ROMANIAN JOURNAL of MORPHOLOGY and EMBRYOLOGY

Vol. 64 No. 2, April-June 2023

1. Short histological kaleidoscope - recent findings in histology. Part III

Anne-Marie Constantin, Carmen Mihaela Mihu, Adina Bianca Bosca, Carmen Stanca Melincovici, Mariana Viorica Marginean, Elena Mihaela Jianu, Madalin Mihai Onofrei, Carmen Maria Micu, Bogdan Catalin Alexandru, Rada Teodora Sufletel, Ioana Maria Moldovan, Andrei Coneac, Andreea Crintea, Roxana Adelina Stefan, Paul-Andrei Stefan, Amina Djouini, Alina Simona Sovrea

The paper provides an overview of the current understanding of different cells biology (e.g., keratinocytes, Paneth cells, myoepithelial cells, myofibroblasts, chondroclasts, monocytes, atrial cardiomyocytes), including their origin, structure, function, and role in disease pathogenesis, and of the latest findings in the medical literature concerning the brown adipose tissue and the juxtaoral organ of Chievitz.

Corresponding author: Adina Bianca Bosca, Associate Professor, DMD, PhD; e-mail: biancabosca@yahoo.com

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2. Morphological aspects and therapeutic options in melanoma: a narrative review of the past decade

Andreea Catalina Tinca, Andrada Raicea, Andreea Raluca Szoke, Iuliu Gabriel Cocuz, Mihaela Cornelia Sincu, Raluca Niculescu, Adrian Horatiu Sabau, Maria Catalina Popelea, Radu Florin Fruntelata, Ovidiu Simion Cotoi

Melanoma is a malignant cancer of the skin, the incidence of which has been increasing year by year. This neoplasm has high aggressivity as well as the potential for invasion and metastases. Multiple factors related to the proliferation of this type of tumor have been identified, such as exposure to ultraviolet (UV) radiation and specific genetic backgrounds. From a histological and cytological point of view, the most common cells that are found in melanoma are epithelioid or spindle cells. To confirm the diagnosis and the melanocytic origin of the tumor, specific and sensitive markers are used. Also, observation of the behavior of this cancer, including its proliferative properties, has led to the development of multiple therapies, each of which is characteristic of the pathological stage at the time of diagnosis. While surgery is the most important therapeutic and curative option in cases of melanoma in situ, chemotherapy has been the main treatment for advanced stages of melanoma for many years. However, recently, targeted therapy and immunotherapy have changed the approach to treatment. At present, multiple studies are attempting to obtain further data about the tumor microenvironment and investigating how targeting particular molecules can change the prognosis of patients.

Corresponding author: Andrada Raicea, MD; e-mail: andradaraicea@gmail.com

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3. Conservative treatment of borderline ovarian tumors: a retrospective study

Constantin-Cristian Vaduva, Carmen Constantinescu, Mihaela Tenovici, Lidia Boldeanu, Anca-Maria Istrate-Ofiteru

Borderline ovarian tumors (BOTs) are a group of tumors with histological aspects and intermediate biological evolution between benign and malignant tumors, characterized by epithelial proliferation, lack of stromal invasion and nuclear atypia. BOTs account for approximately 10-15% of epithelial ovarian carcinomas. The interest in fertility preservation is very important as most BOTs are diagnosed in patients less than 40 years of age. Since borderline tumors occur in young, fertile women, the therapeutic approach depends on both staging and the need to preserve ovarian function and fertility. Treatment of BOT is primarily surgical, but recently fertility-preserving surgery has become more important. If infertility persists, ovarian induction or in vitro fertilization (IVF) may be suggested in selected cases.

Corresponding author: Constantin-Cristian Vaduva, Associate Professor, MD, PhD; e-mail: cristian.vaduva@umfcv.ro; Lidia Boldeanu, Lecturer, MD, PhD; e-mail: lidia.boldeanu@umfcv.ro

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4. S100A8/A9 is a valuable biomarker and treatment target to detect and modulate neutrophil involvement in myocardial infarction

Razvan Gheorghita Mares, Adrian Horatiu Sabau, Iuliu Gabriel Cocuz, Mihaela Elena Tomut, Istvan Adorjan Szabo, Andreea Raluca Szoke, Andreea Catalina Tinca, Gabriel Jakobsson, Ovidiu Simion Cotoi, Alexandru Schiopu

