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• Collagen XVIII deficiency induces blood-brain-barrier leakage. • It also induces other pathological changes typical of early CSVD. • Expression of basement membrane ECM molecules was upregulated. • ECM of perineuronal nets around interneurons was impaired. • Perisynaptic ECM proteins and C1q at excitatory synapses were upregulated. Collagen type XVIII (COL18) is an abundant heparan sulfate proteoglycan in vascular basement membranes. Here, we asked (i) if the loss of COL18 would result in blood-brain barrier (BBB) breakdown, pathological alterations of small arteries and capillaries and neuroinflammation as found in cerebral small vessel disease (CSVD) and (ii) if such changes may be associated with remodeling of synapses and neural extracellular matrix (ECM). We found that 5-month-old Col18a1−/− mice had elevated BBB permeability for mouse IgG in the deep gray matter, and intravascular erythrocyte accumulations were observed brain-wide in capillaries and arterioles. BBB permeability increased with age and affected cortical regions and the hippocampus in 12-month-old Col18a1−/− mice. None of the Col18a1−/− mice displayed hallmarks of advanced CSVD, such as hemorrhages, and did not show perivascular space enlargement. Col18a1 deficiency-induced BBB leakage was accompanied by activation of microglia and astrocytes, a loss of aggrecan in the ECM of perineuronal nets associated with fast-spiking inhibitory interneurons and accumulation of the perisynaptic ECM proteoglycan brevican and the microglial complement protein C1q at excitatory synapses. As the pathway underlying these regulations, we found increased signaling through the TGF-ß1/Smad3/TIMP-3 cascade. We verified the pivotal role of COL18 for small vessel wall structure in CSVD by demonstrating the protein's involvement in vascular remodeling in autopsy brains from patients with cerebral hypertensive arteriopathy. Our study highlights an association between the alterations of perivascular ECM, extracellular proteolysis, and perineuronal/perisynaptic ECM, as a possible substrate of synaptic and cognitive alterations in CSVD. [ABSTRACT FROM AUTHOR]

Histopathological changes associated with dilated cardiomyopathy (CMD) are frequently nonspecific and often only present in the terminal stage of the disease. The study followed the histopathological and morphometric quantification of fibrosis and nuclear pleomorphism in CMD. We analyzed left ventricle myocardial fragments harvested during autopsy, from 35 cases with clinical diagnosis of CMD and 5 cases of normal myocardium. Fibrosis was present in all CMD cases, with higher values compared with control cases. Nuclear pleomorphism was identified in 18 cases (45%), two of the analyzed parameters, respectively the ratio of nuclear diameters and roundness of nucleus, revealing significant differences in CMD compared to the control cases. Myocardial fibrosis present in all cases of CMD represents a major feature of the disease. The nuclear pleomorphism due to the nuclei change in diameters and size was more pronounced in the vicinity of fibrosis areas, possibly related to this alteration. [ABSTRACT FROM AUTHOR]

Aquaporins are a vast family of channel proteins whose main role is the bidirectional transport, depending on the osmotic gradient, of water across the lipid membranes, which have low permeability for this solvent. The aquaporins are also involved in the lipid metabolism, the cell proliferation and migration processes, the transport of glycerol, neuroexcitation and epithelial fluids secretion, having numerous roles such as ensuring the water transport in the central nervous system, the production of CSF, aqueous humor and saliva, epithelial hydration, urine concentration and nervous impulse transmission. Up to now, in mammals there have been identified 13 types of aquaporins, each of them annotated from 0 to 12 (Aqp 0 - Aqp 12). Of these, aquaporin-4 (AQP-4) - located in the astroglia - is the most abundant aquaporin in the brain. Although this type of aquaporin is also present inside the brain parenchyma, especially in the astroglial processes lining the neuronal synapses, AQP-4 is mainly located in the astroglial end-feet adjacent to the ependymocyes or endothelial cells, where its main role is to ensure the bidirectional transport of water between the astroglia and the cerebrospinal fluid (CSF) or blood vessels. The presence of AQP-4 on the surface of astroglial processes promotes, in the initial stages of ischemia, the formation of the cytotoxic oedema, but has a protective role against vasogenic oedema. This review aims to describe the roles of AQP-4, and especially the role that this protein has in maintaining the hydric balance in the brain [ABSTRACT FROM AUTHOR]

While chronic kidney disease seems to be an independent risk factor for cognitive decline, its impact on cerebral amyloid-β (Aβ) depositions, one hallmark of Alzheimer's Disease (AD) pathology, has not been investigated. Utilizing 80 male nontransgenic spontaneously hypertensive stroke prone rats (SHRSP) at various ages (12 to 44 weeks), tubulointerstitial renal damage, prevalence of cerebral microhemorrhages and Aβ accumulations were quantified. Using age-adjusted general linear models we investigated the main and interaction effects of renal damage and cerebral microhemorrhages on cerebral Aβ load. In addition, using post mortem human brain tissue of 16 stroke patients we examined the co-localization of perivascular Aβ deposits and small vessel wall damage. Statistical models revealed an age-independent main effect of tubulointerstitial kidney damage on brain Aβ accumulations, which was reinforced by the consecutive presence of cerebral microhemorrhages. Moreover, cerebral microhemorrhages independently predicted brain Aβ burden in SHRSP. In up to 69% of all human cases perivascular Aβ deposits were detected in the direct vicinity of small vessel wall damage. Our results support the associations between vascular pathology and Aβ deposition, and demonstrate a relationship between chronic kidney disease and cerebral Aβ pathology. Hence, our data suggest that prevention of chronic renal damage may reduce cerebral Aβ pathology. [ABSTRACT FROM AUTHOR]

Background: Calcifying epithelial odontogenic tumor (CEOT) is a rare benign neoplasia, locally aggressive, that tends to invade bone and adjacent soft tissues. This case report describes the thirteenth known case of CEOT with maxillary sinus extension and the second one that also involves the buccal vestibule mucosa with peculiar histopathological and immunohistochemical data. Case presentation: Here we report the case of a 45-year-old female with a CEOT diagnosed and treated at the Oral & Maxillofacial Surgery Department, County Clinical Emergency Hospital of Craiova, Romania. The clinical and imaging investigation revealed an intraosseous tumor developed from the left posterior maxilla with maxillary sinus and buccal vestibule mucosa extension. Histopathology found an epithelium-rich CEOT variant, but with scattered S100 positive clear cells, focal small rounded cementum-like deposits and areas with some degree of nuclear pleomorphism. The immunohistochemical investigations emphasised its local aggressiveness behavior with involvement of multiple molecular mechanisms that underlie tumor invasiveness. A subtotal maxillectomy was performed followed by defect reconstruction. Conclusions: We discuss the relevant clinicopathological features of an aggressive rare case of CEOT with maxillary sinus extension and buccal vestibule mucosa involvement. The immunohistochemical study suggests its utility in attempting to assess the degree of local tumor aggressiveness and thus in adopting the most efficient therapeutic attitude. [ABSTRACT FROM AUTHOR]

Aims: Pancreatic cancer and hepatocellular carcinoma are two of the most aggressive types of cancer with limited therapeutic options in stages of advanced disease. Our objective is to assess the safety and feasibility of injecting iron oxide nanoparticles (IONs) via endoscopic ultrasound (EUS)-guidance, both systemically and locally in the liver and pancreas in order to study new potential therapies for liver and pancreatic tumors.Material and Methods: Six domestic pigs were used for our study design, and divided into three groups: two were injected in the portal vein, and other four were subjected to local exposure of IONs in the liver and pancreas, two each. The pigs were on a 7 days follow-up and necropsy was performed with their organs harvested. A 3T MRI scan was also performed.Results: All animals underwent an endoscopic ultrasound fine needle injection (EUS-FNI) procedure without any complications. EUS-FNI procedure had an average time of 5 minutes and 21 seconds and consisted of 2 ml of ION injection. No perforations and no risk of potential bleeding were recorded. Macroscopic changes were observed only after pancreatic EUS-FNI. A significant amount of IONs was observed in the liver after local injection and after vascular EUS-FNI. The imaging results were confirmed by pathological examination with most of the IONs accumulated in Ito-like cells, Kupfer cells, and sinusoids.Conclusions: IONs have been widely studied for both diagnostic and therapeutic purposes. Their injection through EUS-guidance may develop new diagnosis strategies as well as curative or palliative therapies in pancreatic and liver tumors. [ABSTRACT FROM AUTHOR]

