Volume 66, Number 3, 2020
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Category Archives: Number
Controls in Real-Time Polymerase Chain Reaction Based Techniques
DOI: 10.2478/amma-2020-0025
From its discovery in the 1980s, Polymerase chain reaction was further developed and is nowadays used as the foundation for the various PCR-based techniques used in molecular diagnosis across different species, and numerous types of samples. Real-Time PCR enables the user to monitor the amplification of a deoxyribonucleic acid (DNA) or complementary DNA (cDNA) target during the PCR run, in real-time, and not at the end, as it is the case in conventional PCR. The most frequent types of applications include gene expression analysis, gene silencing, variant analysis, and fusion temperature analysis. Given its vast field of application, a key question remains, and it is related to the controls (negative controls, positive controls, internal exogenous and endogenous controls) and their purpose in a Real-Time PCR experiment. In this paper, we set out to find how and when to use them, and which type of controls are suitable for certain experiment types, since the use of appropriate controls during Real-Time PCR experiments will reduce the effects of variables aside from the independent variable within the sample, therefore yielding accurate results, be it in research or diagnostic purposes.
Keywords: positive control, internal exogenous control, internal endogenous controls, real-time PCR, negative control
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SARS-CoV2 Infection in a Multiple Sclerosis Patient Treated with Natalizumab – A Case Presentation
DOI: 10.2478/amma-2020-0022
Introduction: The novel coronavirus, SARS-CoV2, has rapidly spread worldwide and led to an intense collaboration among both physicians and researchers in order to stop its dissemination. Little is yet known about how this virus behaves, but recent studies have suggested the role of integrins in the viral penetration of target cells. Natalizumab is an anti-α4-β1 integrin monoclonal antibody used in the treatment of multiple sclerosis (MS), a neurodegenerative auto-immune disease affecting primarily young adults. MS patients have a greater susceptibility to develop severe infections especially enhanced by the disease-modifying therapies (DMTs) which are currently recommended for their treatment. Natalizumab is considered the safest high-efficacy DMT in times of COVID-19 outbreak.
Case presentation: We hereby describe the first case from Romania of a MS patient treated with Natalizumab who subsequently acquired SARS-CoV2 infection and whose recovery was excellent, with no functional neurological or respiratory sequelae.
Conclusion: The favourable evolution of our patient supports the potential therapeutic effect Natalizumab might have in SARS-CoV2 treatment by specifically blocking integrins and by its immunosuppressant characteristics.
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The Role of Acceptance and Planning in Stress Management for Medical Students
DOI: 10.2478/amma-2020-0020
Objective: The purpose of this study was to analyze the role of two coping mechanisms, namely Acceptance and Planning, in stress management among medical students.
Methods: This research included two groups, a target group consisting of medical students (N = 100; Mage = 22.34) and a control group which was composed of physical education and sports students (N = 100; Mage = 20.11). For the target group, a low level of stress was induced, the students being informed that their overall behavior during an examination would be analyzed later by a group of psychologists, after which they were filmed while taking the exam. The students from the control group performed a physical exercise while they were filmed, and they were told that the correctness of the exercise would be evaluated by experts in physical education and sports on the basis of the recorded images. After completing the tasks, both the students in the target group and those in the control group completed the COPE questionnaire.
Results: The statistical data interpretation revealed a significant statistical difference regarding the two coping mechanisms, namely Acceptance [M = 10.73; t (19) = 3.79, p <0.001; CI -1.91, -0.60], and Planning [M = 9.47; t (19) = 4.70, p <0.01; CI -1.99, -0.81]. According to statistical data analysis, we did not find another significant statistical difference among the remaining 13 coping mechanisms.
Conclusions: To efficiently manage stress during exams, medical students use Acceptance and Planning coping mechanisms, which may increase their emotional regulation abilities and help them focus on problem-solving.
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Ambulatory Treatment of Pressure Sores in a Patient without Neurological Lesions
DOI: 10.2478/amma-2020-0021
Introduction: Pressure sores have been defined as areas of localized soft tissue ischemic necrosis caused by prolonged pressure related to posture, which usually occur over a bony prominence. The most affected areas are those that undergo a lot of tension such as trochanters, sacrum, malleoli, heels and occiput. Most people that are affected by pressure sores have an underlying pathology that makes them immobile, confining them to chairs or bed for longer periods of time. Pressure ulcers may have direct causes (pressure, shear, friction, immobility, loss of sensation) or indirect causes.