Myocardial infarction (MI) leads to irreversible ischemic damage of the heart muscle and is the leading cause of heart failure. The ischemic cardiac injury triggers a potent local and systemic immune response. In the acute phase post-MI, neutrophils infiltrate the myocardium in large numbers and induce further cardiomyocyte death, expanding the infarcted area. The alarmin S100A8/A9 is a proinflammatory mediator primarily produced by myeloid cells, with an emerging role in MI. We previously demonstrated that short-term inhibition of S100A8/A9 during the inflammatory phase of the immune response to MI improves long-term cardiac function. In the present study, we investigated the effects of S100A8/A9 blockade on myocardial inflammation and post-ischemic myocardial injury in a mouse model of coronary artery ligation. Immunohistochemical (IHC) staining revealed that the presence of S100A9 is strongly correlated with neutrophil infiltration in the myocardium on days 1 and 3 post-MI. A 3-day treatment with the S100A8/A9 blocker ABR-238901 starting immediately after MI decreased the number of neutrophils and S100A9 presence in the myocardium and had a positive impact on cardiac damage, reducing infarction size. These findings promote S100A9 as an IHC biomarker of neutrophil infiltration and a promising immunomodulatory target to regulate neutrophil recruitment, reduce ischemic injury and promote long-term beneficial cardiac recovery after MI.

Corresponding author: Adrian Horatiu Sabau, MD, PhD Student; e-mail: adrian-horatiu.sabau@umfst.ro

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5. The added value of CA125, HE4, and CA72-4 as markers for ovarian endometriosis diagnosis

Romeo Micu, Adriana Maria Ioana Gaia-Oltean, Livia Budisan, Cornelia Braicu, Alexandru Irimie, Ioana Berindan-Neagoe

Objective: This study aimed to evaluate the prognostic value as diagnosis makers of cancer antigen (CA)125, human epididymis 4 (HE4), and CA72-4 serum levels in ovarian endometriosis (OvEndo). Patients, Materials and Methods: The serum levels of CA125, HE4, and CA72-4 were measured using enzyme-linked immunosorbent assay (ELISA) technique for a group of 29 cases of OvEndo and a control (CTR) group of 26 cases. Results: Results were compared between groups and statistical correlation was analyzed between the three biomarkers. (i) For CA125, we found a statistically significant difference in-between the mean serum levels of the two groups: 9.02 U/mL in the OvEndo group versus 7.1 U/mL in the CTR group (p=0.0158). (ii) A similar situation was found for CA72-4 levels in OvEndo group, where the mean serum level was 6.1 U/mL compared to 3.5 U/mL in the CTR group, showing a significant difference (p=0.0185). (iii) The mean serum level of HE4 in the OvEndo group was 7.6 ng/mL versus 7.8 ng/mL in the CTR group, and we found it highly significant (p=0.0001). HE4 levels were highly correlated with CA72-4 levels (p<0.0001), while CA125 levels were not correlated with HE4 and CA72-4. Conclusions: Measurements of CA125 can be used in the diagnosis of OvEndo mainly in association with HE4 serum levels, which are lower in endometriosis patients. CA72-4 levels are highly correlated with HE4 levels in patients with OvEndo, while no correlation with the other two markers was found. This correlation needs further investigation to establish if it may be used as a possible diagnostic tool in clinical practice.

Corresponding author: Ioana Berindan-Neagoe, Professor, MD, PhD; e-mail: ioananeagoe29@gmail.com

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6. Immunoexpression of Claudins -3, -4 and -7 in prostate adenocarcinomas

Tudor Cristian Timotei Popescu, Alex Emilian Stepan, Mirela Marinela Florescu, Mioara Desdemona Stepan, Cristiana Eugenia Simionescu

Claudins are a family of essential tight junction proteins, abnormally expressed in human carcinomas. The studies that indicated the involvement of claudins in tumor biology and progression suggest the possibility of their utility as markers for diagnosis or prognosis, but also as possible targets for therapy. We investigated 50 prostate adenocarcinomas (PAs) for which we followed the expression of Claudins -3, -4 and -7 in relation to International Society of Urological Pathology (ISUP) grades. We observed the positivity for Claudin-3, Claudin-4, and Claudin-7 in 76%, 74% and 46% of cases. Analysis of the immunoexpression pattern revealed the cytoplasmic and nuclear translocation for Claudins -3 and -4, and only cytoplasmic for Claudin-7. For all claudins investigated, we noted a final staining score with significantly higher values or at the limit of statistical significance for PA belonging to ISUP groups 1-4. The internalization of Claudins -3, -4 and -7 expression, regardless of the degree of PA, indicates their involvement in prostate carcinogenesis. In addition, the similar immunoexpression patterns of the three investigated claudins and their positive linear correlation suggest a coordinated regulation and indicate the possibility of a targeted treatment strategy.

