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Background: The effects of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection during pregnancy remain relatively unknown., Aim: We present this original paper where we analyzed 60 parturients, at term, 30 without associated infection (C-) and 30 with associated infection (C+), present at birth., Methods: We analyzed the blood count and placental microscopic structure through classical and immunohistochemical staining and observed the placental areas affected by the presence of SARS-CoV-2., Results: SARS-CoV-2 infection was accompanied by a decrease in the number of lymphocytes, the number of platelets and the presence of placental structural changes, identifying extensive areas of amyloid deposits, placental infarcts, vascular thrombosis, syncytial knots, with a decrease in placental vascular density and the presence of infection in the cells located at decidual level, at syncytiotrophoblast level and at the level of the cells of the chorionic plate, still without overcoming this barrier and without causing any fetal infection in the analyzed cases., Conclusions: This study shows that the invasion of SARS-CoV-2 in the placenta can produce significant structural changes, with a decrease in placental vascular density that can have significant implications on proper fetal perfusion. Also, the presence of immunoreactivity at the level of decidua, the placental villi, as well as the chorionic plate proves that the virus can overcome the maternal-fetal barrier. However, in the analyzed cases there were no fetal infections at birth, which may show that local placental factors can be a protective filter for the fetus.

Hepatocellular carcinoma (HCC) is the main primary liver malignancy, being associated with both health and economic burden worldwide. Recently, novel molecular markers and possible therapeutic targets were identified. Different adhesion molecules, as well as possible angiogenesis-associated targets can be prime candidates when investigating novel therapies. Considering these premises, our goal was to study the co-existence of E-cadherin and aquaporin 1 (AQP1) in a series of HCC diagnosed patients. Utilizing archived tissue fragments from 17 patients diagnosed with well-to-moderate and poorly differentiated HCC, as well as four samples of normal liver tissue and using a highly specific biotin-free tyramide amplification technique, we have assessed here the expression of E-cadherin and AQP1 during HCC carcinogenesis. Moreover, as we have observed that some of the AQP1 expression seems membrane-bound, we have sought to evaluate their co-localization. Our data showed, as expected, that E-cadherin decreases from control tissue to low-grade and respectively, high-grade HCC. AQP1 was expressed, also as already known, at the level of endothelial blood vessels and bile ducts epithelia, however, we have showed here for the first time that this water pore is also expressed in the cytoplasm and membranes of hepatocytes, both in control and HCC tissue. Moreover, AQP1 expression parallels the decrease of E-cadherin expression during carcinogenesis, but together with this downregulation, we have also found a spatial decrease in the colocalization of the two proteins. Altogether, utilizing a biotin-free tyramide signal amplification technique, this study shows for the first time that AQP1 is expressed at the level of liver epithelia, in both control and HCC tissue.

Objective: In this pilot study, we tested the feasibility of cardiac structures reconstruction from histological sections in 12-13 weeks normal fetuses. Conventional autopsy is hampered at this gestational age because of the small size of the heart anatomical structures, while alternative non-invasive methods for pathology examination of the fetus are expensive, rarely available and lack accuracy data regarding the confirmation of first trimester heart defects suspected by early prenatal ultrasound (US) scans., Materials and Methods: Normal hearts from fetuses aged 12-13 gestational weeks (GW) were harvested for histological preparation, virtual reconstruction, and cardiac structures analysis. The normalcy of heart structures was confirmed before pregnancy termination, using a detailed US scan protocol. The fetal heart was routinely processed for formalin fixation and paraffin embedding (FFPE) and 10 μm seriate sections have been cut until finishing the specimen. All sections have been scanned and a three-dimensional (3D) reconstruction of the whole organ has been rendered, based on computer-aided manual tracing. Using the 3D navigation software, the main cardiac structures were searched for a proper and confident visualization., Results: Five cases were investigated. Visualization of the normal heart cavities, including atrioventricular septum was very good in all fetuses. The entire course of right and left ventricle outflow tracts was confidently confirmed, along the branching pattern of aorta and pulmonary artery trunk. Regarding the veno-atrial connections, it was easy to identify the entrance of the inferior and superior caval veins into the right atrium, but a detailed review of the histological sections was necessary for the visualization of the left atrium venous openings. The inherent morphological deformation following heart block sectioning resulted in a lower resolution or quality of the "reconstructed" planes, but these distortions did not represent a significant impediment in any of the cases. The resources involved ordinary histology and information technology (IT) equipment. To further decrease the time involved by the protocol, many steps may be automated: cutting, coloring, and scanning., Conclusions: The results indicate that this method can be implemented to routine clinical practice. The use of 3D reconstruction of fetal heart histological sections in first trimester may serve as an important audit to confirm the normalcy of heart structures. Also, the histological and postprocessed information is retained, and this volume can be stored, reanalyzed, or sent online for a second opinion. The method involves relatively undemanding resources, i.e., hardware, software, competences, and time. The procedure could also benefit from refinements used in other imaging techniques to limit human-computer interactions, such as sections distortion.

