Moldovan Medical Journal

5-11

Authors: Igor Casian, Ana Casian

Background: Thymoquinone (TQ) and thymohydroquinone (THQ) are natural substances with antibacterial, antimycotic, antiviral, anticancer, antioxidant activity. Previously, was developed the method for obtaining Wild bergamot essential oil with high TQ content. The objective of this study was to create methods for obtaining individual substances TQ and THQ from Wild bergamot herb. Material and methods: Aerial parts of Wild bergamot have been collected from the plantation of the Scientific Practical Centre of Nicolae Testemitanu State University of Medicine and Pharmacy. The hydrodistillation method has been used for essential oil isolation and fractionation, and purification of individual substances. The analytical part of study has been performed by the HPLC method. Results: THQ was obtained in crystalline form by reducing TQ directly in the essential oil with ascorbic acid, sulphurous acid, or natural components of the Wild bergamot essential oil. Subsequent oxidation of THQ with hydrogen peroxide or nitrous acid gave TQ. Additionally, the yield of TQ was increased by optimising the parameters of plant material fermentation and using two-stage hydrodistillation with intermediate oxidation of the THQ formed at the first stage. Conclusions: Several technological procedures were created to obtain THQ from the Wild bergamot essential oil with a yield from 68-74% to 93-97% and TQ from THQ with a yield of 87-92%. The procedure of plant material processing has been optimised also to obtain essential oil with 48-56% TQ and not more than 12% residual phenols.

20-23

Authors: Veronica Cotelea, Natalia Corolcova, Luminița Mihalcean, Mihaela Burac, Liliana Profire, Iurie Dondiuc

Background: Preterm birth remains a leading obstetrical complication because of the incomplete understanding of its multifaceted etiology. It is known that immune alterations toward a proinflammatory profile are observed in women with preterm birth, but therapeutic interventions are still lacking because of scarcity of evidence in the integration of maternal and placental interrelated compartments. Objective: To investigate value of Lactoferrin (LF) and Interleukine-6 (IL-6) in the preterm labor. Material and methods: The study comprised 65 women with spontaneous preterm labor and 65 women with term labor. Maternal plasma concentrations of Lactoferrin and Interleukine-6 were detected by standard test system Aeskulisa Lactoferrin and Best-Vector A-8768 for Interleukine-6 Ref 3307 which (GmbH & Co, Germany) gave an analytical sensitive of 1.0 U/ml for Lactoferrin and 0.131 pg/ml for Interleukine-6. Results: Plasma levels of Lactoferrin in women with preterm labor were lower (µmedian = 0.90 U/ml) (p<0.001) than in the control subjects (µmedian = 40.68 U/ml). Plasma levels of Interleukine-6 in the plasma in women with preterm were predictably higher (µmedian = 51.90 pg/ml) than that in the control subjects with term delivery (µmedian = 21.51 pg/ml) (p<0.001). Conclusions: The study findings suggest that plasma levels of Lactoferrin and Interleukine-6 in women with preterm labor may be considered as a promising early biomarker for preterm labor.

29-35

Authors: Vitalie V. Moscalu

Background: Re-evaluation of reconstructive possibilities in the correction of degenerative mitral valve disease is of great clinical necessity nowadays. Material and methods: Analyzing the nature of the pathologies that determined the development mechanism of valve insufficiency, 136 cases of anterior cusp prolapse and 152 cases of posterior cusp prolapse were determined. Results: Cord rupture was established in 79 (58.9%) patients, cusp defects (”Cleft”) were appreciated in positions A1, A2, A3 – 15 cases (5.9%) and in P1, P2, P3 – in 92 (86.6%) cases. The surgical techniques performed were separated into: (1) resection – for the anterior and posterior cusps – 45 cases and accompanied by the slide – in 30 cases; (2) with Gore-Tex neo-chordal implantation – 115 cases, with cord transfer – 30; (3) Cusp enlargement with autologous pericardium – 5 cases, Alfieri procedure – 8. Implantation of a support ring required 130 (97.0%) patients. The correction of the associated valve disease required 125 patients (De Vega – 89.1%, ring – 8). Coronary bypass was required – 16 patients. There were no postoperative fatal cases. Conclusions: Based on the data obtained, reconstructive repair surgery can be can recommend for valves of degenerative, post-traumatic, ischemic, post-endocardial etiology as effective and sustainable techniques over time, being a superior alternative to replacement with prosthetic valves.

41-48

Authors: Olga Macagonova, Adrian Cociug, Tatiana Taralunga, Vladimir Ciobanu, Tudor Braniste, Viorel Nacu

Background: The present work describes the possibility of manufacturing biomaterials from the extracellular matrix for the treatment of the skin wounds. Biomedical collagen-based materials are clinically effective. Collagen is the most abundant and major component of the skin. Porcine collagen is almost similar to the human collagen, it is not immunogenic when used for the therapeutic purposes. Biomaterials can be obtained from the decellularized dermis, being a matrix rich in the collagen and glycoproteins. Material and methods: 3 parallel groups of biomaterials were established and the average value was calculated. To ensure the effectiveness of the decellularization process, the decellularized porcine dermis was compared with the intact sample using qualitative and quantitative criteria. Results: Histologically, the decellularized tissues revealed the presence of fewer cells. As a result, were removed approximately 80.5% of the genetic material from porcine dermal structures, demonstrated by the spectrophotometric quantification of deoxyribonucleic acid. In vitro graft degradation study in 0.01 M phosphate buffer pH 7.4 combined with collagenase, demonstrated a significant (p < 0.05) loss of collagen sponge mass by 100% over one hour in the group II compared to the decellularized dermis in group I which decreased in the weight by 91.3% during 35 hours. Conclusions: Acellular biomaterials are immunologically inert, have hydrophilic and biodegradable properties, thus they can play a key role in the wound care, exerting the transfer of the bioactive molecules and drugs directly into the wound.

