Volume 24 (2016): Issue 1 (March 2016)

Our current clinical doctrine and practice is based upon a classification of diabetes which relies mainly on some clinical manifestations/criteria, rather than markers of the pathophysiological mechanisms of the disease. An improved classification based on such biological markers (i.e. of insulin resistance, beta cell dysfunction, autoimmunity) may assist in clinical decision and may offer the opportunity of an optimized therapeutic strategy. We address here some important questions that have not yet been clarified, e.g. which markers/indicators best define the main pathogenic mechanisms of the disease in a patient with diabetes and what threshold values are relevant for this purpose.

The management of the critically ill polytrauma patient is complex due to the multiple complications and biochemical and physiopathological imbalances. This happened due to the direct traumatic injury, or due to the post-traumatic events. One of the most complex physiopathology associated to the multiple traumas is represented by microvascular damage, subsequently responsible for a series of complications induced through the imbalance of the redox status, severe molecular damage, reduction of the oxygen delivery to the cell and tissues, cell and mitochondrial dead, augmentation of the inflammatory response and finally the installation of multiple organ dysfunction syndrome in this type of patients. A gold goal in the intensive care units is represented by the evaluation and intense monitoring of the molecular and physiopathological dysfunctions of the critically ill patients. Recently, it was intensely researched the use of microRNAs as biomarkers for the specific physiopathological dysfunctions. In this paper we wish to present a series of microRNAs that can serve as biomarkers for the evaluation of microvascular damage, as well as for the evaluation of other specific physiopathology for the critically ill polytrauma patient.

Introduction. Several alternative methods to peripheral blood DNA extraction have been implemented so far. Saliva seems to represent a very advantageous type of sample, easy to harvest and able to generate DNA yields comparable to those extracted from blood mononuclear cells.

Material and methods. 8 patients suspected of ankylosing spondylitis, 9 patients with various hematological malignancies, displaying post-chemotherapy leucopenia and 30 healthy volunteers were included in our study. DNA was extracted with various commercially available kits and used for HLA typing either by PCR amplification, or by PCR followed by hybridization.

Results. Our data regarding HLA typing support already published results regarding the good DNA quality that allows its use in various molecular biology techniques. However, when attempting to use saliva from immunosuppressed patients for DNA extraction we have generated very low yields, comparable again with the ones obtained from peripheral blood. Flow cytometry and immunocytochemistry investigations confirmed the low number of leukocytes present in the saliva of these patients, while the number of epithelial cells was virtually unchanged.

Conclusions. The main source of saliva DNA seems to be represented by leukocytes present in this fluid and not by the epithelial cells. Under these circumstances, for immunosuppressed patients saliva cannot represent an alternative to blood when attempting DNA extraction.

Introduction: Being a multifactorial disease, stroke is one of a major causes of death and disability worldwide. Several genetic polymorphisms have been associated with stroke etiophatology and FGB −455 G>A and GP IIIa PIA1/A2 are among them. In the present study, we investigated the association between FGB −455 G>A and GP IIIa PIA1A2 polymorphisms and the risk of ischemic stroke in a group of Romanian stroke patients.

Subjects and methods: This case-control study included 148 patients with ischemic stroke and 150 healthy age, sex and ethnically matched unrelated controls. FGB −455G>A and GP IIIa PIA1A2 genotyping was carried out using PCR-RFLP. The association of FGB −455G>A and GP IIIa PIA1A2 polymorphisms and cardiovascular risk factors with ischemic stroke was tested using logistic regression analysis.

Results: Molecular analysis did not reveal an increased frequency of the FGB -455 G>A variant allele and GP IIIa PIA1/A2 variant allele in the study group compared to the control group (p = 0.140, OR = 0.750, 95% CI = 0.522 - 1.077; p = 0.823, OR = 0.944, 95% CI = 0.558 - 1.599 respectively). Furthermore, after performing logistic regression analysis adjusted for the known risk factors, a positive association with stroke was found in smokers (p = 0.026, OR = 1.800, 95% CI = 1.071 - 3.024)

Conclusions: No association was found between FGB −455 G>A and GP IIIa PIA1/A2 polymorphisms and ischemic stroke in the studied population.