Skin burns are one of the most common injuries associated with increased morbidity and mortality, especially in the children and the elderlies. Severe burns, especially, result in a systemic immune and inflammatory response, which may reflect in multiple organ insufficiency, and a fast and effective local restorative process is essential for functionality recovering, as well as for interrupting the generalized systemic response. We have aimed here to assess the effect of different wound dressings in what it regards the morphology and clinical restoration after a skin burn. On a rat animal model, we have evaluated the macroscopic and histopathological features of controlled third degree skin burns in control animals versus treatments with local dressings of silver sulfadiazine (SDA) cream, simple gel (G), gel + silver nanoparticles (AgNPs) (G+NPS), gel + exosomes (G+EXO) and gel + AgNPs + exosomes (Gel+NPS+EXO), at 14 days and, respectively, 21 days after the lesion. Tissue fragments were harvested and processed for histopathology and immunohistochemistry. Immunofluorescence was utilized to evaluate the maturity of underlaying granulation tissue based on double stainings for smooth muscle actin (SMA) and cluster of differentiation 31 (CD31). Our study showed variability in what it regards the vessel density and immunoexpression of SMA between the treatments, and image analysis revealed that most SMA reduction and blood vessel density reduction in the maturing granulation tissue occurred for the G+NPS and G+NPS+EXO treatments. A complete re-epithelization was also observed for the G+NPS+EXO treatment. Overall, our results show that improved topic treatments promote faster re-epithelization and reparation of the dermis after skin burn lesions, providing thus an avenue for new treatments that aim both local recuperation and systemic infection prevention.

Schwannomas are well-encapsulated, benign tumors arising from Schwann cell constituting the nerve sheaths. We report a case of a cystic conventional schwannoma, measuring 2.5×3 cm, located in the distal forearm of a 33-year-old female patient with no sign of neurofibromatosis type 2 or type 3. The mass was painless, but Tinel's sign was positive. Ultrasonography showed a soft tissue cystic mass diagnosed as being synovial cyst in the left distal forearm. Surgical resection was performed without any complications. Histopathological examinations oriented the diagnosis to a schwannoma, but immunohistochemical stainings proved the tumor origin in Schwann cells and made the differential diagnosis with other pathologies, like neurofibroma, leiomyoma, epithelial cyst, synovial cyst, or ganglion cyst.

The fundamental discovery of the hepatitis C virus (HCV) in 1989 has led to winning this year's Nobel Prize in Medicine. This achievement guided all the steps in identifying the elements of the virus, in order to develop the treatment and to increase the screening solutions, which have slowed the exposure to the virus. The management of infection started with interferon-alpha (IFN-α), which has later enhanced by adding Ribavirin. Nowadays, HCV treatment is based on direct-acting antiviral agents (DAAs). Currently, HCV infection benefits of curative treatment, with which most patients can be cured. When speaking about hepatitis C future, we can say it is looking bright, considering all the progress that has been made in recent years and all the options that we have for curing all genotypes of HCV infection. The aim of this review is to sum up the historical characteristics of HCV discovery, the evolution of treatment and screening actions, gaps, and stages for achieving the international elimination target of the World Health Organization.

Introduction: While the visual inspection of histopathology images by expert pathologists remains the golden standard method for grading of prostate cancer the quest for developing automated algorithms for the job is set and deep-learning techniques have emerged on top of other approaches., Methods: Two pre-trained deep-learning networks, obtained with transfer learning from two general purpose classification networks - AlexNet and GoogleNet, originally trained on a proprietary dataset of prostate cancer were used to classify 6000 cropped images from Gleason2019 Challenge., Results: The average agreement between the two networks and the six pathologists was found to be substantial for AlexNet and moderate for GoogleNet. When tested against the majority vote of the six pathologists the agreement was perfect and moderate for AlexNet, and GoogleNet, respectively. Despite our expectations, the average inter-pathologist agreement was moderate, while between the two networks it was substantial. Resulted accuracy for AlexNet and GoogleNet when tested against the majority vote as ground truth was of 85.51% and 74.75%, respectively. This result was higher than the score obtained on the dataset that they were trained on, showing their generalization capabilities., Conclusions: Both the agreement and the accuracy indicate a better performance of AlexNet over GoogleNet, making it suitable for clinical deployment thus could potentially contribute to faster, more accurate and with higher reproducibility prostate cancer diagnosis.

Non-alcoholic fatty liver disease (NAFLD) is one of the most common conditions worldwide that targets the liver parenchyma. NAFLD represents an intrahepatic triglyceride accumulation in the absence of excessive alcohol consumption and other diseases that affect the liver parenchyma. The current "gold standard" for evaluating the amount of intrahepatic fat is represented by liver biopsy, but many patients are reluctant and hardly accept undergoing this procedure due to its invasive nature. The current study addresses this aspect by evaluating the reliability of liver magnetic resonance spectroscopy (MRS) in diagnosing NAFLD, compared to the traditional invasive liver biopsy. The present study included a total of 38 patients based on several well-defined inclusion and exclusion criteria. We used the same NAFLD grading system for both liver MRS and liver biopsy: grade 0: <5% hepatocytes are affected; grade I: 5-33% hepatocytes are affected; grade II: 34-66% hepatocytes are affected; grade III: >66% hepatocytes are affected. Regarding the NAFLD grade, over three-quarters of patients were classified as grade I and grade II, with a strong predilection for men. The current results indicated a significant association between the NAFLD grade indicated by liver MRS and the NAFLD grade indicated by liver biopsy. At the end of our study, we recommend using liver MRS for evaluating and grading NAFLD in association with other parameters like serum triglycerides and body mass index grade as this protocol can enhance early detection and provide an accurate grading that will lead to a proper management of this disease.

