Category Archives: Original Research

The effect of postprandial in vivo and experimental in vitro hyperlipidemia on human peripheral blood monocytes

Ion Bogdan Manescu1, Mariuca Manescu1, Elena Cristina Preda2, Doina Ramona Manu1, Minodora Dobreanu1,3

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-2022-0026

Objective: In this study, we aimed to investigate the effect of transient postprandial in vivo and prolonged experimental in vitro hyperlipidemia on human peripheral blood monocytes.
Methods: Peripheral blood was collected from seven healthy subjects after an overnight fast and three hours after a standardized high-fat meal. Both fasting and postprandial samples were stained for surface markers CD14/CD11b and intracellular lipids using BODIPY493/503. Postprandial samples only were used for isolation of peripheral blood mononuclear cells that were further incubated overnight with postprandial hyperlipidemic autologous plasma, then stained as described above. All samples were analyzed on a FACSAria III flow cytometer.
Results: Flow cytometric analysis revealed two monocyte populations (CD14+): CD14low and CD14high. In fasting, these populations show similar morphology (FSC/SSC), but different expressions of CD14, CD11b, and BODIPY493/503. At three hours postprandially, a moment of maximum hyperlipidemia, neither population suffered significant changes. After the 24-hour incubation, cell activation was observed in both populations: similar fold change increase in SSC, increase in FSC for CD14high cells only, similar foldchange increase in CD14, slightly higher foldchange increase in CD11b for CD14low monocytes, and significantly higher foldchange increase in lipid content for CD14high monocytes. CD14high monocytes appear to undergo a more intense activation than CD14low monocytes.
Conclusions: We conclude that all monocytes react after prolonged in vitro exposure to plasma lipids, each subset having its own activation pattern. All monocyte types may play a role in inflammation and the development of plaques. Monocyte assays are a valuable tool for the investigation of atherosclerosis at the cellular level.

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Artificial intelligence based model for establishing the histopathological diagnostic of the cutaneous basal cell carcinoma

Andrei Calin Dragomir1, Iuliu Gabriel Cocuz1, Ovidiu Simion Cotoi2, Leonard Azamfirei3

1. George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
2. Department of Pathophysiology, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
3. Department of Anesthesia and Intensive Care, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania

DOI: 10.2478/amma-2022-0020

Introduction: Artificial intelligence (AI), a component of computer science, has the ability to process the multitude of medical data existing in the medical system around the world. The goal of our study is to build an AI model, based on Machine Learning, capable of assisting pathologists around the world in the diagnosis of the basal cell carcinoma of the skin.
Material and Method: Our study is represented by the development of a Mask-RCNN (Mask Region-based Convolutional Neural Network) model, for the detection of cells with typical basal cell carcinoma tumoral changes. A number of 258 digitized histological images were used. The images emerged from Hematoxylin&Eosin stained pathology slides, diagnosed with cutaneous basal cell carcinoma between January 2018 and December 2021, at the Pathology Service of the Mureș County Clinical Hospital.
Results: All the used images have the unique resolution of 2560×1920 pixels. For the learning process, we divided these images into two datasets: the learning dataset, representing 80% of the total images; and the test dataset, representing 20% of the total images. The AI ​​model was trained using 1000 epochs with a learning rate of 0.00025 and only one classification category: basal cell carcinoma.
Conclusions: The AI model successfully identified in 85% of the cases the areas with pathological changes present in the input images.

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An evaluation of concordance between linear measurements obtained from conventional, digital and reconstructed three-dimensional printed orthodontic models: an in vitro study

Shravan Shetty1, Srikant Natarajan2, Supriya Nambiar1, Prithvi Shetty3, Kavery Chengappa4, Mukul Shetty5

1. Department of Orthodontics and Dentofacial Orthopaedics, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, Karnataka, India
2. Department of Oral Pathology and Microbiology, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, Karnataka, India
3. Department of Public Health Dentistry, A.J.Institute of Dental Sciences, N.H.17, Kuntikana, Mangalore, Karnataka, India
4. Department of Public Health Dentistry, Manipal College of Dental Sciences Mangalore, Manipal Academy of Higher Education, Manipal, Karnataka, India
5. Department of Orthodontics and Dentofacial Orthopedics, A B Shetty Memorial Institute of Dental Sciences; NITTE Deemed to be University, Mangalore, Karnataka, India

