Category Archives: Volume 64

Phenolic Content from Medicinal Plants and their Products Used in Veterinary Medicine

DOI: 10.2478/amma-2018-0013

Objective: The aim of the study was to determine the content of polyphenols and flavonoids from sixteen selected medicinal plants from the spontaneous Romanian flora and fifteen tinctures obtained with propylene glycol.
Methods: The polyphenols were determined by the Folin-Ciocalteu method while the flavonoids by using a colorimetric method from the 10th edition of the Romanian Pharmacopoeia. The antioxidant activities of the most common nine medicinal plants and fifteen tinctures were determined by DPPH and ABTS methods.
Results: The results highlighted that the phenolic compounds and flavonoids have contributed to their antioxidant activities and the medicinal plants and tinctures included in the study are rich sources of natural antioxidants.
Conclusions: There are a wide variety of extraction methods for the determination of phenolics and flavonoids. The study confirms a correlation between phenolic and flavonoid contents obtained by using the DPPH and ABTS tests.

Full text: PDF

Optimization of a Density Gradient Centrifugation Protocol for Isolation of Peripheral Blood Mononuclear Cells

DOI: 10.2478/amma-2018-0011

Objective: Peripheral blood mononuclear cells (PBMC) are extremely important in the body’s immune response. Their isolation represents a major step in many immunological experiments. In this two phase study, we aimed to establish an optimum protocol for PBMC isolation by density-gradient centrifugation.
Methods: During Phase-1, we compared two commercially available PBMC isolation protocols, Stemcell Technologies (ST) and Miltenyi Biotec (MB), in terms of PBMC recovery and purity. Twelve blood samples were assigned to each protocol. Each sample was divided in three subsamples of 1ml, 2ml and 3ml in order to assess the influence of blood sample volume on isolation performance. During Phase-2, a hybrid protocol was similarly tested, processing six blood samples. Additionally, we performed a flow cytometric analysis using an Annexin-V/Propidium-Iodide viability staining protocol.
Results: Phase-1 results showed that, for all subsample volumes, ST had superior PBMC recovery (mean values: 56%, 80% and 87%, respectively) compared to MB (mean values: 39%, 54% and 43%, respectively). However, platelet removal was significantly higher for MB (mean value of 96.8%) than for ST (mean value of 75.2%). Regarding granulocyte/erythrocyte contamination, both protocols performed similarly, yielding high purity PBMC (mean values: 97.3% for ST and 95.8% for MB). During Phase-2, our hybrid protocol yielded comparable results to MB, with an average viability of 89.4% for lymphocytes and 16.9% for monocytes.
Conclusions: ST yields higher cell recovery rates and MB excels at platelet removal, while the hybrid protocol is highly similar to MB. Both cell recovery and viability increase with blood sample volume.

Full text: PDF

Aquaporin 2 Expression in Human Fetal Kidney Development

DOI: 10.2478/amma-2018-0010

Introduction: The metanephrogenic zone, renal cortex and renal pyramids develop into their final form by week 13. The metanephric kidney produces large quantities of diluted urine in order to maintain volumes of amniotic fluid. Aquaporins are transmembrane protein channels that enable water transport through biological membranes. Aquaporin 2 (AQP2) is a water channel found in the supranuclear region and apical area of the cell membrane of the kidneys collecting tubule cells. Its main function is reabsorption of water through vasopressin stimulation.
Materials and methods: Immunohistochemistry was used to study fetal renal tissue of 34 post-mortem fetuses of 9 weeks to 24 weeks gestational age.
Results: AQP2 expression is present in connecting tubules and collecting tubules during the targeted time period. From week 9 to 12, the expression is cytoplasmic. From week 13 to 20 the enhancement of expression in the apical cell membrane occurs with the advancement of fetal age. At the end of the studied period, from week 21 to 24, both cytoplasmic and apical expression were observed. In animal studies AQP2 expression has an increasing trend during development. In contradiction with these results, other authors described low AQP2 levels in the human fetal kidney.
Conclusions: This study helps to understand the amniotic fluid’s homeostasis during pregnancy. In the beginning of the fetal period AQP2 protein is present in the cytoplasm of epithelial cells of the collecting duct and distal connecting duct. During the fetal period, AQP2 expression in collecting ducts becomes more enhanced in the apical membrane of the cells.