We are reporting a case of natural evolution and pathological data from a young person that was diagnosed with coronavirus disease 2019 (COVID-19). All data has been collected from the autopsy of a 30-year-old female, which was performed by the Department of Forensic Medicine from Emergency County Hospital, Drobeta Turnu Severin, Mehedinţi County, Romania. The infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was confirmed by reverse transcription polymerase chain reaction (RT-PCR) on the lung tissue which was obtained during autopsy. This case provides the opportunity to study the natural evolution of COVID-19 pneumonia in a young person with clinical signs of pneumonia but without associated comorbidities. The patient had not received any treatment. The histopathological examination of the lung revealed a process of productive proliferation, proteinaceous and fibrin-macrophagic interalveolar spaces exudate, and lesions consistent with vasculitis. In the heart, we identified a cardiac thrombus. These changes are likely to suggest an advanced natural evolution of SARS-CoV-2 virus infection.

A significant cause of end-stage renal disease in infants (40% to 50% of cases) is represented by the group of renal cystic diseases. Actually, the fourth cause of renal failure in young adults is the autosomal dominant polycystic kidney disease (ADPKD). Moreover, the most common genetically inherited kidney disease was proved to be ADPKD, affecting 1-5 per 10 000 individuals. The study was conducted over a period of three years (July 26, 2015-October 30, 2018) on 22 patients aged between two days and 36 months, diagnosed with polycystic kidneys that presented multiple hospital admissions in the Department of Nephrology, "Maria Skłodowska Curie" Emergency Children's Hospital, Bucharest, Romania. The nephrectomy sections were obtained from the material of the Department of Pathology of the same Hospital. Prenatal ultrasonography results were correlated with positive family history of polycystic kidney disease (PKD), fetal enlarged kidneys and oligohydramnios. Neonatal diagnosis of PKD was considered when some of the neonates presented palpable flank masses that caused fetal dystocia. On the other hand, the pediatric clinical examination of older infants revealed abdominal distention secondary to renal masses. After surgical resection, the overall aspect of the kidneys showed that the normal parenchyma had been mostly replaced by cysts with thin, translucent walls that contained a clear fluid. Microscopy confirmed that the parenchyma was mostly replaced by dilated cysts delineated by simple cuboidal or simple flattened epithelium, with areas of remnant fetal kidney parenchyma separated by an enriched stroma. Immunohistochemistry for blood vessels (CD34) revealed normal fine walled blood vessel arcades in the control kidneys, while in most areas from polycystic disease, the blood vessels exhibited enlarged, thickened endothelium, and less collapsed lumens. Regarding the proliferative capacity of the tissues, our Ki67 immunostaining revealed that the less formed, younger tubules in the pathological state had a higher proliferative index compared to control tissue. There seemed to be less albumin immunostaining in the epithelia of the distal contort tubules but that distinction was present also in our pathology. The overall expression level was reduced in polycystic cases (p<0.05), and it could be that this expression decrease might be related to the reduced function of these kidneys. According to what literature states, we have emphasized in our study that aquaporin 1 (AQP1) showed overall decreased reactivity in PKD along with its expression in proximal tubule epithelia.

Ischemic stroke occurs through embolic or thrombotic obliteration of an artery from cerebral circulation and represents over 80% of all stroke cases. One of the fiercest complications after stroke is edema, which results from imbalanced water diffusion around the blood vessels walls. Water diffusion around blood vessel walls occurs physiologically mainly through two protein-formed pores, namely aquaporins (AQPs) 1 and 4. Here, we compare for the first time the expression patterns and colocalization degrees of the two AQPs in control brain tissue and in peri-ischemic regions, on tissue obtained from eight patients with confirmed ischemic pathology and from five control cases. Our analysis showed that AQP4 is more abundant that AQP1, especially in the cortex and in the organized scar areas. The colocalization of the two markers was high, both located on the astrocytes membranes, but the colocalization degree decreased in the scar peri-ischemic regions. Colocalization with basement membranes was also lower for AQP1 compared to AQP4, in all regions analyzed.

