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Open access

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.

Open access

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.

Open access

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.

Open access

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.

Open access

Anna Snarska, Dominika Wysocka, Liliana Rytel, Sławomir Gonkowski, Hanna Pawelec and Przemysław Sobiech


Introduction: Simvastatin is a substance which is commonly used as a medicine to reduce cholesterol level. Unfortunately, it shows numerous side effects. Simvastatin affects various internal organs, and among other detriments to health may cause persistent muscle weakness, osteolytic processes, headaches, and rashes. Until now knowledge of the influence of simvastatin on bone marrow cells has been rather scant and fragmentary.

Material and Methods: During this experiment the numbers of all types of cells in the leukocytic system of porcine bone marrow were evaluated after 28 and 56 days of oral administration of simvastatin at a dose of 40 mg/day/animal.

Results: Simvastatin caused an increase in the number of all types of cells in the leukocytic system, and the most visible fluctuations concerned promyelocytes.

Conclusion: Observations obtained during the present study indicated that the results of the action of simvastatin on porcine bone marrow differ from those observed in other mammal species, including human. This may be due to various metabolic pathways within the bone marrow in the particular species, but the exact mechanisms of these actions are unknown at the present time.

Open access

Agnieszka Noszczyk-Nowak, Maciej Zacharski and Marcin Michałek


Introduction: In recent years, the high sensitivity and specificity of novel miRNA biomarkers have been utilised for early diagnosis and treatment monitoring of various diseases. Previous reports showed that abnormal expression of miR-208 in mice resulted in the development of an aberrant cardiac conduction system and consecutive arrhythmias. On the other hand, a study on infarcted human heart tissue showed upregulation of miR-208a in subjects with ventricular tachyarrhythmias compared to healthy controls. We prospectively investigated the expression of miR-208a and -208b in the serum of dogs presenting different cardiac arrhythmias.

Material and Methods: A total of 28 dogs with atrial fibrillation (n = 8), ventricular premature contractions (n=6), conduction system disturbances (n = 7), and free of heart conditions (as controls) (n = 7) were enrolled in the study. Total RNA was extracted from serum samples and miR-208a and -b, miR-16 as well as a cel-miR-39-5p spike-in were analysed with qPCR and ddPCR.

Results: miR-208a and miR-208b were not expressed in any of the samples. The calculated ddPCR miR-16 relative expression (normalised with cel-miR-39 spike-in) showed a good correlation (r = 0.82; P < 0.001) with the qPCR results.

Conclusion: This outcome warrants further investigation, possibly focusing on tissue expression of miR-208 in the canine heart.

Open access

Katarzyna Dudek and Dariusz Bednarek


Introduction: Mycoplasma bovis is known as a causative agent of many disorders in cattle. In Europe, there is still a lack of commercial vaccines against M. bovis infection. Acute phase response (APR) is a non-specific host reaction to infection, most seen in changes in production of acute phase proteins. The aim of this study was to analyse APR in calves administered with an experimental M. bovis vaccine.

Material and Methods: Twelve healthy female calves were divided into two equal groups: experimental and control. The experimental vaccine containing the field M. bovis strain and two adjuvants such as saponin and lysozyme dimer was subcutaneously administered to the experimental group. Phosphate buffered saline was taken as the placebo and given to the control group by the same route as the vaccine. Blood samples were collected prior to the study (day 0), then daily up to day 7, and then each seven days until day 84 post vaccination. The concentrations of serum amyloid A (SAA), haptoglobin (Hp), interferon-γ (IFN-γ), and inteleukin-4 (IL-4) were determined using commercial ELISA kits.

Results: Following the vaccination, a significant increase in SAA, Hp, and IFN-γ concentrations was observed when compared to the unvaccinated calves, whereas the IL-4 concentration was not detectable.

Conclusion: The experimental saponin-based M. bovis vaccine containing lysozyme dimer adjuvant visibly stimulated the APR in the calves, and some specific cytokines (Th1-dependent) directly involved in this response.

