Category Archives: Original Research

Study of the anxiolytic effect of propranolol and dextromethorphan in mice using a model of psychogenic stress

DOI: 10.2478/amma-2021-0034

Objective: To find the anxiolytic effects in a psychogenic stress model in mice, the present study has investigated the interaction between propranolol and dextromethorphan.
Methods: 50 Albino Swiss male mice were housed in groups of 10 per cage. The beta-adrenergic receptor blocker propranolol (20 mg/kg bw), the N-methyl-D-aspartate (NMDA) receptor dextromethorphan (30 mg/kg bw), and their combination were administered 10 minutes after exposure to predator odor. The treatments included diazepam as positive control and normal saline as negative control. Anxiety-like behaviors were evaluated using the elevated plus-maze test (EPM) 7 days after stress induction.
Results: Regarding the length of stay (F=25.53; p<0.0001)/ number of entries in the open arms (F=3.533; p=0.0416), time (F=6.127; p=0.0045)/ number of entries in the closed arms (F=5.690; p=0.0141), time in center-point (F=3.577; p=0,0295), and total distance traveled (F= 4.711; p=0.0145), there was a significant difference among the treated groups. Propranolol and dextromethorphan treated groups expressed lower time in the closed arms vs Placebo (p=0.0089, respectively p=0.0111). In addition, the time spent in the open arms was higher in propranolol group vs placebo group (p=0.0215).
Conclusions: Considering obtained data, there was a decrease of symptoms in the sympathetic nervous system and the psychological stress disappeared in mice applying a treatment of 20 mg/kg bw propranolol. Our findings indicated that dextromethorphan partially mediated the anxiolytic-like activity. However, the combination of these two drugs did not express anxiolytic effects.

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Relationship between obesity and psychosocial risk factors in hypertensive patients

DOI: 10.2478/amma-2021-0030

Introduction: Unhealthy lifestyle habits, including sedentary behaviour and improper diet are major risk factors for obesity. Overweight is very frequent in hypertensive and type 2 diabetic patients. Psychosocial factors are associated with multiple health behaviors related to overweight/obesity. The aim of the study was to evaluate the relationship between obesity and psychosocial risk factors in hypertensive patients having different levels of education.
Material and methods: 623 hypertensive patients, admitted to the Cardiovascular Rehabilitation Clinic in Tîrgu Mureș were evaluated by weight, height, waist circumference, laboratory analyses and psychosocial risk based on a validated questionnaire on nine topics. Non-paired Student t test was used for statistical evaluation of the obtained data.
Results: Obesity or overweight affected 86.9% of the studied hypertensive patients. Obesity was more frequent in subjects having type 2 diabetes as comorbidity. Mean BMI was significantly higher in the lower education group (p=0.007) compared to those with higher level of education. Average triglyceride and uric acid levels were significantly higher in obese subjects compared to those having lower BMI (p<0.0001). Patients living alone had significantly lower mean BMI (p=0.006) and waist circumference values (p=0.001) compared to those living with their spouse.
Conclusions: Weight excess in very frequent in the studied hypertensive patients, especially in those with type 2 diabetes as comorbidity and having lower educational level. Hypertriglyceridemia and hyperuricemia occurred more frequently in obese subjects compared to non-obese ones. Overweight is less frequent in patients living alone.

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The antibacterial potential of biosynthesized silver nanoparticles using beech bark and spruce bark extracts

DOI: 10.2478/amma-2021-0043

Introduction: Lately, nanotechnology focuses on the green synthesis of AgNPs, using different plant materials, as this method is accessible, cost-efficient, and ecological. The study aimed to investigate the antibacterial potential of AgNPs synthesized using beech/spruce bark extracts (BBE/SBE) and silver salts (acetate/nitrate).
Method: The growth rates of Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922 were evaluated in the presence of the AgNPs solutions. The checkerboard method was performed to evaluate if these solutions exert synergistic activity with gentamicin.
Results: For E. coli, synergistic effects were observed for the combination of gentamicin 0.25mg/L with AgNP BBE Nit (0.145mg/mL) and with AgNP SBE Ac (0,09mg/mL). For S. aureus, no synergistic effects were observed. Overall, the AgNP BBEs solutions combined with gentamicin presented lowest values of fractional inhibitory concentration than the ones registered for the combination of AgNP SBEs with gentamicin, for both bacterial strains. The growth rate of S. aureus was inhibited by all the tested AgNPs at the measured time points. For E. coli, after 24 hours of incubation, the growth rate was inhibited only in the presence of AgNP SBE Ac. After 6 hours of incubation, the growth rate of E. coli was almost stationary in the presence of AgNP BBE Nit.
Conclusions: The biosynthesis of AgNPs is a valuable choice for obtaining substances with antibacterial potential.