Adenomyosis is a benign pathology, common to both women at reproductive age as well as later during menopause. This condition is accompanied by a strong symptomatology, which has induced intense research on this topic. From a morphological point of view, it is represented by the existence of endometrial glands and, sometimes, of the periglandular stroma (endometriosis) in the structure of the myometrium, at a significant distance from the normal endometrium. Various inflammatory, vascular and mechanical factors accentuate the symptoms and evolution of this pathology. Our study included a total number of 32 patients, eight cases for each of the following histopathological subtypes: endometrium - proliferative phase, endometrium - secretory phase, myometrium with endometrial glands (adenomyosis), and myometrium with hyperplastic transformation of endometrial glands (hyperplastic adenomyosis), respectively. We have conducted clinical, morphological and morphopathological studies of the structures in question. Using the classical histological technique (Hematoxylin-Eosin), we identified the glandular structures; utilizing immunohistochemistry, we have labeled the endometrial epithelium with the anti-cytokeratin 7 (CK7) antibody and we analyzed the periglandular cell types of the immune system: T-lymphocytes using anti-cluster of differentiation (CD) 3 antibody, macrophages using anti-CD68 antibody, mast cells using anti-tryptase antibody, periglandular vascularization with the reaction using anti-CD34∕anti-CD31 antibodies, thus demonstrating their involvement in the development of adenomyosis. The interesting aspect of this study is the technique of simultaneously labeling of the inflammatory, vascular and epithelial elements.

Pancreatic ductal carcinoma is the most common type of pancreatic cancer, and currently represents the fourth cause of death by cancer, worldwide. Among classical pancreatic markers that ascertain the histopathology, new emerging targets have been proposed for both diagnostic and prognostic purposes. In the present study, utilizing a group of 28 confirmed resected pancreatic ductal carcinomas, we have assessed the immunoexpression and correlation ratios of mothers against decapentaplegic homolog 4 (Drosophila) (SMAD4)∕transforming growth factor beta receptor 2 (TGFβR2), and vimentin∕cluster of differentiation 105 (CD105). SMAD4 showed an overall increase in tumors versus pancreatic control tissue, but a decrease from G1 towards poorly differentiated tumors, while TGFβR2, vimentin and CD105 showed higher expression values in the tumor areas. Vimentin-CD105 colocalization degree decreased in tumor tissues compared to controls, illustrating a desynchronization of these two markers, both of them being negative in the tumor epithelia. Altogether, it is highly plausible that all these key players revolve around the epithelial-to-mesenchymal transition phenomenon, and this itself modulates the clinical outcome of the patient.

Cutaneous melanoma is the most aggressive type of skin cancer, with high invasive potential. Lentigo maligna melanoma (LMM) is a relatively rare type, accounting for about 10% of all melanomas, while the most common subtype of melanoma on the face, typically on chronically sun-exposed skin of elderly people. Its in situ stage is lentigo maligna (LM). During the process of transformation from LM to LMM, tumor cells secrete or induce the release from neighboring cells of large amounts of matrix metalloproteinases (MMPs) that degrade the extracellular matrix. Some MMPs, as MMP3 and MMP9 expressed melanoma cells is associated with statistical significance in both in vitro and in vivo studies, with an invasive phenotype. Unfortunately, there is scarce data published about MMPs expression in LM∕LMM, as majority of research on melanoma refer to superficial spreading and nodular melanoma. Our personal, unpublished yet fully data is an attempt to complete a specific panel of immunohistochemical markers that could explain the slow growing rate of LMM.

Endometriosis is a benign pathology, commonly found in women at reproductive age. It is represented by the ectopic presence of the endometrial glandular epithelium in several tissues and organs. This ectopically located tissue can display premalignant or even malignant changes under the influence of certain factors that affect cell structure, function and proliferation. Our study includes a total of 28 patients, with endometriosis of different localizations: ovarian or pelvic endometriosis, adenomyosis or endometriosis of the abdominal wall. We performed a clinical and statistical analysis upon the collected clinical and laboratory data, together with the results obtained by using classical histological and immunohistochemical (IHC) profiling. The classical staining revealed the existence of the ectopic glandular epithelium, while the IHC reactions obtained with the anti-cytokeratin (CK) 7∕anti-CK20, anti-estrogen receptor alpha (ERα)∕anti-progesterone receptor (PR) antibodies, ascertained that these tissues were of endometrial origin. The environmental, hormonal or inflammatory factors influence these areas, so that the ER∕PR scores may be modified, the cellular proliferation might be increased (Ki67+ marker), the anti-apoptotic B-cell lymphoma 2 (BCL2) protein expression and phosphatase and tensin homolog (PTEN) may also be modified. Moreover, tumor protein 53 (p53) was positive in cases with atypia, density of inflammatory cells clearly increased compared to the adjacent normal endometrium, respectively with cluster of differentiation (CD) 3+, CD20+, CD68+, CD79a+, and tryptase+ cells, all of which may influence the cellular structure, histological architecture of the surrounding microenvironment and cause premalignant or even malignant changes in endometriosis outbreaks.

Meningitis and encephalitis are inflammatory diseases in which acute and chronic inflammatory cells infiltrate leptomeninges, especially the arachnoid, and migrate through the subarachnoid space and by diapedesis, in order to extend around blood vessels and into the brain parenchyma. To what extent migrated/resident inflammatory cells participate to these interactions, or what are exactly the initial steps by which these cells reach the brain interstitium, it is not yet completely known. Recent years have brought new insights into the description of water flow circuits in the brain, suggesting that the cerebrospinal fluid enters the brain within the perivascular spaces of arteries, while interstitial fluid drains along perivascular venous sector. Moreover, it has been showed that vascular basement membranes have a complex multi-layered architecture that originates with epithelial, endothelial, smooth muscle cells and glial cells, and that the virtual space between these layers might be in fact an essential component of these perivascular spaces. Starting from a patient that presented with active pulmonary tuberculosis and with consecutive purulent-hemorrhagic meningitis and encephalitis, we have characterized here the compartments in which immune cells can be found in the brain tissue. Besides the classical histopathological description, what was of interest here, was that we identified for the first time mononucleated inflammatory cells that seemed to be present in pockets of the vascular basement membranes, small spaces devoid of red blood cells. Although this is mere a morphological observation, future high-resolution studies should clarify it this is a possible route for the immune cells entering the brain.

