Category Archives: Original Research

A Standardized Dissection Protocol to Generate Aortic Valvular Scaffolds from Porcine Hearts

Ionela Movileanu1, Marius Harpa1, Klara Branzaniuc1, Horatiu Suciu1, Ovidiu S. Cotoi1, Peter Olah1, Dan Simionescu2

1 University of Medicine and Pharmacy of Tirgu Mureş, Romania
2 University of Medicine and Pharmacy of Tirgu Mureş, Romania , Biocompatibility and Tissue Regeneration Laboratories, Department of Bioengineering, Clemson University, Clemson, USA

DOI: 10.1515/amma-2017-0029

Objective: To describe a particular harvesting procedure for isolating intact porcine aortic heart valve roots as potential sources for biologic scaffolds.
Methods: Fresh porcine hearts were brought to the Tissue Engineering and Regenerative Medicine Laboratory at the University of Medicine and Pharmacy in Targu Mures. The aortic roots were extracted from the porcine hearts by anatomical dissection. For this purpose, we used a basic surgical instrument kit. This initial phase was the first step in obtaining acellular extracellular matrix as a biologic scaffold material.
Results: Aortic roots were isolated with preservation of the ascending aorta as well as the intact aortic sinus and coronaries together with the adjacent myocardial tissue and anterior leaflet of the mitral valve. This approach allowed for safe mounting of roots into mounting rings for perfusion decellularization.
Conclusions: The described procedure is a feasible protocol for obtaining intact biological valvular scaffolds from porcine hearts. Reduced requirements regarding tools and personnel underline the easiness of aortic root harvesting using this particular procedure.

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Ultrasound Guidance Versus Peripheral Neurostimulation for Brachial Plexus Block Anesthesia with Axillary Approach and Multiple Injection Technique

Alexandra Lazar, János Szederjesi, Elena Iftenie, Leonard Azamfirei

University of Medicine and Pharmacy Tîrgu Mureș, Romania

DOI: 10.1515/amma-2017-0025

Introduction: There are several approaches for brachial plexus anesthesia: supraclavicular, infraclavicular, interscalenic and axillary. Out of these, the axillary approach is considered to be the safest because of the low risk of lesioning the adjacent structures, low risk of phrenic nerve blockade or of producing an iatrogenic pneumothorax. The block can be performed by one single injection at the site, by two injections or by several injection, among each nerve of the plexus. Ultrasound was introduced in regional anesthesia since 1978, being used initially as an auxiliary method to peripheral neurostimulator.
Objectives: The evaluation of ultrasound efficiency as an auxiliary method for brachial plexus block performance, in terms of success rate, vascular punctures. The influence of obesity on performing time, total duration of the block, and success rate of brachial plexus block.
Material and method: Prospective, randomized study which enrolled adult patients, scheduled for surgical emergency or elective surgical intervention on upper limb with brachial plexus block by axillary approach, using either the peripheral nerve stimulation or the ultrasound guidance.
Results: We enrolled 160 patients, grouped in two sets- the ultrasound group= 82 patients (US) the neurostimulation group = 78 patients (NS). Vascular punctures were statistically significant different p= 0, 04. The success rate was not influenced by the obesity.
Conclusions: Ultrasound guidance makes axillary brachial plexus block safer, we can recommend ultrasound guidance as routine for axillary brachial plexus block. The obese patient can beneficiate by both methods of brachial plexus blockage.