Corresponding author: Alex Emilian Stepan, Professor, MD, PhD; e-mail: astepan76@yahoo.com; Mirela Marinela Florescu, Teaching Assistant, MD, PhD; e-mail: mirelaflorescu88@gmail.com

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7. In vitro evaluation of Axitinib and Sorafenib treatment in glioblastoma cell viability and morphology

Alexandru Oprita, Mihaela Amelia Dobrescu, Elena Victoria Manea, Stefana Oana Popescu, Ani Simona Sevastre, Andreea Silvia Pirvu, Iuliana Mihaela Buzatu, Daniela Elise Tache

The formation, proliferation, and evolution of glioblastoma (GB) are significantly influenced by pathological angiogenesis. This is supported by several growth factor receptors, such as the vascular endothelial growth factor receptor (VEGFR). In this experiment, we examined how the Food and Drug Administration (FDA) approved VEGFR blockers Sorafenib and Axitinib affect the viability of GB cells in vitro. Cells were cultivated in 96-well culture plates for the experiments, afterwards Sorafenib and Axitinib were administered at doses ranging from 0.3 microM to 80 microM. 2,5-Diphenyl-2H-tetrazolium bromide (MTT) assay was used to assess the impact of VEGFR inhibition on high-grade glioma (HGG) cell lines. To observe the morphological changes in cell shape, we used a 10x magnification microscopy. Our results showed that both Axitinib and Sorafenib retarded GB1B culture proliferation in a dose- and time-dependent manner in comparison to control cohorts that had not received any treatment. The half maximal inhibitory concentration (IC50) value for Axitinib was 3.5839 microM after three days of drug administration and 2.2133 microM after seven days of drug administration. The IC50 value for Sorafenib was 3.5152 microM after three days of drug administration and 1.6846 microM after seven days of drug administration. After the treatment with Axitinib or Sorafenib, very few cells became rounded and detached from the support, others remained adherent to the culture substrate, but acquired a larger, flatter shape. Our results indicate that VEGFR might serve as a key target in the treatment of GB. Although it is known that in vitro some drugs block the VEGFR more potently, clinical evidence is required to show whether this actually translates to better clinical outcomes.

Corresponding author: Iuliana Mihaela Buzatu, MD, PhD Student; e-mail: buzatu_iuliana@yahoo.com

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8. Involvement of TLR9 in priming the immune response in oral papillomatosis induced by low-risk HPV

Elena Cristina Andrei, Maria Cristina Munteanu, Cristina Jana Busuioc, Catalina Gabriela Pisoschi, Garofita Olivia Mateescu, Sanda Amelia Dracea, Ileana Monica Banita

Oral papillomatosis represents a benign lesion of the oral mucosa often induced by human papillomavirus (HPV) or having a non-infection local or general etiology. HPVs are very well adapted and efficient viruses able to produce changes in the immune system, endowed with the ability to replicate in the keratinocytes and to remain silent. The natural evolution of HPV infection is different, depending on the efficiency of the innate immune system. The purpose of this study was to explore Toll-like receptor 9 (TLR9) immunohistochemical expression in low-risk (LR)-HPV oral infection and its ability to facilitate an efficient immune response by activating the macrophages, which serve as main antigen-presenting cells. Samples of two groups of oral mucosae - LR-HPV-positive and HPV-negative - were processed for immunohistochemistry technique and incubated with antibody against TLR9 and cluster of differentiation 68 (CD68). Image analysis and morphometry were conducted to assess the intensity of TLR9 immune signal in the epithelium and the number of macrophages labeled by CD68. We found a statistically significant difference between macrophage count for the subjects in HPV-positive and HPV-negative groups; thought no significant differences of TLR9 immune signal was noted, which demonstrates a diminished immune response in HPV-positive group, probably influencing the time of lesion s clearance.