Introduction. Dawn phenomenon could have deleterious effect on overall glycemic control. Glycemic variability may be an independent risk factor for the development of diabetes chronic complications. The study aimed to evaluate any correlations between the dawn phenomenon and parameters of glycemic variability in a cohort of type 2 diabetes patients (T2DM). Material and methods. This retrospective observational study included 131 T2DM patients. Continuous glucose monitoring (CGM) has been performed. Data from the first 24h of full recording were used for analysis of glycemic variability indices: mean level of 24h interstitial glucose value and standard deviation; % coefficient of variation; J index; mean amplitude of glycemic excursion - MAGE; continuous overall net glycemic action (CONGA) at 1, 2, 4 and 6 hours; mean of daily differences (MODD) index. Results. Mean age was 56.04 ± 9.91 years, 35.9% women, 17.6% on diet, 53.4% on oral therapy and 29% on insulin. Dawn phenomenon was more frequent in patients below 60 years (70%) and in oral therapy group (72.85%). Significant correlations between the dawn phenomenon and j-index, MAGE, CONGA-4 and CONGA-6 have been found in T2DM patients on diet therapy alone. The amplitude of dawn phenomenon was 46.10 ± 24.40 mg/dl and significantly correlated (p<0.05) after adjustment for age, gender and treatment with % CV, MAGE, CONGA-1, CONGA-2, CONGA-4, CONGA-6 and MODD. Conclusions. The dawn phenomenon significantly increases the glycemic variability parameters in drug-naive T2DM patients, with no impact in T2DM on oral or insulin therapy.

Background and Aim: Type 2 diabetes (T2DM) has been associated with hypertension (HTN) and elevated high-sensitivity C-reactive protein (hsCRP), but the possible implication of blood pressure (BP) variability in increasing hsCRP in T2DM are incompletely understood. We aimed to assess the association between hsCRP and BP variability during 24-hour ambulatory BP monitoring in T2DM and healthy control subjects.

Material and Method: The cross-sectional study included data from T2DM patients with normal BP (n=9), controlled HTN (n=46), uncontrolled HTN (n=20), and healthy controls (n=11). HsCRP was assessed using ELISA technique. All subjects underwent 24-hour ambulatory BP monitoring; BP variability was calculated using standard deviation.

Results: We found that hsCRP was associated with daytime and 24-hours systolic and diastolic BP variability. Higher hsCRP were observed in T2DM patients with uncontrolled HTN and high BP variability compared to the other three groups. In multiple regression analysis, hsCRP was predicted by daytime and 24-hour diastolic BP variability.

Conclusions: Our findings suggest that high hsCRP was associated with increased ambulatory BP variability in T2DM and control subjects. The contribution of both hsCRP and BP variability to cardiovascular risk stratification in T2DM needs to be evaluated in prospective studies.

Introduction. With age, sex hormone deficiency leads to reduced bone mineral density (BMD) in men. The aim of our research is to analyze the role of serum sex steroids in assessing BMD in the men included in this study. Materials and methods. This cross-sectional study included 146 men aged 65-85 years old with osteopenia or osteoporosis (study group) and 121 men with normal BMD (control group). Serum levels of total testosterone (Tt) and free testosterone (Tf) were measured by immunoassay, and estradiol (E2) levels were measured by the immunoenzymatic method. Femoral neck and lumbar spine BMD was examined by dual-energy X-ray absorptiometry. Results. Tf and E2 deficiency was significantly associated with low BMD (p=0.007). No association was found between Tt deficiency and reduced BMD. Tf levels (p<0.001) and E2 levels (p=0.003) were significantly lower in patients with reduced BMD compared to those with normal BMD. Significant more cases with low levels of Tf (p=0.015) and E2 (p<0.001) were found in patients with osteoporosis compared to those with osteopenia. Conclusions. Sex hormones deficiency in men was significantly correlated with the decrease of BMD. Determination of serum Tf and E2 levels is important in assessing the risk of osteoporosis in male subjects.