56-65

Authors: Nadejda Diaconu, Irina Civirjic, Galina Sorici, Aurel Grosu, Tatiana Cuzor, Sabina Racila-Iatco

Background: Few studies have evaluated the thrombolytic treatment in patients with intermediate-high risk pulmonary embolism, making this study more valuable. Material and methods: It was a prospective, non-randomized, open-label, single-center study. Eligible patients at the age of 18 or older with an acute pulmonary embolism (PE) confirmed by CT pulmonary angiography with onset until 14 day and signs of right ventricular (RV) overload on echocardiography took part in the study. Pulmonary Arterial CT Obstruction Index Rate (PACTOIR) was used to define the localization and the expansion zone of thromboembolism. This study included 18 patients with intermediate risk and acute submassive pulmonary thromboembolism. In thrombolysis (TT) group (n=9) were used 50 mg of tissue-plasminogen activator (t-PA) administered in infusion as 0.4 mg/h for 2 hours. In the standard anticoagulation group, unfractioned heparin (UFH) was administered as a bolus of 70 units/kg or a maximum of 5000 units, followed by continuous infusion at an initial rate of 16 units/kg or a maximum of 1000 units/h. Results: The mean age for TT group was 69 vs 63 for the UFH group. PACTOIR was 100% in 3 patients in the half-dose rt-PA group and in 2 patients in the UFH group. RV/LV diameter ratio decreased from baseline to 48 h post-procedure (1.55 vs. 1.13; mean difference, -0.42; p < 0.0001). Mean pulmonary artery systolic pressure was 55 mm Hg in both groups (p < 0.05), with 53 [43–60] in TT group vs. 41.5 [37–45] mmHg in UFH group, P< 0.05). The rate of secondary endpoints was significantly higher in the UFH group with a high rate of pulmonary hypertension (0 vs. 19%, p=0.003). Conclusions: Half-dose thrombolytic therapy in patients diagnosed with submassive pulmonary embolism significantly reduced death and hemodynamic decompensation in the first 7 days compared to anticoagulant therapy only. With all that being said, it can be concluded that patients with high-intermediate risk PE could benefit from reduced-dose TT.

71-77

Authors: Ecaterina Carpenco, Veaceslav Fulga, Lilian Saptefrati, Lilia Sinitina, Valeriu David

Background: The angiopoietin/TIE system is one of the signalling pathways that regulate vascular development and remodelling during morphogenesis of the placental complex. Deregulation of the vascularization process is associated with primary placental insufficiency through destabilization of the maternal-fetal functional system. Disruption of expression of pro-angiogenic factors is associated with reduction of the microcirculatory bed and installation of circulatory dysfunction. The aim: Evaluation of Ang2 immunoexpression in early term compromised uterine pregnancies in the context of chorio-villous circulatory dysfunction in primary placental insufficiency. Material and methods: Abortion product from 67 patients with early term compromised pregnancies (stagnant pregnancies – 43 cases, early miscarriages – 11 cases, control batch that included 13 cases of pregnancies solved on social indications) were immunohistochemically evaluated with the marker for anti-Ang2 and anti-CD31. Results: The majority of sites analyzed were Ang2 negative, with the exception of syncytiotrophoblast, that reached the highest score (+3) in most cases. A maximum placental vascularisation index was achieved in the control group. The stagnant pregnacies batch has registered the lowest vascular density mean. Conclusions: The placentation period is characterized by a weak Ang2 cellular environment in the chorio-villous germinal site in the group of short-term compromised pregnancies, except a highly positive syncytiotrophoblast. The immunoexpression profile in vascular endothelium correlates statistically significantly with placental vascularization index and chorio-villous vascular density in stagnant pregnancies.

83-90

Authors: Victor Ojog

Background: Portal vein is the most enigmatic vessel of our body because it regulates own contractile performances using a special pace-maker mechanism represented by cells of Cajal. The contribution of various metabolic mediators and natural vasotropic agents in the control of the portal blood circuit is much less studied compared to the arterial system in general and the hepatic system in particular. The studies designed on the structure, function, and reactivity of the portal vein in different preconditioning have brought some common but also distinct evidence of the arterial system. Nitric oxide production is higher partly due to reduced arginase expression, but muscular media is thinner. Periodic spontaneous contractions directed towards the liver gate are characteristic for portal vein (PV), and the longitudinal muscle fibers are considered to be responsible for this phenomenon. Spontaneous rhythmic oscillations of the cells of Cajal are triggered by increasing calcium ion concentration leading to their depolarization. PV constrictor effect of phenylephrine is dependent on the activity of receptors to ET-1. For PV is characterized the acetylcholine induced contraction either in vivo or in vitro, and this effect is thought to be dependent on ET-1. Conclusions: The establishment of main particularities of portal vein reactivity of action of different paracrine, endocrine, and hemodynamical stimuli represents an important tool for prediction of contractile disorders leading plausible to portal hypertension. Likewise, a well proven interplay between cholinergic and adrenergic stimulations and on the other hand between Ang II and ET-1 actions must be a support for pharmacological modulating of portal vein reactivity disorders.