Corresponding author: Catalina Gabriela Pisoschi, Professor, PhD; e-mail: catalina.pisoschi@umfcv.ro; Maria Cristina Munteanu, Lecturer, PhD; e-mail: cristina.munteanu@umfcv.ro

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9. Otosclerosis under the magnifying glass

Claudia Raluca Balasa Virzob, Raluca Maria Closca, Marioara Poenaru, Raluca Morar, Nicolae Constantin Balica, Cristian Andrei Sarau, Ioana Ionita, Flavia Baderca

Otosclerosis is a bone condition affecting the stapes bone within the otic capsule, and its exact cause is still unknown. It is characterized by a lack of proper remodeling of newly formed vascular and woven bone, leading to the development of abnormal osteons and the formation of sclerotic bone. Bilateral otosclerosis is seen in 80% of patients and 60% of otosclerosis patients have a family history of the condition. The etiology of this disease is still unknown, there are lots of theories to explain it. The histopathological (HP) studies of otosclerosis showed that osteoblasts, osteoclasts, vascular proliferation, fibroblasts, and histiocytes were observed in the stapes footplate. The onset of the symptoms occurs by the early third decade of life, usually it doesn t start later. In otosclerosis, the energy exerted by sound at the level of the tympanic membrane is reduced in the inner ear due to the fixation and rigidity of the ossicular chain, leading to hearing loss, especially for low frequencies. The primary clinical symptom of otosclerosis is conductive hearing loss but it is important to note that sensorineural hearing loss and mixed hearing loss can also occur as secondary symptoms of the condition. Another symptom present in patients with otosclerosis is tinnitus. The paper carried out a retrospective study of 70 patients diagnosed with otosclerosis in the Department of Otorhinolaryngology of Emergency City Hospital, Timisoara, Romania, between January 2021 to December 2022. Tissue fragments were processed at Service of Pathology by standard Hematoxylin-Eosin staining. The HP diagnosis was completed using Masson s trichrome staining, Giemsa histochemical staining, and immunohistochemical (IHC) reactions with anti-cluster of differentiation (CD)20, anti-CD3, anti-CD4, anti-CD8, anti-CD34, and anti-CD31 antibodies. The microscopic examination showed a chronic diffuse inflammatory infiltrate that consisted predominantly of mature T-lymphocytes, immunohistochemically positive for CD3, CD4 and CD8. There were also present rare CD20-positive B-lymphocytes. Among the lymphocytes, relatively numerous mast cells were identified, highlighted histochemically by the Giemsa staining. They had numerous purple-violet intracytoplasmic granules. In the connective tissue support, a relatively rich vascular network was identified, consisting of hyperemic capillaries, highlighted immunohistochemically with anti-CD31 and anti-CD34 antibodies. Bone tissues trabeculae showed extensive areas of fibrosis. The collagen fibers were highlighted by Masson s trichrome staining, being stained in green, blue, or bluish green.

Corresponding author: Ioana Ionita, Associate Professor, MD, PhD; e-mail: ionita.iulia@umft.ro

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10. Clinical and morphofunctional identity of the nasal SMAS

Marius Valeriu Hinganu, Delia Hinganu, Octavian Dragos Palade, Iuliana Eva, Simona Ruxandra Volovat, Ramona Paula Cucu, Victor Vlad Costan

The fascial system of the face (superficial musculo-aponeurotic system, SMAS) in the nasal part is a sustained layer that connects the nearby regions. In this paper, we aimed to emphasize the presence of SMAS in different areas of the nasal region: ala nasi, nasolabial fold, nasal dorsum and radix. We performed three studies (anatomical, histological, and radiological) to demonstrate the existence of nasal SMAS. The study group consisted of cadaveric analyses and retrospective analysis of the patient radiological data. The nasal SMAS was identified as a superficial fascia and a subcutaneous adipose layer. The anatomical dissection study together with histological and radiological evaluations demonstrated the presence of SMAS in the nasal region. We identified peculiarities of the nasal SMAS in two areas: in the ala nasi where it is thinner, and the deep part of the dermis does not adhere to the underlying structures and at the radix and dorsum nasi, where the adipose layer is very thin. The results of our research define nasal SMAS as a unit of great value in facial surgeries, such as facial rejuvenation, the resolution of malformations, or tumor removal.

