Daniel Emil Albu, Monica Copotoiu, Peter Szmuk and Sanda-Maria Copotoiu
Irina Rosca, Andra-Cristina Bostanaru, Bogdan Minea, Valentin Nastasa, Iulian Gherghel, Carmen-Valentina Panzaru, Mihai Mares and Valentina Ruxandra Moroti-Constantinescu
Background: During the last two decades a major increase in the proportion of severe fungal infections has been noted due to the excessive use of broad-spectrum antibiotics, catheters, and a growing number of immunocompromised patients.
Objectives: This is the first investigation providing complete data regarding the phenotypic and genotypic profiles of Candida albicans (C. albicans) isolates in Romanian patients.
Methods: We investigated 301 isolates in terms of genotype determination (G), resistogram (R), phospholipase activity (Pl), haemolysis (Hl), proteinase activity (Pt), and biofilm formation (BF).
Results: The analyzed isolates of C. albicans showed low values for Pt (61.73%), Hl (95.49%), and BF (60.71%), and did not present any Pl activity (92.23%). More than half of the investigated samples were genotype A with 450 bp (52.92%) and the majority (86.19%) were resistant to sodium selenite (A), boric acid (B), sodium periodate (D) and silver nitrate (E), but sensitive to cetrimide (-). One-way ANOVA analysis revealed significant effects of the infection site on biofilm formation (p = 0.0137) and no significant correlation was found between the genotype (A, B, C) and the infection site (p =0.449).
Conclusions: Based on the obtained results it can be concluded that C. albicans isolates in Romanian patients exhibit different genotypic and phenotypic patterns, and no significant correlations between genotype and infection site could be observed.
George Andrei Crauciuc, Florin Tripon, Alina Bogliş, Amalia Făgărăşan and Claudia Bănescu
Small supernumerary marker chromosome (sSMC) is a rare chromosomal abnormality and is detected in about 0.3% in cases with multiple congenital anomalies (MCA) and/or developmental delay. Different techniques for investigation of cases with MCA and/or developmental delay are available ranging from karyotyping to molecular cytogenetic technique and ultimately multiplex ligation dependent probe amplification (MLPA). Here we present a patient with multiple congenital anomalies for which classical cytogenetic technique was used as a first step in diagnosis and the results being confirmed by MLPA. The karyotype disclosed a sSMC considered to be a fragment of chromosome 22. The MLPA analysis using SALSA MLPA probemix P064-C2 Microdeletion Syndromes-1B confirmed the karyotype results, and according to the manufacturer’s recommendation we performed another confirmation analysis with MLPA probemix P311-B1 Congenital Heart Disease and MLPA probemix P250-B2 DiGeorge. We also suspected an Emanuel syndrome and performed another MLPA analysis with SALSA MLPA probemix P036-E3 Subtelomeres Mix 1 and probemix P070-B3 Subtelomeres Mix 2B for investigation of subtelomeric region that revealed a duplication of 11q25 region and the confirmation was performed using SALSA MLPA probemix P286-B2 Human Telomere-11.
In conclusion, we consider that MLPA is a valuable method for identification of sSMC in children with developmental delay and congenital anomalies. Genetic diagnosis using different molecular techniques, such as MLPA, for increasing accuracy in identification of chromosomal structural aberrations has an important role in clinical diagnosis and in genetic counselling and our case explain the importance of using a specific laboratory technique for each stage of diagnosis.
Elena-Carmina Drăgulescu, Mihaela Oprea, Cătălina Zorescu, Roxana Şerban and Irina Codiţă
A prolonged outbreak of Healthcare-Associated Infections (HCAIs) evolved since December 2013, in a Newborns Unit from Hospital A, sited in the North-Eastern development region, Romania. A first cluster consisted of 19 cases, of which 18 infections in newborns and 1 labour infectious complication in a mother. Except for five cases declared and treated in the Neonatology Unit as hospital-acquired infections, the other cases were discharged and further required rehospitalisation and treatment.
Eight of these innitialy discharged cases were readmitted to the Pediatric Surgery Unit and two others to the Pediatrics Unit of Hospital B, while three others were readmitted to three hospitals: one to the Pediatrics Unit of Hospital C, and other two to Hospital A and Hospital D, respectively. The mother with the labour infectious complication was readmitted to the Gynecology Unit of the Hospital A.
