadic ovarian cancers do not qualify for BRCA testing according to inclusion criteria, and a complete BRCA screening of all cancers is neither feasible nor recommended. Despite the large diversity of BRCA mutations worldwide, some recurrent mutations have higher frequencies in diverse populations. Precisely screening for recurrent mutations in a target population allows to rapidly identifying mutation carriers without sequencing the entire BRCA genes. In Romanian population and neighboring countries, several recurrent mutations have already been described. In a consecutive series of 50 sporadic ovarian cancer patients, not qualifying for BRCA complete testing, we screened for 9 most common BRCA mutations, by multiplex-PCR, RFLP and targeted Sanger sequencing. Our results revealed 6 different BRCA mutations in 8 unrelated patients, with a frequency of 16%, much higher than expected. We further recommend screening for the identified mutations in larger series of cancer patients. The results are highly beneficial to cancer patients, healthy relatives, and overall, considering prevention in cancer a priority, to public health system and future of oncogenetics in Romania

Parole chiave

  • ovarian cancer
  • hereditary predisposition
  • BRCA genes
  • recurrent mutations
Accesso libero

SH2B3 (LNK) rs3184504 polymorphism is correlated with JAK2 V617F-positive myeloproliferative neoplasms

Pubblicato online: 27 Jul 2020
Pagine: 267 - 277

Astratto

Abstract

Background: Pathogenesis and phenotypic diversity in myeloproliferative neoplasms (MPN) cannot be fully explained by the currently known acquired mutations alone. Some susceptible germline variants of different genes have been proved to be associated with the development of these diseases. The goal of our study was to evaluate the association between the rs3184504 polymorphism of SH2B3 (LNK) gene (p.R262W, c.784T>C) and the risk of developing the four typical MPN - polycythemia vera (PV), essential thrombocythemia (ET), primary myelofibrosis (PMF), and chronic myeloid leukemia (CML).

Material and methods: We investigated the SH2B3 rs3184504 T>C polymorphism by real-time PCR in 1901 MPN patients (575 with PV, 798 with ET, 251 with PMF, and 277 with CML), all of them harboring one of the specific driver mutations - JAK2 V617F or CALR in case of PV, ET and PMF, or BCR-ABL1 in case of CML, and 359 controls.

Results: Overall, the TT homozygous genotype was significantly associated with BCR-ABL1-negative MPN (OR = 1.34; 95% CI = 1.03-1.74; crude p-value = 0.02; adjusted p-value = 0.04). The most significant association was seen in case of PV (OR = 1.54; 95% CI = 1.14-2.06; crude p-value = 0.004; adjusted p-value = 0.024). Also, SH2B3 rs3184504 correlated significantly with JAK2 V617F-positive MPN (OR = 1.36; 95% CI = 1.04-1.77; crude p-value = 0.02; adjusted p-value = 0.08), but not with those CALR-positive. ET (regardless of molecular subtype) and CML were not correlated with SH2B3 rs3184504.

Conclusions: The SH2B3 rs3184504 polymorphism is associated with risk of MPN development, especially PV. This effect is restricted to JAK2 V617F-positive PV and PMF only.

Parole chiave

  • myeloproliferative neoplasms
  • somatic mutations
  • genetic predisposition
Accesso libero

Mucopolysaccharidosis type I - Clinical and genetic characteristics of Romanian patients

Pubblicato online: 27 Jul 2020
Pagine: 279 - 286

Astratto

Abstract

Background: Mucopolysaccharidosis type I (MPS I) is an autosomal recessive lysosomal storage disorder caused by a deficiency of α-L-iduronidase (IDUA), which leads to the accumulation of partially digested glycosaminoglycans (dermatan sulfate and heparan sulfate) in the lysosomes and induces multisystemic alteration. Hurler (severe), Scheie (mild), and Hurler/Scheie (intermediate) syndromes are clinical subtypes of MPS-I. To date, more than 290 IDUA mutations have been reported. The purpose of this study was to present the clinical and genetic characteristics of Romanian MPS I syndrome patients and their genotype-phenotype correlation.

Patients and methods: Seven patients (5 girls and 2 boys) with MPS type I, belonging to 4 unrelated families, aged 0,75-17.9 years, were enrolled. The study methods consisted in: clinical and standard auxological assessment, bone radiographs, joint ultrasonography, goniometry, neurological and psychological evaluation, hepatic and splenic ultrasonography, cardiological evaluation, otorhinolaryngology examination, ophthalmological examination, spirometry, α-L-iduronidase enzyme activity assay and molecular analysis.

