Tom 29 (2021): Zeszyt 1 (January 2021)

MicroRNAs (miRNAs) are a group of non-coding RNA molecules that have an important role in modulating the expression of genes involved in regulating cellular functions. A growing number of studies suggest the abnormal expression of microRNAs in different types of cancer cells. MiRNA-124 is a microRNA that is down-regulated in many types of cancer cells, including bladder cancer. Our objective is to provide a review of the key publications that studied the effect of miR-124 on bladder cancer. This review focus on the targets and different pathways of miR-124 that were identified in various studies and differences between their expressions in normal urothelium and tumor tissues. We also include data regarding urinary methylations levels of miR-124 and their role in bladder cancer diagnosis and prognosis. Subsequently, we establish future perspectives of miR-124 research and its promising role in bladder cancer.

Background: Hepatitis C virus can be eradicated with antiviral therapy, thus reducing the risk of disease progression and death associated with the final stage of liver disease.

Methods: 241 patients received PrOD+RBV for 12 weeks. Clinical and laboratory data were assessed at baseline, week 4, 8, 12 (end of treatment, EOT), and 12 weeks after therapy (sustained virological response, SVR). Subsequently, biological and virological measurements were performed at least 48 weeks after obtaining SVR12 in responder patients.

Results: Per protocol SVR12 rate was 97,6%. Severe adverse events were reported in 3 patients (1.24%) and led to treatment discontinuation (liver decompensation). One 58-year-old patient who completed the treatment died before SVR evaluation due to acute mesenteric ischemia (not related to antiviral therapy). Baseline total bilirubin above 2 mg/dl can be considered a predictive factor for non-response to PrOD+RBV treatment (p = 0.004). Of the 30 patients evaluated at least 48 weeks after SVR no one presented relapses, with no statistically significant differences in biological parameters changes and no adverse events were noted during the 48-week follow up period.

Conclusion: Our study revealed the high effectiveness and good safety profile of PrOD +RBV in patients with genotype-1b HCV compensated cirrhosis (Child Pugh A) which were maintained during a 48-week period after treatment finalization.

Background: Tumor characterization through the study of molecular biology has become an invaluable tool in understanding cancer development and evolution due to its relationship with chromosomal mutations, alterations or aberrations. The purpose of this study was to investigate the involvement of genes such as TLR-4 and DNA repair pathways (XRCC1 and XPD) in laryngeal cancer susceptibility in a Romanian population. Method: We performed a case-control study on 157 laryngeal cancer patients and 101 healthy controls. Genetic testing was carried out using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism. Results: We identified the Gln allele of the XPDLys751Gln polymorphism as an individual risk factor in laryngeal cancer development (Gln vs Lys, adjusted OR=1.65, 95%CI=1.13–2.40, P=0.008). Subjects with the mutant homozygote variant (Gln/Gln) had a two fold increase in cancer risk (adjusted OR=2.18, 95%CI=1.06–4.47, p=0.028) when compared to the reference wild type genotype (Lys/Lys). Stratification by sex and age, identified males under 62 years as the most susceptible group with an almost three fold risk (adjusted OR=2.94, 95%CI=1.31–6.59, p=0.007) for the dominant model (Lys/Gln+Gln/Gln). No associations were found for TLR-4Thr399Ile, XRCC1Arg194Trp and XRCC1Arg399Gln. Conclusion: The results of the study show that the XPDLys751Gln polymorphism may be among other independent risk factors for developing laryngeal cancer where as TLR-4Thr399Ile, XRCC1Arg194Trp and XRCC1 Arg399Gln show no such association. However, we consider the relative small number of the subjects selected for this analyses a possible limitation towards the real influence the obtain results may pertain in laryngeal cancer evolution.