Multiple sclerosis (MS) is a complex chronic neurodegenerative disease that involves an abnormal autoimmune response directed against the brain, nerves and spinal cord; it is considered the most frequent cause of neurological disability, because MS-associated inflammatory lesions can affect a wide range of systems to a varying degree and may cause a plethora of neurological comorbidities and symptoms. The symptoms are quite variable from patient to patient and depend on the spatial distribution of the central nervous system (CNS) lesions, but usually involve sensory disturbances, cognitive deficits, unilateral vision loss, bladder dysfunction, ataxia, fatigue, double vision, weakness of the limbs and intestinal disorders. Experimental autoimmune encephalitis (EAE) mouse model reproduces the pathological features of the human disease, and is a widely used model used for studying the pathology and different treatment options in the preclinical stage. In this study, we aimed to evaluate the motor function, as well as the degree of demyelination and inflammatory changes in the brains of mice immunized for the myelin oligodendrocyte glycoprotein (MOG)35-55, and treated with Cerebrolysin. Animals were randomly assigned to one of the three groups: (i) EAE untreated group (n=10), (ii) EAE treated group (n=10), and (iii) control group (n=5), and their motor dysfunction was followed until the clinical pathology begun to improve. We also analyzed histologically and immunohistochemically the lesions in the optical nerves, cervical spinal cord and medulla. Our results showed higher deficit scores for untreated animals compared to treated animals. After harvesting the tissue, we have first evaluated the density of myelin in the optical nerves, cervical spinal cord and medulla and we found significant differences between treated and untreated groups of animals. We continued to investigate the structure of the CNS parenchyma by evaluating the intensity and morphology of the neuronal cytoskeleton and microglia by immunohistochemical stainings. Although larger animal groups are necessary, this is the first pilot study to investigate the use of a neurotrophic factor as a putative treatment option for a MS model.

Cholangiocarcinoma is an extremely rare and highly aggressive primary malignancy of the biliary tract. The current report illustrates a rare case of distal common bile duct adenocarcinoma encountered in a 65-year-old male patient who was thoroughly investigated after presenting with a two weeks history of pain in the right hypochondrium, jaundice and unintentional weight loss (6 kg in two weeks). The medical team opted for a pancreaticoduodenectomy (Whipple procedure) managing to obtain negative resection margins of the tumor with a favorable immediate postoperative evolution. However, the surgical team was forced to reintervene twice due to complications caused mainly by the patient's disregard.

Introduction: Hepatocellular carcinoma (HCC) currently represents a major health concern, mainly for its shifting pre-existing conditions that in turn lead to late diagnosis, thus increasing the mortality rate. An improved training of medical personnel involved in diagnosis, staging and the management of treatment is required., Aim: We thus aimed to transition tumor view from two-dimensional, on-screen, methods to real, palpatory three-dimensional (3D) representations that can be printed using generally available tools, thus approachable in virtually any medical setting worldwide., Materials and Methods: After obtaining ethical clearance, we included imaging contrast-enhanced data from 10 confirmed cases of HCC that we translated into a 3D computer render of the tumor with as much morphological data as possible. In addition, we simulated the inner structure of each tumor, simulating different stiffness levels across their respective surfaces, in order to better gauge possible necrosis or vascular particularities. This translated into 3D printed models that were obtained by using commercially available materials, experimenting with different filling methods in order to better simulate the stiffness of the lesion., Results: We administered a structured questionnaire to 43 students and 12 resident doctors (gastroenterologists and surgeons) that manage HCC cases. We assessed tumor morphology and the usefulness of the proposed model in everyday practice and evaluated their use in an academic environment., Conclusions: The proposed method provides a cheap alternative to costly medical simulators, providing both curricular advantages as well as integrating well into normal HCC medical management.

Stroke is one of the most important cause of death and disability, especially when considering the increasing life expectancy worldwide, with ischemic stroke being much more common than hemorrhages. In the physiopathological chain of ischemic stroke, brain edema is one first element that if attended to might reduce tissue necrosis, penumbra, and increase functional recovery. Aquaporin 4 (AQP4) has been found to be the most important water channel in the brain, and its inhibition before inducing focal ischemia in an animal model of stroke proved to alleviate the pathology and improve clinical recovery. In this study, we have treated a rat model of ischemic stroke with the AQP4 inhibitor TGN-020 after inducing the lesion, and performed for the first time a direct histopathological evaluation of the brain and infarct's volume. Besides utilizing immunohistochemistry targeting the growth-associated protein-43 (GAP-43) in order to delineate the infarct regions, we have used the Cavalieri's principle of tissue volume measurements starting form seriate sections, and for the first time in neuropathology, we have utilized a high-resolution object scanner to assess global brain volume changes. Our results showed that TGN-020 clearly reduces infarct volume in TGN-020 treated animals compared to untreated animals, as well as the volume of the brain hemispheres. Although reduced, the effect was also present in the contralateral hemisphere. Given these data, a more in-depth characterization of the histopathological and molecular changes induced by AQP-4 are needed for considering it as a bona fide treatment option.