Results: The seven patients originated from 4 unrelated families, three patients with severe, two patients with intermediate and two with attenuated clinical phenotype. Each patient presented the classical picture of MPS type I picture, represented by: variable coarse facial features, arthropathy, hepatosplenomegaly, cardiac involvement, respiratory dysfunction and neurological impairment. Five patological variants, three point mutations (p.Q70 *, p.I238Q and p.K324R), two deletion c.1045_1047delGAC, c.46_57delTCGCTCCTG) and an insertion (c.1389 insC) were identified in both alleles of the ADUA gene in homozygous or heterozygous form. Two novel mutations (p.K324R and c.1389 insC) were reported. The p.Q70*(c.208C>T) variant was identified in 2 families with severe form of disease (Hurler syndrome) in homozygous status in one family and in compound heterozygous status in the other family.

Conclusion: The p.Q70* missense variant was the most frequent, correlated in all the cases who presented it with severe form, Hurler syndrome, the other mutations being usually isolated and particular for each patient, associated in our patients with less severe MPS I phenotype, as Hurler-Scheie or Scheie syndrome. The results of this study indicated the mutational heterogeneity of the IDUA gene and the difficulty to indicate some correlation between the genotype and phenotype in MPS I patients.

Parole chiave

  • MPS I
  • IDUA
  • mutations
Accesso libero

Comparative Evaluation of RNAlater Solution and Snap Frozen Methods for Gene Expression Studies in Different Tissues

Pubblicato online: 27 Jul 2020
Pagine: 287 - 297

Astratto

Abstract

Introduction: Freezing of tissues with liquid nitrogen is the most common method in studies performed at the RNA level. However, the use of RNA stabilization solutions has become a popular alternative method. The aim of this study is to investigate the effectiveness of RNAlater on RNA stabilization in different tissues.

Material and Methods: In this study, RNA were isolated from the lung, heart, liver and skeletal muscle tissues of rats that were frozen with liquid nitrogen (snap frozen, SF group) or stored in RNAlater solution (RL group), and the changes in concentration, purity, reference genes expression, and fold-change levels between groups were analyzed.

Results: In the RL group, the concentration of RNA isolated from the liver tissues was higher (P<0.05), whereas the A260/280 ratio was lower in the heart and liver tissues (P<0.05). PPIA and SRP72 genes were found to have lower Ct values in the heart tissues of rats in the RL group (P<0.05 and P<0.001, respectively) than the SF group. Expression levels of PPIA, ACTB, and SRP72 genes across the tissues were found to be different between the groups (P<0.05). The gene expression level examined in terms of fold-change was significantly different in the RL group (upregulated up to 4 folds and downregulated about 0.5 fold) (P< 0.05).

Conclusions: The results showed that RNAlater can maintain the RNA integrity and can also change the results of gene expression because it does not inhibit biological activity. The snap freezing method is more reliable because gene expression is more stable in tissues frozen with liquid nitrogen.

Parole chiave

  • RNAlater
  • Snap Frozen
  • RNA Quality
  • Reference Gene
  • Gene Expression
Accesso libero

The association of anthropometric parameters with markers of insulin and leptin secretion and resistance in type 2 diabetes mellitus

Pubblicato online: 27 Jul 2020
Pagine: 299 - 314

Astratto

Abstract

Aim: We evaluated the association between anthropometric parameters and markers of insulin and leptin secretion/resistance in patients with type 2 diabetes mellitus (T2DM).

Material and methods: This post-hoc data analysis from a cross-sectional study included 176 T2DM patients. Laboratory tests (serum leptin, soluble form of leptin receptor (sObR), C peptide, glycemic and lipid parameters) and anthropometric parameters were obtained, adiposity indexes (including body adiposity index (BAI), visceral adiposity index (VAI)), indicators of insulin resistance, β-cell function, and leptin resistance (Free Leptin Index, FLI) were calculated.