101-110

Authors: Lilia Rotaru, Oxana Grosu, Stanislav Groppa

Background: Parkinson’s disease (PD) affects more than 1% of the population aged over 65 years and manifests with both motor symptoms – bradykinesia, rest tremor, rigidity, and non-motor symptoms. Cognitive impairment and dementia are recognized non-motor symptoms that can significantly affect the quality of life of both the patient and caregivers and are a risk factor for institutionalization in nursing homes and a risk factor for early mortality. Cognitive impairment is frequent in Parkinson’s disease (PD) that can develop even before the diagnosis of Parkinson’s disease based on its motor features. Conclusions: There are several clinical, molecular, and imaging factors that constitute risk factors for the development of Parkinson’s disease dementia, in which basal cholinergic and prefrontal dopaminergic systems are involved. Histological changes are Lewy-body, Alzheimer, but also vascular pathology. Clinically can be distinguished subjective cognitive decline, mild cognitive impairment and, subsequently, Parkinson’s disease dementia. There are no remedies with a proven effect to prevent the occurrence of cognitive decline in PD. The only approved drug for already developed D-PD is the cholinesterase inhibitor – donepezil. Non-pharmacological interventions are thought to be beneficial. A multidisciplinary approach to cognitive impairment is recommended, with specific pharmaceutical treatment of the cognitive disorder and comorbidities, and appropriate rehabilitation.

5-12

Authors: Viorel Prisacari, Iana Baranetchi, Marcela Chilianu, Cristina Rarancean

Background: Septic nosocomial infections are a major public health issue. Both the risk of contracting and the specificity of nosocomial pathology directly depend upon the type of inpatient settings, as well as on the institution-specific risk factors. Material and methods: The study used a descriptive observation method based on a cross-sectional study. The present research documented and analysed retrospectively 687 follow-up records of patients admitted to different intensive care units. Results: The study results found that the incidence of septic nosocomial infection within various intensive care units (ICU) differs, ranging between 24.68% up to 34.8%. The structure of nosological forms was dominated by severe infections as pneumonia – 50.7%, septicemia – 12.68%, surgical site infections – 12.60%, urinary tract infections – 8.45%. The polyetiological structure of pathogens varied depending on the types of ICU. Microorganisms of the genus Staphylococcus, Acinetobacter, Clebsiella, Pseudomonas and Enterobacter predominated in most gram-negative (87.25%) cases, being multi-drug resistant to antibiotics. The following risk factors for the development of nosocomial septic infections were identified: the widespread use of invasive devices in the treatment process, patient’s comorbidities, polytraumas, vasopressors administration, the length of hospital stay within the ICU, etc. The clinical and economic effect is also important; hence the hospital stay length of patients with nosocomial infections was 2.2-2.5 times, the hospital stay cost per patient was 4.56 times, and the mortality rate was 4.55-8.43 higher compared to patients with no purulent nosocomial infections. Conclusions: Septic nosocomial infections are an urgent issue for ICU admission, which requires the implementation of comprehensive programs to prevent morbidity and reduce microbial antibiotic resistance.

18-23

Authors: Oxana Grosu, Lilia Rotaru, Stela Odobescu, Marina Sangheli, Svetlana Plesca, Ghenadie Carausu, Ion Moldovanu

Background: Chronic pain is a public health problem due to its high prevalence, disability, and associated comorbidities. In the Republic of Moldova, there are no health policies or strategies regarding chronic pain, and the burden on the population is not known because it is not registered nor monitored by the state. The aim was to analyze the knowledge, attitudes, and practices of neurologists regarding the management of chronic pain in the Republic of Moldova. Material and methods: Mixed observational study (qualitative and quantitative) was designed and carried out using a Knowledge, Attitudes, Practices research questionnaire. The study includes 50 neurologists, average age of 47.4±9.52 years, women (82%), urban area (80%), interact with chronic pain patients on a daily basis (86%). Results: Know about pain measurement tools (42%), use (40%), know clinical guidelines (40%), and pharmacological and non - pharmacological treatments (92%). The neurologists (62%) presented negative attitudes about chronic pain patients. Just 18 % have sufficient knowledge and skills to deal with such a patient. Received training on chronic pain issues 64% of neurologists. Conclusions: Neurologists have little knowledge of pain measurement tools and guidelines and don’t use them, which makes chronic pain poorly addressed, evaluated, and treated at the national level. Recognize the right of the patient to live without suffering and to benefit from quality services focused on their needs, but consider patients difficult to approach, communicate and work with.

31-34

Authors: Lilia Rotaru, Oxana Grosu, Stela Odobescu, Ion Moldovanu

Background: Pain is frequent in Parkinson's disease (PD) and has a great impact on life quality. The aim of the study was to establish the presence of the pain in a cohort of PD patients. Material and methods: Study sample consist of 102 consecutive PD patients, (mean age 61.51 ± 8.87 y.o.; disease duration 65.78 ± 41.34 mo.; 55 women and 47 men) evaluated for pain presence and divided into groups with and without pain: "PD pain +" and "PD pain -" Results: The groups were similar according to ages (61.02 ± 9.61 vs. 62.51 ± 7.69 y.o.) and levodopa dose (729.85 ± 483.29 vs. 708.31 ± 357.50). Pain was present in 64 patients (62.7%) of all study group; more frequent in women (56.3% vs. 43.8%, p> 0.05), with motor fluctuations (72.4% vs. 27.6%, p> 0.05), with dyskinesia (64.0 % vs. 36.0%, p> 0.05) and restless leg syndrome patients (72.7% vs. 27.3%, p> 0.05). Akinetic-Rigid (0.83 ± 0.80 vs. 0.64 ± 0.56, p> 0.05), and quality of life scores (59.70 ± 25.46 vs. 53.84 ±35.76, p> 0.05) were insignificant higher in "BP pain + "patients. They had longer disease duration (74.19 ± 39.99 y.o. vs 53.29 ± 41.06 y.o. p=0.017), higher depression (16.36 ± 11.97 vs. 8.09 ± 6.42, p=0.000), psychological (10.28 ±6.20 vs. 4.77 ± 2.82, p=0.000) and non-motor symptoms (66.27 ±39.25 vs. 46.68 ± 32.56, p=0.015) scores. Conclusions: Pain is common in PD, especially in long disease associated with motor complications, depression and other non-motor symptoms.

