Category Archives: Original Research

Online pharmacy: customer profiling

Marius Călin Cherecheș1, Cristian Olimpiu Popa2

1. Faculty of Pharmacy, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
2. Faculty of European Studies, Babeș-Bolyai University, Cluj-Napoca, Romania

DOI: 10.2478/amma-2021-0031

Introduction: Nearly three out of four European citizens purchase goods online. The main advantages of using an online pharmacy are lower prices and convenience. In addition, patients appreciate a higher degree of privacy and access to a wider selection of health products. Study shows that the number of online users is on the rise in major countries like the USA, EU member states or India.
Methods: The study is based on data collected in April 2020 at the country level (Romania) from a sample of 789 adults over the age of 18, using CATI. A simple random sampling procedure was applied, with a confidence interval of 3.5% and a confidence level of 95%.
Results: About 17.7% of Romanians have purchased healthcare products through online pharmacies or drugstores, of those 60% are women, between 30 – 64 years old (84 %), have a monthly income of more than 2700 lei (90%), and a university degree (76%). The most common reasons for choosing an online pharmacy are the ability to compare prices, shop quickly, and have prompt access to items and information, together with a wider range of products.
Conclusions: Respondents’ motivation to use online pharmacies appears to be more related to the benefits of the online media, rather than to some unmet needs in brick-and-mortar pharmacies. Search capabilities, access to more information, price comparison, as well as a larger range of products and availability 24/7 are some reasons why patients utilise online pharmacies to purchase medicines.

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Breastfeeding and diversification attitudes among Romanian mothers

Anastasia Simion1, Maria Simion2, Geanina Moldovan3

1. County Emergency Clinical Hospital Targu Mures, Romania
2. George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
3. Department of Community Nutrition and Food Safety, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania

DOI: 10.2478/amma-2021-0029

Introduction: Recommended by the World Health Organization as the optimal way of infant feeding, maternal breast milk represents the best nourishment for the newborn baby during its first six months. The purpose of this study was to evaluate some of the Romanian mothers’ characteristics that can influence their attitude towards breastfeeding and food diversification.
Methods: A questionnaire about 32 questions, including demographics items and breastfeeding attitudes, was sent online (socializing platforms) in 2020 to mothers from all Romania districts. Our sample included at the end 1768 subjects, who fully completed the questionnaire. Statistical analysis was carried out using the GraphPad statistical software.
Results: The prevalence of breastfeeding for more than six months was only 32.18% in our group, and most of them were educated mothers who lives in urban areas (OR=2.76), were married (OR=1.98), had over 30 years old (OR=1.43) and have more than one child (OR=1.74).
Conclusions: We underline the importance of tackling in our future community interventions some of the socio-demographic characteristics of pregnant women (like groups education, good and accessible information about breastfeeding, young age, first pregnancy, or mothers from rural areas as well) in developing good habits of breastfeeding or complementary feeding, in order to improve their children health status and proper development.

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The benefits of manual therapy and active cervical exercises in pacients with cerviogenic headache

Manuela Lalu1, Petru Mihancea1, Olivia Andreea Marcu2

1. Doctoral School, University of Oradea, Romania
2. Preclinics department, Faculty of Medicine and Pharmacy, University of Oradea, Romania

DOI: 10.2478/amma-2021-0022

Introduction: Cervicogenic headache stems from a dysfunctional mechanism that frequently affects the upper region of the cervical spine, often involving multiple tissues connected with the occipital, submandibular, and orofacial region.
Objectives: The main objective of this study is to demonstrate the therapeutic effectiveness of a personalized program of manual therapy and specific exercises in patients with cervicogenic headaches.
Material and Methods: The study included 44 patients. Specific functional tests of the cranio-cervical-mandibular complex, namely the evaluation of the cranio-cervical-mandibular complex (according to Rocabado), cervical flexion-rotation test, cervical flexor endurance test, cervical extensors endurance test, and palpation of trigger points and cervical spine dynamics, were employed.
Results: The analysis of demographic data shows a significantly increased prevalence of headache among women. The symptomatology that almost always accompanies episodes of cervicogenic headache is localized neck pain. The intensity and frequency of the symptoms are strongly associated with alterations in the dynamics of the upper cervical spine and chronic sleep disorders.
Conclusion: Cranio-cervical manual therapy has proven effective in patients with cervicogenic headaches. The implementation of a strategy based on manual therapy and active exercise had a superior statistical and clinical result compared to the exclusively manual therapeutic approach.