Corresponding author: Delia Hinganu, Associate Professor, MD, PhD; e-mail: delia_f24@yahoo.com; Octavian Dragos Palade, Associate Professor, MD, PhD; e-mail: drpalade@gmail.com

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11. Effects of Coriandrum sativum on distant organ inflammation and apoptosis due to liver ischemia/reperfusion injury

Aysel Kukner, Pelin Toros, Gizem Soyler, Selin Isik, Gozde Ogutcu, Oguzhan Edebal, Filiz Mericli

Liver ischemia/reperfusion (IR) often affects distant organs, such as small intestine, kidney, and lung. Coriandrum sativum (CS) has an antioxidant and anti-inflammatory effect on liver damage. The aim of this study was to investigate the anti-inflammatory and antiapoptotic effects of CS extract on small intestine, lung, and kidney after the liver IR injury. Small intestine, lung, and kidney tissues were evaluated and scored in terms of cell degeneration, inflammation, and congestion, as well as caspase-3 (Cas-3) and cluster of differentiation 31 (CD31) immunostainings were carried out. Renal enzymes, creatinine and urea levels were measured biochemically in serum. After IR, a decrease in villi size, diffuse degeneration, epithelial cell shedding and extensive congestion in the capillaries were observed. Meanwhile, the number of degenerated villi and congestion decreased in the IR+CS group. Due to IR, increased congestion was detected in the interalveolar septum of the lungs and in the capillaries between the kidney tubules. It was also observed that the positively stained cells with Cas-3 and CD31 were increased in the lung, kidney, and small intestine tissues of the IR group, and decreased in the IR+CS group. Kidney enzymes, urea and creatinine levels were significantly increased in the IR group and decreased in the IR+CS group. In conclusion, it was observed that liver IR caused changes in distant organs, especially in the small intestine, lung, and kidneys. Damaging effects of IR as well as apoptosis and inflammation were found to be decreased in the groups treated with CS.

Corresponding author: Aysel Kukner, Professor, PhD; e-mail: akukner@hotmail.com

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12. Placental pathology in early-onset fetal growth restriction: insights into fetal growth restriction mechanisms

Daniela-Loredana Bujorescu, Adrian Claudiu Ratiu, Andrei Gheorghe Marius Motoc, Ioan Cosmin Citu, Ioan Sas, Ion Florin Gorun, Oana-Maria Gorun, Roxana Folescu, Daniela Gurgus

Background: Early-onset fetal growth restriction (FGR), an identifiable variant of FGR, exhibits divergences in its severity, management, and placental pathologies when juxtaposed with late-onset FGR. The objective of this cross-sectional investigation was to scrutinize placental pathologies in pregnancies afflicted by early-onset FGR, emphasizing a comparative analysis between cohorts with and without preeclampsia (PE). Patients, Materials and Methods: The study encompassed a cohort of 85 expectant mothers who received a diagnosis of early-onset FGR. Rigorous histopathological (HP) and immunohistochemical (IHC) assessments were conducted on the placentas. Comparative analyses were performed, distinguishing between individuals diagnosed with both PE and early-onset FGR, and those presenting normotensive early-onset FGR. Results: HP analysis unveiled a multitude of shared placental lesions, encompassing retroplacental hemorrhage, expedited villous maturation, infarctions, and calcification-associated fibrin deposits. IHC investigations displayed affirmative immunoreactivity for anti-hypoxia-inducible factor (HIF) and anti-vascular endothelial growth factor (VEGF) antibodies within the placental infarcted villitis. Moreover, noteworthy variances in placental measurements and distinctive lesions were discerned when comparing the PE and early-onset FGR cohort with the normotensive group. Conclusions: Maternal malperfusion emerged as a pivotal determinant linked to placental lesions in pregnancies affected by early-onset FGR. Remarkably, the occurrence of infarctions, specifically delayed infarctions, exhibited a noteworthy correlation with PE. These findings accentuate the significance of pursuing additional research endeavors aimed at unraveling the intricate mechanisms governing maternal malperfusion and its consequential influence on placental health in the context of early-onset FGR, with particular attention to the interplay with PE.