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Assessment of Post-Occlusive Reactive Hyperaemia in the Evaluation of Endothelial Function in Patients with Lower Extremity Artery Disease

Cosmin Carasca1, Annamaria Magdas2, Ioan Tilea3, Alexandru Incze2

1 University of Medicine and Pharmacy of Tirgu Mureş, Romania
2 Department of Internal Medicine IV, University of Medicine and Pharmacy of Tirgu Mureş, Romania
3 Cardiovascular Rehabilitation Clinic, Department of Family Medicine, Institute of Cardiovascular Diseases and Transplantation Tîrgu-Mures, Romania, University of Medicine and Pharmacy of Tirgu Mureş, Romania

DOI: 10.1515/amma-2017-0024

Background: The aim was to assess endothelial function with photoplethysmography (PPG), by post-occlusive reactive hyperaemia (PORH) combined with alprostadil challenge test in patients with peripheral artery disease (PAD).
Methods: Forty-nine PAD patients stage II-III Fontaine (39 male, 10 female, mean age 68.45±5.86 years) and a control group of 49 healthy individuals (24 male, 25 female; mean age 25.1±3.8 for a young subgroup; 71.0±0.16 years for an elderly subgroup) were included. Ankle-brachial index (ABI) was assessed at baseline, peripheral perfusion (PP) and PORH were assessed at baseline and after the 30 minutes administration of parenteral alprostadil.
Results: After 3 minutes of arterial occlusion, peripheral perfusion increased from 0.69±0.94 mV/V to 2.27±2.42 mV/V (p<0.0001). After alprostadil challenge, peripheral perfusion increased from 0.84±1.24 mV/V to 4.52±3.52 mV/V (p<0.0001). In controls PP was 2.4±1.7 mV/V versus 3.8±1.5 mV/V, p<0.0001.
Conclusion: In patients with PAD, an increase in PORH after alprostadil challenge due to the release of nitric oxide (NO), provides information on the endothelial function and could reflect the presence of collaterals. In the healthy control group, the increase in PORH could reflect the integrity of main arterial branch. In PAD patients with an increase in PORH, conservative therapy should be preferred over surgical revascularisation.

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Letrozole Determination by Capillary Zone Electrophoresis and UV Spectrophotometry Methods

Aura Rusu1, Maria-Alexandra Sbanca2, Nicoleta Todoran3, Camil-Eugen Vari4

1 Department of Pharmaceutical Chemistry, University of Medicine and Pharmacy of Tîrgu Mureş
2 University of Medicine and Pharmacy of Tîrgu Mureş
3 Department of Pharmaceutical Technology, University of Medicine and Pharmacy of Tîrgu Mureş
4 Department of Pharmacology and Clinical Pharmacy, University of Medicine and Pharmacy of Tîrgu Mureş

DOI: 10.1515/amma-2017-0022

Objective: Letrozole is a highly potent oral nonsteroidal aromatase inhibitor triazole derivative. The aim of this study was to quantify letrozole from bulk, pharmaceutical formulation, and spiked urine samples by developing a simple, rapid and cost effective capillary electrophoresis method. Methods: A capillary zone electrophoresis method was optimized and validated. Additionally, an UV spectrophotometry method was used for comparing results. Results:The capillary zone electrophoresis method using a 90 mM sodium tetraborate background electrolyte proved to be an efficient method for determination of letrozole in a very short time, less than 2 minutes, using 20 kV voltage, 50 mbar/2 seconds pressure and 50°C temperature as optimum parameters. Additionally, the UV spectrophotometry method proved to be simple and efficient to quantify letrozole from bulk material and pharmaceutical formulation with linearity of response between 5 to 20 µg·mL-1 concentrations. For both methods, validation parameters, including linearity, detection and quantification limits were determined. Also we proved that our electrophoretic method has potential in analyzing letrozole from biological samples, obtaining encouraging results on estimation of letrozole from spiked urine samples without any special treatment. Conclusions: To quantify letrozole from bulk material, pharmaceutical preparations, and spiked urine samples the capillary zone electrophoresis method using a tetraborate sodium background electrolyte has proven to be simple and appropriate. Also a simple UV spectrophotometric method has been developed and validated for the same purposes.