Background & Aims: Colorectal cancer represents the third most common malignancy and the fourth most common cause of cancer death worldwide. The existence of drug-resistant colon cancer stem cells is thought to be one of the most important reasons behind treatment failure in colon cancer, their existence putatively leading to metastasis and recurrences. The aim of our study was to investigate the immunoexpression patterns of CD133 and CD166 in colon carcinoma, both individually and in combination, assessing their significance as prognostic markers. Methods. A total of 45 retrospective colon adenocarcinoma cases were investigated by enzymatic and multiple fluorescence immunohistochemistry for their CD133 and CD166 expression and colocalization. Results. Both CD133 and CD166 were expressed to diffierent extents in all cancer specimens, with a predominant cytoplasmic pattern for CD133 and a more obvious membranous-like pattern for CD166. Overall, when comparing their reactivity for the tumoral tissue, CD166 expression areas seemed to be smaller than those of CD133. However, there was a direct correlation between CD133 and CD166 expression levels throughout the entire spectrum of lesions, with higher values for dysplastic lesions. Colocalization of CD133/CD166 was obvious at the level of cells membranes, with higher coefficients in high grade dysplasia, followed by well and moderate differentiated tumours. Conclusions. CD133/CD166 colocalization is an early event occurring in colon tumorigenesis, with the highest coefficients recorded for patients with high grade dysplasia, followed by well differentiated tumours. Thus, we consider that the coexpression of these two markers could be useful for further prognostic and therapeutically stratification of patients with colon cancer. [ABSTRACT FROM AUTHOR]

Matrix metalloproteinases (MMPs) are well-recognized denominators for extracellular matrix remodeling in the pathology of both ischemic and hemorrhagic strokes. Recent data on non-nervous system tissue showed intracellular and even intranuclear localizations for different MMPs, and together with this, a plethora of new functions have been proposed for these intracellular active enzymes, but are mostly related to apoptosis induction and malign transformation. In neurons and glial cells, on human tissue, animal models and cell cultures, different active MMPs have been also proven to be located in the intra-cytoplasmic or intra-nuclear compartments, with no clear-cut function. In the present study we show for the first time on human tissue the nuclear expression of MMP-9, mainly in neurons and to a lesser extent in astrocytes. We have studied ischemic and hemorrhagic stroke patients, as well as aged control patients. Age and ischemic suffering seemed to be the best predictors for an elevated MMP-9 nuclear expression, and there was no evidence of a clear-cut extracellular proteolytic activity for this compartment, as revealed by intact vascular basement membranes and assessment of vascular densities. More, the majority of the cells expressing MMP-9 in the nuclear compartment also co-expressed activated-caspase 3, indicating a possible link between nuclear MMP-9 localization and apoptosis in neuronal and glial cells following an ischemic or hemorrhagic event. These results, besides showing for the first time the nuclear localization of MMP-9 on a large series of human stroke and aged brain tissues, raise new questions regarding the unknown spectrum of the functions MMPs in human CNS pathology. [ABSTRACT FROM AUTHOR]

Abstract: The varied morphological and biochemical forms in which amyloid deposits in brain of Alzheimer''s disease (AD) patients are complex and their mechanisms of formation are not completely understood. Here we investigated the ability of fractal dimension (FD) to differentiate between the textures of commonly observed amyloid plaques in sporadic and familial AD patients and aged-control individuals as well as in transgenic mouse models of amyloidosis. Studying more than 6000 amyloid plaques immunostained for total Aβ (Aβt), Aβ40 or Aβ42, we show here that Aβ40 FD could efficiently differentiate between (i) AD patients and aged-control individuals (P <0.001); (ii) sporadic and familial AD due to presenilin-1 or APP (A692G) mutations (P <0.001); and (iii) three transgenic mouse models of different genotypes (P <0.001). Furthermore, while diffuse and dense-core plaques present in humans and transgenic mice had comparable FDs, both Aβt and Aβ42 FD could also differentiate diffuse plaques from other plaque types in both species (P <0.001). Our data suggest that plaque FD could be a valuable tool for objective, computer-oriented AD diagnosis as well as for genotype-phenotype correlations of AD. [Copyright &y& Elsevier]

ngiogenesis is involved in tumor progression of oral squamous cell carcinoma (OSCC). In this study, we have investigated by immunohistochemistry vascular endothelial growth factor (VEGF) expression in tumor cells and we have correlated VEGF expression to microvessel area, evaluated by using CD105 as a marker of endothelial cells, in bioptic specimens of 54 human OSCC. Results demonstrated that VEGF is highly expressed in OSCC tumor specimens when compared to pre-neoplastic and normal tissues, without differences between the edge and inside the tumor. Moreover, VEGF expression is reduced in poor differentiated OSCC tumors when compared to moderate and good differentiated forms, and tumor microvessel area is higher in tumors when compared to pre-neoplastic lesions and normal tissues. Finally, VEGF and CD105 may be considered as reliable markers of tumor angiogenesis and progression in OSCC, even if we did not demonstrate any correlation between VEGF expression, tumor microvascular area, clinical stage, and lymph node status. [ABSTRACT FROM AUTHOR]

Ex-vivo freshly surgical removed pancreatic ductal adenocarcinoma (PDAC) specimens were assessed using pCLE and then processed for paraffin embeding and histopathological diagnostic in an endeavour to find putative image analysis algorithms that might recognise adenocarcinoma. Methods: Twelve patients diagnosed with PDAC on endoscopic ultrasound and FNA confirmation underwent surgery. Removed samples were sprayed with acriflavine as contrast agent, underwent pCLE with an experimental probe and compared with previous recordings of normal pancreatic tissue. Subsequently, all samples were subjected to cross-sectional histopathology, including surgical resection margins for controls. pCLE records, as well as corespondant cytokeratin-targeted immunohistochemistry images were processed using the same morphological classifiers in the Image ProPlus AMS image analysis software. Specific morphometric classifiers were automatically generated on all images: Area, Hole Area (HA), Perimeter, Roundness, Integrated Optical Density (IOD), Fractal Dimension (FD), Ferret max (Fmax), Ferret mean (Fmean), Heterogeneity and Clumpiness. Results: After histopathological confirmation of adenocarcinoma areas, we have found that the same morphological classifiers could clearly differentiate between tumor and non-tumor areas on both pathology and correspondand pCLE (area, roundness, IOD, ferret and heterogeneity (p < 0.001), perimeter and hole area (p < 0.05). Conclusions: This pilot study proves that classical morphometrical classifiers can clearly differentiate adenocarcimoma on pCLE data, and the implementation in a live image-analysis algorithm might help in improving the specificity of pCLE in vivo diagnostic. [ABSTRACT FROM AUTHOR]

Background: Hepatic steatosis has been identified as an independent risk factor for post-operative complications. The aim of our research was to assess how inflammation and neoangiogenesis associated with different stages of hepatic steatosis are related to post-operative complications in patients who undergo hepatic resection. Methods: Our study included 19 patients with hepatic steatosis undergoing liver resection for primary or secondary tumors. For every patient we performed immunostaining using a panel of 5 primary antibodies (CD3, CD20, CD68, CD31, CD34) to highlight inflammation and neoangiogensis in the non-tumoral hepatic parenchyma. Results: Taking into consideration the number of vessels as well as the signal area and integrated optical density (IOD) forCD3, CD20, CD68, and also the degree of steatosis, the univariate analysis with a log-rank (Mantel-Cox) test revealed that patients with higher values of CD31 and CD34 had a higher rate of post-operative complications on a 30-day follow-up period. Also, we used a Mann-Whitney U and Kruskal-Wallis H tests for group distributions. We noticed thatCD34 was significantly increased in patients diagnosed with steatosis compared to the control group and there was a statistically significant difference between CD31 median values of S0 (27.6) and S1 (55.8) grades. Conclusion: Patients with steatosis that presented higher values of CD31 and CD34 had a higher rate of post-operative complications. Further studies should assess the value of pre-operative evaluation of angiogenesis in patients with liver steatosis submitted to liver surgery. [ABSTRACT FROM AUTHOR]

Patients with primary colorectal cancer can present with obstructions, tumor bleeding, or perforations, which represent acute complications. This paper aimed to analyze and compare the clinical and pathological profiles of two patient groups: one with colorectal cancer and a related complication and another without any specific complication. We performed a five-year retrospective study on colorectal cancer patients admitted to a surgery unit and comparatively explored the main clinical and pathological features of the tumors belonging to the two groups. A total of 250 patients with colorectal cancer were included in the analysis. Of these, 117 (46.8%) had presented a type of complication. The comparative analysis that examined several clinical and pathological parameters showed a statistically significant difference for unfavorable prognosis factors in the group with complications. This was evident for features such as vascular and perineural invasion, lymph node involvement, pathological primary tumor stage, and TNM stage. Colorectal cancers with a related complication belonged to a group of tumors with a more aggressive histopathologic profile and more advanced stages. Furthermore, the comparable incidence of cases in the two groups of patients warrants further efforts to be made in terms of early detection and prognosis prediction of colorectal cancer. [ABSTRACT FROM AUTHOR]