Corresponding author: Adrian Claudiu Ratiu, Lecturer, MD, PhD; e-mail: dr.ratiu@gmail.com

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13. Epidemiological and pathological characteristics of spinal metastases from gastrointestinal cancers - a series of 40 cases

Stefan Selaru, Anca Sava, Dragos Viorel Scripcariu, Claudia Florida Costea, Ana Maria Dumitrescu, Bogdan Costachescu, Gabriela Florenta Dumitrescu, Corina Ciupilan, Ruxandra Vatavu, Raluca Maria Haba, Vladimir Poroch, Lucia Corina Dima-Cozma, Valeriana Vornicu, Cristinel Ionel Stan

Spinal metastases from gastrointestinal (GI) cancer are rare and as a result there are only case reports or small series in the literature. The aim of our work was to identify the demographic aspects, the location, and the histopathological aspects of spinal metastases from GI cancers diagnosed and treated in a reference Hospital in Romania over a period of nine years, and comparing the data obtained with those from the recent literature. This is a retrospective case series study on spinal metastases from GI cancers, developed in patients older than 18 years that were surgically treated between January 2013 and December 2021 within three Neurosurgery Clinics from Prof. Dr. Nicolae Oblu Emergency Clinical Hospital, Iasi, Romania, which is a tertiary Hospital in Romania regarding the surgical treatment of spinal metastases. We included in our study the patient s demographic data (age and gender), clinical data (location of spinal metastases), radiological investigations and pathological features of the lesions. Regarding the immunohistochemical stainings, the following antibodies were used: anti-cytokeratin (CK)7, anti-CK20, anti-CK19, anti-caudal-type homeobox 2 (CDX2), anti-human epidermal growth factor receptor 2 (HER2), and anti-Ki67. Our series included 40 adult patients (>=18 years old) with a male:female ratio of 3:1, in favor of male patients. The mean age of all patients was 66.42 years. The primary sites of spinal metastases from GI cancers were from all segments of the GI system: the most frequent, however, was from the colorectal level (40%) and the least from the oral cavity level (2.5%). The most common site of spinal metastases from GI cancer was predominantly lumbar region (47.5%), and the most frequent histological type was adenocarcinoma (57.5%), followed by hepatocellular carcinoma (27.5%), pancreatic ductal carcinoma (5%) and squamous cell carcinoma (2.5%). Our results have important clinical implications because they suggest that there are certain subsets of patients with certain types of GI cancers that cause metastases in certain regions of the spine.

Corresponding author: Anca Sava, Professor, MD, PhD; e-mail: dr.anca.sava.68@gmail.com; Dragos Viorel Scripcariu, MD, PhD; e-mail: dscripcariu@gmail.com

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14. Nasal polyposis from cystic fibrosis in children - the experience of a single center

Gheorghe Iovanescu, Liviu Laurentiu Pop, Daniela Roxana Vintila, Karina Cristina Marin, Carmen Aurelia Mogoanta, Dorela Codruta Lazureanu, Veronica Madalina Boruga, Ioana Mihaela Ciuca

Background: Cystic fibrosis (CF) is the most common monogenic disease, characterized by clinically notable polymorphism. Respiratory disease is the main factor that influences the disease outcome and prognosis of the patient with CF, bacterial infections being responsible for severe exacerbations and rhinosinusitis a difficult complication, besides lung disease. Aim: The aim of the paper was to present a case series of CF-associated nasal polyposis and our management experience, providing new data for nasal and sinus complications. Patients, Materials and Methods: Patients attending the National Cystic Fibrosis Center, Timisoara, Romania, were evaluated for nasal polyposis. Besides clinical examination, endoscopy, and computed tomography (CT) was performed for comprehensive evaluation. Patients with persistent symptoms or with complicated sinusitis underwent surgical approach. Results: Fourteen (18.18%) children were diagnosed with nasal polyposis and had surgery, with positive outcomes. One patient received Omalizumab for an associated, uncontrolled asthma with a subsequent substantial effect with the significant polyp reduction and lack of recurrence. Conclusions: Even if extremely difficult to manage, complicated nasal polyposis CF related might have an improved outcome and better life quality.