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Vascular injury in macroscopically normal skin of patients with severe COVID-19 infection: clinical-pathologic correlations

Mircea-Catălin Coșarcă, Vasile Adrian Mureșan, Emoke Horvath, Eliza Russu, Janos Szederjesi, Matild Keresztes

George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania

DOI: 10.2478/amma-2021-0027

Objectives: Taking into account that the documentation of the histopathological features in severe disease caused by SARS-CoV-2 has been scarce due to the avoidance of performing autopsies, the aim of the study was to detect the microscopic changes associated with severe COVID-19 infection in normal-appearing skin, without prominent dermatologic signs of a generalized microvascular thrombotic disorder, in accordance with the clinical evolution of disease.
Methods: In this morphological and immunohistochemical study we included cutaneous biopsy samples from 12 symptomatic patients with severe and critical type SARS-CoV-2 infection (with the admission date between February and June 2020), treated in the Intensive Therapy Unit Care of Emergency County Hospital Targu-Mures, Romania.
Results: The average age of our patients was 65.18 ± 14.21 years (range 41 to 83), and 66.67% of the patients were male. The histological and immunohistochemical assessment of cutaneous biopsies: in 4 cases the histological examination revealed small fibrin thrombi in deep-seated venules and small veins of subcuticular adipose tissue, and also 4 cutaneous biopsies showed occlusive vascular thrombosis in association with massive perivascular inflammatory infiltrate destroying and compromising the integrity of the vessel wall. The immunohistochemical examination of the composition of perivascular inflammatory infiltrate showed a predominance of CD3 positive lymphocytes, admixed with CD68 positive Mo/MF, some of them activated with FXIII expression. In the perivascular infiltrate, the presence of granulocytes and B lymphocytes was not characteristic.
Conclusion: According to our observations, in severe COVID-19, the cutaneous tissue is involved even in the absence of clinically obvious changes. Due to the relatively easy accessibility of skin samples, these could be applied to determine the severity of the patient’s clinical status, and to predict the necessity for anti-complement or anticoagulant treatments in the early stages of a severe SARS-CoV-2 infection.

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The utility of MLPA in Familial Hypercholesterolemia diagnosis

George Valeriu Moldovan1, Adina Huțanu2, Liliana Demian3, Laszlo Hadadi4, Bogdan Mănescu2, Minodora Dobreanu2

1. Microbiology Department, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
2. Laboratory Medicine Department, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
3. Center for Advanced Medical and Pharmaceutical Research, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
4. Emergency Institute for Cardiovascular Diseases and Transplantation Targu Mures, Romania

DOI: 10.2478/amma-2021-0021

Background: Familial Hypercholesterolemia (FH) is an inherited disease, associated with an increased risk of atherosclerosis, manifested clinically as premature coronary heart disease. FH is biochemically characterized by increased Cholesterol and Low-density Lipoprotein Cholesterol serum levels. The diagnosis is often made using clinical scores however, the definitive FH diagnosis should point out the underlying molecular change, which can be: a point mutation within the three major genes, a number of single nucleotide polymorphisms determining the polygenic etiology, or copy number variations in the Low-density lipoprotein receptor gene.
Objective: In the present study we investigated copy number variations as a possible etiological factor for FH in a cohort of patients with documented premature coronary heart disease.
Methods: The study population consisted of 150 patients with premature coronary heart disease documented by angiography, all being under lipid-lowering therapy, and 20 apparently healthy controls. Serum lipids were assessed using the Cobas Integra 400 plus and commercial reagents. Copy number variations were evaluated with the SALSA MLPA Probemix P062 LDLR kit.
Results: Cholesterol, Triglycerides, Low-density Lipoprotein Cholesterol and High-density Lipoprotein Cholesterol showed no difference between patients and controls. No copy number variations were detected in the investigated regions, namely all 18 exons and the promoter region of the Low-density lipoprotein receptor gene.
Conclusions: Even in the presence of negative results, the Familial Hypercholesterolemia genetic diagnosis has to be further pursued in the presence of a clinical diagnosis, as the identification of the molecular etiology may bring additional clinical and therapeutical benefits, as well as open the possibility for “cascade screening”.

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Cytokine production in ex-vivo stimulated fresh and cryopreserved T-cells

Monica Vuță1, Ionela-Maria Cotoi1, Ion Bogdan Mănescu2, Doina Ramona Manu3, Minodora Dobreanu1,2,3

1. George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
2. County Emergency Clinical Hospital of Targu Mureș, Romania
3. Center for Advanced Medical and Pharmaceutical Research, Targu-Mures, Romania