39-43

Authors: Rodica Bilba, Valeriu Cusnir, Liliana Dumbraveanu, Gheorghe Necula

Background: Myopia is labeled as one of the most common eye disorders, one of the most effective methods of its treatment being orthokeratological treatment. Anisometropic amblyopia continues to be treated by optical correction applied separately or in combination with occlusion or other therapies. Material and methods: The study included 32 patients (64 eyes), who were divided by 8 patients (16 eyes) into 4 groups (2 primary and 2 control), depending on the presence or absence of amblyopia and the degree of myopia, small and medium). Subjects in the baseline group underwent combined treatment between orthokeratology and physiotherapy. Results: The combined treatment resulted in the decrease of myopia according to the spheroequivalent by 60% in patients with amblyopia and mild myopia and from 90% in those without amblyopia. The values of the antero-posterior axis had a similar dynamic (p <0.001). In patients with moderate myopia its evolution decreased by 95% compared to patients with amblyopia – by 60%. The degree of anisometropia decreased by 10% (p > 0.05). Corrected visual acuity depending on the degree of amblyopia increased by 50% in cases with mild amblyopia and 150% in cases with moderate amblyopia (p < 0.001). The absolute volume of accommodation increased by 70% in patients with mild amblyopia and by 300% in patients with moderate amblyopia (p <0.001). Conclusions: It is rational to apply refractive therapy in the treatment and prevention of acquired uncomplicated myopia, with an average index of quality of life of 93.1%, versus 39.3% for optical correction.

52-57

Authors: Svetlana Plesca, Oxana Grosu, Marina Sangheli

Background: Low back pain (LBP) for many years is considered one of the most common conditions causing work absenteeism and long-term disability, with important implications for public health systems and economies. Pain generators of LBP are various, being distinguished specific and nonspecific causative mechanisms. The term „nonspecific” LBP remains ambiguous as potential sources of pain are supposed to be muscles or joints, but supplementary investigations do not correlate enough to explain the pain intensity and disability. The nociceptive and/or neuropathic mechanisms characteristic for acute pain tend to be influenced by central sensitization while pain chronification occurs, leading to new descriptor as nociplastic pain. Chronic low back pain, considered mostly non-specific, was mechanistically referred to primary musculoskeletal low back pain, the concept introduced in the new ICD-11 classification. The process of acceptance by the scientifical medical community raised debates and discussions. The aim of the study was to analyze the evolving concept of nonspecific low back pain to chronic primary musculoskeletal low back pain. A narrative literature review was carried out. Conclusions: The term non-specific low back pain is used when the pain generators have not been accurately determined or cannot fully explain the existing symptomatology. Chronic primary musculoskeletal low back pain is better explained by central sensitization mechanisms and altered nociception, named nociplastic pain. Because of raised ambiguities regarding this concept further studies are expected to shed light on the problem.

66-76

Authors: Natalia Cernei, Ruslan Baltaga, Serghei Sandru, Oleg Arnaut, Ivan Civirjic, Victoria Moghildea, Olga Gherasim

Introduction: The recorded studies suggest that there is clear evidence-based association between various laboratory biomarkers and COVID-19 disease severity. These marker levels reflect the intensity of the cytokine-mediated hyperinflammatory response, which is strongly associated with a poor outcome of SARS-CoV-2 infection. Conclusions: C-reactive protein is not only a systemic inflammatory marker, but also an important regulator of inflammatory processes. The level of this protein is positively correlated and can be widely used to predict the severity, prognosis and mortality in COVID-19 patients, additionally to vital signs monitoring, supportive care, oxygen therapy, ventilation and circulatory support. Procalcitonin is an indicator of disease severity, which can facilitate timely clinical decision-making, and determination of procalcitonin levels during COVID-19 patients’ follow-up, as well as being used in assessing risk, predicting prognosis, and improving patient survival. The assessment of hematological laboratory parameters upon admission, which help in differentiating between severe and non-severe cases, high-risk and low-risk cases of mortality, allows raising awareness, monitoring and timely treatment of patients with COVID-19, as well as their early improvement of clinical condition. Inflammatory biochemical and hemocytometric measures are feasible, easily interpretable, and widely available biomarkers in most healthcare settings, favorable for being used in treatment and severity determination, in predicting clinical outcomes, and in the prognosis of patients with COVID-19. However, the assessment of the accuracy of these biomarkers needs to be determined in further more relevant worldwide studies, showing a more precise design, more accurate performance, and having larger sample sizes.