Results: The body mass index (BMI), diabetes duration, VAI and leptin correlated independently with HOMA-IR, while BMI, diabetes duration and HbA1c with HOMA-B. The total body fat mass (TBFM), C peptide, diabetes duration, BMI and BAI correlated with leptin concentrations, while the first three with FLI. VAI was an indicator of insulin resistance (β=0.166, p=0.003), while BAI of leptin secretion (β=0.260, p=0.010). TBFM strongly associated with leptin resistance and secretion (β=0.037, r=0.688, p<0.0001, and β=0.521, r=0.667, p<0.0001), and BMI correlated weakly with insulin secretion and resistance. While insulin and leptin secretion increased progressively with BMI, leptin and insulin resistance became significant only in case of obesity. The sObR was significantly associated with C peptide concentrations (β=-0.032; p=0.044), but not with HOMA-B or -IR. A strong positive correlation between the C peptide/leptin ratio and non-fat mass /TBFM ratio was noted (r=0.62 [0.52, 0.71], p<0.0001).

Conclusions: Parameters of peripheral adiposity correlated better with markers of leptin system, and those of visceral adiposity with markers of insulin secretion/resistance. The sObR correlated independently and negatively with C peptide.

Parole chiave

  • type 2 diabetes
  • insulin resistance
  • leptin system
  • anthropometric parameters
Accesso libero

Evaluation of oxidative stress in bladder in urethral injuries: an experimental model in rats

Pubblicato online: 27 Jul 2020
Pagine: 315 - 323

Astratto

Abstract

Background: The aim of this study is to determine the changes of some oxidative stress parameters following an experimental urethral injury.

Materials and Methods: 24 Wistar-Albino male rats were used and 18 of them were exposed to urethral injury. Total oxidant status (TOS) and total antioxidant capacity (TAC) values in bladder and penile tissues and sera of rats during creation and healing stages of injury (control, day-1, 7 and 21) were examined and evaluated histopathologically.

Results: TOS levels in both tissue and serum groups increased nearly 4x fold in first day following injury. On the seventh day, TOS levels of bladder and penile tissue showed a nearly 50% decrease according to first day (p=0.005; p=0.025, respectively). At the end of third week, while bladder TOS levels decreased to control values, penile tissue TOS levels remained at high levels. The bladder TAC values mildly increased in first day (p=0.026) and then returned to normal levels in seventh day and continued to decrease down by half of control levels at the end of third week (p=0.005). TAC levels of penile tissue increased twofold in first day (p=0.021) and remained at same high levels for three weeks.

Conclusion: We observed that first day after traumatic urethral injury seems to be very important for evaluation of traumatic injuries and wound healing. We believe when an urethral injury was occured, both urethra and bladder should be handled with care and without delay during urethral healing, because of the presence of oxidative changes in bladder.

Parole chiave

  • urethral injury
  • bladder
  • total oxidant status
  • total antioxidant capacity
  • oxidative stress index
Accesso libero

Total Errors of TSH results by fluorescent immunoassay technique from dried blood spots in a newborn screening program for congenital hypothyroidism - variations in time

Pubblicato online: 27 Jul 2020
Pagine: 325 - 330

Astratto

Abstract

Objective:the aim of this study was to evaluate the imprecision of the method used to determine TSH (Thyroid Stimulating Hormone) levels from dried blood spots in a newborn screening program and how the value of the coefficient of variation influences the total error of the method.

Methods:A short-term evaluation of imprecision was compared with the imprecision of the method assessed over five years. The coefficient of variation from the best quality control period and the worst quality control period were used. For Bias assessment mean results from the external quality program were used. Total Error was calculated with Bias and Coefficient of Variation values. A freely available software was used for standard deviation and coefficient of variation profiling.

Results:The values of the coefficient of variation for the short-term were lower than values obtained in worst quality control period but higher than in the best quality control period. Total error was higher than the accepted value for low-level control in the worst quality control period. Images obtained with the software showed that for high-level control coefficient of variation is concentration-dependent but this finding is not similar for low-level control.

Conclusions:Total Error of the Method may be subject to change in time. Initial evaluation of imprecision must be performed on a short term analysis but a continuous evaluation should be performed as the performance of a method may change in time. The evaluation should be performed on clinically significant levels for each parameter.