Open access

R. E. Antia and J. Ogunsola


A modified agyrophil technique was applied to peripheral blood smears to determine the mean AgNOR counts (MAC) of lymphocytes and ultimately assess the state of the lymphoid system in various clinical conditions of dogs. Fifty dogs, from clinically normal to pets with leukaemia, presented to the Veterinary Teaching Hospital, were recruited. Blood smears from each dog were stained with routine Romanowsky and modified agyrophil stains. Signalment, clinical diagnoses and hematologic parameters of the dogs were related to the MAC. An AgNOR proliferative index (AgPI) — percentage of lymphocytes with 3 or more AgNORs, was determined, and correlated with MAC. The statistical significance was determined at P < 0.05. MAC ranged from 1.17 in clinically healthy patients to 6.00 in leukaemic patients. The MAC was 2.00 in patients (n = 26) with lymphocyte counts within reference intervals (900—2400 per microliter); 2.23 in patients (n = 4) with lymphopenia; 2.18 in patients with lymphocytosis (n = 18) and 4.73 in patients (n = 4) with lymphocytic leukemia. Also, the MAC was 2.00 in non-anemic dogs while it was 2.47, 2.49 and 3.06 in patients with mild, moderate and severe anaemia, respectively. The MAC correlated strongly with AgPI (r = 0.91). The ancillary AgNOR technique provides a cheaper, more rapid and sensitive tool than routine lymphocyte counts in assessing the state of lymphoid proliferation in a variety of conditions in the dog.

Open access

T. A. Ajadi and M. O. Olaniyi


A nine weeks old female Large White piglet which was presented to the Veterinary Teaching Hospital, Federal University of Agriculture, Abeokuta, with a complaint of extra limbs was diagnosed with pygomelia and concurrent true hermaphroditism based on gross morphologic features, radiography, exploratory laparotomy and histopathology of the malformed organs. The piglet had two well-developed extra hind limbs consisting of the femur, tibia, fibula and the phalanges. Radiographically, the accessory limbs were attached to the ischium through a rudimentary pelvic bone. The supernumerary limbs were smaller than the normal appendages, but contained equal digits. The anal orifice was observed cranial to the right supernumerary limb. Caudal to the left supernumerary limb a rudimentary penis was observed. Two oval shaped fibrous masses were palpated in the inguinal canal of the piglet. In addition, there was a transparent tubular tract measuring 24 cm in length which contained serous fluid. The right kidney was rudimentary measuring 2.10 cm, while the left kidney appeared hypertrophied measuring 6.10 cm. The histology of the left kidney showed dysplastic areas of undifferentiated mesenchymal stroma in the cortex and medulla with the presence of groups of immature glomeruli in the cortex. The tubules in the medulla were scanty in number and had atypical epithelium. The adrenal glands had normal architecture with ectopic adrenal tissue in the adrenal capsule, while the ovaries and uterus were normal. It was concluded that the complex anomalies in the piglet might be as a result of a complex mode of inheritance.

Open access

Marcin Weiner, Wioletta Żukiewicz-Sobczak, Małgorzata Tokarska-Rodak, Dorota Plewik, Anna Pańczuk, Marta Siłuch, Jerzy Zagórski, Paweł Sobczak, Tomasz Chmielewski, Stanisława Tylewska-Wierzbanowska, Mariia Shkilna, Mykhailo Korda, Ivan Klisch, Mykhailo Andreychyn and Mariana Pavliuk


Introduction: Lyme borreliosis/Lyme disease is caused by Borrelia burgdorferi and is one of the most common vector-borne diseases transmitted by ticks.

Material and Methods: A total of 136 Ixodes ricinus ticks, collected in the Ternopil (Ukraine) region, including 126 adults (70 females and 56 males), and 10 nymphs were examined. The identification of the species and their developmental form was based on morphological characteristics.

Results: PCR with B5S-Bor and 23S-Bor primers resulted in Borrelia burgdorferi sensu lato DNA amplification among six ticks (4.4%). The detailed analysis based on the DNA sequencing showed the presence of DNA of Borrelia afzelii in four samples; the remaining two represented Borrelia burgdorferi sensu lato complex, although their genospecies were not determined. The research confirmed the dominance of Borrelia afzelii genospecies in the ticks from Ukraine.

Conclusion: It seems reasonable to undertake similar research in ticks from other regions of Ukraine. Knowledge in this field can be useful for public health and planning the prevention of tick-borne diseases.