Aim: The aim of our study was to assess glial fibrillary acidic protein (GFAP) glial cell phenotype in the enteric nervous system (ENS) in colorectal adenocarcinoma of different tumor grading and, also, to establish correlations between these changes and the tumor proliferative activity and the tumor-infiltrating leukocytes., Patients, Materials and Methods: We ran an observational, prospective study on a group of 52 patients diagnosed with colorectal adenocarcinoma. They were surgically treated in the 1st Surgery Clinic of the Emergency County Hospital of Craiova, Romania. From the surgically resected pieces, after pathological confirmation and tumor grading, 3-μm thick seriate sections were cut and processed for immunohistochemistry for detecting GFAP, S100, CD45 and Ki-67., Results: Evaluation of GFAP glial cell type in the ENS of colorectal cancer with different stages of differentiation showed that the density of these nervous elements is higher in well-differentiated (G1) colorectal tumors compared to moderately differentiated (G2) and poorly differentiated (G3) colorectal tumors. For well-differentiated colorectal adenocarcinoma, we did not find any correlations between GFAP glial cell type in the ENS and the tumor proliferative activity or with tumor-infiltrating leukocytes. In what the moderately and poorly differentiated adenocarcinoma are concerned, we found a high inverse variation between GFAP glial cell type in the ENS and the proliferative activity, on one hand, and, between GFAP glial cell type in the ENS and the tumor-infiltrating leukocytes, on the other hand., Conclusions: The decrease in the density of GFAP glial cell type in the ENS with tumor grading of colorectal cancer and the inverse variation with the tumor proliferative activity and with the tumor-infiltrating leukocytes might serve as putative prognostic factors in colorectal cancer.

The aim of this study is to assess the status of synapses in normal colorectal tissue compared to neoplastic colorectal tissue, and to correlate this status with survival in patients with colorectal neoplasia. Our study included 61 patients diagnosed with colorectal adenocarcinoma, representing the study group, and 53 patients diagnosed with benign conditions, that required a resection of a colorectal segment, representing the control group. We performed the immunohistochemical staining by using anti-synaptophysin antibody, which identifies synaptic vesicles and, so, we managed to analyze the expression of synapses in colorectal adenocarcinoma. Regarding both the signal area and integrated optical density (IOD) of the synaptophysin, the univariate analysis with a log-rank (Mantel-Cox) test indicated that patients with a low level of synaptophysin had a better overall survival rate than those with a high-level synaptophysin. Also, we noticed that tumor size, tumor invasion and lymph node metastasis were significantly associated with the overall survival rate, whereas the other clinicopathological features were not. In conclusion, the status of synaptic vesicles evaluated via synaptophysin expression in patients with colorectal cancer positively correlates with the survival rate and it can play a role in the neoplastic therapy process.

Squamous cell carcinoma (SCC) of the lip represents 15-30% of SCC of cephalic extremity, located on the lower lip in about 90% of cases. The present paper aimed to define the profile of SCC of the lip with major risk factors. The study included 20 selected cases diagnosed with lower lip SCC, using a panel of antibodies which addressed cell proliferation (Ki67), perturbation of the cell cycle (p53), angiogenesis (VEGF - vascular endothelial growth factor), factors related to tumor cell interaction with the extracellular matrix (CD44). Ki67 immunoexpression was identified in all the cases. Poorly differentiated (PD) SCC presented a mean value of Ki67 positivity index (PI) significantly higher compared to well-differentiated (WD) and moderately differentiated (MD) SCC. We found significantly higher mean values of Ki67 PI in pT3 lesion, compared with pT2 and pT lesions, and with no statistically significant differences in lip SCC with associated lymph node metastasis (pN1), compared to those with no lymph node metastasis (pN0). PD SCC presented a higher mean value of p53 PI compared to WD and MD SCC, but without significant differences. Analysis indicated significantly higher values in pT3 lesions and in pT2 and pT1 and in pN1 SCC. In WD SCC, CD44 immunoexpression had a higher intensity. For PD and MD SCC the immunolabelings presented low÷moderate heterogeneous intensity. WD lip SCC presented a statistically significant higher mean value for CD44 PI compared to MD and PD SCC, and not statistically significant higher in pT1, pT2 then in pT3 and in pN0 cases then in pN1. WD lip SCC presented statistically significant higher mean value of VEGF PI related to those with MD and PD SCC. VEGF PI values were higher in pT1, pT2 then in pT3 and in the pN0 SCC, but without statistically significant differences. We found a positive linear correlation for Ki67÷p53, although statistically not significant and for CD44÷VEGF statistically significant (p=0.001). Also, the analysis identified negative a linear statistically significant correlation for Ki67÷CD44 and for Ki67÷VEGF statistically significant as well (p=0.001). Immunohistochemical investigations in lip SCC, regarding the expression of p53, Ki67, CD44 and VEGF, revealed results that suggest their ability to assess the prognosis and progression of tumor evolution.