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The Influence of CYP2D6 Phenotype on the Pharmacokinetic Profile of Atomoxetine in Caucasian Healthy Subjects

Ioana Todor1, Dana Muntean1, Maria Neag2, Corina Bocsan2, Anca Buzoianu2, Laurian Vlase1, Daniel Leucuta3, Ana-Maria Gheldiu1, Adina Popa4, Corina Briciu4

1 University of Medicine and Pharmacy “Iuliu Hatieganu”, Faculty of Pharmacy, Department of Pharmaceutical Technology and Biopharmaceutics, Cluj-Napoca, Romania
2 University of Medicine and Pharmacy “Iuliu Hatieganu”, Faculty of Medicine, Department of Pharmacology, Toxicology and Clinical Pharmacology, Cluj- Napoca, Romania
3 University of Medicine and Pharmacy “Iuliu Hatieganu”, Department of Medical Informatics and Biostatistics, Cluj-Napoca, Romania
4 University of Medicine and Pharmacy “Iuliu Hatieganu”, Faculty of Pharmacy, Department of Clinical Pharmacy, Cluj-Napoca, Romania

DOI: 10.1515/amma-2017-0023

Objective: To analyze a potential phenotypic variation within the studied group based on the pharmacokinetic profile of atomoxetine and its active metabolite, and to further investigate the impact of CYP2D6 phenotype on atomoxetine pharmacokinetics.
Methods: The study was conducted as an open-label, non-randomized clinical trial which included 43 Caucasian healthy volunteers. Each subject received a single oral dose of atomoxetine 25 mg. Subsequently, atomoxetine and 4-hydroxyatomoxetine-O-glucuronide (glucuronidated active metabolite) plasma concentrations were determined and a noncompartmental method was used to calculate the pharmacokinetic parameters of both compounds. Further on, the CYP2D6 metabolic phenotype was assessed using the area under the curve (AUC) metabolic ratio (atomoxetine/ 4-hydroxyatomoxetine-O-glucuronide) and specific statistical tests (Lilliefors (Kolgomorov-Smirnov) and Anderson-Darling test). The phenotypic differences in atomoxetine disposition were identified based on the pharmacokinetic profile of the parent drug and its metabolite.
Results: The statistical analysis revealed that the AUC metabolic ratio data set did not follow a normal distribution. As a result, two different phenotypes were identified, respectively the poor metabolizer (PM) group which included 3 individuals and the extensive metabolizer (EM) group which comprised the remaining 40 subjects. Also, it was demonstrated that the metabolic phenotype significantly influenced atomoxetine pharmacokinetics, as PMs presented a 4.5-fold higher exposure to the parent drug and a 3.2-fold lower exposure to its metabolite in comparison to EMs.
Conclusions: The pharmacokinetic and statistical analysis emphasized the existence of 2 metabolic phenotypes: EMs and PMs. Furthermore, it was proved that the interphenotype variability had a marked influence on atomoxetine pharmacokinetic profile.

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Silver Cation Coordination Study to AsW9 Ligand – A Trilacunar Arsenotungstate Compound

Lavinia Berta, Andrei Gâz, Francisc Boda, Augustin Curticapean

University of Medicine and Pharmacy of Tirgu Mures, Romania

DOI: 10.1515/amma-2017-0018

Objective: The main objective of this research is to find the coordination ratio between AsW9 and Ag+, as a preliminary study for synthesizing a new silver-arsenotungstate complex.
Material and method: The ligand:cation molar ratio in complexes was determined by conductometric and potentiometric titrations of AsW9 with silver salts: CH3COOAg, AgNO3.
Results: The ratio was obtained from the inflexion points of the curves when molar ratio was plotted versus conductivity, or from the equivalence point when silver added volume was plotted versus pH value. Each graphic shows one point of inflexion corres ponding to 1:1.54 ratio of AsW9:Ag+. In the same manner, the equivalent volumes determined by graphical method gave the ratio 1:1.53. The spectral results confirmed that a AsW9:Ag+ complex was formed since the ligand absorption maxima values have been changed from 190 nm to 197 nm in the case of using AgNO3 and 196 nm for CH3COOAg corresponding to the W=Od bond, and from 246,5 nm to 274 nm (AgNO3) and 270 nm (CH3COO-Ag+) for the W-Ob,c-W bond.
Conclusions: Silver cation exhibit a preference for AsW9 in a ratio of 3 to 2. This ratio can be associated to a sandwich type arrangement, with two trilacunary Keggin building blocks incorporating 3 metal cations in a tetrahedral geometry.