Parole chiave

  • Six Sigma
  • TSH
  • Imprecision
  • dried blood spot (DBS)
Accesso libero

Characterization of antibiotic resistance integrons harbored by Romanian Escherichia coli uropathogenic strains

Pubblicato online: 27 Jul 2020
Pagine: 331 - 340

Astratto

Abstract

Because little is known about the integrons which constitute an important means of spreading resistance in bacteria circulating in Romania, this study aimed to detect antibiotic resistance gene cassettes embedded in integrons in a convenient collection of 60 ciprofloxacin-resistant Escherichia coli isolates of various phylogroups, associated with community-acquired urinary tract infections. Characterization of the integrons was accomplished by PCR, restriction fragment length polymorphism typing, and DNA sequencing of each identified type. More than half of the tested E. coli strains were positive for integrons of class 1 (31 strains) or 2 (1 strain). These strains derived more frequently from phylogenetic groups A (15 of 21 strains), B1 (10 of 14 strains), and F (3 of 4 strains), respectively. While 20 strains carried class 1 integrons which could be assigned to nine types, eleven strains carried integrons that lacked the 3’-end conserved segment. The attempts made to characterize the gene cassettes located within the variable region of the various integrons identified in this study revealed the presence of genes encoding resistance to trimethoprim, aminoglycosides, beta-lactams or chloramphenicol. The evidence of transferable resistance determinants already established in the autochthonous E. coli strains highlights the need for improved control of resistance-carrying bacteria.

Parole chiave

  • Escherichia coli
  • urinary tract infections
  • integrons
  • antibiotic resistance
Accesso libero

Carbapenemase Producing Enterobacterales in Romania: Investigating the Origins

Pubblicato online: 27 Jul 2020
Pagine: 341 - 348

Astratto

Abstract

Carbapenemase-producing Enterobacterales (CPE) have emerged and spread in Romania since 2010. According to the reports of the EuSPACE (European survey of carbapenemase-producing Enterobacteriaceae) the epidemio-logical stage of the CPE expansion in Romania has shifted from sporadic occurrence in 2013 directly to inter-regional spread in 2014-2015. In this study we aimed to provide data from the timeframe when the dissemination of the carbapenemase genes in Romania began, by retrospectively analyzing CPE strains in a tertiary care university hospital. During the period of November 2012 – October 2013 we found 107 CPE (8.78%) out of 1219 non-duplicate Enterobacterales strains. 26 isolates of various Enterobacterales species carried blaNDM-1, 83 Klebsiella pneumoniae strains were positive for blaOXA-48-like and 2 of these co-harboured blaNDM-1. The increased incidence of OXA-48 producing K. pneumoniae was linked to a two-peaked hospital outbreak during February and May 2013. The percentage of 24.3% of NDM-1 producers was alarming due to the diversity of involved species and the higher resistance levels to carbapenems compared with blaOXA-48-like gene carriers. Plasmid replicon typing revealed a great diversity of plasmids in NDM-1-positive strains, belonging to incompatibility groups A/C, FII, FIIk, HI2, L and M. The strong connection between certain plasmid groups and host species suggests that the transfer of broad host-range plasmids through conjugation does not play the main role in the successful spread of blaNDM-1 among Enterobacterales species.

Parole chiave

  • carbapenem resistance
  • carbapenemase genes
  • plasmid
  • NDM-1
  • minimum inhibitory concentration
10 Articoli
Accesso libero

Metabolomic biomarkers of polycystic ovary syndrome related-obesity: a review of the literature

Pubblicato online: 27 Jul 2020
Pagine: 241 - 255

Astratto

Abstract

Background and objectives: Polycystic ovary syndrome (PCOS) displays a phenotype-dependent cardio-metabolic risk. By performing a systematic search of the literature, we aimed to summarize metabolomic signatures associated with obesity in PCOS women.

Data sources and study eligibility criteria: We conducted a comprehensive search including: Embase, PubMed, and Web of Science until 31st of May 2019. We used the terms: metabolomics and polycystic ovary syndrome. We excluded the following papers: animal studies, studies that included only lean PCOS women, reviews, meta-analyses, results of interventional studies, those that did not apply metabolomic techniques.

Results: The lipid signature in obese women with PCOS showed increased levels of free fatty acids (carnitine, adipic acid, linoleic acid, oleic acid) and lower levels of lysophosphatidylcholines and glycerolphosphocholine compared with non-obese PCOS women. Regarding carbohydrate metabolism, a decrease in citric and lactic acid levels characterized obese PCOS women. Decreased lactic acid in obese PCOS women suggests augmented insulin stimulated glucose muscle use in lean, but not in obese women. Considering amino acid metabolomic markers, valine, glycine, serine, threonine, isoleucine and lysine were higher in obese PCOS women. Patients with visceral obesity presented a diminished uptake of essential amino acids, BCAA, leucine and serine in the skeletal muscle. α-ketoglutarate was significantly higher in obese women with PCOS in comparison with lean women with PCOS, distinguishing these 2 subgroups of PCOS with high ‘predictive accuracy’.