DOI: 10.2478/amma-2021-0012

Objective: In vitro cytokine production by peripheral blood mononuclear cells (PBMCs) is an important and reliable measure of immunocompetence. PBMC can be stimulated directly after isolation or frozen for later use. However, cryopreservation may affect cell recovery, viability and functionality. This study aims to investigate cytokine synthesis in ex-vivo stimulated fresh and cryopreserved CD4+ and CD4- T cells.
Methods: PBMCs were obtained by Ficoll gradient centrifugation from heparinized peripheral blood of 6 middle-aged clinically healthy subjects. Half of these cells (labeled “Fresh”) was further processed and the other half (labeled “Cryo”) was cryopreserved at -140°C for up to 3 months. Fresh-PBMCs were activated with Phorbol-Myristate-Acetate/Ionomycin/Monensin for 5 hours immediately after isolation while Cryo-PBMCs were identically activated after thawing and cell resting. Activated cells were fixed, permeabilized and intracellular cytokine staining was performed using Phycoerythrin (PE)-conjugated antibodies for Interleukin-2 (IL-2), Tumor Necrosis Factor-alpha (TNF-a), and Interferon-gamma (IFN-g). All samples were analyzed within 24 hours by flow cytometry.
Results: Both Fresh and Cryo CD3+CD4+/CD3+CD4- subpopulations partially produced each of the three cytokines. A higher percentage of CD4+ T cells produced IL-2 and TNF-a and a greater percentage of CD4- T cells were found to produce IFN-g. A significantly higher percentage of Cryo-lymphocytes was shown to produce TNF-a in both CD3+CD4+ (31.4% vs 24.9%, p=0.031) and CD3+CD4- (22.7% vs 17.9%, p=0.031) subpopulations. No notable difference was found for IL-2 and IFN-g production between Fresh and Cryo T cells.
Conclusion: Cryopreservation for up to 3 months significantly increases TNF-a production of T-cells in clinically healthy middle-aged subjects.

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Evaluation of arterial stiffness in systolic heart failure

Lajos Fehérvári1, István Adorján Szabó2, Lóránd Kocsis1, Attila Frigy3

1. Department of Cardiology, Clinical County Hospital Mures, Targu Mures, Romania
2. Department of Pathophysiology, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
3. Department of Internal Medicine IV, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania

DOI: 10.2478/amma-2021-0009

Objective: Micro- and macrovascular changes can occur in heart failure, and could influence its prognosis and management. In a prospective study, we proposed the evaluation of arterial stiffness (macrovascular function) and its correlations in patients with systolic heart failure.
Methods: 40 patients (32 men, 8 women, mean age 63±2.9 years), with hemodynamically stable systolic heart failure (left ventricular ejection fraction, EF<40%) were enrolled in the study. In every patient, beyond routine explorations (ECG, cardiac and carotid ultrasound, laboratory measurements), arterial stiffness was assessed by measuring pulse wave velocity (PWV). The correlations of PWV with clinical and echocardiographic characteristics were studied using t-test and chi-square test (p<0.05 being considered for statistical significance).
Results: The average PWV was 8.55±2.2 m/s, and 16 patients had increased PWV (>10 m/s). We found significantly higher PWV values in patients older than 65 years (p<0.001), in patients with eGFR <60 ml/min/1.73 m2 (p<0.001), hypertension (p=0.006), and increased (>1 mm) carotid intima-media thickness (p=0.016). PWV was found to be significantly lower when EF was <30% (p=0.049). Furthermore, the presence of an increased PWV was correlated significantly with age (p<0.001), and (with borderline significance) with eGFR <60 ml/min/1.73 m2 and, inversely, with EF<30%.
Conclusions: Increased arterial stiffness reflected by high PWV is frequently present in patients with systolic heart failure, and is mainly correlated with general risk factors of arterial involvement. Low EF, due to low stroke volume and decreased systolic arterial wall tension can influence the values and the interpretation of PWV.

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Formulation and Evaluation of Sustained Release Ibuprofen Matrix Tablets Using Starch from Maize Genotypes as Polymer

Lateef Gbenga Bakre1, Emmanuel Akinyele1, Oluyemisi Bamiro1, Olutayo Adeleye1, Olobayo Kunle2

1. Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmacy, Olabisi Onabanjo University, Nigeria
2. Department of Pharmaceutical Technology and Raw Materials Development, National Institute for Pharmaceutical Research and Development, Nigeria

DOI: 10.2478/amma-2021-0018

Objective: Maize plants have been genetically engineered to produce genotypes with agriculturally desirable traits such as high starch content, pest resistance and increased nutritional value. Maize starch has been widely used as an excipient in pharmaceutical formulations. This study aims to produce sustained release ibuprofen tablets using starch obtained from different maize genotypes as polymers.
Methods: Ibuprofen matrix tablets were prepared with the starches isolated from the maize genotypes and the unmodified plant. The mechanical properties of the tablets were evaluated using the crushing strength (CS), friability (FR) and CSFR. A 32 factorial design was applied using the time taken for 50 % (T50) and 90 % (T90) drug release as dependent variables while the polymer-drug ratio and polymer types were the independent variables.
Results: The CSFR was significantly higher (p<0.05) in tablets formulated with the starches obtained from the modified cultivars. Drug release for all the formulations fitted the Higuchi model while the mechanism of release was generally by super case transport. The polymer-drug ratio and polymer type strongly interacted to increase the dissolution times (T50 and T90) and CSFR. Starches isolated from the genetically modified cultivars provided a more sustained release of ibuprofen from the tablet matrix through erosion and polymer relaxation.
Conclusion: The results indicate that the genetic modification of maize can quantitatively affect the drug release modifying effects of maize starch in drug formulation.