Full text: PDF

MTHFR – Ala222Val Effects on Metabolic Syndrome Progression

DOI: 10.2478/amma-2018-0009

Objective: Methylene-tetrahydrofolate reductase (MTHFR) is involved in adapting metabolism to environmental challenges by various mechanisms, including the control of gene expression by epigenetic and post-translational changes of transcription factors. Though a metabolic syndrome candidate gene, association studies of its common polymorphism rs1801133 (MTHFR-Ala222Val) remain inconclusive with important ethnic differences, and the effect on disease progression was not addressed.
Methods: 307 middle-aged metabolic syndrome patients in a central Romanian hospital setting were investigated metabolically, and genotyped by PCR-RFLP. Disease progression was assessed by the age of onset of metabolic components, as well as development of non-alcoholic fatty liver disease and atherosclerotic complications.
Results: The minor allele frequency of rs1801133 was 30.13%. Metabolic parameters showed no statistically significant differences according to genotype, but variant carriers developed dysglycemia and dyslipidemia earlier (53.28±10.8 vs 59.44±9.31 years, p<0.05 and 58.57±11.31 vs 64.72±10.6 years, p<0.1).While the polymorphism did not influence hepatic complications, an inverse association was found for manifest atherosclerosis (OR=0.49, p=0.006, 95%CI:0.29-0.81), which may be folate-status dependent, and needs further investigations. Simultaneous analysis with transcription factor polymorphisms (rs1801282, rs8192678) showed that the more protective genotypes were present the later metabolic disturbances developed, and in the presence of the other two variants the apparent protective cardiovascular effect disappeared.
Conclusions: The common functional polymorphism rs1801133 may influence metabolic syndrome progression, the age of onset of components and development of atherosclerotic complications. Besides simple additive effects, complex mitigating and aggravating variant interactions may exist, and the protective or predisposing outcome may depend on modifiable environmental factors.

Full text: PDF

The Value of Perioperative ECG Recordings in the Cardiovascular Monitoring and Risk Assessment of Asymptomatic Patients with Total Hip Arthroplasty

DOI: 10.2478/amma-2018-0008

Aim: To evaluate the value of a more closed perioperative ECG follow-up in the investigation and forecasting of cardiovascular events during and after hospital admission.
Methods: We included in the study 30 asymptomatic patients from cardiac point of view (14 women, 16 men, mean age 63 years), who underwent total hip arthroplasty. ECGs were performed on the day before surgery, on the day of surgery, after the intervention, and on the 3rd day. We looked for the correlations between ECG changes and clinical events.
Results: The commonest postoperative new ECG changes were QTc prolongation (14 patients – 46,6%) and sinus tachycardia (8 patients – 26,6%). There were no major cardiac events during hospitalization, and we found possible correlations in two cases between perioperative ST-T changes and postdischarge clinical events: one exitus and one case of ischemic heart disease.
Conclusions: Our data do not support the efficacy of a more closed, routine ECG follow-up in the studied, low risk patient population, but new perioperative ST-T changes always have to be taken into account.

Full text: PDF

Erratum

Due to an editing error of the authors, the article „Laparoscopic Greater Curvature Plication for Morbid Obesity: Indications, Results, Perspectives” published in Acta Medica Marisiensis, 2015;61(2):142-144 (DOI: 10.1515/amma-2015-0041) was published without the financial support statement. By the request of corresponding author Borz C, we publish the following erratum:
Disclosure. None of the authors reported conflict of interest regarding this study. The study was financially supported by the University of Medicine and Pharmacy Tirgu Mureș, grant no. 8918/2015.

Full text: PDF

Mediators of Inflammation as a Link Between Diabetes Mellitus and Periodontal Breakdown

DOI: 10.2478/amma-2018-0005

Our objective was to investigate immunological changes that occur in saliva of subjects with type 2 diabetes mellitus (T2DM) without signs of periodontal disease and to establish if salivary inflammatory cytokines are a possible link between diabetes mellitus and periodontal breakdown. Material and methods. Twenty T2DM subjects with no periodontal disease and twenty healthy controls were registered for the present study. TNF-α and IL-6 level from saliva and serum were measured. Periodontal tissue samples were histologically examined.

Results: TNF-α and IL-6 levels were higher in T2DM subjects compared to controls, with an extremely significant difference in saliva (p<0.001). Significant inflammation, affecting both epithelial and connective tissues was present in periodontal biopsies. Conclusions: The subjects showed an increased TNF-α and IL-6 levels, both in serum and -mostly in -saliva of diabetics without signs of periodontal disease, confirming the hypothesis of immunological implication, as a correlation between periodontal disease incidence and diabetes mellitus. Histologic alterations, suggesting a local inflammatory state, were present in periodontal tissue of diabetics, confirming the above hypothesis. The study reveals that saliva analysis is a quite efficient method in testing the periodontal breakdown progression in the subjects with T2DM.

Full text: PDF

A winter song in the black forest

At the beginning of a new year, the editors decided to publish an article dedicated to the emergency rescue system in Romania. It is basically not only a brief recall of the history of the airborne medical system in Romania, but also an update on the assets and the performances of the medicalized helicopters. Twenty-eight years following the establishment of the SMURD in Romania, and fifteen years from the birth of the helicopter fleet in Romania, airborne crews experienced the leapfrog from debutants to experienced, consolidated medical teams. [More]

Full text: PDF

Emergency Air Rescue System in Romania

DOI: 10.2478/amma-2018-0007

The helicopter, as a means of transport, has facilitated a significant decrease in intervention time at the site of request, increasing the chances of survival of the critical patient. Since 2003, SMURD has managed to form a fleet composed of nine helicopters and two airplanes. From an operational and strategic point of view, the SMURD intervention unit, set up seven Aeromedical Operational Bases (A.O.B.) equipped with helicopters and materials necessary for their operation. There is a dynamic increase in the number of air rescue missions in Romania, with most missions being carried out by the air rescue bases in Târgu Mureş and Bucharest. Specialty literature has clearly demonstrated the positive impact on the survival of critical patients assisted by airborne crews, so it is necessary for the Romanian air rescue system to grow up. It is necessary to increase the number of air bases, purchase new helicopters and to continue the training programs of both pilots and medical personnel.

Full text: PDF