5-12

Authors: Valeriu David, Vergil Petrovici, Lilian Saptefrați, Veaceslav Fulga, Ecaterina Carpenco, Elena Frant

Background: The molar and non-molar lesions are determined during histomorphological examination and treated as inconclusive. The establishing of a marker by immunohistochemical investigations could influence the accuracy of the morphopathological diagnosis. The aim is evaluation of p57 immunoexpression in the trophoblastic germ compartment in molar and non-molar pregnancies. Material and methods: Abortion products from 15 patients with hydatidiform mole, 18 pregnancies solved on social indications and 16 short-term disordered pregnancies were evaluated depending on the immunoexpresion of p57. Results: The hydatidiform mole was classified based on anti-p57 immunoexpression into: complete hydatidiform mole – 8 cases (negative immunoexpression or expression in ˂10% of villous cytotrophoblast) and partial hydatidiform mole – 7 cases (positive expression in ˃10% of the villous cytotrophoblast). Basal deciduous and extravillous cytotrophoblasts were positive in 100% of cases and served as internal control. Hepatocytes were used as negative control. In the control group, the positive immunoexpression was attested in >10% of cases in the villous trophoblast. Conclusions: Differential immunoexpression of p57 protein in the germinal cytotrophoblast allows subclassification of molar pathology into complete and partial forms, while not allowing the differentiation between partial hydatidiform mole and non-molar lesions. Immunohistochemical evaluation of p57kip2 protein in molar and non-molar pathology is useful in differential diagnosis as a complementary method.

20-29

Authors: Vitalie Cobzac, Mariana Jian, Tatiana Globa, Viorel Nacu

Background: The existing surgical techniques used to regenerate articular cartilage fail. Utilisation of hierarchical, biphasic structures obtained from osteochondral tissue, through demineralisation, decellularization, longitudinal perforation and combination with chondroprogenitor cells, presents a high potential in cartilage defects regeneration. Material and methods: The research was performed on 36 rabbits, separated equally in two experimental and one control group. In the experimental groups, the experimental osteochondral defects of 4-4.5 mm in depth, were performed with a 3.7 drill bit at the level of weight bearing surface of the medial femoral condyle. In the 1st group the defects were treated with grafts combined with autologous chondrocytes, and in the 2nd group with grafts combined with autologous mesenchymal stem cells. In the control group, cartilaginous defects were treated by transferring the osteochondral plugs taken from the trochlear groove. The rabbits were removed from the experiment at 6 and 12 weeks. The results were evaluated by Unified Histological Score of Regenerated Cartilage (UHSRC). Results: At 6 weeks, according to UHSRC, the 1st group had 28.33±1.53 points, the 2nd group –27.67±2.08 points and the control group –26.33±1.53 points (p˃0.1; p˃0.2). At 12 weeks the 1st group had 18.68±5 points, the 2nd group –14.89±3.76 points and the control group –17.22 ±4.84 points (p˃0.5; p˃0.2). Conclusions: According to UHSRC, the experimental groups don’t show a significant difference compared to the control group at 6 and 12 weeks, also the quality of regenerated cartilage is poor.

36-40

Authors: Olga Macagonova, Adrian Cociug, Viorel Nacu

Introduction: Decellularized biomaterials derived from the biological tissues are ideal for tissue engineering applications because they mimic the biochemical composition of the native tissue. The physical and structural properties of the scaffold are important in the fields of tissue engineering and regenerative medicine. Material and methods: Study material was 20 decellularized dermal grafts. 10 samples were obtained from piglets slaughtered in the slaughterhouse. Other tissues (n=10) were received from the donor from the Human Tissue and Cell Bank of the Republic of Moldova. Extracellular matrices were obtained by decellularization with 0.5% sodium dodecyl sulfate/0.1% EDTA solution. The evaluation of the structural characteristics was carried out by the histological examination with hematoxylin and eosin, scanning electron microscopy and the quantification of the amount of deoxyribonucleic acids. Assessment of the physical characteristics included analysis of extracellular matrix volume porosity, density, and swelling rate. Results: Histological examination revealed fewer cells in decellularized tissues compared to non-decellularized ones. More than 80.5% of nucleic acids were removed from porcine matrix and 82.5% of genetic material – from decellularized human dermal structures. A mean correlation and inverse dependence of -0.43 was shown between porosity and swelling rate of decellularized dermis. Conclusions: The decellularization process significantly (P<0.05) removed the cellular components while preserving the connective three-dimensional structure of the dermal matrices clearly shown by quantification of the amount of DNA and microscopic examination of the structures.

47-50

Authors: Chiril Voloc, Aliona Rotaru, Alexandru Voloc, Eliane Kamgaing Kuissi, Joel Fleury Djoba Siawaya, Simon Jonas Ategbo

Background: Vitamin D plays an important role in the prevention of many diseases. More than 1 billion people worldwide suffer from vitamin D deficiency. Vitamin D deficiency can contribute to the development of 16 types of cancer (breast, colon, prostate, etc.), cardiovascular diseases, stroke, autoimmune diseases, periodontal pathologies, transplant failure in endo-alveolar surgery, etc. There are several risk factors that would prevent the achievement of treatment objectives. The national protocol for deficiency rickets prevention privileges vitamin D 2 versus vitamin D 3 , which creates a medical risk factor compromising oral health in both children and adults. The article provides sufficient arguments in favour of vitamin D3 vs vitamin D2 prescription for prophylactic and treatment purposes. Conclusions: Considering that vitamin D deficiency is currently a global public health problem, it can be proposed to declare vitamin D deficit/deficiency a priority public health problem at the national level. Vitamin D3 should be elective in preventing deficit. Taking into account the multitude of acute and chronic diseases related to vitamin D deficiency, in order to improve the status of vitamin D in all population categories, it is necessary to include vitamin D3 in the list of molecules fully subsidized by the state and distributed free of charge at least to children under the age of 5 years and adolescents in the period of intensive growth.