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Flow Cytometry Assessment of Bacterial and Yeast Induced Oxidative Burst in Peripheral Blood Phagocytes

Floredana-Laura Șular1,2, Minodora Dobreanu1,2

1 Discipline of Laboratory Medicine, University of Medicine and Pharmacy of Tîrgu Mureș, Romania
2 Central Laboratory, Emergency Clinical County Hospital of Tîrgu Mureș, Romania

DOI: 10.1515/amma-2017-0020

Objective: The aim of this study was to verify in our laboratory conditions the performance criteria of a commercial kit (PhagoburstTM, Glycotope Biotechnology) as described by the producers. We have also partially altered the use of the available kit by introducing a non-opsonized Candida albicans stimulus, in addition to the opsonized Escherichia coli stimulus provided by the manufacturer.
Material and methods: The peripheral blood samples of 6 clinically healthy adults were tested in triplicate according to the manufacturer recommendations. The intra-assay imprecision, as well as the ranges of neutrophil and monocyte burst activation triggered by various stimuli, were assessed.
Results: The activation range of granulocytes and monocytes was similar to the one described by the producer in the presence of E. coli (granulocytes: 78.45-99.43% versus 99.6-99.95%, average %CV of 1.53% versus 0.1%, monocytes: 54.63-92.33% versus 81.80-96.67, average %CV 6.92% versus 1.1%). The leukocyte range of activation in the presence of non-opsonized C. albicans was comparable to the one triggered by the fMLP (N-formyl-methionyl-leucyl-phenylalanine) stimulus.
Conclusion: The intra-assay precision obtained in our laboratory conditions, as well as the ranges of activated leukocytes, are comparable to the ones described by the producer when using E. coli as a stimulus. The present study shows that introducing an extra fungal stimulus for burst oxidation assessment could provide additional information regarding the non-specific cellular immune response, particularly in patients at risk for candidemia.

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Changes in Knee Joint Space Width in Treatment with a New Hyaluronic-Based Hydrogel

Octav Marius Russu, Andrei Marian Feier, Tudor Sorin Pop, Marcela Todoran, István Gergely

University of Medicine and Pharmacy Tirgu-Mures, Romania

DOI: 10.1515/amma-2017-0017

Objective: Our purpose was to assess the effect of a new hyaluronic acid-based (Hymovis®) injections on joint space width narrowing in patients diagnosed with knee osteoarthritis.
Methods: A prospective clinical trial was conducted in the Department of Orthopedics and Traumatology II from the Clinical County Hospital, Tîrgu Mureș, Romania. Thirty-five patients diagnosed with idiopathic knee osteoarthritis received two intraarticular injections with hyaluronic acid-based hydrogel (24 mg of hyaluronic acid/3 ml) at one-week interval. Anteroposterior radiographs were obtained before the injections, at six and twelve months after. Minimum joint space width was measured by two senior orthopaedics surgeons at each follow up. Each radiograph was measured again by the same evaluators two weeks apart.
Results: Thirty-one patients were present at the final follow-up. A minor reduction in mean weight was noticed (from 82.2 kg ± 16.2 kg to 80.9 kg ± 16.0, p > 0.398) without any correlation with joint space width narrowing. There were no major changes at the first follow up (6 months) regarding joint space narrowing. A reduction in joint space width was observed however at 12 months varying from 4.4 mm (SD ± 1.64, range 1.8-7.1) at the first assessment to 4.3 mm (SD ± 1.26, range 0.0-6.8) at the final follow-up but with no statistical difference (p=0.237).
Conclusion: No significant modification in joint space width at the final follow-up secondarily proved that two injections of Hymovis® may slow down narrowing in the knee joint space over a one-year period.