FMS-like tyrosine kinase 3 gene internal tandem (FLT3-ITD) mutations represent one of the most frequent genetic lesions in acute myeloid leukemia (AML) and imparts a negative prognostic. For an optimal patient management, current clinical guidelines recommend the evaluation of the allelic ratio (AR), expressed as the DNA FLT3-ITD/WT mutational burden. We sought to evaluate the differences between the AR and FLT3-ITD/WT mRNA ratio (RR) and their respective impact on the biological characteristics of AML patients. A total of 32 DNA and mRNA samples from AML patients with FLT3-ITD were evaluated. There was a good correlation between the AR and RR (Spearman’s rho= 0.652, P <0.001). None of the biological characteristics were influenced by the RR values, whereas patients with high AR values (≥0.5) had higher WBC counts (Mann-Whitney, P= 0.01), LDH levels (Mann-Whitney, P= 0.037), and circulating blasts levels (Mann-Whitney, P= 0.023) than patients with low AR values (<0.5). Also, there was a good correlation between AR values and WBC count (Spearman’s correlation, P= 0.001), and LDH levels (Spearman’s correlation, P= 0.007). In our study population the AR, but not the RR, influenced the biological characteristic of patients suggesting a dose-independent effect of FLT3-ITD mutations.

Sexually transmitted infections (STIs) are among the most common infections in Romania. Infection with Ureaplasma urelyticum is one of the major causes of STIs and can cause serious complications. Although tetracycline is the drug commonly used to treat infections caused by U. urealyticum, several studies indicate the emergence and rapid development of strains resistant to these antibiotics in the United States or Europe. Tetracycline resistance in bacteria is encoded by a number of different genetic determinants but in mycoplasmas the only tetracycline resistance determinant that has been reported is the tetM gene. Tetracycline resistance among Ureaplasma spp. is associated with the presence of the horizontally acquired tetM resistance gene. Our study on bacterial DNA aimed to determine the presence of tetracycline-resistant U. urealyticum strains, by identifying the presence of the tetM gene. We used first void urine samples from 622 STI-suspected subjects. DNA was extracted, purified and amplified via PCR for the simultaneous detection of 6 STIs. 68 patients were diagnosed with U. urealyticum. DNA obtained from these samples was amplified using the tetM gene and U. urealyticum - specific urease gene primers. The urease gene was amplified in all samples, confirming the presence of U. urealyticum. The tetM gene was amplified in 2 samples considered tetracycline-resistant strains. The study confirmed the presence of U. urealyticum strains resistant to tetracycline in Romania. The employed technique can produce quick results both for U. urealyticum detection and determination of its resistance to tetracycline using a single easy-to-collect biological sample.

Introduction: Resistance to first-line antibiotics of the Proteeae strains within the difficult-to-treat (DTR) phenotype is a cause of limitation of therapeutic options. The study aimed to characterize these strains, to identify the factors that influence their acquisition and the predictive factors for the patient’s evolution.

Material and methods: Between July 2017 and January 2019, 400 of Proteeae strains were isolated from samples of patients admitted to intensive care units (ICUs) and surgical wards of a university hospital in Romania. The identification and testing of antibiotic sensitivity was performed using the Vitek 2 Compact system. The DTR phenotype was defined as the resistance (or intermediate resistance) to all categories of β-lactams, carbapenems and fluoroquinolones.

Results: Out of 400 Proteeae strains, 21% were of the DTR type, most of them from the species Providencia stuartii and Proteus mirabilis, identified predominantly on the ICUs. The excess fatality in the DTR subsample compared to the non-DTR subsample was 16.37%. The multivariate analysis identified as independent risk factors: the number of antibiotics administered, the number of days of urinary catheterization, the presence of tracheostomy, nasogastric nutrition, respectively belonging to the species P. stuartii. The probabilities of survival were reduced by the presence of the central venous catheter (CVC), tracheostomy, by the increase of the number of hospitalization days respectively of the number of antibiotics administered.

Conclusion: The DTR phenotype in the case of Proteeae strains has been associated especially with the species P. stuartii, with invasive exogenous factors and with an increased fatality.

Introduction: Community-acquired pneumonia (CAP) is the primary cause of severe sepsis. Severity assessment scores have been created, in order to help physicians decide the proper management of CAP. The purpose of this study was to examine the correlations between different CAP severity scores, including qSOFA, several biomarkers and their predictive value in the 30 day follow-up period, regarding adverse outcome.

Materials and methods: One hundred and thirty nine adult patients with CAP, admitted in the Teaching Hospital of Infectious Diseases, Cluj-Napoca, Romania from December 2015 to February 2017, were enrolled in this study. Pneumonia Severity Index (PSI), CURB-65, SMART-COP and the qSOFA scores were calculated at admittance. Also, C-reactive protein (CRP), procalcitonin (PCT) and albumin levels were used to determine severity.