59-63

Authors: Daria Ribacova, Olga Cheptanaru, Diana Uncuta

Background: The dysfunction of the temporomandibular joint (TMJ) becomes a significant and serious problem for modern society. TMJ disorder is the pathology of the stomatognathic system that is encountered more and more often every year, because the etiology is different and the pathogenesis is not well investigated. This disease is the result of the action of many factors that are constantly related to each other and over time can intensify and aggravate each other. A large number of studies have been done, articles and scientific papers have been published on the subject to establish the degree of prevalence and severity of this disease. Every year there are more and more patients with this pathology, which would mean that an effective solution in solving this problem has not yet been found and the topic remains relevant to this day. Insufficient knowledge of the mechanisms of this disease is one of the main reasons for the ineffectiveness of the treatment methods used. The objective of the work was to identify the clinical relevance of temporomandibular joint diseases. Conclusions: TMJ dysfunction is a topic that does not lose its relevance, because it is still a problem to identify the causes of pathologies of the TMJ and their treatment. TMJ pathology can be caused by various factors, the identification of this problem should be approached with all seriousness, taking into account all diagnostic methods and a high-quality anamnesis taken from the patient.

5-9

Authors: Olga Penina, Galina Obreja, Elena Raevschi

Background: Disregarding the recent rise in life expectancy in Moldova, the gap with Western countries is very high. The aim of the study is to identify the causes of death and the age groups responsible for excess mortality in Moldova in relation to the European average model in 2001-2019. Mortality beyond the European model set as a threshold was considered excessive. Material and methods: Data were retrieved from the Human Cause-of-Death Database and the WHO mortality database. Multiple decrement life tables by cause were computed for Moldova and the model (Germany, England and Wales, Czech Republic, Poland). Results: 27% of all deaths in males and 13% of all deaths in females under the age of 70 are excessive compared with the model. 80% of excess deaths were attributable to cardiovascular and digestive diseases (both males and females) and external causes (males). Excess deaths were mainly concentrated between the ages of 40 and 70 in men and 50 and 80 in women. Over the study period, cardiovascular diseases contributed the most to the decline in excess mortality in females, but not males. For the latter, excess mortality increased because of the cardiovascular component, completely compensating for moderate progress in external causes of death. Conclusions: Narrowing the life expectancy gap between Moldova and Western countries should be possible through better control of the key risk factors behind the identified causes of death. Key words: causes of death, excess deaths, life expectancy, cardiovascular component.

16-23

Authors: Victoria Verejan, Eugeniu Bendelic, Jana Bernic

Background: Visual disturbances may result in a long-term complication after mild traumatic brain injury (mTBI) in children. These problems may affect both near work and reading, and thus affect activities of daily life and the child’s return to school activity. The purpose of the study was to assess the visual acuity disturbances and refractive status in children with persisting symptoms after mild traumatic brain injury. Material and methods: Forty-eight patients with persisting symptoms after mild traumatic brain injury and anomalies of visual acuity were included. Visual symptoms and refractive status were assessed during the eye examination. Results: Thus, in the mTBI group the visual acuity for the right eye was of 0.09-0.5 in 83.7% (40 patients), in 16.3% (8 patients) – right eye 0.6-0.8, comparing to the control group, where 62% patients had the visual acuity ranged almost in 1.0, just 14% (7 patients) ranged 0.09-0.5 and in 24% (12 patients) – 0.6-0.8. The visual acuity for the left eye in the research group was of 0.09-0.5 in 89.8% (43 patients), in 10.2% (5 patients) – for the left eye was 0.6-0.8, comparing to the control group, where 66% patients had the visual acuity ranged almost in 1.0, just 24% (12 patients) it ranged 0.09-0.5 and in 14% (5 patients) – 0.6-0.8. Conclusions: Visual acuity (VA) is affected primary after head trauma although it has big chances to get better with a vision therapy in a time period ranged between 3 and 6 months after the trauma. In most of the cases, we speak of a blurred vison in the near work and relative unclear perception at far. Autorefraction data usually will reveal a slight hyperopia with a possible astigmatic component ranged between 1D to 3D, and in 4.1%-8.2% of cases a slight myopia referring to the spherical compound and 18.4%-32.7% astigmatic compound, also ranged between 1D and 3D.

31-35

Authors: Andrei Scureac, Serghei Cumpata, Evghenii Gutu

Background: Gastroesophageal reflux disease (GERD) is nowadays a highly prevalent, chronic condition, with 10% to 30% of Western populations affected by weekly symptoms. The patient who does not respond to the empiric antisecretory treatment, with alarming symptoms, or referred to surgery should undergo an esophagogastroduodenoscopy (EGD). Material and methods: This was a retrospective and descriptive study of patients with GERD admitted for antireflux laparoscopic surgery from 2012 to 2019. All endoscopic data were analyzed with the following variables: age, gender, reflux esophagitis and its severity, esophageal ulcers and strictures, Shatzky’s ring, Barrett’s esophagus (BE), incompetence of the esogastric junction; hiatal hernia. Results: A total of 152 patients were included in the study. The age of the patients ranged from 19 to 76 years, averaging 52 years. Among them, 97 (63.8%) were women and 55 (35.38%) men. A wide variety of endoscopic features has been found: non-erosive GERD (6.57%); reflux esophagitis (Savary-Miller) – I (21.05%), II (44.07%), III (23.68%); esophageal ulcer (1.31%), BE (1.97%). The majority of patients present axial hiatal hernia (92.76%) corresponding to Hill grade IV incompetence of the flap valve. Hill grade III was present in 4.6% of cases, grade II – 2.63%. Conclusions: The patients with GERD may have a wide range of endoscopic features (from normal to esophagitis, hiatal hernia, strictures and EB). Considering the multitude of data provided by endoscopic examination in patients with GERD, it can be certainly stated that EGD is one of the most important investigations in these patients, and is mandatory in those selected for surgical treatment.