After pregnancy termination or loss, histologic three-dimensional imaging can be used to confirm the presence of fetal cardiac malformations detected during first-trimester ultrasound examination. BACKGROUND: We aimed to evaluate the usefulness of three-dimensional (3D) reconstruction of histology slides to confirm congenital heart disease (CHD) detected by first-trimester fetal cardiac ultrasonography. Conventional autopsy is hindered by the small size of the first-trimester fetal heart, and current CHD confirmation studies employ the use of highly specialized and expensive methods. TECHNIQUE: An extended first-trimester ultrasound examination protocol was used to diagnose fetal heart anomalies. Medical termination of pregnancies was followed by fetal heart extraction. The specimens were sliced, and the histology slides were stained and scanned. The resulting images were processed, and volume rendering was performed using 3D reconstruction software. The volumes were analyzed by a multidisciplinary team of maternal–fetal medicine subspecialists and pathologists and compared with ultrasound examination findings. EXPERIENCE: Six fetuses with heart malformations were evaluated using histologic 3D imaging: two with hypoplastic left heart syndrome, two with atrioventricular septal defects, one with an isolated ventricular septal defect, and one with transposition of the great arteries. The technique allowed us to confirm ultrasound-detected anomalies and also identified additional malformations. CONCLUSION: After pregnancy termination or loss, histologic 3D imaging can be used to confirm the presence of fetal cardiac malformations detected during first-trimester ultrasound examination. Additionally, this technique has the potential to refine the diagnosis for counseling regarding recurrence risk and retains the advantages of standard histology. [ABSTRACT FROM AUTHOR]

Dilated cardiomyopathy is the most common form of cardiac muscle disease, accounting for approximately 60% of all cardiomyopathies. We proposed to identify histopathological changes of the myocardium in dilative cardiomyopathy. This study comprised a total of 19 cases, represented by myocardial fragments from deceased patients with diagnosis of dilated cardiomyopathy. Histopathological analysis allowed changes to be observed for both myocytes and myocardial interstitial components. We have found a combination of hypertrophic, atrophic and normal myocardocytes, or associated with the presence of hydropic changes. We rarely identified the aspect of myocytosis, cytoplasmic accumulation of lipofuscin pigment or mucinous material, and variable nuclear pleomorphism. At the interstitial level we noticed changes in fibrosis, lipomatosis and rarely the presence of inflammatory infiltrate. Histopathological characteristics of the myocardium in dilated cardiomyopathy are numerous but nonspecific, similar to those in the terminal stages of other cardiac diseases. [ABSTRACT FROM AUTHOR]

Non-amyloid cerebral small vessel disease (CSVD) and cerebral amyloid angiopathy (CAA) may be interrelated through the damaged basement membranes (BMs) and extracellular matrix changes of small vessels, resulting in a failure of β-amyloid (Aβ) transport and degradation. We analyzed BM changes and the pattern of deposition of Aβ in the walls of blood vessels in spontaneously hypertensive stroke-prone rats (SHRSP), a non-transgenic CSVD model. In 45 SHRSP and 38 Wistar rats aged 18 to 32 weeks: (i) the percentage area immunostained for vascular collagen IV and laminin was quantified; (ii) the capillary BM thickness as well as endothelial and pericyte pathological changes were analysed using transmission electron microscopy (TEM); and (iii) the presence of vascular Aβ was assessed. Compared with controls, SHRSP exhibited a significantly higher percentage area immunostained with collagen IV in the striatum and thalamus. SHRSP also revealed an age-dependent increase of the capillary BM thickness and of endothelial vacuoles (caveolae) within subcortical regions. Endogenous Aβ deposits in the walls of small blood vessels were observed in the cortex (with the highest incidence found within fronto-parietal areas), striatum, thalamus and hippocampus. Vascular β-amyloid accumulations were frequently detected at sites of small vessel wall damage. Our data demonstrate changes in the expression of collagen IV and of the ultrastructure of BMs in the small vessels of SHRSP. Alterations are accompanied by vascular deposits of endogenous Aβ. Impaired β-amyloid clearance along perivascular and endothelial pathways and failure of extracellular Aβ degradation may be the key mechanisms connecting non-amyloid CSVD and CAA. [ABSTRACT FROM AUTHOR]

Background and purpose: In the central nervous system, a multitude of changes have been described associated with severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection, such as microglial activation, perivascular lymphocyte cuffing, hypoxic‐ischaemic changes, microthrombosis, infarcts or haemorrhages. It was sought here to assess the vascular basement membranes (vBMs) and surrounding perivascular astrocytes for any morphological changes in acute respiratory syndrome (coronavirus disease 2019, COVID‐19) patients. Methods: The light microscopy morphology of the vBMs and perivascular astrocytes from brains of 14 patients with confirmed SARS‐CoV‐2 infection was analysed and compared to four control patients utilizing fluorescent immunohistochemistry for collagen IV and astrocytes (GFAP), endothelia (CD31), tight junction 1 (TJ1) adhesion protein, as well as the aquaporin 4 (AQP4) water channel. On 2D and 3D deconvoluted images from the cortex and white matter, vessel densities, diameters, degree of gliosis, collagen IV/GFAP and GFAP/AQP4 colocalizations were calculated, as well as the fractal dimension of astrocytes and vBMs viewed in tangential planes. Results: Fractal dimension analysis of the GFAP‐stained astrocytes revealed lower branching complexities and decreased GFAP/collagen IV colocalization for COVID‐19 patients. Interestingly, vBMs showed significantly increased irregularities (fractal dimension values) compared to controls. Vessel diameters were increased in COVID‐19 cases, especially for the white matter, TJ1 protein decreased its colocalization with the endothelia, and AQP4 reduced its co‐expression in astrocytes. Conclusions: Our data on the irregularity of the basement membranes, loss of endothelial tight junction, reduction of the astrocyte end‐feet and decrease of AQP4 suggest subtle morphological changes of the blood–brain barrier in COVID‐19 brains that could be linked with indirect inflammatory signalling or hypoxia/hypercapnia. [ABSTRACT FROM AUTHOR]

Digestive cancer represents a severe public health problem, being one of the main causes of death. It is considered a multifactorial disease, with hereditary predisposition, environmental factors, and other factors involved in carcinogenesis. Both the evolution and the pathogenesis of digestive neoplasms remain incompletely elucidated. As a multifactorial disease, it can be approached by taking into account the biopsychosocial influences via enteric nervous system. Many peptides and non-peptides having a neurotransmitter role can be found in the enteric nervous system, which can influence the neoplastic process directly or indirectly by affecting some angiogenic, growth, and metastasis factors. However, neurotransmitters can also cause directly, through intercellular signalizing, the angiogenesis, the proliferation, and the digestive neoplasms’ metastasis. This new approach to neoplasms of the digestive tube assumes broader psychosocial factors can play an important role in the understanding the ethiopathogenie, the evolution of the disease, and determination of possible molecular targeted therapies; it also suggests that behavioral strategies may be important for maintaining a healthy state with respect to the digestive tract. [ABSTRACT FROM AUTHOR]

Angiogenesis plays a key role in the development and dissemination of colorectal cancer. In advanced stages this malignancy has a poor prognosis due to the inefficacy of the existent treatment and at this point the outcome cannot be completely predicted by common prognostic factors such as lymph node involvement, tumor size or local extension of the disease. For the management of these patients, a vast number of studies about appropriate biomarkers have been reported and they have been classified into prognostic biomarkers, predictive biomarkers, pharmacodynamic biomarkers and surrogate endpoints. The evaluation of prognostic biomarkers using immunohistochemistry, is increasingly used in the histopathologic diagnosis, having the potential to predict responsiveness to standard chemotherapy or novel molecular targeted therapy. The aim of this article is to address current immunohistochemical angiogenesis-based prognostic markers for patients with colorectal cancer, to summarize their current state and to discuss future perspectives. [ABSTRACT FROM AUTHOR]

In this pilot study, we tested the feasibility of cardiac structures reconstruction from histologic sections in 12-13 weeks 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 accurate data regarding the confirmation of first trimester heart defects suspected by early prenatal ultrasound scans. Hearts from fetuses aged 12-13 gestational weeks were harvested for histological preparation, virtual reconstruction, and cardiac structures analysis. A detailed ultrasound scan protocol was used for heart evaluation. 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 proper and confident visualization. This study included normal and abnormal fetal hearts. Visualization of the normal heart cavities, including atrioventricular septum, was very good in all fetuses. The pathological aspects of abnormal heart specimens were successfully identified and confirmed through this method. A detailed review of the histological sections was necessary for confirmation of some structures. The results demonstrate that this method can be applied to routine clinical practice. The use of 3D reconstruction of fetal heart histological sections in the first trimester may serve as an important audit to confirm the normalcy of heart structures. Also, the histological and post processed 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. [ABSTRACT FROM AUTHOR]