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E-cigarette and alternative tobacco products consumption among adolescents in Chisinau, Republic of Moldova – A repeated cross-sectional study

DOI: 10.2478/amma-2021-0039

Objective: The main objective of the study was to evaluate e-cigarette and alternative tobacco product use in ninth-grade students from Chisinau.
Methods: The repeated cross-sectional study collected data using a questionnaire administered in 2015 and 2018 among ninth-grade students in Chisinau, Republic of Moldova. Absolute and relative frequencies of electronic cigarettes and alternative tobacco products ever use were calculated and 2015 data were compared with those collected in 2018.
Results: The percentage of participants ever using electronic cigarettes in 2015 and 2018 respectively was 42.9% and 59.5% (p=0.001), ever using cigars was 14.9% and 15.9% (p=0.684), ever using pipes was 3.3% and 4.9% (p=0.205), ever using hookah was 17.4% and 21.7% (p=0.086), ever using chewing tobacco was 11.6% and 3.9% (p=0.039), ever using snus was 4.1% and 6.2% (p=0.135), and ever using snuff was 0.5% and 1.1% respectively (p=0.518). Exposure of participants to messages promoting electronic cigarettes was 31.0% in 2015 and 65.8% in 2018 (p<0.0001). Consumption of electronic cigarettes was significantly associated with sex (boys 62.8% vs. girls 43.6%), school performance (low grades 61.5% vs. high grades 50.0%), exposure to pro e-cigarette messages (exposed 61.4% vs. not exposed 44.0%), perceived e-cigarettes harmfulness (less harmful 66.9% vs. more harmful 39.7%), and use of conventional cigarettes (ever tried 79.8% vs. never tried 24.6%).
Conclusions: The study showed an important, statistically significant increase in the consumption of electronic cigarettes among ninth-grade students in Chisinau city, from 2015 to 2018 and an increased exposure to electronic cigarette advertising.

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Mediation of Candida species growth and virulence by the proinflammatory cytokine IL-6

DOI: 10.2478/amma-2021-0036

Interleukin-6 (IL-6) is a cytokine with pleiotropic effects that might also influence the virulence traits of some microorganisms, but its direct influence over Candida spp. is currently unknown. The objective of the study is to determine the influence of IL-6 (250 pg/ml) on the growth rate and biofilm formation of C. albicans, C. parapsilosis, C. krusei, C. auris, and C. guilliermondii, as well as to analyze the influence of this citokine on the expression of three virulence genes (ALS3, HSP70, SAP2), respectively on the germ tube formation ability of C. albicans. The influence of IL-6 on growth rate was assessed by incubating the fungal cells in presence of IL-6 for 48 hours and assessing the optical density of the samples at five timepoints. The biofilm production in presence of IL-6 was studied in microtiter plates, using crystal-violet assay, the intensity of biofilms being evaluated by spectrophotometry. The expression of ALS3, HSP70, and SAP2 in C. albicans was studied by RT-PCR, reported to ACT1 housekeeping gene. The germ-tube test was performed to assess the influence of IL-6 on the filamentation rate of C. albicans. All test results were normalized against control, without added IL-6. The results showed that IL-6 influences the growth rate of C. albicans, C. parapsilosis, C. krusei, C. auris, and C. guilliermondii in a time-dependent way. Also, IL-6 inhibited the formation of biofilms for C. albicans and C. guilliermondii. In C. albicans cells, IL-6 induced upregulation of ALS3 and HSP70, while it down-regulated the SAP2 gene. IL-6 did not influence the germ-tube formation in C. albicans. In conclusion, IL-6 might exert, in vitro, direct effects on the virulence traits of Candida spp., and its influence is dependent on the exposure time. Non-albicans Candida species presented particular responses to IL-6.

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Formulation and evaluation of controlled release ocusert of gatifloxacin and prednisolone

DOI: 10.2478/amma-2021-0032

Objective: Ocuserts are sterile, solid, or semisolid dosage forms prepared to attain increased contact time between the drug and the conjunctival tissue to keep up a constant release of drug when placed in the lower cul-de-sac or conjunctival sac of the eye. The aim of formulating this delivery system was to treat both inflammatory and infectious conditions of the eye with increased ocular residence time by releasing drugs at a slow and constant rate.
Method: Gatifloxacin, and prednisolone ocuserts were prepared by solvent casting method, and evaluated for physical appearance, uniformity of weight, thickness, folding endurance, drug content, surface pH, in-vitro, and ex-vivo release profile.
Results: All formulated inserts exhibited positive results in terms of their evaluation parameters. Ocuserts were sterile, with no turbidity in selected media during the study, and they were stable throughout six months.
Conclusion: Results suggested that prepared optimized ocusert formulation would be a suitable alternative to eye drops for treating conjunctivitis and other bacterial infections.

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Online pharmacy: customer profiling

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

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

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

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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