40-47

Authors: Mihail Popovici, Lucia Ciobanu, Ion Popovici, Victoria Ivanov, Mihaela Munteanu, Ion Ion Popovici, Tatiana Danila, Valeriu Cobet

Background: Coronary thrombosis is the key pathogenic mechanism of acute heart attack, including non-ST segment elevation (NSTEMI). Given that, the detection of reliable markers of hemostasis disorders is important in the process of optimizing the diagnosis of NSTEMI. Material and methods: The study was conducted on 54 patients with NSTEMI (average age 69.7±1.5 years). In 60% of cases, 3-vessel disease was noted; 56% of patients had ejection fraction >50%, and Killip class I of heart failure was revealed in 78% of patients. With the help of the STA-Liatest (France) equipment, the blood tests determined the following hemostasis markers: fibrin monomers (FM), thrombotic complex activity of factors II, VII and X. Additional markers like Procoag, the coagulation indicator dependent on circulating phospholipids or SPA, D-dimers, as well as factors C, S and antithrombin III were appreciated. The values of these markers determined by the same method in 20 healthy persons (control group) were used as normal values. Results: Circulating level of FM on admission was increased twice, while the values of Procoag and SPA were significantly decreased by 35.3% compared to the control. Factors C, S and antithrombin III were 54-80% of the control value range, and D-dimers were within the permissible values. In the acute phase of the heart attack, a deterioration of hemostasis indicators was noted, excepting the D-dimers. The levels of FM determined 24 and 72 hours after revascularization were consistently increased (up to 3.8 times) compared to the control, while Procoag and SPA decreased by 54-57%. Further reduction of factors C, S and antithrombin III accounted for 42-54% of normal indicators. After 5 days, an improvement in hemostasis markers was observed, but a significant difference still remained comparing to the control group. Conclusions: The hemostasis particularities discovered in patients with NSTEMI indicate the features of an activated prothrombotic status, and FM could be an important diagnostic marker of NSTEMI, due to its most significant deviation from the normal value (>100%). It can reliably reflect the thrombin level, which triggers the last enzymatic phase of thrombus formation.

56-60

Authors: Stela Vudu, Gouri Durga Krishna

Background: Hypothyroidism is the deficiency in the production of thyroid hormones to meet the requirements of peripheral tissues. Subclinical hypothyroidism (SCH) is defined biochemically as normal serum free thyroxin concentration in the presence of an elevated serum thyroid-stimulating hormone (TSH) concentration. SCH may be associated with low-grade systemic inflammation (increased high sensitivity-C reactive protein (hs-CRP), one possible explanation may be that TSH in adipocytes promotes the release of interleukin-6 (IL-6). Studies have confirmed inflammatory biomarkers like hs-CRP and IL-6 to be a predictor of cardiovascular (CV) events. However, the treatment of SCH remains subject to debate. Conclusions: It is increasingly evident that SCH has prognostic values and crucial clinical effects, which leads to the view of SCH not being a compensated biochemical change sensu strictu. Even a modest but consistent fluctuation in the circulating thyroid hormone levels can create a response from the human heart. Well-timed treatment should be considered as a precaution to avoid the unfavourable CV diseases. The inflammatory biomarkers, namely CRP and IL-6 are exceptionally robust markers of cardiovascular risk. Thus, using these biomarkers may be helpful in assessing the cardiovascular risk in patients with subclinical hypothyroidism.

5-9

Authors: Lilia Andronache, Valeriana Pantea, Aurelian Gulea, Inna Svet, Vasilii Graur, Valerii Matcovschi, Marina Gamaniuc, Valentin Gudumac

Background: Currently, there is a growing interest in new copper (Cu2+) heterocyclic coordination compounds (CC), isothiosemicarbazide derivates, which demonstrated multiple beneficial properties, but their effect on reactions with free radicals such as the superoxide radical has not been investigated. Material and methods: The action of new micromolecular complexes of copper (Cu2+) chloride and bromide with methyl n- (prop-2-en-1-yl) -2- (pyridin- 2-ylmethylidene) hydrazine carbimidothioate on capturing activity of the superoxide radical was determined by the spectrophotometric method in vitro experiments. Results: It was established that the micromolecular complexes of copper (II) chloride and bromide with methyl n-(prop-2-en-1-yl)-2-(pyridin-2- ylmethylidene) hydrazine carbimidothioate have been found to possess strong superoxide radical inhibitor properties when interacting with a superoxide radical. In addition to this, the IC50 of the studied compounds depends on the nature of the acid-ligand in the internal sphere of the complex and increases in the following sequence: Cl-–Br-. Conclusions: The established property of mentioned compounds is new, because their use as micromolecular inhibitors of superoxide radicals has not been described so far. The synthesized CC expand the arsenal of superoxide radical inhibitors with high biological activity. Their possible significance for the development of new treatment strategies for diseases associated with the overproduction of superoxide radicals is discussed.