Case presentation: This paper will present the case of a 53-year-old male. After about 3 weeks of being immobilized in bed in dorsal decubitus due to an acute respiratory distress which required intubation and mechanical ventilation, the patient developed very large pressure sores on the left sacral and buttock area and left sole. He was treated in the outpatient department using multiple therapeutic protocols based on the alternation of antibiotic creams and modern absorbent dressings.
Conclusions: The evolution of this patient was long but favorable, the left sacral and buttock area eschar closing completely while the sole eschar had a good evolution, but is still undergoing treatment.
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The Anti-tobacco Legislation’s Impact on Air Pollution in Romanian Foster Care Homes
DOI: 10.2478/amma-2020-0019
Objective: The research aimed to investigate the impact of the anti-tobacco legislation (2016) on air quality in Romanian foster care homes.
Methods: The measurements took place in foster care homes situated in three Romanian counties. We recorded data three times (in 2014, in 2016 – six months after the implementation of the anti-tobacco legislation, in 2019). In 41 foster care homes were measured the indoor and outdoor Particulate Matter (PM2.5) level in the air, using the TSI SidePak™ Personal Aerosol Monitor device. Descriptive statistics and t-test were used for data analysis (significant difference if p < 0.05).
Results: The indoor PM2.5 values were higher in every year than outdoor values. Analyzing the anti-tobacco legislation’s impact on air quality, we found no difference between the indoor data measured in 2014 and in 2016, but there was a significant difference in the outdoor values (they were higher in 2016). Comparing the values measured after the implementation of the legislation (2016 vs. 2019) we found no difference in either indoor or outdoor values.
Conclusions: The anti-tobacco legislation has not influenced the indoor air pollution (PM2.5) level. The increased outdoor PM2.5 values suggest that the residents of the foster care homes are smoking more in front of the house after the introduction of the anti-tobacco legislation in 2016.
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Asymptomatic Myxomatous Degeneration of the Mitral Valve, in a Young Male
DOI: 10.2478/amma-2020-0018
Background: In young patients, the most common cause of mitral valve regurgitation is prolapse or flail of the valve, which is morphologically reflected by myxomatous degeneration. In rare cases, such this presented in this paper, this lesion might be incidentally identified, in young people.
Case presentation: A 43-year-old male, with familial history of hypertension, presented in our Department of Cardiology for a routine check-up. Despite of lack of symptoms, an intense murmur was detected at mitral auscultation area. The echocardiography revealed a severe mitral regurgitation caused by flail of posterior mitral valve. The patient was transferred to the Cardiac Surgery Department for surgical intervention consisting in mitral valve repair. The histopathological examination revealed severe myxomatous degeneration of the spongiosa and fibrosa layers. The recovery after surgery was very good with no postoperative complications. The patient was discharged seven days after surgery, with no significant mitral regurgitation at the three-month follow-up.
Conclusions: In young, asymptomatic patients, the proper time for mitral valve repair should be decided by an interdisciplinary team. For a better understanding of myxomatous degeneration particularities, histopathological assessment should be done in any replaced valve. Certification of such lesions, under microscope, might be an indicator for familial cardiovascular screening, in first degree relatives.
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Establishing the Diagnostic and Prognostic Value of Serum Interleukin 6 Levels in Sepsis
DOI: 10.2478/amma-2020-0014
Objective: Establishing a serological profile of interleukin 6 (IL-6) in order to evaluate its usefulness as a biological marker for the diagnosis and early prognosis in sepsis.
Materials and Methods: The study included 246 individuals, divided into 2 groups: 131 in the septic subgroup (S) and 115 in the septic shock (SS) subgroup. Inflammatory markers, bacteriological examinations and laboratory samples were determined within 24 hours of the first signs of infection. Severity scores were also calculated within the first day of the onset of sepsis.