Limitations: Overall, a small number of studies have focused on the impact of obesity on the metabolic fingerprints of PCOS women. There is need for properly controlled, high-quality studies.

Conclusions: There is compelling evidence of significant alterations in carbohydrate, lipid, and amino acid metabolism in women with PCOS and obesity. Metabolomics may identify new metabolic pathways involved in PCOS and improve our understanding of the complex relation between PCOS and obesity in order to personalize PCOS therapy.

Parole chiave

  • untargeted metabolomics
  • polycystic ovary syndrome
  • obesity
  • biomarkers
  • targeted metabolomics
Accesso libero

High frequency of BRCA recurrent mutations in a consecutive series of unselected ovarian cancer patients

Pubblicato online: 27 Jul 2020
Pagine: 257 - 266

Astratto

Abstract

Hereditary predisposition to breast and ovarian cancer (HBOC) is diagnosed by molecular analysis of deleterious mutations in BRCA genes, allowing oncogenetic follow-up of patients and of their families. BRCA testing addresses only to HBOC families, using restrictive inclusion criteria based on familial history of cancer and age at diagnosis. Sporadic ovarian cancer has high incidence and mortality in Romania, with low median age of diagnosis and possibly a higher magnitude of hereditary contribution comparing to othe populations. However, sporadic ovarian cancers do not qualify for BRCA testing according to inclusion criteria, and a complete BRCA screening of all cancers is neither feasible nor recommended. Despite the large diversity of BRCA mutations worldwide, some recurrent mutations have higher frequencies in diverse populations. Precisely screening for recurrent mutations in a target population allows to rapidly identifying mutation carriers without sequencing the entire BRCA genes. In Romanian population and neighboring countries, several recurrent mutations have already been described. In a consecutive series of 50 sporadic ovarian cancer patients, not qualifying for BRCA complete testing, we screened for 9 most common BRCA mutations, by multiplex-PCR, RFLP and targeted Sanger sequencing. Our results revealed 6 different BRCA mutations in 8 unrelated patients, with a frequency of 16%, much higher than expected. We further recommend screening for the identified mutations in larger series of cancer patients. The results are highly beneficial to cancer patients, healthy relatives, and overall, considering prevention in cancer a priority, to public health system and future of oncogenetics in Romania

Parole chiave

  • ovarian cancer
  • hereditary predisposition
  • BRCA genes
  • recurrent mutations
Accesso libero

SH2B3 (LNK) rs3184504 polymorphism is correlated with JAK2 V617F-positive myeloproliferative neoplasms

Pubblicato online: 27 Jul 2020
Pagine: 267 - 277

Astratto

Abstract

Background: Pathogenesis and phenotypic diversity in myeloproliferative neoplasms (MPN) cannot be fully explained by the currently known acquired mutations alone. Some susceptible germline variants of different genes have been proved to be associated with the development of these diseases. The goal of our study was to evaluate the association between the rs3184504 polymorphism of SH2B3 (LNK) gene (p.R262W, c.784T>C) and the risk of developing the four typical MPN - polycythemia vera (PV), essential thrombocythemia (ET), primary myelofibrosis (PMF), and chronic myeloid leukemia (CML).

Material and methods: We investigated the SH2B3 rs3184504 T>C polymorphism by real-time PCR in 1901 MPN patients (575 with PV, 798 with ET, 251 with PMF, and 277 with CML), all of them harboring one of the specific driver mutations - JAK2 V617F or CALR in case of PV, ET and PMF, or BCR-ABL1 in case of CML, and 359 controls.

Results: Overall, the TT homozygous genotype was significantly associated with BCR-ABL1-negative MPN (OR = 1.34; 95% CI = 1.03-1.74; crude p-value = 0.02; adjusted p-value = 0.04). The most significant association was seen in case of PV (OR = 1.54; 95% CI = 1.14-2.06; crude p-value = 0.004; adjusted p-value = 0.024). Also, SH2B3 rs3184504 correlated significantly with JAK2 V617F-positive MPN (OR = 1.36; 95% CI = 1.04-1.77; crude p-value = 0.02; adjusted p-value = 0.08), but not with those CALR-positive. ET (regardless of molecular subtype) and CML were not correlated with SH2B3 rs3184504.