13-19

Authors: Vitalie Cobzac, Liliana Verestiuc, Mariana Jian, Viorel Nacu

Background: Articular cartilage has poor regenerative capacities. Numerous cartilage repair techniques are known, including implantation of autologous chondrocytes. Material and methods: From 18 rabbits pieces of cartilage were harvested from femoral condyle. Minced cartilage was treated with 0.25% trypsin-EDTA. In the 1st group (n=9) the cartilage was digested with 0.6% collagenase in 15 ml tubes by shaking in incubator at 37°C, 5%CO2. In the 2nd group (n=9) digestion was performed in 25cm2 cell culture flasks placed on the lateral side, monitoring the process under a microscope after 120 minutes. The isolated cells were cultured to a 80-90% confluence. The chondrocytes were identified using histochemical staining after culturing for 16 days in overconfluence. Results: Chondrocytes isolation in the 1st group lasted a fixed 360 minutes, in the 2nd group – 140±10 minutes. In the 1stgroup were isolated 9.2x104±3.1x104 chondrocytes with a viability of 85.36±16.41%, but in the 2nd group – 1.6x105±3.4x104 chondrocytes with a viability of 98.09±3.85%. The mean period of cell culture in the 1st group was 15±2 days, in the 2nd group – 11±3 days. In first passage of the 1st group were obtained – 1.2x106 ±4.3x105 chondrocytes and in the 2nd group – 2.92x106 ±3.6x105 chondrocytes. The secreted extracellular matrix by chondrocytes was stained specifically for cartilaginous tissue. Conclusions: The method used for chondrocytes isolation has a direct impact on the number of isolated cells, their viability, but also upon the culture period and the number of cells obtained during the first passage.

26-32

Authors: Elena Pavlovschi, Alina Stoian, Grigore Verega, Viorel Nacu

Background: The use of bone graft has been a successful step in the treatment of a large number of diseases of the osteoarticular system. But a massive bone defect remains a dilemma for modern reconstructive surgery. Present methods used have a high level of morbidity and complication. Literature indicates the absence of an optimal solution in massive bone defects healing. The aim of this study: to perform an in vivo preliminary study of vascularization of the hind limb in the rabbit model, for obtaining a graft able for further inclusion in the host blood circulation, without immunosuppression by decellularization. Material and methods: The study was performed on the 12 laboratory rabbits. After euthanasia of the rabbit, the femoral and tibiofibular bone was collected without soft tissue, only with the vascular pedicle, and keeping the passage through the vessels. In the abdominal aorta was injected contrast material, with the subsequent preparation of the arterial vessels, succeeded by anatomical, morphological, radiography, and microangiography study of this vascularized bone segment. Results: The principal nutrient artery of the rabbit femur springs from the lateral circumflex femoral artery. The optimal segment for vascularized allografting (the rabbit model) was determined the upper third of the femur with the up to the level of the internal iliac artery. So, it could be used as a bone graft for further conservation and decellularization. Conclusions: The vascularized allogeneic bone without immunosuppression would be a perfect alternative in the treatment of the massive bone defects.

37-40

Authors: Victor Schiopu, Vasile Turcanu, Nicolai Ghidirim

Background: Timely diagnosis of primary retroperitoneal tumours is one of the current challenges of clinical oncology. This is due to the rarity, polymorphism and diagnostic difficulties of primitive retroperitoneal tumours. Material and methods: The study is cross-sectional, prospective and retrospective. The study group is represented by 118 patients with abdominal and retroperitoneal space tumours. Using the receiver operating characteristic (ROC) analysis curve and calculating the average quality of the diagnostic model, the informativeness of ultrasonography in the diagnosis of primary retroperitoneal tumours (PRT) was appreciated. Results: For tumour localization, the ultrasonography (USG) as a diagnostic model demonstrated an appropriate use criteria (AUC) of 0.641 (95% CI 0.541, 0.740, p <0.001), and the mean quality of the diagnostic model was 0.54. Following the statistical analysis, was found a partial correlation between the size of the tumour and the dimensions estimated at USG of 0.540 (95% CI 0.295, 0.737, p <0.001), which represents a high positive correlation. To determine the uni- or multicentric character of the tumour, the USG demonstrated an integrative value of sensitivity and specificity of 0.644 (95% CI 0.415, 0.873, p <0.001. In assessing the proximity ratio of retroperitoneal tumours, the highest AUC was recorded in the assessment of the ratio of tumour to pancreas – 0.838 (95% CI 0.705.0.971, p <0.001) and kidney – 0.861 (95% CI 0.699, 1.024, p <0.001). Conclusions: Ultrasonography is a fairly informative imaging diagnostic method in the diagnosis of retroperitoneal tumours. The characteristics of the tumours obtained after the ultrasound examination provide indirect information about the malignant or benign nature of the primitive tumour, which allows the assessment of the next stages of diagnosis and treatment.

49-52

Authors: Victoria Rusu

Background: Sevoflurane is the inhalational anesthetic agent that is used widely in operating room. It is currently the most commonly used inhalation anesthetic in operating rooms. A series of studies on animal and human model detected the association of intraoperative use of sevoflurane and postoperative cognitive dysfunction (POCD) manifestation. On the other hand other studies demonstrate the same POCD associated with intravenous agents. Relevant multicentric trials got the reasons to suspect other key factors in developing postoperative cognitive dysfunction. Conclusions: The intra-anesthetic use of sevoflurane has been associated for a long time with the higher incidence of POCD. The mechanism was not identified, and the theory of neuroinflammation remained the main key of pathophysiological reaction that leads to cognitive dysfunction. Recent multicentre trial gives reliable information that the use of intravenous anesthetic agents is associated with the same POCD. Neuroinflammation remains to be the mediator of cognitive disorders, and apparently IL-6 keeps a major role in them. Future studies are needed to be conducted to identify the role of anesthetic agents in determining the neuroinflammation.