Gastric cancer continues to be a significant malignancy worldwide, accounting for approximately one million new cases in 2020. Scientists are focusing on the cancerous cells' plasma membrane (PM) as a potential therapeutic target in cancer because it functions as the cell's interface with its environment through a variety of mechanisms. The capacity of membrane shape and its structures to influence biological processes frequently occurs through the regulation of enzymes or preferential protein binding to membranes via membrane shape changes. We aimed here to assess the morphological irregularities of the cellular membranes in gastric adenocarcinoma tumors, and to find any putative differences from normal gastric mucosae epithelial cells. We analyzed the pattern of E-cadherin at the level of the cell membrane using the fractal dimension (FD) analysis on fluorescence immunohistochemistry samples labeled with E-cadherin in gastric well/moderate and solid gastric adenocarcinoma from patients without any associated chemotherapeutic treatment or radiotherapy. Images were binarized based on a fixed threshold of the E-cadherin fluorescence channel, and then the FD of the binarized image outlines has been calculated in order to assess the ruggedness of the cellular membranes. Overall assessment of the FD revealed that the subtle membrane variations were evident enough to deem a statistically significant difference and the complexity of the membrane roughness was clearly higher for adenocarcinoma cases. We intended to evaluate if separating adenocarcinoma cases as low grade (G1 and G2) and high grade (G3 and solid), FD analysis could still differentiate membrane patterns and check if the available clinical parameters like age, gender, tumor location, lymph ganglia involved might correlate with FD values for adenocarcinoma patients. Altogether, the morphological analysis of a simple marker for the cell membrane can identify and distinguish tumor cells. Although there was a limited correlation between this analysis and the main clinical and pathological indicators of the disease, it will be very useful in the future for automatic computer-assisted diagnosis on slides, as well as for evaluating cellular adhesion and inter-cellular trafficking in cancer cells. [ABSTRACT FROM AUTHOR]

Endoscopic ultrasound (EUS) gained wide acceptance as the diagnostic and minimally invasive therapeutic approach for intra-luminal and extraluminal gastrointestinal, as well as various non-gastrointestinal lesions. Since its introduction, EUS has undergone substantial technological advances. This multi-centric study is a retrospective analysis of a prospectively maintained database of patients who underwent EUS for the evaluation of lesions located within the gastrointestinal tract and the proximal organs. It aimed to extensively assess in dynamic the dual-center EUS experience over the course of the past 20 years. Hence, we performed a population study and an overall assessment of the EUS procedures. The performance of EUS-FNA/FNB in diagnosing pancreatic neoplasms was evaluated. We also investigated the contribution of associating contrast-enhanced ultrasound imaging (CE-EUS) with EUS-FNA/FNB for differentiating solid pancreatic lesions or cystic pancreatic lesions. A total of 2935 patients undergoing EUS between 2002–2021 were included, out of which 1880 were diagnostic EUS and 1052 EUS-FNA/FNB (80% FNA and 20% FNB). Therapeutic procedures performed included endoscopic transmural drainage of pancreatic fluid collections, celiac plexus block and neurolysis, while diagnostic EUS-like CE-EUS (20%) and real-time elastography (12%) were also conducted. Most complications occurred during the first 7 days after EUS-FNA/FNB or pseudocyst drainage. EUS and the additional tools have high technical success rates and low rates of complications. The EUS methods are safe, cost effective and indispensable for the diagnostic or therapeutic management in gastroenterological everyday practice. [ABSTRACT FROM AUTHOR]

Tumor vascular perfusion pattern in gastric cancer (GC) may be an important prognostic factor with therapeutic implications. Non-invasive methods such as dynamic contrast harmonic imaging endoscopic ultrasound (CHI-EUS) may provide details about tumor perfusion and could also lay out another perspective for angiogenesis assessment. Methods: We included 34 patients with GC, adenocarcinoma, with CHI-EUS examinations that were performed before any treatment decision. We analyzed eighty video sequences with a dedicated software for quantitative analysis of the vascular patterns of specific regions of interest (ROI). As a result, time-intensity curve (TIC) along with other derived parameters were automatically generated: peak enhancement (PE), rise time (RT), time to peak (TTP), wash-in perfusion index (WiPI), ROI area, and others. We performed CD105 and CD31 immunostaining to calculate the vascular diameter (vd) and the microvascular density (MVD), and the results were compared with CHI-EUS parameters. Results: High statistical correlations (p < 0.05) were observed between TIC analysis parameters MVD and vd CD31. Strong correlations were also found between tumor grade and 7 CHI-EUS parameters, p < 0.005. Conclusions: GC angiogenesis assessment by CHI-EUS is feasible and may be considered for future studies based on TIC analysis. [ABSTRACT FROM AUTHOR]

Ectopic endometrial epithelium associates a wide spectrum of symptomatology. Their evolution can be influenced by inflammatory and vascular changes, that affect not only the structure and cell proliferation rate, but also symptoms. This prospective study involved tissue samples from surgically treated patients, stained using classical histotechniques and immunohistochemistry. We assessed ectopic endometrial glands (CK7+, CK20−), adjacent blood vessels (CD34+), estrogen/progesterone hormone receptors (ER+, PR+), inflammatory cells (CD3+, CD20+, CD68+, Tryptase+), rate of inflammatory cells (Ki67+) and oncoproteins (BCL2+, PTEN+, p53+) involved in the development of endometriosis/adenomyosis. A CK7+/CK20− expression profile was present in the ectopic epithelium and differentiated it from digestive metastases. ER+/PR+ were present in all cases analyzed. We found an increased vascularity (CD34+) in the areas with abdominal endometriosis and CD3+−:T-lymphocytes, CD20+−:B-lymphocytes, CD68+:macrophages, and Tryptase+: mastocytes were abundant, especially in cases with adenomyosis as a marker of proinflammatory microenvironment. In addition, we found a significantly higher division index-(Ki67+) in the areas with adenomyosis, and inactivation of tumor suppressor genes-p53+ in areas with neoplastic changes. The inflammatory/vascular/hormonal mechanisms trigger endometriosis progression and neoplastic changes increasing local pain. Furthermore, they may represent future therapeutic targets. Simultaneous-multiple immunohistochemical labelling represents a valuable technique for rapidly detecting cellular features that facilitate comparative analysis of the studied predictors. [ABSTRACT FROM AUTHOR]

Objectives: Epidemic methicillin-resistant S. aureus (MRSA) clones cause infections in both hospital and community settings. As a biofilm phenotype further facilitates evasion of the host immune system and antibiotics, we compared the biofilm-forming capacities of various MRSA clones. Methods: Seventy-six MRSA classified into 13 clones (USA300, EMRSA-15, Hungarian/Brazilian etc.), and isolated from infections or from carriers were studied for biofilm formation under static and dynamic conditions. Static biofilms in microtitre plates were quantified colorimetrically. Dynamic biofilms (Bioflux 200, Fluxion, USA) were studied by confocal laser-scanning and time-lapse microscopy, and the total volume occupied by live/dead bacteria quantified by Volocity 5.4.1 (Improvision, UK). Results: MRSA harbouring SCCmec IV produced significantly more biomass under static conditions than SCCmec I–III (P = 0.003), and those harbouring SCCmec II significantly less than those harbouring SCCmec I or III (P<0.001). In the dynamic model, SCCmec I–III harbouring MRSA were significantly better biofilm formers than SCCmec IV (P = 0.036). Only 16 strains successfully formed biofilms under both conditions, of which 13 harboured SCCmec IV and included all tested USA300 strains (n = 3). However, USA300 demonstrated remarkably lower percentages of cell-occupied space (6.6%) compared to the other clones (EMRSA-15 = 19.0%) under dynamic conditions. Time-lapse microscopy of dynamic biofilms demonstrated that USA300 formed long viscoelastic tethers that stretched far from the point of attachment, while EMRSA-15 consisted of micro-colonies attached densely to the surface. Conclusions: MRSA harbouring SCCmec types IV and I–III demonstrate distinct biofilm forming capacities, possibly owing to their adaptation to the community and hospital settings, respectively. USA300 demonstrated abundant biofilm formation under both conditions, which probably confers a competitive advantage, contributing to its remarkable success as a pathogen. [ABSTRACT FROM AUTHOR]

Gastric cancer remains a health problem, with treatment indications varying with the TNM stage. We aimed in this study to highlight the role of EUS in GC patients and also to calculate the accuracy, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of EUS for T and N staging in our group of patients with this disease. In this study, we included 41 GC patients, and individual values for every T stage accuracy, sensitivity, specificity, PPV, NPV, correct staging, understaging, and overstaging were calculated. EUS overall accuracy for T staging was 58.53%, with the highest sensitivity reached for the T4 stage, 95.83%. For N+vs. N-staging, EUS accuracy was 68.29%, with a sensitivity of 75% and a specificity of 44.44%. The positive and negative predicted values for the presence or absence of nodal disease were 82.75%, respectively 33.33%. In conclusion, this study confirmed the importance of EUS for the assessment of GC T and N stage and highlighted the role of this tool in the detection of liver micrometastasis unrevealed by other imaging techniques like abdominal ultrasound or MSCT. [ABSTRACT FROM AUTHOR]