Results: The mean PSI of all patients was 93.30±41.135 points, for CURB-65 it was 1.91±0.928 points, for SMART-COP it was 1.69±1.937 points. The mean qSOFA was 1.06±0.522 points, 21 (14.9%) were at high risk of in-hospital mortality. In the group of patients with qSOFA of ≥2, all pneumonia severity scores and all biomarkers tested were higher than those with scores <2. We found significant correlations between biomarkers and severity scores, but none regarding adverse outcome.

Conclusion: The qSOFA score is easier to use and it is able to accurately evaluate the severity of CAP, similar to other scores. Biomarkers are useful in determining the severity of the CAP. Several studies are needed to assess the prediction of these biomarkers and severity scores in pneumonia regarding adverse outcome.

Introduction: Perforated appendicitis (PA) in children is associated with a considerable risk for postoperative complications (POCs) such as wound infection and intra-abdominal abscess.

The aim of this study was to determine the diagnostic accuracy of hematological parameters in the early POC detection in children after PA surgery.

Materials and Methods: The study enrolled 71 patients with PA divided into two groups: 14 patients with POC (POC+ group) and 57 patients without POC (POC− group). Clinical and hematological parameters were followed preoperatively, prior to the surgery (PRO) and postoperatively on day 2 (POD2) and day 4 (POD4).

Results: The POC+ group had longer duration of higher axillar temperature as well as extended intensive and inpatient care. This group also had a significantly lower absolute neutrophil count ratio between POD2 and POD4. According to the receiver operating characteristic curve analysis, relative neutrophil count on POD4 higher than 71.8% and the ratio of absolute neutrophil count between POD2 and POD4 lower than 44.5% were found to be useful for predicting POC.

Conclusion: Absolute neutrophil count ratio between POD2 and POD4 and relative neutrophil count at POD4 could be efficient in identifying children at higher risk of developing POC after PA surgery.

Background: Coronavirus disease 2019 (COVID-19) has spread rapidly in China and globally. In order to control the spread of the epidemic, it is important to find an efficient diagnostic method.

Objectives: The aim of this study was to assess the responses of antibodies during SARS-CoV-2 infection in relation to disease severity and to evaluate the association between the positive rate of antibody detection and nucleic acid test.

Methods: Ninety patients with SARS-CoV-2 infection were recruited in this retrospective observational study. Demographic, clinical data, and SARS-CoV-2 IgM and IgG antibodies in serum specimens were detected at 4 and 6 weeks after diagnosis.

Results: IgM and IgG antibody levels showed a decreased tendency, the titers at week 4 were higher than the titers at week 6: The positive rates of IgM at week 4 and 6 were 92.9% and 67.9%, respectively. The positive rates of IgG at week 4 and week 6 were 100%. No association was found between the positive rate of antibody detection at week 4 or 6 and that of nucleic acid test (P>0.05). No difference between the positive rate of antibodies against SARS-CoV-2 in severe and non-severe COVID-19 patients was observed.

Conclusions: Antibody detection is an effective means in the diagnosis of COVID-19. The titer and positive rate of IgM are lower than those of IgG in the first six weeks after infection. Positive rate of antibodies was not different between severe and non-severe patients.

Reactivation of hepatitis B virus (HBV) infection has been described in patients with HBsAg negative and antiHBc positive (occult hepatitis B infection -OBI) receiving immunosuppressive therapy (IST). The lack of proper monitoring of patients with this HBV infection during IST can result in viral reactivations with high level of transaminases, jaundice and even acute liver failure. In these situations, it is mandatory to start antiviral therapy with nucleot(s) ide analogs (NA) which produce a strong viral suppression. We report a series of five cases of OBI patients with severe HBV reactivation during IST. One patient was diagnosed with hematologic malignancy (non-Hodgkin lymphoma), two with rheumatoid arthritis, one with psoriasis and one patient with renal transplant. All the patients were evaluated and treated for the reactivation of HBV in the Prof. Dr. Matei Bals National Institute of Infectious Diseases, a tertiary care hospital from Bucharest, Romania. At the time of HBV reactivation diagnosis, 3 patients were asymptomatic and two developed jaundice. All had acute ALT flares (more than 10 times the upper limit of normal range - ULN), very high HBV viral loads and anti-HBc serum IgM antibodies. All patients were immediately treated with ETV 0.5 mg/day and if it was possible, IST was stopped. In all cases was obtained quickly HBsAg loss under antiviral therapy.