Results: The SS subgroup (median 121.2 pg/ml, 18.59-10235 pg/ml; SD = 1920) shows significantly higher values of IL-6, compared to the S subgroup (median 43.49 pg/ml, 13, 27-6566 pg/ml; SD = 1367) (p = 0.0026). The SS subgroup has a significantly higher death rate than S subgroup (p = 0.001). The cut-off values of the mortality prediction degree were 184.74 pg/ml. The area under the curve of the cytokine IL-6 for the differentiation of sepsis from septic shock was 0.693 (95% CI 0.582-0.790, p = 0.002). The optimal value of the cut-off that allows the differentiation of the septic subgroup from the one with organ dysfunction, was 52.72 pg / ml.
Conclusion: Serum IL-6 values are significantly higher in the septic shock group. All deceased patients had higher IL-6 serum values.
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Effect of Isoflurane Anesthesia on the Heart Rate and Blood Pressure Response to Autonomic Nervous System Stimulation and Inhibition in Rats
DOI: 10.2478/amma-2020-0016
Objective: Many experimental studies aim to assess the autonomic nervous system (ANS), but this is often hampered by interactions with anesthetic drugs. We aimed to evaluate whether isoflurane anesthesia is suitable for ANS evaluation in rats.
Methods: Six Wistar rats were anesthetized with isoflurane (4 L/min, 2.5%). Systolic blood pressure (SBP) and heart rate (HR) were measured at baseline and 20 min after sympathetic inhibition (propranolol, 5 mg/kg) and stimulation (isoproterenol, 2.5 mg/kg), and parasympathetic inhibition (atropine nitrate, 2 mg/kg) and stimulation (carbamylcholine, 0.4 mg/kg; acetylcholine, 0.1 mg/kg). Six additional rats were used to assess the effects of isoproterenol, carbamylcholine, and atropine nitrate in the absence of anesthesia.
Results: Propranolol significantly decreased the HR and the SBP, whereas isoproterenol significantly increased the HR (all p≤0.01) in the isoflurane-anesthetized rats. However, the HR response to sympathetic stimulation was significantly reduced in the anesthetized compared to the non-anesthetized rats (p=0.03). Carbamylcholine and acetylcholine significantly decreased both the HR and SBP (all p<0.05) in the anesthetized rats, but the response to carbamylcholine administration was significantly more pronounced in the non-anesthetized rats (p=0.03). Atropine nitrate significantly increased the HR (p<0.001) in the non-anesthetized rats, but it had no effect on either the HR or the SBP in the presence of isoflurane anesthesia (both p>0.05).
Conclusions: Isoflurane anesthesia appears to interfere with both components of the ANS and is therefore not an optimal approach for experimental ANS evaluation. Our data indicate autonomic receptors and/or post-receptor mechanisms as the most likely site for isoflurane-ANS interactions.
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Left Ventricular Function and Morphology after Cardiac Surgery for Severe Mitral Insufficiency – A Single Center Experience
DOI: 10.2478/amma-2020-0015
Introduction: Mitral insufficiency is a common valvular disease affecting 10% of the general population. The main treatment of the severe mitral regurgitation is surgical. We have analyzed the impact of cardiac surgery on the left ventricular performance and morphology, in patients treated for severe mitral insufficiency accordingly to the type of intervention, ischemic time and type of cardioplegia.
Methods: Ninety patients diagnosed with isolated severe mitral insufficiency that benefited from mitral valvular replacement or mitral valvuloplasty were retrospectively enrolled. The left ventricle, the left atrium, the right ventricle diameters and the left ventricle ejection fraction were measured by two-dimensional (2D) echocardiography before and after surgery. The influence of the myocardial ischemia time and the type of cardioplegia administered on the ventricular systolic function were also analyzed.
Results: Regardless the surgical technique chosen, after surgery we noticed a decreased size of the left ventricle (preoperative mean 54.91mm ±8.18 vs postoperative mean 51.94mm±8.15, p=0.035), right ventricle (preoperative mean 33.49mm±5,87 vs postoperative mean 32.41mm±6.03, p=0.0001), as well as the ejection fraction (preoperative mean 51.29%±8.51 vs postoperative mean 46.57%±8.71, p=0.0001).
Conclusions: Immediately after surgery, a decrease in the size of cardiac cavities as well as a decrease of the left ventricle ejection fraction is noticed.
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