Abstract: Transgenic mouse models of Alzheimer''s disease (AD) expressing high levels of amyloid precursor protein (APP) with familial AD (FAD) mutations have proven to be extremely useful in understanding pathogenic processes of AD especially those that involve amyloidogenesis. We earlier described Austrian APP T714I pathology that leads to one of the earliest AD age-at-onsets with abundant intracellular and extracellular amyloid deposits in brain. The latter strikingly was non-fibrillar diffuse amyloid, composed of N-truncated Aβ42 in absence of Aβ40. In vitro, this mutation leads to one of the highest Aβ42/Aβ40 ratios among all FAD mutations. We generated an APP T714I transgenic mouse model that despite having 10 times lower transgene than endogenous murine APP deposited intraneuronal Aβ in brain by 6 months of age. Accumulations increased with age, and this was paralleled by decreased brain sizes on volumetric MRI, compared to age-matched and similar transgene-expressing APP wild-type mice, although, with these levels of transgenic expression we did not detect neuronal loss or significant memory impairment. Immunohistochemical studies revealed that the majority of the intraneuronal Aβ deposits colocalized with late endosomal markers, although some Aβ inclusions were also positive for lysosomal and Golgi markers. These data support earlier observations of Aβ accumulation in the endosomal–lysosomal pathway and the hypothesis that intraneuronal accumulation of Aβ could be an important factor in the AD pathogenesis. [Copyright &y& Elsevier]

Frontotemporal lobar degeneration (FTLD) is a clinically, pathologically and genetically highly complex disorder. In the last few years enormous progress has been made in dissecting the genetic etiology of FTLD. Mutations have been identified in the progranulin gene (PGRN), the charged multivesicular body protein 2B gene (CHMP2B) and the valosin-containing protein gene (VCP). Mutations in these genes all lead to FTLD pathology characterized by ubiquitin-immunoreactive neuronal cytoplasmic and intranuclear lentiform inclusions (FTLD-U). The similar pathology suggests that these genes may be connected trough a common disease pathway leading to neurodegeneration and the formation of these pathognomic inclusions. This review focuses on the molecular genetic processes underlying FTLD-U pathology. Copyright © 2007 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Gastric cancer represents the third most frequent cause of death worldwide, with the treatment being impaired also by the fact that patients present in the late stages of disease progression. We have aimed here to evaluate the main clinical and pathological features of all recorded cases of gastric tumor patients presented between January 2015 and December 2020 within the Emergency County Hospital of Craiova. Our retrospective analysis identified a total number of 745 cases, and showed a relative homogenous distribution of the age of the patients / year, with the peak age at presentation of 70-80 years old, and with males having a slightly higher prevalence compared to females. There was no correlation of the number of hospitalization days with the localization of the tumor, but the patients in the age group 60-70 years of age tended to show longer hospitalization times compared to the rest of the age groups. Also, pyloric/ antral tumors tended to present at younger ages compared to other localizations, and interestingly, these patients also represented most of the casuistry. Altogether, the distribution of gastric cancer patients' features did not change significantly in the last 5 years despite treatment advances (especially chemo-and radiotherapy), and the advanced stage of presentation call for a more aggressive detection and increased awareness of the population for this frequent pathology. [ABSTRACT FROM AUTHOR]

Pancreatic ductal adenocarcinoma (PDAC) is characterized by an intense fibrosis which enhances the growth of tumor cells and drives an epithelialmesenchymal transition. The consequence is the uncontrolled proliferation of cancer cells, including pancreatic cancer stem cells (PCSC), which can enhance the metastatic potential. Objective. The aim of this study is to assess the expression of pancreatic cancer stem cells, on pancreatic endoscopic ultrasound-guided fine-needle biopsy (EUS-FNB) specimens. Material and Methods. A total of 21 patients diagnosed with PDAC after EUS-FNB within the Research Center of Gastroenterology and Hepatology of Craiova, were included in the study, and the samples were sent for immunohistochemical assessment. Anti-Human CD 24 (policlonal, Abcam, ab 199140) and EpCAm (Epr 20532-225, Rb monoclonal, Abcam, ab 223582) were used. Results. All cases were positive for CD24 and EpCAM. In NOS ductal adenocarcinomas, CD24 immunomarking was heterogeneous, with variable intensity and a variable number of positive cells within the same case. Well and moderately differentiated adenocarcinomas that formed tubular or papillary structures showed a higher intensity CD24 expression compared to poorly differentiated or undifferentiated adenocarcinomas. EpCam showed a stronger immunolabeling intensity in the analyzed samples. PDAC EpCam expression was of moderate and high intensity and with a heterogeneous appearance in NOS G1 adenocarcinomas, but the intensity of the immunomarking tended to decrease with increasing histological grade. Conclusions. EUS-FNB PCSC immunohistochemistry is feasible. At the subcellular level, the basolateral membranous and cytoplasmic staining of EpCam in PDAC could be correlated with tumor invasion and possibly with the unfavorable evolution of this type of carcinoma, while CD24 frequently presents an apical immunostaining pattern and could suggest the presence of an epithelial-type phenotype of tumor cells, with a lower potential for invasiveness and metastasis. [ABSTRACT FROM AUTHOR]

Gastric cancer currently represents one of the most important public health problems. Recent studies have demonstrated the existence of strong correlations between the vegetative nervous system and the role it plays in the initiation of the oncogenetic process and the progression of cancer. The purpose of this paper is to demonstrate the involvement of the sympathetic and parasympathetic vegetative nervous system in the evolution of gastric cancer, according to the stage of tumor differentiation. In this current paper we have included a number of four patients diagnosed with gastric cancer post UGI (Upper Gastrointestinal Endoscopy) and have analyzed relations that exist between the tumor differentiation degree and the metanephrine and normetanephrine serum level in the blood of the patients. Following the research, we have observed an increased value of the metanephrine and normetanephrine serum level in the patient which displayed the lowest degree of differentiation. [ABSTRACT FROM AUTHOR]

Methotrexate (MTX) is a folic acid analog with anti-proliferative (anti-neoplastic, cytotoxic), immunosuppressive and anti-inflammatory properties, which has been used in the treatment of various cutaneous disorders, such as psoriasis, keratoacanthoma, pityriasis rubra pilaris, atopic dermatitis, mycosis fungoides, bullous skin diseases, systemic sclerosis, morphea, lupus erythematosus, dermatomyositis and crusted scabies. Inhibition of cell proliferation is explained through its role in blocking DNA/RNA synthesis, by inhibiting dihydrofolate reductase, necessary for the production of pyrimidine and purine nucleotides. An anticancer effect can be related to α-oxoaldehyde metabolism (MTX increases methylglyoxal levels). Its anti-inflammatory property is based on the inhibition of 5-aminoimidazole-4-carboxamide ribonucleotide transformylase, thus increasing intracellular and extracellular adenosine, a purine nucleoside with anti-inflammatory effect. This drug can limit inflammation by scavenging free radicals and decreasing malondialdehyde-acetaldehyde protein-adduct production. Moreover, the anti-proliferative and anti-inflammatory effects can also be related to inhibition of the DNA methylation pathway, thus inhibiting methionine formation. The aim of the present study was to report various dermatological cases from our daily practice that demonstrate the efficacy of MTX in the treatment of cutaneous diseases, highlighting different mechanisms of action: its anti-inflammatory effect in psoriasis and its anti-proliferative, and anti-neoplastic effect in well-differentiated squamous cell carcinoma or in keratoacanthoma. Moreover, different administration pathways and doses are addressed. Assessment of the treatment plan, clinical improvement of cutaneous lesions, biologic evaluation, final aesthetic result, quality of life, as well as potential adverse effects and drug tolerance related to each case mentioned. [ABSTRACT FROM AUTHOR]

Melanoma is the most severe type of skin cancer and its incidence has increased in the last decades. In the United States, it is the 6th most common cancer in both men and women. Prognosis for patients with melanoma depends on the stage of the disease at the time of diagnosis and it can be influenced by the immunologic response. Melanoma has been historically considered an immunogenic malignancy. It often contains great amount of immune cells (different subsets of T-cells, dendritic cells, macrophages, neutrophils, mast cells, B lymphocytes), which may reflect a continuous intercommunication between host and tumor. It is not established if tumor-infiltrating lymphocytes (TILs) are induced by tumor cells or by other components of the microenvironment or when they are a host direct immunologic reaction. It has been observed that in many cases, the presence of a dense TIL is associated with good prognosis. The pattern and activation state of the cells which constitute TIL is variable and modulates the clinical outcome. An important step in the understanding of tumor immunobiology is the analysis of the populations and subsets of immune cells that form TIL. Besides its prognostic significance, after approval of cytotoxic T lymphocyte antigen 4, programmed cell death-1 and programmed death-1 ligand antibodies for the treatment of melanoma, the assessment of immune infiltrate composition has become even more captivating, as it could provide new target molecules and new biomarkers for predicting the effect of the treatment and disease outcome in patients treated with immunotherapy. In this review we discuss current state of knowledge in the field of immune cells that infiltrate melanoma, resuming the potential of TIL components to become prognostic markers for natural evolution, for response to drugs or valuable targets for new medication. [ABSTRACT FROM AUTHOR]