DOI: 10.2478/amma-2022-0028

Objective: To evaluate the potential use of digital and reconstructed three-dimensional printed models as an alternative to conventional plaster models by assessing the accuracy of their linear measurements.
Methodology: Pre-treatment plaster models of 45 patients were selected from the archives of the Department of Orthodontics. Each physical plaster model was scanned and digitized using a three-dimensional (3D) laser surface scanning system (inEOS X5, Dentsply Sirona, Bensheim, Germany). The scanned STL files were later used to reconstruct models by 3D printing using Figure4® standalone 3D printer (3D systems, Rock Hill, South Carolina). Measurements of teeth 11 and 16, the transverse width of the upper jaw between the first molars (MM – intermolar width) and canines (CC – intercanine width) were done manually using a digital vernier caliper (Mitutoyo, Kawasaki, Japan), and the CAD Assistant software (Open cascade, Guyancourt, France). Intra examiner data, Intraobserver variability, and measurement accuracy were evaluated using Intraclass Correlation Coefficient (ICC) analysis was done using SPSS 20.0.
Results: The intraclass correlation coefficients were >0.8 indicating high reproducibility and reliability. Significant differences were found between the physical and the digital models but to a small proportion which were deemed not clinically relevant.
Conclusion: Both the digital models and reconstructed three-dimensional printed models using Figure4® technology were clinically permissible in terms of accuracy and reproducibility. The digital storage, transmission, and treatment planning in an environmentally friendly manner should promote digital over conventional records.

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

Elena Cristina Preda1, Valeriu Moldovan1, Oana Roxana Oprea2

1. County Emergency Clinical Hospital, Târgu Mureș, Romania
2. George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania

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

Oana Liliana Atomei1, Paula Paraschiva Monor1,2, Bogdan Aurelian Stana3,4, Monica Tarcea5

1. Faculty of Medicine and Biological Sciences, „Ștefan cel Mare” University of Suceava, Romania
2. Emergency County Hospital „Sfântul Ioan cel Nou” Suceava, Romania
3. Physiology Department, Faculty of Medicine, „Gr. T. Popa” University of Medicine and Pharmacy, Iași, Romania
4. 2nd Pediatrics Clinic – Emergency Clinical Hospital for Children „St. Maria”, Iași Romania
5. 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-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

Radu Ovidiu Togănel, Răzvan Lucian Coșeriu, Camelia Vintilă, Anastasia Simion

County Clinical Hospital Mures, 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

Oana Elena Branea1, AnaMaria Romina Budeanu2, Răzvan Gabriel Budeanu3, Adrian Ștefan Chiuzan4, Ioana Lăcrămioara Nazaret5, Sanda Maria Copotoiu1, Alexandra Elena Lazăr1

1. Department of Anesthesia and Intensive Care, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Târgu Mureș, România
2. Department of Anesthesia and Intensive Care, Emergency Clinical County Hospital of Târgu Mureș, România
3. Department of Radiology, Emergency Clinical County Hospital of Târgu Mureș, România
4. Department of Radiology, Emergency Clinical County Hospital of Bistrița, România
5. Department of Anesthesia and Intensive Care, Emergency Clinical County Hospital of Piatra Neamț, România

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

Anithab Alex, Keerthan Shashidhar, Kuttappa M N, Krishna U S Nayak, Sai Kalyan Menta, Anushree A

Department of Orthodontics and Dentofacial Orthopedics, A B Shetty Memorial Institute of Dental Sciences; NITTE Deemed to be University, Mangalore, Karnataka, India

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

Ika Rahmawati Sutejo, Arifah Nur Hasanah, Faizah Ramadhani Sudarko

Faculty of Medicine, University of Jember, East Java, Indonesia

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

Karina-Alexandra Cojocaru1, Oana Roxana Oprea2,3, Minodora Dobreanu2,3

1. Municipal Emergency Hospital Roman, Romania
2. Emergency Clinical County Hospital of Targu-Mures, Romania
3. Laboratory Medicine Department, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania

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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