Category Archives: AMM 2018, Volume 64, Number 2

Emergency Technology at Our Reach Might Make a Change – Instead of an Editorial

DOI: -

A recent study published in the Chest showed that often enough community-acquired pneumonia was associated with a clear chest radiograph (1). In 3% of the case, the clinical profile of these patients with CT-only pneumonia did not differ from those with pulmonary infiltrates on chest radiograph also in terms of comorbidities, vital signs, and length of stay, etiology, mechanical ventilation and admission in the Intensive Care Unit (ICU), septic shock or inhospital mortality. Often enough critically ill patients need CT scans for diagnostic purposes or reassessments. To comply with the indications for CT scans, the patients must be transported in remote locations, or several floors down. Transport of the critically ill is by no means a trifle, for it could jeopardize the outcome. When the risks associated with transportation overcome the benefits, alternative methods of diagnoses, equally efficient need to be used. Technology helps a lot. Thoracic bioimpedance CT scans at the bedside may be the answer for patients who cannot wait for time-consuming investigations, who need a better monitoring of their respiration at the bedside, who could benefit from in depth monitoring of their respiratory pressions, an assessment of their work of breathing (WOB), or a change of their treatment. [More]

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Characterization of Inclusion Complexes between Miconazole and Different Cyclodextrin Derivatives

DOI: 10.2478/amma-2018-0012

Objective: Miconazole, an imidazole antifungal derivative, is a very hydrophobic compound, a major drawback in obtaining topical pharmaceutical formulations with optimal bioavailability. Cyclodextrins (CDs) may increase local drug delivery by enhancing the drug release and/or permeation. The aim of the study is the characterization of inclusion complexes between miconazole and different CD derivatives.
Methods: Several CD derivatives were tested in the experiments. The binary systems between miconazole and different CDs were prepared in 1:1 molar ratios by physical-mixture and kneading methods. Differential scanning calorimetry (DSC) and Fourier transformed-infrared spectroscopy (FT-IR) methods were used to characterize solid state interactions between miconazole and CDs in their binary systems.
Results: The FT-IR analysis suggests the formation of a new solid phase, indicating a molecular interaction between the components. The DSC analysis sustains the hypothesis of formation of partial inclusion complexes between miconazole nitrate and CD. Conclusion: The thermic behaviour of the complexes depends both on the preparation method and the composition of the products.

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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.

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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.

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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.

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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.

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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.

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