Sinonasal papilloma is a benign tumor, derived from Schneiderian sinonasal epithelium. There have been described three histological subtypes: inverted, oncocytic and exophytic. The case presented here is A 66-yearold male patient, which was hospitalized in our Otolaryngology Department for a giant tumor, that was exteriorized from the left nostril, repeated epistaxis, nasal obstruction and anosmia. The computed tomography scan revealed an iodophilic and non-homogeneous tumor, with areas of necrosis, which included the entire left nasal cavity, with extension to the rhinopharynx and the left maxillary sinus. We completely removed the tumor by an endoscopic medial maxillectomy, with the subsequent histopathological examination revealing an inverted papilloma, with areas of low grade dysplasia and also areas with oncocytic Schneiderian papilloma. At the six-months postoperative control, there was no tumor recurrence. Major issues of this type of tumor is fast invasion capacity and numerous local recurrence. More recent studies have shown that these relapses are often overdue tumors. [ABSTRACT FROM AUTHOR]

The aquaporins (AQP), a protein family, were first discovered in the early 1990s. The primary role of aquaporins is to facilitate water transport across multiple cell types. In the spinal cord and brain responsible for most of the water diffusion are AQP4 and AQP1. In this paper, we describe the structure, localization and role of this water channel family, especially AQP4 and AQP1. AQP4 is involved in various pathologies such as: stroke, brain tumors, Neuromyelitis optica (NMO), Alzheimer's Disease (AD), traumatic brain injury, Parkinson's Disease, hydrocephalus, schizophrenia, epilepsy, major depressive disorder, autism. Brain edema is the most important acute complication of the hypoxic-ischemic and it has no pathogenic treatment. Imaging and histopathology studies have shown that inhibition of AQP4 reduces brain edema. [ABSTRACT FROM AUTHOR]

Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) is a glycoprotein belonging to the carcinoembryonic antigen (CEA) family that is expressed on a wide variety of cells and holds a complex role in inflammation through its alternate splicing and generation of various isoforms, mediating intricate mechanisms of modulation and dysregulation. Initially regarded as a tumor suppressor as its expression shows considerable downregulation within the epithelia in the early phases of many solid cancers, CEACAM1 has been linked lately to the progression of malignancy and metastatic spread as various papers point to its role in tumor progression, angiogenesis, and invasion. We reviewed the literature and discussed the various expression patterns of CEACAM1 in different types of tumors, describing its structure and general biologic functions and emphasizing the most significant findings that link this molecule to poor prognosis. The importance of understanding the role of CEACAM1 in cell transformation stands not only in this adhesion molecule's value as a prognostic factor but also in its promising premise as a potential new molecular target that could be exploited as a specific cancer therapy. [ABSTRACT FROM AUTHOR]

Polycystic kidney disease represented by autosomal dominant polycystic kidney disease (ADPKD) and autosomal recessive polycystic kidney disease (ARPKD) have a major impact of mortality in children. We conducted a study of a premature infant with an estimated gestation date of 32 weeks with a presumptive prenatal diagnosis of right polycystic kidney. A 28-year-old primigravida with pre-eclampsia was admitted at the gynecology unit of Clinical Emergency County Hospital of Craiova. The clinical examination revealed a large abdominal distention due probably to the right polycystic kidney, suspected on prenatal ultrasound and radiography. The preterm neonate undergone right nephrectomy 5 days after birth. Histopathology of the kidney was performed in the Pathology Department of the Emergency County Hospital of Craiova and in the Center for Microscopic Morphology and Immunology of U.M.F. of Craiova. Microscopy revealed dilated cysts lined by simple cuboidal or flattened epithelium, and islets of remnant kidney parenchyma separated by edematous stroma. Immunohistochemistry for CD34 revealed incomplete blood arcades which did not seem to be in contact with all the tubular elements of the parenchyma, when compared to a control age-matched kidney. The patient had a favorable postoperative evolution, she was clinically stable on discharge from the hospital with a follow-up strategy including genetic testing. [ABSTRACT FROM AUTHOR]

Classical or M1 activity of microglia/macrophages has been described in several neurodegenerative and brain inflammatory conditions and has also been linked to expansion of ischemic injury in post-stroke brain. While different pathways of M1 polarization have been suggested to occur in the post-stroke brain, the precise underlying mechanisms remain undefined. Using a transient middle cerebral artery occlusion (MCAO) rat model, we showed a progressive M2 to M1 polarization in the perilesional brain region with M1 cells becoming one of the dominant subsets by day 4 post-stroke. Comparing key receptors involved in M1 polarization (CD8, IFNγR, Clec4, FcγR, TLR3 and TLR4) and their signal transducers (Syk, Stat1, Irf3, and Traf6) at the day 4 time point, we showed a strong upregulation of CD8 along with SYK transducer in dissected perilesional brain tissue. We further showed that CD8 expression in the post-stroke brain was associated with activated (CD68+) macrophages and that progressive accumulation of CD8+CD68+ cells in the post-stroke brain coincided with increased iNOS (M1 marker) and reduced Arg1 (M2 marker) expression on these cells. In vitro ligand-based stimulation of the CD8 receptor caused increased iNOS expression and an enhanced capacity to phagocytose E. coli particles; and interestingly, CD8 stimulation was also able to repolarize IL4-treated M2 cells to an M1 phenotype. Our data suggest that increased CD8 signaling in the post-stroke brain is primarily associated with microglia/macrophages and can independently drive M1 polarization, and that modulation of CD8 signaling could be a potential target to limit secondary post-stroke brain damage. [ABSTRACT FROM AUTHOR]

Aquaporin-4 (AQP4) is the most abundant water channel in the brain, and its inhibition before inducing focal ischemia, using the AQP4 inhibitor TGN-020, has been showed to reduce oedema in imaging studies. Here, we aimed to evaluate, for the first time, the histopathological effects of a single dose of TGN-020 administered after the occlusion of the medial cerebral artery (MCAO). On a rat model of non-reperfusion ischemia, we have assessed vascular densities, albumin extravasation, gliosis, and apoptosis at 3 and 7 days after MCAO. TGN-020 significantly reduced oedema, glial scar, albumin effusion, and apoptosis, at both 3 and 7 days after MCAO. The area of GFAP-positive gliotic rim decreased, and 3D fractal analysis of astrocytic processes revealed a less complex architecture, possibly indicating water accumulating in the cytoplasm. Evaluation of the blood vessels revealed thicker basement membranes colocalizing with exudated albumin in the treated animals, suggesting that inhibition of AQP4 blocks fluid flow towards the parenchyma in the paravascular drainage pathways of the interstitial fluid. These findings suggest that a single dose of an AQP4 inhibitor can reduce brain oedema, even if administered after the onset of ischemia, and AQP4 agonists/antagonists might be effective modulators of the paravascular drainage flow. [ABSTRACT FROM AUTHOR]

Despite their commensal status, staphylococci can become problematic pathogens expressing multiple and redundant virulence factors. This study aimed to evaluate aggressiveness markers comparatively in staphylococcal strains isolated from severe infections versus asymptomatic carriage in order to identify clinically relevant bacterial traits that could easily be detected in clinical practice and could be suggestive for particular host-pathogen interactions such as cyto-adhesion or biofilm formation, ultimately orienting the clinical decision-making process. We have used in vitro phenotypic methods to assess adhesion to and invasion of eukaryotic cells, biofilm development, and expression of soluble virulence factors in 92 Staphylococcus spp. strains. The adhesion index, invasion capacity, biofilm formation and expression of soluble factors did not differ significantly between clinical and commensal strains. The major bacterial traits we found to be significantly more prevalent in clinical staphylococci were the aggregative adhesion pattern ( P = 0.012), cluster adhesion ( P = 0.001) and tetrad morphology ( P = 0.018). The aggregative adhesion pattern was correlated with higher cyto-adhesion ( P < 0.001), higher invasion capacity ( P = 0.003) and lower Carmeli scores ( P = 0.002). Three major bacterial traits, namely tetrad morphology, aggregative adhesion pattern, and resistance to methicillin (acronym: TAM), can be used to compute an aggressiveness score (SAS) predictive of the staphylococcal strain's virulence and capacity to initiate and develop a biofilm-driven chronic infectious process versus a fulminant acute infection, in a susceptible host. [ABSTRACT FROM AUTHOR]