A number of fifteen Staphylococcus aureus (S. aureus) strains isolated from the HCAI first episode and 8 strains from 7 HCWs were received by „Cantacuzino” Institute, Nosocomial Infections and Antibiotic Resistance Laboratory from the County Public Health Directorate, for confirmation and molecular typing.
After a first round of interventions for infection control, a second episode bursted in Hospital A and our laboratory received six other S. aureus isolates from newborns, hospital environment, and HCWs.
Public Health interventions based on epidemiologic data and molecular microbiology results were finally successful. The evolution of all cases was favorable.
An important factor favoring the outbreak was the moving of the Birth Unit of Hospital A to an innapropriate location for an 18-month interval, more than innitially estimated, in relation to rehabilitation of the ward.
We considered to report this episode taking into account the unusual evolution, the risk of multiresistant bacterial strains spreading, and multiple unwanted consequences caused by shortcomings in providing appropriate hygiene conditions.
Asad Hamad, Muhammad Arfan, Shujaat Ali Khan, Nighat Fatima, Arshad Mehmood Abbasi and Abdul Mannan
Artemisia vulgaris L. (Mugwort or Afsantin) has been used to treat various diseases since ancient times by the inhabitants of Himalayan region-Pakistan. Methanolic fractions (HA1-HA9) obtained from the aerial parts of A. vulgaris were evaluated for their antioxidant, antimicrobial and brine shrimp cytotoxic activities. Fraction HA8 showed substantial phenolics content with value of 26.29±1.4μgEQ/mg and DPPH scavenging (82.84±3.01%). Conversely, total flavonoids content of 7.32±0.07μgEQ/mg was determined in HA1 fraction. Fraction HA1 also showed significant cytotoxic effect with the value LD50 of 144.94μg/mL. Fractions HA7 and HA9 depicted maximum total antioxidant activity and ferric ion reduction (96.25±3.29 and AAE/mg and 176.91±8, respectively). All fractions showed encouraging results against bacterial strains Bordetella bronchiseptica and Micrococcus luteus, while HA2 fraction showed the highest percentage inhibition Mucor species with zone of inhibition of 13.25±0.35mm. A total of 7 fractions showed significant antileishmanial activity with survival percentage ranging 0.00 to 19. To sum up, results of the current study indicated that the plant can be further explored for isolation of antileishmanial and antimicrobial compounds, which could be used for drug development.
Sorin Dinu, Grațiela Țârdei, Emanoil Ceaușu, Simin Aysel Florescu, Laurențiu Micu, Alina Monica Ecobici, Mariana Mihăilă and Gabriela Oprișan
Background: Severe complications of chronic hepatitis C – i.e. cirrhosis and hepatocellular carcinoma – are important causes of morbidity and mortality worldwide. Despite the overwhelming rates of sustained virologic response achieved after therapy with different combinations of direct-acting antiviral drugs (DAAs), treatment failure is still recorded, and is due to the mutations harboured by hepatitis C virus (HCV) resistance associated variants (RAVs) selected during therapy. Baseline RAVs testing was found significant for guiding treatment in the cases of treatment failure and, sometimes, in naïve patients.
Methods: Romanian chronic hepatitis C patients unexposed to DAAs and infected with subtype 1b HCV were studied. Serum samples were used for Sanger population sequencing of a fragment containing NS3 viral protease, known to harbour resistance mutation against protease inhibitors (PIs).
Results: Catalytic triad and zinc-binding site in the studied sequences were conserved. Low-intermediate resistance mutations to first generation PIs were detected either alone or in conjunction with resistance substitutions associated with second generation PIs. Cross-resistance and reduced susceptibility to certain DAAs were observed.
Discussion: This study focused on HCV patients infected with subtype 1b strains, the most prevalent in Romania. The rate of RAVs found in this work is consistent with the results reported by similar studies from other countries. Noticeably, numerous polymorphisms of unknown significance to DAAs resistance, but reflecting the high genetic variability of HCV, were found in the studied sequences. Testing for RAVs can be a useful method for guiding treatment in a cost-efficient manner in developing countries where access to DAAs is limited.
Octavia Sabin, Ioana Corina Bocșan, Adrian Trifa, Zoltan Zsigmond Major, Simona Codruta Heghes, Emanuela Brusturean Bota and Anca Dana Buzoianu
Aim: A possible molecular mechanism of clinically defined multidrug-resistant epilepsy involves drug efflux transporters such as P glycoprotein (P-gp), a member of the ATP-binding cassette subfamily B1 (ABCB1). We have investigated the prevalence of the C3435T, G 2677T/A, and T129C single-nucleotide polymorphisms in the promoter region of MDR1 gene, in Romanian epileptic patients.