Background: Beta-thalassaemia (β-thalassaemia) major patients are severely anaemic and require life-long blood transfusions for survival. These patients require iron-chelation therapy as a result of iron overload due to the monthly blood transfusions. The iron over load can cause oxidative damage and pro-inflammation and therefore, hasten mortality. Thus, regular monitoring of the oxidative stress and pro-inflammation status may be useful in these patients.

Methods: Measurement of biomarkers is usually performed on serum samples but the evaluation in non-invasive samples such as saliva would be more favourable in paediatric cases. In this study, the levels of pro-inflammatory cytokines such as tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) as well as oxidative indices such as lipid hydro peroxide, advanced oxidation protein products (AOPP), ferric reducing antioxidant power (FRAP), uric acid (UA) and glutathione peroxidase (GPx) activity in a total of 65 β-thalassaemia major patients (all on iron chelation) and 55 healthy control subjects were assessed. All the above biochemical parameters, measured using well established assay techniques, were detectable in saliva samples.

Results: Non parametric analyses showed that lipid hydroperoxide (LH) and glutathione peroxidase (GPx) activities were significantly higher in β-thalassaemia major patients. All other parameters were not significantly different between patient and control groups implying that iron chelation therapy was successful in attenuating oxidative stress. Strong positive correlation was observed between FRAP and UA levels. There was also a notable difference in tumour necrosis factor-α (TNF-α) between the patients and healthy controls when analysed according to ethnicity and age. AOPP level in β+-thalassaemia homozygous patients were significantly higher than β+/β0-compound heterozygous and β0-thalassaemia homozygous patients.

Conclusion: Saliva may serve as a reliable, non-invasive sample which can be used to assess oxidative indices and pro-inflammatory cytokines in β-thalassaemia major patients.

Acquired von Willebrand disease (AvWD) represents a rare, potentially severe and most likely underdiagnosed category of hemorrhagic syndromes determined by quantitative, qualitative or functional, nonhereditary, alterations of von Willebrand factor (vWF) that occur in the context of various underlying diseases. It is diagnosed mainly in adults, without any personal or familial history of bleeding. The etiopathogeny of AvWD is complex, marked by the intervention of multiple mechanisms, occuring in the evolution of neoplasia, autoimmune disorders, cardiovascular diseases and other conditions. The clinical and laboratory manifestations are similar to the congenital form with mucocutaneous hemorrhage in patients without bleeding history and demonstration of quantitative and/or functional anomalies of vWF. Treatment has two major objectives: control of bleeding and therapy of the underlying condition. As a practical illustration of the theoretical aspects we present 3 clinical cases of AvWD diagnosed in the Colţea Hospital Department of Hematology during the last 10 years.

The surveillance of shigellosis is carried out under the auspice of the European Centre for Disease Prevention and Control which requires a reliable laboratory-based surveillance at national level. To date, little information is published about the members of Shigella spp. responsible for Romanian cases of shigellosis which hinders the understanding of the current epidemiology of shigellosis. Consequently, this retrospective study aimed to assess the diversity of virulence profiles displayed by the strains circulating in our region, by using key chromosome- and plasmid-associated markers, and to document the prevalence of pHS-2-like plasmid previously proposed as a potential marker for reactive arthritis.

The study focused on 65 Shigella sonnei and 49 Shigella flexneri clinical isolates, originated from local patients, recovered through the national surveillance system in 2009 - 2013. PCR assays were performed for the detection of ipaH, ipaBCD, ial, sen, set1A, set1B, sat, and pic genes, and a PCR-sequencing approach on plasmid preparations was used for identifying pHS-2-specific sequences.

Overall, the virulence markers ranged in prevalence from 21% (set1A, set1B, pic) to 100% (ipaH). S. flexneri isolates displayed a higher content of virulence markers than S. sonnei, the most common genotype, detected exclusively in S. flexneri serotype 2a isolates, being defined by the association ipaH+ipaBCD+ial+sen+sat+set1A+ set1B+pic. pHS-2-like plasmids were found in S. flexneri isolates of various serotypes suggesting the potential to trigger postinfection complications in specific subjects.

This study provided baseline data regarding the molecular characteristics of the Shigella strains from Romania, useful for defining the picture of shigellosis in our region.