Conclusions: The SH2B3 rs3184504 polymorphism is associated with risk of MPN development, especially PV. This effect is restricted to JAK2 V617F-positive PV and PMF only.

Parole chiave

  • myeloproliferative neoplasms
  • somatic mutations
  • genetic predisposition
Accesso libero

Mucopolysaccharidosis type I - Clinical and genetic characteristics of Romanian patients

Pubblicato online: 27 Jul 2020
Pagine: 279 - 286

Astratto

Abstract

Background: Mucopolysaccharidosis type I (MPS I) is an autosomal recessive lysosomal storage disorder caused by a deficiency of α-L-iduronidase (IDUA), which leads to the accumulation of partially digested glycosaminoglycans (dermatan sulfate and heparan sulfate) in the lysosomes and induces multisystemic alteration. Hurler (severe), Scheie (mild), and Hurler/Scheie (intermediate) syndromes are clinical subtypes of MPS-I. To date, more than 290 IDUA mutations have been reported. The purpose of this study was to present the clinical and genetic characteristics of Romanian MPS I syndrome patients and their genotype-phenotype correlation.

Patients and methods: Seven patients (5 girls and 2 boys) with MPS type I, belonging to 4 unrelated families, aged 0,75-17.9 years, were enrolled. The study methods consisted in: clinical and standard auxological assessment, bone radiographs, joint ultrasonography, goniometry, neurological and psychological evaluation, hepatic and splenic ultrasonography, cardiological evaluation, otorhinolaryngology examination, ophthalmological examination, spirometry, α-L-iduronidase enzyme activity assay and molecular analysis.

Results: The seven patients originated from 4 unrelated families, three patients with severe, two patients with intermediate and two with attenuated clinical phenotype. Each patient presented the classical picture of MPS type I picture, represented by: variable coarse facial features, arthropathy, hepatosplenomegaly, cardiac involvement, respiratory dysfunction and neurological impairment. Five patological variants, three point mutations (p.Q70 *, p.I238Q and p.K324R), two deletion c.1045_1047delGAC, c.46_57delTCGCTCCTG) and an insertion (c.1389 insC) were identified in both alleles of the ADUA gene in homozygous or heterozygous form. Two novel mutations (p.K324R and c.1389 insC) were reported. The p.Q70*(c.208C>T) variant was identified in 2 families with severe form of disease (Hurler syndrome) in homozygous status in one family and in compound heterozygous status in the other family.

Conclusion: The p.Q70* missense variant was the most frequent, correlated in all the cases who presented it with severe form, Hurler syndrome, the other mutations being usually isolated and particular for each patient, associated in our patients with less severe MPS I phenotype, as Hurler-Scheie or Scheie syndrome. The results of this study indicated the mutational heterogeneity of the IDUA gene and the difficulty to indicate some correlation between the genotype and phenotype in MPS I patients.

Parole chiave

  • MPS I
  • IDUA
  • mutations
Accesso libero

Comparative Evaluation of RNAlater Solution and Snap Frozen Methods for Gene Expression Studies in Different Tissues

Pubblicato online: 27 Jul 2020
Pagine: 287 - 297

Astratto

Abstract

Introduction: Freezing of tissues with liquid nitrogen is the most common method in studies performed at the RNA level. However, the use of RNA stabilization solutions has become a popular alternative method. The aim of this study is to investigate the effectiveness of RNAlater on RNA stabilization in different tissues.

Material and Methods: In this study, RNA were isolated from the lung, heart, liver and skeletal muscle tissues of rats that were frozen with liquid nitrogen (snap frozen, SF group) or stored in RNAlater solution (RL group), and the changes in concentration, purity, reference genes expression, and fold-change levels between groups were analyzed.

Results: In the RL group, the concentration of RNA isolated from the liver tissues was higher (P<0.05), whereas the A260/280 ratio was lower in the heart and liver tissues (P<0.05). PPIA and SRP72 genes were found to have lower Ct values in the heart tissues of rats in the RL group (P<0.05 and P<0.001, respectively) than the SF group. Expression levels of PPIA, ACTB, and SRP72 genes across the tissues were found to be different between the groups (P<0.05). The gene expression level examined in terms of fold-change was significantly different in the RL group (upregulated up to 4 folds and downregulated about 0.5 fold) (P< 0.05).