Pathological diagnosis of prostate adenocarcinoma often requires complementary methods. On prostate biopsy tissue from 39 patients including benign nodular hyperplasia (BNH), atypical adenomatous hyperplasia (AAH), and adenocarcinomas, we have performed combined histochemical-immunohistochemical stainings for argyrophilic nucleolar organizer regions (AgNORs) and glandular basal cells. After ascertaining the pathology, we have analyzed the number, roundness, area, and fractal dimension of individual AgNORs or of their skeleton-filtered maps. We have optimized here for the first time a combination of AgNOR morphological denominators that would reflect best the differences between these pathologies. The analysis of AgNORs' roundness, averaged from large composite images, revealed clear-cut lower values in adenocarcinomas compared to benign and atypical lesions but with no differences between different Gleason scores. Fractal dimension (FD) of AgNOR silhouettes not only revealed significant lower values for global cancer images compared to AAH and BNH images, but was also able to differentiate between Gleason pattern 2 and Gleason patterns 3-5 adenocarcinomas. Plotting the frequency distribution of the FDs for different pathologies showed clear differences between all Gleason patterns and BNH. Together with existing morphological classifiers, AgNOR analysis might contribute to a faster and more reliable machine-assisted screening of prostatic adenocarcinoma, as an essential aid for pathologists. [ABSTRACT FROM AUTHOR]

Aquaporin-4 (AQP4) and glutamate transporter-1 (GLT-1) represent the major water and glutamate astrocyte buffering gateways in the brain. Utilizing perilesional ischemic human cortices, we have performed here for the first time an integrative assessment on both AQP4 and GLT-1, and on their proximity to blood vessels and neurons. Counting the relative number of AQP4±/GLT-1±/glial fibrillary acidic protein± cells showed that doublepositive variants were overall most frequent, and their number tended to decrease from organized and recent perilesional cortices to controls. AQP4/GLT-1 colocalization showed higher coefficients for the perilesional cortices compared with controls, suggesting an increased water/glutamate-buffering capability. Distance frequency analysis of AQP4/ GLT-1 in relationship to neurons showed that both markers were concentrated at 20-40 μm around the perikarya; with AQP4 being more abundant in close proximity, these differences were not being driven by changes in neuropil density alone. Our study suggests a dual, simultaneous astrocytic function depending on the relative distance to neurons and vessels, with increased water and glutamate-buffering capability in the mid perineuronal space, and an increased water-buffering capability in the immediate perineuronal space, even higher than around vessels. Thus, adding specific AQP4/GLT-1 modulator agents selectively depending on the acute/chronic phase of stroke might increase the efficacy of existing treatments. [ABSTRACT FROM AUTHOR]

Aquaporin-4 ( AQP4) and glutamate transporter-1 ( GLT-1) represent the major water and glutamate astrocyte buffering gateways in the brain. Utilizing perilesional ischemic human cortices, we have performed here for the first time an integrative assessment on both AQP4 and GLT-1, and on their proximity to blood vessels and neurons. Counting the relative number of AQP4±/ GLT-1±/glial fibrillary acidic protein± cells showed that double-positive variants were overall most frequent, and their number tended to decrease from organized and recent perilesional cortices to controls. AQP4/ GLT-1 colocalization showed higher coefficients for the perilesional cortices compared with controls, suggesting an increased water/glutamate-buffering capability. Distance frequency analysis of AQP4/ GLT-1 in relationship to neurons showed that both markers were concentrated at 20-40 μm around the perikarya; with AQP4 being more abundant in close proximity, these differences were not being driven by changes in neuropil density alone. Our study suggests a dual, simultaneous astrocytic function depending on the relative distance to neurons and vessels, with increased water and glutamate-buffering capability in the mid perineuronal space, and an increased water-buffering capability in the immediate perineuronal space, even higher than around vessels. Thus, adding specific AQP4/ GLT-1 modulator agents selectively depending on the acute/chronic phase of stroke might increase the efficacy of existing treatments. [ABSTRACT FROM AUTHOR]

The tumor microcirculation is characterized by an abnormal vascular network with dilated, tortuous and saccular vessels. Therefore, imaging the tumor vasculature and determining its morphometric characteristics represent a critical goal for optimizing the cancer treatment that targets the blood vessels (i.e. antiangiogenesis therapy). The aim of this study was to evaluate new vascular morphometric parameters in colorectal cancer, difficult to achieve through conventional immunohistochemistry, by using the confocal laser endomicroscopy method. Fresh biopsies from tumor and normal tissue were collected during colonoscopy from five patients with T3 colorectal carcinoma without metastasis and were marked with fluorescently labeled anti-CD31 antibodies. A series of optical slices spanning 250 µm inside the tissue were immediately collected for each sample using a confocal laser endomicroscope. All measurements were expressed as the mean ± standard error. The mean diameter of tumor vessels was significantly larger than the normal vessels (9.46±0.4 µm vs. 7.60±0.3 µm, p = 0.0166). The vessel density was also significantly higher in the cancer vs. normal tissue samples (5541.05±262.81 vs. 3755.79±194.96 vessels/mm3, p = 0.0006). These results were confirmed by immunohistochemistry. In addition, the tortuosity index and vessel lengths were not significantly different (1.05±0.016 and 28.30±3.27 µm in normal tissue, vs. 1.07±0.008 and 26.49±3.18 µm in tumor tissue respectively, p = 0.5357 and p = 0.7033). The daughter/mother ratio (ratio of the sum of the squares of daughter vessel radii over the square of the mother vessel radius) was 1.15±0.09 in normal tissue, and 1.21±0.08 in tumor tissue (p = 0.6531). The confocal laser endomicroscopy is feasible for measuring more vascular parameters from fresh tumor biopsies than conventional immunohistochemistry alone. Provided new contrast agents will be clinically available, future in vivo use of CLE could lead to identification of novel biomarkers based on the morphometric characteristics of tumor vasculature. [ABSTRACT FROM AUTHOR]

Background and Aims. Hepatocellular carcinoma (HCC) remains a leading cause of death by cancer worldwide. Computerized diagnosis systems relying on novel imagingmarkers gained significant importance in recent years. Our aimwas to integrate a novel morphometric measurement-the fractal dimension (FD)-into an artificial neural network (ANN) designed to diagnose HCC. Material and Methods. The study included 21 HCC and 28 liver metastases (LM) patients scheduled for surgery. We performed hematoxylin staining for cell nuclei and CD31/34 immunostaining for vascular elements. We captured digital images and used an in-house application to segment elements of interest; FDs were calculated and fed to an ANN which classified them as malignant or benign, further identifying HCC and LM cases. Results. User intervention corrected segmentation errors and fractal dimensions were calculated. ANNs correctly classified 947/1050 HCC images (90.2%), 1021/1050 normal tissue images (97.23%), 1215/1400 LM (86.78%), and 1372/1400 normal tissues (98%). We obtained excellent interobserver agreement between human operators and the system. Conclusion. We successfully implemented FD as a morphometric marker in a decision system, an ensemble of ANNs designed to differentiate histological images of normal parenchyma from malignancy and classify HCCs and LMs. [ABSTRACT FROM AUTHOR]

Introduction: Numerous anti-angiogenic agents are currently developed to limit tumor growth and metastasis. While these drugs offer hope for cancer patients, their transient effect on tumor vasculature is difficult to assess in clinical settings. Confocal laser endomicroscopy (CLE) is a novel endoscopic imaging technology that enables histological examination of the gastrointestinal mucosa. The aim of the present study was to evaluate the feasibility of using CLE to image the vascular network in fresh biopsies of human colorectal tissue. For this purpose we have imaged normal and malignant biopsy tissue samples and compared the vascular network parameters obtained with CLE with established histopathology techniques. Materials and Methods: Fresh non-fixed biopsy samples of both normal and malignant colorectal mucosa were stained with fluorescently labeled anti-CD31 antibodies and imaged by CLE using a dedicated endomicroscopy system. Corresponding biopsy samples underwent immunohistochemical staining for CD31, assessing the microvessel density (MVD) and vascular areas for comparison with CLE data, which were measured offline using specific software. Results: The vessels were imaged by CLE in both normal and tumor samples. The average diameter of normal vessels was 8.560.9 mm whereas in tumor samples it was 13.5±0.7 mm (p = 0.0049). Vascular density was 188.7±24.9 vessels/mm2 in the normal tissue vs. 242.4±16.1 vessels/mm2 in the colorectal cancer samples (p = 0.1201). In the immunohistochemistry samples, the MVD was 211.2±42.9/mm2 and 351.3±39.6/mm2 for normal and malignant mucosa, respectively. The vascular area was 2.960.5% of total tissue area for the normal mucosa and 8.5±2.1% for primary colorectal cancer tissue. Conclusion: Selective imaging of blood vessels with CLE is feasible in normal and tumor colorectal tissue by using fluorescently labeled antibodies targeted against an endothelial marker. The method could be translated into the clinical setting for monitoring of anti-angiogenic therapy. [ABSTRACT FROM AUTHOR]

