Category Archives: Original Research

Clear or White? A RT-PCR plate comparison for SARS-CoV-2 diagnosis

DOI: 10.2478/amma-2022-0024

Objective: During the COVID-19 pandemic, laboratories have used various extraction and amplification kits, associated with different auxiliary materials. This study aims to investigate how different types of plates may affect RT-PCR performance.
Methods: Data on the positive controls (PCs) of RT-PCR runs for SARS-CoV-2 detection between December 2020 and April 2022 was collected retrospectively in the Molecular biology department of the County Emergency Clinical Hospital of Târgu Mureș’s clinical laboratory. The materials used consisted in MOLgen SARS-CoV-2 (Adaltis) and EliGene COVID19 CONFIRM 500 R amplification kits, 96-well full-skirted white and clear plates, and clear films. Microsoft Excel was used for the database and it included information about Cycle threshold (Ct) and maximum fluorescence. Statistical analysis, performed in MedCalc, consisted of Grubbs test, Kolmogorov–Smirnov Test, F test, T student test, and Mann-Whitney test to compare central tendencies. The significance threshold was set at p<0.05.
Results: The Ct comparison for MOLgen kit white plates vs clear plates: FAM channel- U=1052.5, Z=2.07, p=0.038, medians for white plates and clear plates were 22.80 and 23.25, respectively; ROX channel- U=784, Z=3.21, p=0.001, medians 21.93 and 21.43, respectively; Cy5 channel- U=1028.5, Z=1.95, p=0.518, medians 22.12, 21.75, respectively. For EliGene kit: U=848.5, Z=3.27, p=0.001, medians 28.26 (white plates) and 28.0 (clear plates). Comparison of the maximum fluorescence reached on both kits with white and clear plates computed p values <0.0001.
Conclusions: Between white and clear plates there are statistically significant differences considering Ct values and maximum read fluorescence, but with no impact on test outcome.

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Nutrition in pregnancy – impact on anaemia in pregnant women

DOI: 10.2478/amma-2022-0016

Aim: Most pregnant women can achieve their caloric and nutritional needs by careful food choices during pregnancy, but there are fairly common situations when pregnant women develop anaemia, which needs to be managed properly to avoid maternal-foetal complications. The main objective of the study was to identify the nutritional factors favoring the occurrence of iron deficiency anaemia in pregnant women.
Methods: Data collection for the cross-sectional study was carried out using a questionnaire that provided information on nutrition and risk factors during pregnancy in a group of patients from Suceava. The questionnaire was given to pregnant women admitted to the Emergency County Hospital „Sfântul Ioan cel Nou” Suceava, Romania, between March and May 2017.
Results: The anaemia occured in 46.3% of pregnant women. Although 22.2% of pregnant women received oral iron supplementation, effects on anaemia correction were not observed (p=0.02). We found that weight status did not influence anaemic status (p=0.51), and the presence of anaemia in pregnancy was independent of risk factors for pregnancy (smoking, p=0.05; alcohol consumption, p=0.66), iron absorption inhibitors in the diet (coffee, p=0.33; tea, p=0.53), water intake (p=0.52) or night eating (p=0.27).
Conclusions: The results of dietary survey showed no direct link between the prevalence of anaemia and nutritional factors. Lower pre-pregnancy BMI was not associated with higher risk of iron deficiency anaemia, and daily iron supplementation during pregnancy did not meet the needs to counteract the anaemic syndrome. Nutrition education is particularly important for the prophylaxis of iron deficiency anaemia in pregnant women.

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An overview of the microflora of the patients admitted in an Ear Nose and Throat tertiary hospital from Romania

DOI: 10.2478/amma-2022-0017

Objective: The purpose of this retrospective study is to describe clinical, epidemiological and bacteriological findings on the Ear, Nose and Throat infections from patients admitted in a tertiary hospital from Romania, which might bring supportive data for the management of the ENT patients.
Methods: Clinical data from 146 patients admitted in a clinic from Romania, between February 2016 and April 2018 were collected from the official registries of the Microbiology Laboratory, where the collected samples were processed by classical microbiological methods.
Results: From 164 analyzed biological samples, 45.1% were suggestive for infection. Methicillin-sensitive Staphylococcus aureus was the most prevalent bacterial isolate (28.72%), followed by Pseudomonas aeruginosa (12.77%). There were 26 different species of microbes. Out of a total of 77 bacterial isolates, 62.33% were Gram positive and 37.66% Gram negative. The study showed that 11 ear discharge samples were positive in female and male patients. Upper respiratory tract exudates were positive on 3 females (21.4%) and on 17 males (28.8%). There were 7 positive pus samples from females, and from 25 males.
Conclusions: Men were more affected by Ear Nose and Throat infections, caused by bacteria mostly from the genus Staphylococcus and Pseudomonas. More attention at the samples collection and a rigorous clinical examination should reduce the load on bacteriology laboratory. In specific cases, a negative bacteriological result can prove as an exclusion diagnostic.

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Computed tomography evaluation of diaphragm alterations in 20 critically ill COVID-19 positive patients

DOI: 10.2478/amma-2022-0014

Objective: Diaphragmatic dysfunctions are multiple and critical illnesses often lead to the muscular atrophy that affects respiratory and peripheral muscles. The primary objective was to investigate diaphragm thickness in hospitalized patients. Secondary objectives were to assess clinical evolution and outcome. Methods: In a mean time period of 7.9 days, two different chest computed tomographies were used in order to examine diaphragm alterations of right and left diaphragm in 20 critically ill patients tested Real-Time Polymerase Chain Reaction positive to Severe Acute Respiratory Syndrome Coronavirus-2. Patients were divided in two groups (one group <5% decrease in diaphragm thickness and another group ≥5% decrease in diaphragm thickness).
Results: Results showed that patients presented low 10 years predicted survival rate (Charlson Comorbidity Index > 7.7±3.08), marked inflammatory status (C-Reactive Protein = 98.22±73.35, Interleukine-6 = 168.31±255.28), high physiologic stress level (Neutrophil/Lymphocyte Ratio = 31.27±30.45), respectively altered acid-base equilibrium. Half of the investigated patients had decrease in diaphragm thickness by at least 5% (right diaphragm = -7.83%±11.11%, left diaphragm = -5.57%±10.63%). There were no statistically significant differences between those with decrease of diaphragm thickness and those without diaphragm thickness, regarding length of stay in Intensive Care Unit and in hospital, inflammatory markers, and acid-base disorders.
Conclusions: Patients were admitted in Intensive Care Unit for acute respiratory failure and half of the investigated patients displayed diaphragm alterations at CT scan.

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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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