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Beneficial effects of metformin on haloperidol-induced motor deficits in rats. A behavioral assessment

George Jîtcă1,2, Zsolt Gáll1, Camil E Vari1, Bianca E Ősz1, Amelia Tero-Vescan3, Alexandra Groșan1, Maria T. Dogaru1

1. Department of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
2. Doctoral School of Medicine and Pharmacy, I.O.S.U.D., George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
3. Department of Biochemistry, Faculty of Pharmacy, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania

DOI: 10.2478/amma-2021-0017

Objective: One of the most common side effects of haloperidol is the extrapyramidal syndrome, resulting from inhibition of nigrostriatal dopaminergic circuits and mitochondrial dysfunction due to structural similarities to pyridinium derivative, MPP+ that induce oxidative stress. In exchange, the use of metformin appears to enhance neurogenesis, energy metabolism, and oxidative status, so these properties can be speculated in the context of drug-induced pseudoparkinsonism by haloperidol.
Methods: To assess motor coordination and activity, rodents were divided into four groups: CTR (n = 10) – animals that received distilled water, METF (n = 10) – animals that received metformin 500 mg / kgbw, HAL (n = 10) – animals that received haloperidol 2mg / kgbw, HALMETF (n = 10) – haloperidol 2mg / kgbw and metformin 500 mg / kgbw. The treatment was administered for 34 days at the same time by gastric gavage, during which time behavioral tests, rotarod (days 7, 14, 21, 28), catalepsy (day 30), open field (day 32) and novel object recognition (day 34) were performed.
Results: The monitored parameters, showed significant differences between the groups of interest (HAL and HALMETF respectively), so that the administration of metformin at the beginning of treatment reduces the cataleptic behavior. The HALMETF group shows an attenuation of the motor deficit during the rotarod test and the freezing period from the Open Field test, is diminished.
Conclusions: Metformin treatment has a beneficial effect in haloperidol-treated rats, demonstrated by decreased cataleptic behavior, improved motor performance and reduced haloperidol-induced anxiety behavior.

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High-risk morphological features are less prevalent among small (<5mm) papillary thyroid microcarcinomas compared to larger (≥5mm) tumors: a study of 206 cases

Emőke – Andrea Szász1, Adela Nechifor-Boilă1,2, Ancuța Zahan1, Angela Borda1,3

1. Department of Histology, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
2. Department of Pathology, Targu Mures County Hospital, Targu­ Mures, Romania
3. Department of Pathology, Targu Mures Emergency County Hospital, Targu Mures, Romania

DOI: 10.2478/amma-2021-0016

Introduction: Papillary thyroid microcarcinoma (PTMC) is defined as a PTC measuring 1 cm or less, incidentally discovered. The aim of this study was to determine whether small (<5mm) tumors by contrast with large (≥5mm) ones are less frequently associated with high-risk morphological features, predictive of tumor aggressiveness.
Materials and methods: All consecutive PTMC cases registered at the Department of Pathology, Târgu-Mureş Emergency County Hospital between 2003-2014 were reviewed. The following have been assessed: tumor size, subcapsular versus nonsubcapsular location, extrathyroidal extension/invasion into the perithyroidal adipose tissue, multifocality, resection margins, lymph node involvement, histological variant, tumor border, stromal reaction (fibrosis/desmoplasia/sclerosis), presence of plump pink cells, nuclear features of the tumor cells, intratumoral lymphocytic infiltrate, multinucleated giant cells, psammoma bodies and stromal calcification. The cases were split in two categories: small (< 5mm) and large (≥ 5mm) PTMCs and the pathological features were evaluated in comparison.
Results: Our study included 206 cases, 91 large and 115 small PTMCs, respectively. Large PTMCs were significantly associated with the presence of plump pink cells (p=0.002), well developed PTC nuclear features (p=0.003), stromal reaction (fibrosis/desmoplasia/sclerosis) (p<0.001), infiltrative tumor border (p=0.011), subcapsular location (p<0.001), positive resection margins (p=0.022), stromal calcifications (p<0.001) and intratumoral multinucleated giant cells (p<0.001). Small PTMCs were generally well circumscribed and nonsubcapsular.
Conclusions: Our results have shown that small (<5mm) PTMCs are less frequently associated with high-risk morphological features, predictive of tumor aggressiveness compared with large (≥5mm) tumors and could thus be considered as low-risk cancers.

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