Frontotemporal dementia (FTD) with ubiquitin-immunoreactive neuronal inclusions (both cytoplasmic and nuclear) of unknown nature has been linked to a chromosome 17q21 region (FTDU-17) containing MAPT (microtubule-associated protein tau). FTDU-17 patients have consistently been shown to lack a tau-immunoreactive pathology, a feature characteristic of FTD with parkinsonism linked to mutations in MAPT (FTDP-17). Furthermore, in FTDU-17 patients, mutations in MAPT and genomic rearrangements in the MAPT region have been excluded by both genomic sequencing and fluorescence in situ hybridization on mechanically stretched chromosomes. Here we demonstrate that FTDU-17 is caused by mutations in the gene coding for progranulin (PGRN), a growth factor involved in multiple physiological and pathological processes including tumorigenesis. Besides the production of truncated PGRN proteins due to premature stop codons, we identified a mutation within the splice donor site of intron 0 (IVS0 + 5G > C), indicating loss of the mutant transcript by nuclear degradation. The finding was made within an extensively documented Belgian FTDU-17 founder family. Transcript and protein analyses confirmed the absence of the mutant allele and a reduction in the expression of PGRN. We also identified a mutation (c.3G > A) in the Met1 translation initiation codon, indicating loss of PGRN due to lack of translation of the mutant allele. Our data provide evidence that PGRN haploinsufficiency leads to neurodegeneration because of reduced PGRN-mediated neuronal survival. Furthermore, in a Belgian series of familial FTD patients, PGRN mutations were 3.5 times more frequent than mutations in MAPT, underscoring a principal involvement of PGRN in FTD pathogenesis. [ABSTRACT FROM AUTHOR]

Background: We previously identified increased tissue localization of monomeric C-reactive protein (mCRP) in the infarcted cortical brain tissue of patients following ischaemic stroke. Here, we investigated the relationship of mCRP expression in haemorrhagic stroke, and additionally examined the capacity of mCRP to travel to or appear at other locations within the brain that might account for later chronic neuroinflammatory or neurodegenerative effects. Methods: Immunohistochemistry was performed on Formalin-fixed, paraffin-embedded archived brain tissue blocks obtained at autopsy from stroke patients and age-matched controls. We modelled mCRP migration into the brain after haemorrhagic stroke by infusing mCRP (3.5 µg) into the hippocampus of mice and localized mCRP with histological and immunohistochemistry methods. Results: On human tissue in the early stages of haemorrhage, there was no staining of mCRP. However, with increasing post-stroke survival time, mCRP immunostaining was associated with some parenchymal brain cells, some stroke-affected neurons in the surrounding areas and the lumen of large blood vessels as well as brain capillaries. Further from the peri-haematoma region, however, mCRP was detected in the lumen of micro-vessels expressing aquaporin 4 (AQP4). In the hypothalamus, we detected clusters of neurons loaded with mCRP along with scattered lipofuscin-like deposits. In the peri-haematoma region of patients, mCRP was abundantly seen adjacent to AQP4 immunoreactivity. When we stereotactically injected mCRP into the hippocampus of mice, we also observed strong expression in distant neurones of the hypothalamus as well as cortical capillaries. Conclusions: mCRP is abundantly expressed in the brain after haemorrhagic stroke, directly impacting the pathophysiological development of the haematoma. In addition, it may have indirect effects, where the microcirculatory system appears to be able to carry it throughout the cortex as far as the hypothalamus, allowing for long-distance effects and damage through its capacity to induce inflammation and degenerate neuronal perivascular compartments. [ABSTRACT FROM AUTHOR]

Scop. Evaluarea participării componentei imune în procesele de remaniere la nivelul mucoasei rinosinusale interesate de procesul infecţios cronic de etiologie bacteriană. Material şi metodă. Studiul nostru s-a desfăşurat în SCJU Craiova pe o perioada de 3 ani şi a inclus 856 de pacienţi' diagnosticaţi cu rinosinuzită cronică bacteriană. 324 dintre aceştia au fost supuşi cel puţin unei intervenţii chirurgicale, obţinându- se fragmente de mucoasa sinusală, ce au fost prelucrate în nistologia clasică şi tehnici imunohistochimice pentru evidenţierea componentelor celulare imune, implicate în răspunsul inflamator local, precum şi a fenomenelor de remaniere prin evaluarea componentei angiogenetice. Am folosit markerii CD20, CD3, CD68 şi CD34 pentru evidenţierea limfocitelor B, T, macrofagelor, respectiv a celulelor endoteliale. Rezultat. Limfocitele B au avut o distribuţie neomogenă în infiltratul inflamator, fiind mai abundente perivascular şi în zonele de eroziune epitelială. Uneori, infiltratul inflamator a avut tendinţă de organizare foliculară, zone în care limfocitele B au fost mai abundente în zona centrală a acestor structuri. Limfocitele T au apărut a fi mai abundente decât limfocitele B, cu localizare preponderentă subepitelial şi perivascular. Macrofagele au fost identificate atât în epiteliul de acoperire, cât şi subepitelial. în ceea ce priveşte procesul de angiogeneză, tehnicile de imunohistochimie au evidenţiat existenţa unui număr mult mai mare de vase sanguine tinere. Concluzii. Reacţia inflamatorie este o reacţie de apărare complexă, care include atât fenomene alterative, cât şi fenomene reacţionale, vasculo-exsudative, proliferative şi reparatorii ineficiente. în aceste cazuri avansate, neglijate, rezecţia completă de mucoasă sinusală pare a fi singura soluţie viabilă. [ABSTRACT FROM AUTHOR]

Introducere. Simptomatologia clinică în cancerul de amigdală palatină a fost ştearsă şi confundată în primele stadii ale afecţiunii cu o simptomatologie de tip inflamator, motiv pentru care majoritatea pacienţilor s-au prezentat tardiv la medic, în stadiile III şi IV, conform clasificării TNM. Acuzele subiective au fost de tipul disfagiei şi odinofagiei unilaterale, otalgie reflexă, expectoraţii serosanghinolente, scădere ponderală marcată, halenă fetidă şi tulburări fonatorii de tipul stomatolaliei. Material şi metodă. Studiul s-a efectuat pe 132 de cazuri de tumori maligne amigdaliene (98,2% carcinoame scuamoase şi 1,8% limfoame), diagnosticate în clinica ORL prin biopsie excizională la Spitalul Clinic Judeţean de Urgenţă Craiova, între anii 2010 şi 2014. Piesele prelevate au fost supuse prelucrării în tehnici histologice clasice, respectiv imunohistochimice, utilizând ca markeri de proliferare celulară ate PCNA şi p53. Rezultate. Studiul histologic a evidenţiat faptul că numai 10% au fost carcinoame scuamoase bine diferenţiate, 37,27% carcinoame moderat diferenţiate, iar 52,7% slab diferenţiate. Carcinoamele epidermoide bine diferenţiate au apărut ca fiind formate din celule de aspect epitelial, poliedrice, joncţionate între ele prin desmozomi, amintind de celulele epiteliale ale stratului spinos. Celulele neoplazice au prezentat dimensiuni variate, cu nudei mari, cu inversarea raportului nucleu/citoplasmă. Nucleii celulelor carcinomatoase au apărut cel mai adesea hipercromi, cu contur neregulat, cu invaginări, lobulări sau înmuguriri nucleare, cu cromatină dispusă inegal, determinând astfel o coloraţie neomogenă a nucleului. Frecvent s-au înregistrat monstruozităţi nucleare sau celule poliploide. Nucleii au apărut cel mai adesea voluminoşi, multipli, bine evidenţiaţi. Frecvent s-a observat mitoze atipice, asimetrice, bipolare sau chiar'tripolare. Variaţiile celulare şi nucleare au determinat un aspect pleomorf al parenchimului tumoral, celulele superficiale carcinomatoase formând adesea perle cheratozice. Studiul imunohistochimic al factorilor de proliferare celulară PCNA şi p53 a arătat o imunomarcare intensă a celulelor din stratul profund şi absenţa imunomarcajului în perlele cheratozice. Imunocolorarea cu PCNA a marcat neuniform celulele carcinomatoase, inclusiv pe cele cu nudei monstruoşi, dovedind că acestea se găsesc în diverse stadii ale ciclului celular. Concluzii. Carcinoamele epidermoide bine diferenţiate au reprezentat doar 10% din studiul cancerelor de amigdală palatină. Parenchimul tumoral s-a caracterizat prin variat pleomorfism celular şi nuclear. Imunomarcarea PCNA şi p53 a fost intens pozitivă în celulele tumorale, inclusivîn celulele cu nudei monstruoşi, dovedind capacitatea proliferativă mare a celulelor carcinomatoase. [ABSTRACT FROM AUTHOR]