Corresponding author: Carmen Aurelia Mogoanta, Associate Professor, MD, PhD; e-mail: carmen_mogoanta@yahoo.com

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15. A clinical and statistical study on enamel hypomineralization of the first permanent molar in the period of mixed dentition

Stephanie-Nicole Stoica, Victor Nimigean, Simona Andreea Moraru, Ioan Sirbu, Vanda Roxana Nimigean

Background: The first permanent molar (FPM) is the first tooth in the permanent dentition that emerges in the oral cavity, at around the age of six and behind the deciduous teeth, thus converting the primary dentition into a mixed dentition. Its early formation represents a risk factor for the onset of hard dental tissues disorders, the most common being the molar incisor hypomineralization (MIH). Aim: The aim of the study was to assess the hypomineralization type of developmental defects of enamel at FPM level during the mixed dentition stage, in a group of Romanian children. Patients, Materials and Methods: The research was conducted on 87 patients, 44 females and 43 males, divided into the following age groups: 5-6 years, 6-7 years, 7-8 years, 8-9 years, 9-10 years, 10-11 years, 11-12 years and older than 12 years. All children were examined in the dental office by a pediatric dentist. Results: Of the subjects, 40.2% presented hypomineralization lesions on the FPM. Among the affected children, 62.9% were girls and 37.1% were boys. Mild forms were the most prevalent, being found in 71.4% of the affected subjects. Moderate forms were present in 22.9% and severe forms in 5.7% of the affected children. MIH prevalence significantly decreased with increasing age. Only two children belonging to the group 8-9 years and 10-11 years, respectively, showed severe MIH lesions. Statistically significant differences were determined in what regards the distribution of MIH children according to the severity of the lesions in relation to age (p=0.007). Conclusions: Children 6- to 7-year-old have been the most affected by FPM hypomineralization, which indicates the need for early management of the disease, through a thorough diagnosis, and preventive and interceptive therapeutic approaches.

Corresponding author: Victor Nimigean, Professor, DMD, MD, PhD; e-mail: victornimigean@yahoo.com; Simona Andreea Moraru, Assistant Professor, DMD, MD, PhD; e-mail: simona.gaspar@yahoo.com

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16. Assessment of tumor microenvironment in gastric adenocarcinoma

Marius Ionut Stancu, Alexandru Giubelan, George Mitroi, Anca-Maria Istrate-Ofiteru, George Popescu, Sorina Octavia Hontaru, Oana Badea-Voiculescu, Denisa Floriana Vasilica Pirscoveanu, Stelian Stefanita Mogoanta, Laurentiu Mogoanta

Gastric cancer (GC), despite the current possibilities of early diagnosis and curative treatment, remains a major public health problem, being one of the main causes of cancer, due to its detection in advanced stages. Screening programs applied in Western countries led to low incidence rates in these countries. Helicobacter pylori bacterial infection is considered to be the highest risk factor for the onset of GC because it causes chronic inflammation of the gastric mucosa and damages hydrochloric acid secretory glands, eventually leading to atrophic gastritis, which has a potential to progress to GC. In our study, we aimed at assessing the tumor microenvironment in gastric adenocarcinomas as approximately 90% of GCs are adenocarcinomas. Our study showed that the tumor microenvironment has an extremely complex morphological structure, totally different from the microscopic structure of the gastric wall, consisting of stromal cells, lymphocytes, plasma cells, macrophages, blood vessels, collagen fibers, extracellular connective matrix, other cells. The tumor microenvironment presents phenotypic, cellular and molecular heterogeneity; therefore, the microscopic aspect differs from one tumor to another and even from one region to another in the same tumor. Poorly or moderately differentiated adenocarcinomas show a more intense desmoplastic reaction than well-differentiated ones. Alpha-smooth muscle actin (alpha-SMA)-positive stromal cells (tumor-associated fibroblasts) and tumor macrophages were the most numerous cells of the tumor microenvironment. The tumor microenvironment is the result of cooperation between tumor cells, cancer-associated fibroblasts, immune system cells and blood vessels. It allows tumor cells to multiply, grow and metastasize.

Corresponding author: Stelian Stefanita Mogoanta, Professor, MD, PhD; e-mail: ssmogo@yahoo.com; George Mitroi, Associate Professor, MD, PhD; e-mail: gmitroi@yahoo.com

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17. The role of gut dysbiosis in endometriosis diagnosis and treatment approaches - case report

Alexandra Irma Gabriela Bausic, Sanda Maria Cretoiu, Vasilica Bausic, Daniela Roxana Matasariu, Ruxandra Viorica Stanculescu, Elvira Bratila