E-cadherin and β-catenin are two molecules implicated in cellular adhesion. The reduced expression of β-catenin and E-cadherin is associated with the tumoral epithelial-mesenchymal transition process, a key event in the development of endometrial carcinoma. The aim of our study was to investigate the association between the immunoexpression of β-catenin÷E-cadherin and the tumor differentiation degree, presence of lymph nodes, depth of tumor invasion and pTNM stage of endometrioid endometrial carcinomas in order to enhance the potential aggressiveness of these tumors. Our results revealed significant differences in the expression of β-catenin, when grouping for the tumor stage, invasion in the myometrium and degree of differentiation, as well as significant differences in the expression of E-cadherin for tumor degrees of differentiation. E-cadherin and β-catenin expression was stronger in well-differentiated tumors, superficial myometrium invasion and early tumor stages I or II, thus was associated with better prognostic forms of endometrioid endometrial carcinoma. Our study indicated that decreased of the E-cadherin and β-catenin expression is useful for the assessment of tumor aggressivity.

Colorectal cancer represents a severe public health issue. Recent studies have shown the essential role played by nerves and their neurotransmitters in tumor initiation and progression. The aim of this study is to asses the expression of beta 2-adrenergic receptors (β2A) for adrenaline and noradrenaline, and the expression of M3 muscarinic receptors (M3R) for acetylcholine (neurotransmitters produced and released by sympathetic and parasympathetic afferents of the digestive tract and also by the enteric nervous system) in different tumor gradings of colorectal adenocarcinoma, and also the tropomyosin receptor kinase A (TrkA) for the nerve growth factor produced by the cells of colorectal adenocarcinoma. Beta 2-adrenergic receptors were expressed both in normal colic tissue and in the tumor tissues, from the three patients included in the study. It was observed that both area and integrated optical density (IOD) are more elevated for this type of receptor in tumor tissues than in normal colic tissue. For the M3 muscarinic receptors, similarly to beta 2-adrenergic receptors, it was observed a growth both of the area and of the IOD with the tumor grading. The presence of TrkA receptors was also observed both in the normal colic mucosa and in the tumor tissues examined, but with a significant reduction of the signal in the poorly differentiated tumor tissue. Understanding the neurobiology of cancer in this context becomes necessary for establishing much more complex and targeted molecular targeted therapies.

Unlabelled: G-protein coupled estrogen receptor 1 (GPER), a particular extranuclear estrogen receptor (ER), seems not to be significantly involved in normal female phenotype development but especially associated with severe genital malignancies. This study investigated the GPER expression in different types of normal and abnormal proliferative endometrium, and the correlation with the presence of ERα. GPER was much highly expressed in cytoplasm (than onto cell membrane), contrary to ERα, which was almost exclusively located in the nucleus. Both ERs' densities were higher in columnar epithelial then in stromal cells, according with higher estrogen-sensitivity of epithelial cells. GPER and ERα density decreased as follows: complex endometrial hyperplasia (CEH) > simple endometrial hyperplasia (SHE) > normal proliferative endometrium (NPE) > atypical endometrial hyperplasia (AEH), ERα' density being constantly higher. In endometrial adenocarcinomas, both ERs were significant lower expressed, and widely varied, but GPER÷ERα ratio was significantly increased in high-grade lesions., Conclusions: The nuclear ERα is responsible for the genomic (the most important) mechanism of action of estrogens, involved in cell growth and multiplication. In normal and benign proliferations, ERα expression is increased as an evidence of its effects on cells with conserved architecture, in atypical and especially in malignant cells ERα's (and GPER's) density being much lower. Cytoplasmic GPER probably interfere with different tyrosine÷protein kinases signaling pathways, also involved in cell growth and proliferation. In benign endometrial lesions, GPER's presence is, at least partially, the result of an inductor effect of ERα on GPER gene transcription. In high-grade lesions, GPER÷ERα ratio was increased, demonstrating that GPER is involved per se in malignant endometrial proliferations.

The alteration of epithelial stability, which includes changes in the expression of E-cadherin and CD44, is one of the complex biomolecular mechanisms involved in the tumoral epithelial-mesenchymal transition (EMT) process. We followed the E-cadherin÷CD44 immunophenotype by single and double detections in 25 cases of bladder urothelial carcinomas. Our study investigated simultaneously the differences in expression of the two markers, in different tumoral compartments, according to the prognostic parameters of the lesions. The study indicated significant differences in the expression of E-cadherin in relation to tumor grade, depth of invasion, tumor stage and Ki-67 proliferation index (PI), both intratumoral and at the advancing edge. For CD44, expression differences were found between the tumor grades in intratumoral sites, while for both intratumoral and advancing edge compartments the differences occurred for the depth of tumor invasion, tumor stage and Ki-67 PI. The only differences in the expression of the two markers in relation with the presence of lymph node metastasis were for E-cadherin at the advancing edge. In this study, the intratumoral E-caderin-÷CD44- immunophenotype, respectively E-caderin-÷CD44+ at the advancing edge were associated with the tumor aggressiveness analyzed parameters. The maintenance of CD44 expression at the advancing edge represents a negative prognostic factor for bladder urothelial carcinoma and supports the implication of EMT process, through the existence at this level of a cell population with particular properties.