Methods: 70 epileptic patients evaluated in the Neurology Department of Cluj County Hospital were included in the study. The response to treatment was assessed by reviewing the seizure diaries and the patients were classified as responders or non-responders. Antiepileptic drug (AED) plasmatic concentrations were measured and the patients were divided into 2 groups: first group with AED concentrations in therapeutic range and the second one with sub-optimal AED concentrations. Genotyping the DNA samples, we investigated MDR1 gene polymorphism by polymerase chain reaction (PCR). Results were expressed as genotype and allele frequencies per response group and compared between subgroups.
Results: 33 patients (47.14%) were classified as responders, while the remaining 37 patients (52.86%) were classified as non-responders. A comparison of responders and non-responders revealed no significant difference in genotype frequency for any of the three mutations studied. The CT heterozygote for ABCB1 T129C had significantly lower AED concentrations (p=0.041), with no significant difference for the other polymorphisms studied.
Conclusions: In our study we found an association of CT variant in ABCB1 C129T with lower AED plasmatic concentrations and no association between ABCB1 variants and the drug responsiveness.
Vaida Petrauskiene, Ruta Vaiciuniene, Vytautas Kuzminskis, Edita Ziginskiene, Saulius Grazulis, Egle Jonaitiene, Erika Skrodeniene and Inga Arune Bumblyte
Background and objectives: Vascular calcification (VC) is one of the factors associated with mortality in hemodialysis (HD) patients. The purpose of the study was to assess associations between prevalent VC and disturbances of calcium-phosphate metabolism as well as changes in vitamin D (25(OH)D), FGF 23 and MGP levels and to evaluate the possible impact of VC and changes of these biomarkers on survival in HD patients.
Methods: The study population consisted of 81 prevalent patients in the hemodialysis unit of Hospital of Lithuanian University of Health Sciences Kaunas Clinics. A simple vascular calcification score (SVCS) was evaluated as it is described by Adragao et al. 25(OH)D (nmol/L), FGF 23 (ng/L) and MGP (ng/mL) were measured and analysed.
Results: Patients were divided into two groups: SVCS<3 (31 patient (38.3%) and SVCS ≥3 (50 patients (61.7%)). In multivariate logistic regression, age (odds ratio 1.062, 95% CI [1.024-1.1] p=0.001) and diabetes (odds ratio 6.9, 95% CI [1.5-31], p=0.012) were associated with SVCS ≥3. The multivariate logistic regression revealed the highest negative impact of SVCS ≥3, age and 25(OH)D level for death risk.
Conclusion: VC in HD patients is highly influenced by age and presence of diabetes and associated with higher risk of death. No significant association was found between MGP and FGF 23 and VC as well as between these two biomarkers and risk of death. Lower 25(OH)D levels were associated with mortality in this dialysis patients cohort.
Marcin Michałek, Piotr Frydrychowski, Jakub Adamowicz, Agnieszka Sławuta, Urszula Pasławska and Agnieszka Noszczyk-Nowak
Introduction: Ventricular rhythm disturbances are a common pathology in human and veterinary medicine. In humans, the algorithmic approach is used to differentiate wide QRS complex tachycardia. The most commonly used are the aVR and Brugada algorithms as well as the ventricular tachycardia (VT) score developed by Jastrzębski and coworkers. In veterinary medicine, no such algorithms are available and the only parameter used to describe VT abnormalities is the duration of the QRS complexes. The aim of this analysis was determining whether human medicine algorithms for VT are applicable in veterinary medicine to differentiate wide QRS complex tachycardia in dogs.
Material and Methods: A retrospective analysis was performed on 11 dogs of both sexes and various breeds and age diagnosed with VT. The diagnosis was based on ambulatory ECG, further established based on the reaction to lidocaine or adenosine or an invasive electrophysiological study.
Results: Of the 11 tracings passed through the aVR algorithm, 10 met the VT criteria. The most common criterion was the Vi/Vt ratio (8 out of 11 tracings). Based on the VT score, seven out of eight dogs had a high probability of VT.
Conclusion: Retrospective analysis of ECGs by aVR and VT score indicates that the applied algorithms may be useful in differentiating wide QRS complex tachycardia as a quick, easy, and non-invasive alternative to cardiac electrophysiology.