Conclusions: The results showed that RNAlater can maintain the RNA integrity and can also change the results of gene expression because it does not inhibit biological activity. The snap freezing method is more reliable because gene expression is more stable in tissues frozen with liquid nitrogen.

Parole chiave

  • RNAlater
  • Snap Frozen
  • RNA Quality
  • Reference Gene
  • Gene Expression
Accesso libero

The association of anthropometric parameters with markers of insulin and leptin secretion and resistance in type 2 diabetes mellitus

Pubblicato online: 27 Jul 2020
Pagine: 299 - 314

Astratto

Abstract

Aim: We evaluated the association between anthropometric parameters and markers of insulin and leptin secretion/resistance in patients with type 2 diabetes mellitus (T2DM).

Material and methods: This post-hoc data analysis from a cross-sectional study included 176 T2DM patients. Laboratory tests (serum leptin, soluble form of leptin receptor (sObR), C peptide, glycemic and lipid parameters) and anthropometric parameters were obtained, adiposity indexes (including body adiposity index (BAI), visceral adiposity index (VAI)), indicators of insulin resistance, β-cell function, and leptin resistance (Free Leptin Index, FLI) were calculated.

Results: The body mass index (BMI), diabetes duration, VAI and leptin correlated independently with HOMA-IR, while BMI, diabetes duration and HbA1c with HOMA-B. The total body fat mass (TBFM), C peptide, diabetes duration, BMI and BAI correlated with leptin concentrations, while the first three with FLI. VAI was an indicator of insulin resistance (β=0.166, p=0.003), while BAI of leptin secretion (β=0.260, p=0.010). TBFM strongly associated with leptin resistance and secretion (β=0.037, r=0.688, p<0.0001, and β=0.521, r=0.667, p<0.0001), and BMI correlated weakly with insulin secretion and resistance. While insulin and leptin secretion increased progressively with BMI, leptin and insulin resistance became significant only in case of obesity. The sObR was significantly associated with C peptide concentrations (β=-0.032; p=0.044), but not with HOMA-B or -IR. A strong positive correlation between the C peptide/leptin ratio and non-fat mass /TBFM ratio was noted (r=0.62 [0.52, 0.71], p<0.0001).

Conclusions: Parameters of peripheral adiposity correlated better with markers of leptin system, and those of visceral adiposity with markers of insulin secretion/resistance. The sObR correlated independently and negatively with C peptide.

Parole chiave

  • type 2 diabetes
  • insulin resistance
  • leptin system
  • anthropometric parameters
Accesso libero

Evaluation of oxidative stress in bladder in urethral injuries: an experimental model in rats

Pubblicato online: 27 Jul 2020
Pagine: 315 - 323

Astratto

Abstract

Background: The aim of this study is to determine the changes of some oxidative stress parameters following an experimental urethral injury.

Materials and Methods: 24 Wistar-Albino male rats were used and 18 of them were exposed to urethral injury. Total oxidant status (TOS) and total antioxidant capacity (TAC) values in bladder and penile tissues and sera of rats during creation and healing stages of injury (control, day-1, 7 and 21) were examined and evaluated histopathologically.

Results: TOS levels in both tissue and serum groups increased nearly 4x fold in first day following injury. On the seventh day, TOS levels of bladder and penile tissue showed a nearly 50% decrease according to first day (p=0.005; p=0.025, respectively). At the end of third week, while bladder TOS levels decreased to control values, penile tissue TOS levels remained at high levels. The bladder TAC values mildly increased in first day (p=0.026) and then returned to normal levels in seventh day and continued to decrease down by half of control levels at the end of third week (p=0.005). TAC levels of penile tissue increased twofold in first day (p=0.021) and remained at same high levels for three weeks.

Conclusion: We observed that first day after traumatic urethral injury seems to be very important for evaluation of traumatic injuries and wound healing. We believe when an urethral injury was occured, both urethra and bladder should be handled with care and without delay during urethral healing, because of the presence of oxidative changes in bladder.

Parole chiave

  • urethral injury
  • bladder
  • total oxidant status
  • total antioxidant capacity
  • oxidative stress index
Accesso libero

Total Errors of TSH results by fluorescent immunoassay technique from dried blood spots in a newborn screening program for congenital hypothyroidism - variations in time

Pubblicato online: 27 Jul 2020
Pagine: 325 - 330

Astratto

Abstract

Objective:the aim of this study was to evaluate the imprecision of the method used to determine TSH (Thyroid Stimulating Hormone) levels from dried blood spots in a newborn screening program and how the value of the coefficient of variation influences the total error of the method.

