Category Archives: Original Research

Comparative analysis of mandibular changes after orthodontic treatment with and without extraction of four premolars – A digital cephalometric study

DOI: 10.2478/amma-2022-0012

Objective: To compare and evaluate the cephalometric hard and soft tissue changes of mandible after orthodontic treatment with and without extraction of four first premolar.
Methodology: 60 individuals with Class I skeletal pattern were selected as per inclusion criteria. They were divided into two groups: 30 in the extraction group and 30 in the non-extraction group. Each group consists of 15 males and 15 females. Pre-treatment and post-treatment lateral cephalograms of the selected subjects were obtained, analysed and compared to assess the changes might have occurred. Descriptive statistics of the explanatory and outcome variables were calculated by mean and standard deviation for quantitative variables. Paired t test was applied to check the statistical difference of pre-treatment and post-treatment cephalometric parameters within the group (Extraction and non-extraction). Unpaired t test was used to check the statistical difference between extraction and non-extraction groups. The level of significance is set at 5%.
Results: B point moved backward with lower incisor retraction. N-A-Pog shows a statistically significant decrease in the extraction group (P=0.003). B-NP shows a statistically significant decrease in extraction group (P=0.001). N- A’-Pog’ shows a statistically significant increase in non-extraction group (P=0.046). Soft tissue thickness in chin increases in extraction group and decreases in non-extraction group.
Conclusion: The results indicate that premolar extraction will have a flattening effect on the midface. With proper diagnosis and treatment planning, premolar extractions have high potential to improve the facial profile.

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Ethanolic Extract of Edamame (Glycine max L. merril) Enhance Second Degree Burn Wound Healing trough Modulating of Hydroxiproline Levels and Increasing Epithelial Thickness

DOI: 10.2478/amma-2022-0007

Objective: Burns has become a global health problem because it causes about 180.000 deaths worldwide every year. In developing countries, silver sulfadiazine cream is usually used as drug management for second-degree burns, but it is expensive, so several herbal treatments have developed recently. The seed of edamame has genistein compounds that can increase collagen synthesis. The antioxidant component also enhances the burn wound healing process. The study aims to prove the effectiveness of ethanolic extract of edamame’s seed in the second-degree burn wound healing process.
Methods: Burn wound was made by applying hot aluminum (70 °C) coin on the skin rat. The negative control group (C-) was given Na-CMC 0.5%, the positive control group (C+) was given cream silver sulfadiazine, the treatment groups T1, T2, T3, and T4 were given an ethanolic extract of edamame with the concentration of 20%, 40%, 60%, and 80% for 15 days. Hydroxyproline levels were evaluated by hydroxyproline biochemistry assay, the epithelial thickness was observed on histopathological preparations with HE staining.
Results: The results showed that the hydroxyproline levels on the 16th were modulated, while epithelial thickness was higher in the treatment group than in the control group (P<0.05).
Conclusions: We conclude that the certain concentration of ethanolic extract of edamame (Glycine max L. Merril) was effectively enhanced the second-degree burn wound healing by modulating hydroxyproline levels and increasing epithelial thickness.

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The influence of transport condition and processing time on plasma ammonia results

DOI: 10.2478/amma-2022-0005

Objective: Ammonia is extremely unstable in blood specimens and has special requirements during transport, processing and storage. The aim of our study was to determine the stability of ammonia in EDTA K3 blood samples and to establish a protocol for sample handling.
Methods: In this study, 36 healthy subjects and 47 inpatients diagnosed with type 2 diabetes mellitus were enrolled. Two peripheral blood samples were collected from healthy volunteers (Sample A1 and A2) and one peripheral blood sample was collected from the inpatients diagnosed with type 2 diabetes mellitus (Sample B). Sample A1 and B were transported in ice bath within 15 minutes of blood collection, centrifuged immediately and processed. The sample was re-centrifuged after 15 minutes and a second ammonia result was obtained. Sample A2 was transported at room temperature and stored between 2 and 4 hours, centrifuged and plasma ammonia measurement was performed. The sample was re-spun after 15 minutes and a fourth ammonia result was obtained.
Results: In our study, in healthy group the difference between sample A2 and set point value (on ice, 15 minutes) is 25.08 µg/dl, showing an increase of 55.29%. After another 15 minutes, an increase of 82.02% was observed compared with the standard value. In diabetes mellitus group, after 30 minutes of blood collection, an increase of 11% over the set point value was observed.
Conclusions: The blood specimen should be transported on ice to the laboratory and analyzed within 15 minutes of blood collection due to plasma ammonia spontaneously increase.

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Unified protocols for transdiagnostic treatment versus the Beckian’s approach of cognitive behavioral therapy for medical students with emotional disorders

DOI: 10.2478/amma-2022-0002

Background: Emotional disorders are highly prevalent among medical students, impacting their academic performance and general quality of life. It was shown that cognitive-behavioral and emotion regulation strategies are efficient for reducing anxiety and depressive symptoms in young people.
Objective: This study aims to compare the efficiency of standard individual Cognitive-Behavioral Therapy (CBT) to a group intervention based on the Unified Protocol for Transdiagnostic Treatment (UPTT) of emotional disorders in two groups of medical students with similar baseline characteristics.
Method: The sample of this research was composed of 64 medical students in total, of which 34 were included in the CBT group (Mage= 19.5 years, SD= 0.96) and 30 were included in the UPTT group (Mage= 19.4 years, SD= 0.93). The level of irrational beliefs and emotional distress were measured before and after the interventions, which were carried out by certificated psychotherapists.
Results: Statistically significant differences were observed between pre-test and post-test in both groups. The level of irrational beliefs dropped after the CBT (M=62.91, SD=21.30, p<0.05) and the UPTT intervention (M=67.33, SD=34.89, p<0.05), respectively. Similarly, the profile of emotional distress changed after the CBT (M=77.62, SD=16.97, p<0.05) and the UPTT intervention (M=68.77, SD=20.06, p<0.01).
Conclusion: Psychological interventions relying on cognitive-behavioral approaches, including standard, individual psychotherapy and a group intervention designed according to a transdiagnostic protocol, are suitable methods for reducing emotional disorders among medical students.

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Risk Assessment of Failures in Generic Drug Development and Approval Procedure under Competitive Generic Drug Therapy and Patent Challenge Exclusivities Provided by the United States Food and Drug Administration

DOI: 10.2478/amma-2022-0004

Objective: The United States Food and Drug Administration implemented two exclusivity programs Competitive generic therapy and Patent Challenge exclusivity to develop generic drugs, which provide a 180-day monopoly market for first generic applicants in the United States of America. The aim of the present study is to find the root cause of failures in developing and filing the first generic drugs under these exclusivities and to compare both the exclusivities to find the merits and demerits.
Methods: We used descriptive statistics for data analysis of both the exclusivities and Risk assessment was conducted on 14 industries to find the root cause of failures in every stage of the approval procedure by FMECA (Failure mode, Effects and Criticality Analysis).
Results: We found 44% of rejections in competitive generic therapy drugs and 30% of rejections in patent challenge exclusivity drugs. The risk analysis conducted on failures found that, in drug selection, 6% of failures are occurred due to rare diseases. In drug development, 9% of failures are occurred due to formulation failures. In pre-approval, 10% of failures are occurred due to secondary patents. In post-approval, 6% of failures are occurred due to product changes after approval.
Conclusion: We hope this study can give an idea for small and medium companies in developing countries for the early development of generic drugs for life-threatening diseases.

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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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