Background: Endometriosis (EM) is a chronic multifactorial disease characterized by the presence of endometrial tissue outside the uterus. The clear etiopathogenesis of EM is unclear. Increasing evidence was gathered about the crucial involvement of gut microbiota in early stages of the disease, and in its progression. Case presentation: We report the case of a 33-year-old Caucasian woman diagnosed with EM, that presented with painful pelvic (dysmenorrhea, dyspareunia) and gastrointestinal (GI) symptomatology. The patient underwent an intestinal microbiota analysis before the surgical treatment was performed. Discussions: The GI microbiome culture identified high levels of non-pathogen bacteria Escherichia coli, Bifidobacterium, hemolytic E. coli and potential pathogens: Hafnia alvei and Enterobacter cloacae. The mycology culture performed identified the presence of potential pathogens: Candida albicans and C. glabrata. Microscopic examination and polymerase chain reaction (PCR) analysis showed Giardia lamblia in moderate amounts. These findings were compared with the information available in the literature of specialty and they imply that the patient intestinal microbiome is heavily disrupted. Conclusions: There are changes in the microbiota of EM patients in comparison to those not suffering from this disease. The findings addressed in this article characterize the intricate bilateral connection between the microbiota and EM. The goal of future studies ought to be to establish how the microbiome and EM are interconnected by implementing breakthrough diagnostic and treatment strategies.

Corresponding author: Vasilica Bausic, Associate Professor, MD, PhD; e-mail: valibausic@gmail.com; Sanda Maria Cretoiu, Associate Professor, MD, PhD; e-mail: sanda.cretoiu@umfcd.ro

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18. A rare subcutaneous periorbital zoonotic infection in a 42-year-old female patient

Lucian Mihai Florescu, Rossy Vladut Teica, Cristina Mihaela Ciofiac, Loredana Elena Stoica, Ioana Andreea Gheonea

Human dirofilariasis represents a zoonotic infectious disorder caused by parasites belonging to the Dirofilaria genus, which includes numerous species with a large variability regarding the host specificity, life cycle, and clinical manifestations. This disease appears to be a vector-borne parasitosis that is spread through insects - intermediate hosts (usually mosquitos) - and affects several carnivores as definitive hosts. Humans represent an unsuitable host for the parasite to complete its life cycle, being unable to release microfilariae in the blood as the inoculated larvae fail to reach sexual maturity. Therefore, humans are unable to transmit the infection to other humans, neither directly nor through an intermediate host. The current case report indicates a rare encounter of a Dirofilaria repens infection in a 42-year-old female patient living in an urban area (Craiova, Romania), who developed intermittent right periorbital edema after a previous trip to Greece. Over the course of one month, the right periorbital edema gradually remitted, and a firm, round lump developed in the external inferior right periorbital region. There were no similar clinical manifestations among other family members. Her medical history showed no other complaints or any serious general health problems. Also, the patient denied having any pets at home. The blood tests did not reveal any abnormalities. The exact source of infection could not be identified, but the chances for this infection to be related to the history of traveling to Greece are high, given that this region is one of the most important endemic areas in Europe.

Corresponding author: Rossy Vladut Teica, MD, PhD Student; e-mail: rossy.teica@gmail.com

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19. A rare find of a maxillary third molar with five roots: a case report of an unusual morphology

Oana Cella Andrei, Gabriela Ciavoi, Magdalena Natalia Dina, Liana Todor, Daniela Ioana Tarlungeanu, Ruxandra Margarit

Third molars are frequently absent or impacted; their unpredictable prevalence and morphology, with frequent anatomical variations, is a challenge for the practitioner. The number of roots of the upper third molars vary from one to five, but five roots upper third molars cases are extremely rare. The case reported here is of a normotrophic male patient, with no significant general medical history, having extracted an impacted maxillary upper molar, with an unusual position and dimensions and five completely separated formed roots, orientated in diverse directions. It highlights the importance of using the correct surgical technique, without excessive pressure, for avoiding the fracture of the maxillary tuberosity, the displacement of the molar into the pterygopalatine fossa or the opening of the maxillary sinus, especially in the absence of good quality imagistic information like cone-beam computed tomography. The case presented in this paper is very rare and highlights the significance of anatomical knowledge, which is mandatory for managing these cases, to avoid trauma, accidents, and complications and to maintain an optimal time of extraction, since such variations occur, and the conventional radiographs do not offer enough data to clarify the diagnosis.

Corresponding author: Liana Todor, Associate Professor, DMD, PhD; e-mail: liana.todor@gmail.com; Daniela Ioana Tarlungeanu, Assistant Professor, DMD, PhD; e-mail: ioanatarlungeanu@gmail.com

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