Oral cancers still represent a major health problem; regional lymph node metastases occur in 30-40% of head and neck squamous cell carcinomas and are associated with unfavorable prognosis and decreased survival. The study included 35 cases of oral squamous cell carcinomas (OSCC), which were analyzed by double reactions to determine the proliferative activity (anti-human D2-40/Ki67) and the maturity degree (anti-human D2-40/α-SMA) of lymphatic vessels, both intratumoral (IT) and in the advancing edge (AE), and in relation to clinicopathological prognostic parameters. The mean values of D2-40 lymphatic vessel density (LVD) were higher in AE then in IT level. Poorly differentiated carcinomas, T3/T4, presented the highest LVD values, both IT and in the AE. LVD was higher in advanced stages and metastasizing carcinomas. Ki67 was positive in all cases, Ki67 proliferation index (IP) indicated higher values in poorly differentiated carcinoma, T3/T4, metastasizing ones, both IT and in the AE. LVD and IP Ki67 showed a positive linear correlation. D2-40/Ki67-positive vessels were identified only at the AE or close to it. D2-40/Ki67 LVD had highest values in advanced stages carcinoma, with metastases. D2-40/α-SMA-positive vessels were identified only in the neighborhood of the tumor and LVD highest values were present in early-stage carcinomas and without metastases. A negative linear correlation between proliferation and maturity of the lymphatic vessels was found. The study indicated a strong association between lymphatic proliferative activity and lymph node metastases, suggesting the need for targeted antilymphangiogenic therapies in OSCC.

Dupuytren's disease is a progressive fibroproliferative disorder that impairs hand function by altering the normal structures of the palmar fascial bands. Nodules composed almost entirely of myofibroblasts and cords are pathognomonic of Dupuytren's disease. The myofibroblasts express alpha-smooth muscle actin that is especially involved in development of the disease. We aimed to evaluate whether the xenograft of Dupuytren's fibromatosis taken from operating room and transplanted on chick chorioallantoic membrane (CAM) survives with its histological and immunohistological features. Fresh samples obtained from eight patients with Dupuytren's disease were minced and immediately inoculated onto 24 CAMs of 8-day-old chick embryos. The implanted CAMs were examined daily by stereomicroscopy and finally the xenografts were examined and characterized in histological sections using a panel of antibodies. The xenografts were incorporated into the CAMs 6-7 days after transplantation, continued to grow and stimulated angiogenesis in the chick embryo CAMs. The CAMs vessels entered the xenografts and anastomosed with the newly formed xenografts vessels (CD34+ and CD105+) those containing nucleated chick erythrocytes. Myofibroblasts (α-SMA+) and macrophages (CD68+) were readily recognized in the xenograft thickness. We concluded that the xenografts of Dupuytren's fibromatosis transplanted onto chick embryo CAMs continued to develop and preserved the histological and immunohistological features.

Corpora amylacea (CA) are glycoprotein-based depositions that accumulate in the normal aging brain or consecutively to different neuro-degenerative diseases. Although controversies still exists in what it regards their origins and functions, their morphology is described simply as round basophilic entities based on bi-dimensional observations. The aim of the present study was to evaluate the three-dimensional morphology of these bodies in the brain in normal aging. We utilized here brain tissue from six aged patients, and performed an in-depth stereological analysis of CAs based on series of thin serial sections, and 50 μm-thick sections. The thin seriate sections have been counter-stained with Hematoxylin and Eosin, while the thick sections have been immunostained in fluorescence for ubiquitin and GFAP/collagen IV or aquaporin 4. Three-dimensional renderings have been obtained after aligning the serial sections, while high-resolution z-stacks have resulted after deconvolution on the thick sections. More than 70% of all the identified CAs proved to be in fact parts of larger aggregates, where the flattened individual spheroids branched and communicated with other bodies in a complex pattern, and budding of small CAs from larger CAs could be observed. There was a direct correlation between the diameter of the vessels and the number of associated CAs. Astrocyte GFAP and aquaporin 4 signals surrounded CAs, but without any colocalization with the ubiquitin areas, while perivascular CAs were sometimes enclosed in pockets of the basement membranes. In conclusion, as far as we know, this is the first study to describe the three-dimensional branching complexity of corpora amylacea in the brain.

Basal cell carcinoma is the most common malignant tumor of the skin, and it develops most frequently on the head and neck regions. Although most of these tumors are slow growing and with a limited evolution, the existence of some aggressive variants accompanied by a complete neglect from the patient may occasionally lead to invasion of the face and organs of the head and neck. Even though, intracranial invasion of basal cell carcinoma of the scalp is a rare presentation. We describe here the case of a woman who developed an aggressive and neglected morpheiform basal cell carcinoma (ulcus terebrans), which showed a complete invasion through the skull, but with an apparent self-limitation to the pia mater.