Methods:A short-term evaluation of imprecision was compared with the imprecision of the method assessed over five years. The coefficient of variation from the best quality control period and the worst quality control period were used. For Bias assessment mean results from the external quality program were used. Total Error was calculated with Bias and Coefficient of Variation values. A freely available software was used for standard deviation and coefficient of variation profiling.

Results:The values of the coefficient of variation for the short-term were lower than values obtained in worst quality control period but higher than in the best quality control period. Total error was higher than the accepted value for low-level control in the worst quality control period. Images obtained with the software showed that for high-level control coefficient of variation is concentration-dependent but this finding is not similar for low-level control.

Conclusions:Total Error of the Method may be subject to change in time. Initial evaluation of imprecision must be performed on a short term analysis but a continuous evaluation should be performed as the performance of a method may change in time. The evaluation should be performed on clinically significant levels for each parameter.

Parole chiave

  • Six Sigma
  • TSH
  • Imprecision
  • dried blood spot (DBS)
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Characterization of antibiotic resistance integrons harbored by Romanian Escherichia coli uropathogenic strains

Pubblicato online: 27 Jul 2020
Pagine: 331 - 340

Astratto

Abstract

Because little is known about the integrons which constitute an important means of spreading resistance in bacteria circulating in Romania, this study aimed to detect antibiotic resistance gene cassettes embedded in integrons in a convenient collection of 60 ciprofloxacin-resistant Escherichia coli isolates of various phylogroups, associated with community-acquired urinary tract infections. Characterization of the integrons was accomplished by PCR, restriction fragment length polymorphism typing, and DNA sequencing of each identified type. More than half of the tested E. coli strains were positive for integrons of class 1 (31 strains) or 2 (1 strain). These strains derived more frequently from phylogenetic groups A (15 of 21 strains), B1 (10 of 14 strains), and F (3 of 4 strains), respectively. While 20 strains carried class 1 integrons which could be assigned to nine types, eleven strains carried integrons that lacked the 3’-end conserved segment. The attempts made to characterize the gene cassettes located within the variable region of the various integrons identified in this study revealed the presence of genes encoding resistance to trimethoprim, aminoglycosides, beta-lactams or chloramphenicol. The evidence of transferable resistance determinants already established in the autochthonous E. coli strains highlights the need for improved control of resistance-carrying bacteria.

Parole chiave

  • Escherichia coli
  • urinary tract infections
  • integrons
  • antibiotic resistance
Accesso libero

Carbapenemase Producing Enterobacterales in Romania: Investigating the Origins

Pubblicato online: 27 Jul 2020
Pagine: 341 - 348

Astratto

Abstract

Carbapenemase-producing Enterobacterales (CPE) have emerged and spread in Romania since 2010. According to the reports of the EuSPACE (European survey of carbapenemase-producing Enterobacteriaceae) the epidemio-logical stage of the CPE expansion in Romania has shifted from sporadic occurrence in 2013 directly to inter-regional spread in 2014-2015. In this study we aimed to provide data from the timeframe when the dissemination of the carbapenemase genes in Romania began, by retrospectively analyzing CPE strains in a tertiary care university hospital. During the period of November 2012 – October 2013 we found 107 CPE (8.78%) out of 1219 non-duplicate Enterobacterales strains. 26 isolates of various Enterobacterales species carried blaNDM-1, 83 Klebsiella pneumoniae strains were positive for blaOXA-48-like and 2 of these co-harboured blaNDM-1. The increased incidence of OXA-48 producing K. pneumoniae was linked to a two-peaked hospital outbreak during February and May 2013. The percentage of 24.3% of NDM-1 producers was alarming due to the diversity of involved species and the higher resistance levels to carbapenems compared with blaOXA-48-like gene carriers. Plasmid replicon typing revealed a great diversity of plasmids in NDM-1-positive strains, belonging to incompatibility groups A/C, FII, FIIk, HI2, L and M. The strong connection between certain plasmid groups and host species suggests that the transfer of broad host-range plasmids through conjugation does not play the main role in the successful spread of blaNDM-1 among Enterobacterales species.

Parole chiave

  • carbapenem resistance
  • carbapenemase genes
  • plasmid
  • NDM-1
  • minimum inhibitory concentration

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