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Reduced Analgesics Consumption and Pain Intensity after Injections with a New Hyaluronic Acid in Patients with Knee Osteoarthritis

Octav Marius Russu, Andrei Marian Feier, Tudor Sorin Pop, Marcela Todoran, István Gergely

University of Medicine and Pharmacy Tîrgu Mures

DOI: 10.1515/amma-2017-0016

Objective: To determine the influence of a new intraarticular hyaluronic acid based hydrogel (Hymovis®) injections on the amount of analgesics consumption in patients diagnosed with primary knee OA.
Methods: A prospective, single-center study that included 35 patients, aged 45-80 years was conducted in our orthopaedics department. Patients received two intraarticular injections of hyaluronic acid (24 mg/3 ml; 500–730 kDa; Hymovis®) at one week apart. Follow-up was scheduled at 2 and 6 months after the injections. Assessment tools included Visual Analogue Scale (VAS) and an in-house designed questionnaire regarding analgesic consumption (quantity, period and product) during the follow-up.
Results: Compared to baseline, a significant amelioration in visual analogue scale was observed at six months’ follow-up (74.2mm ± 11.7 vs. 57.3mm ± 12.1; p < .0001). 28% (n=10) of the patients reduced their total analgesic consumption at two months after the injections. At final follow-up, the analgesic intake was reduced by more than 50% in almost every case.
Conclusions: Intraarticular administered injections with a novel hyaluronan-based hydrogel (Hymovis®) may reduce the amount of analgesic consumption and self-reported pain intensity in patients with knee OA.

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Evaluation of Early Postoperative Cognitive Dysfunction Incidence and Involved Neurocognitive Functions in Patients with Cardiac and Noncardiac Surgery Under General Anesthesia

Carmen Sircuţa1, Tunde Lucza2, Mihaly Veres1, Ildiko Szomoru3, Leonard Azamfirei1

1 University of Medicine and Pharmacy Tirgu Mures, Romania
2 Emergency Institute for Cardiovascular Diseases and Transplantation Tirgu Mures, Romania
3 Emergency Clinical County Hospital of Tirgu Mures, Romania

DOI: 10.1515/amma-2017-0012

Objective: To analyse postoperative cognitive dysfunction’s (POCD) incidence and cognitive areas involved, in patients with cardiac and general surgery.
Material and Methods: Prospective observational study on 130 patients undergoing general or heart surgery on cardiopulmonary bypass, under general anesthesia. Two groups, 65 members each. Group A had a heart surgery and group B a noncardiac surgery. The same type of anesthetic drugs were used. All patients completed the Montreal Cognitive Assesment (MoCA) questionnaire: preoperative, 24 hours after stopping any medicine acting on central nervous system and 7 days postoperative. We compared the MoCA scores obtained on different cognitive domains in this moments for each group of patients, for neurocognitive functions: visuo-spatial executive, naming, attention, verbal fluency, abstraction, recall, orientation, final score. We compared the scores between the two groups at 24 hours and 7 days postoperatively for the same domains.
Results: POCD was found at 24 hours testing in both groups. At 7 days postoperatively POCD was not found in any of the groups. There was no statistically significant difference in total final score between two groups at 24 hours nor at 7 days postoperative testing. There are significant differences between the two groups, with lower score in cardiac group in 5 of 7 fields at 24 hours testing, with the persistence of difference in 2 of 7 fields at 7 days.
Conclusions: Overall POCD was present at 24 hours but was not found at 7 days testing for none of the groups. POCD is present in some neurocognitive domains and this depends on surgery type.

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