The Impact of Juvenile Microglia Transcriptomics on the Adult Brain Regeneration after Cerebral Ischemia.

Microglial cells play a pivotal role in the brain's health and operation through all stages of life and in the face of illness. The contributions of microglia during the developmental phase of the brain markedly contrast with their contributions in the brain of adults after injury. Enhancing our understanding of the pathological mechanisms that involve microglial activity in brains as they age and in cerebrovascular conditions is crucial for informing the creation of novel therapeutic approaches. In this work we provide results on microglia transcriptomics in the juvenile vs injured adult brain and its impact on adult brain regeneration after cerebral ischemia. During fetal brain development, microglia cells are involved in gliogenesis, angiogenesis, axonal outgrowth, synaptogenesis, neurogenesis and synaptic reorganization by engulfing neuronal extensions. Within the mature, intact brain, microglial cells exhibit reduced movement of their processes in response to minimal neuronal activity, while they continuously monitor their surroundings and clear away cellular debris. Following a stroke in the adult brain, inflammation, neurodegeneration, or disruptions in neural equilibrium trigger alterations in both the genetic blueprint and the structure and roles of microglia, a state often described as "activated" microglia. Such genetic shifts include a notable increase in the pathways related to phagosomes, lysosomes, and the presentation of antigens, coupled with a rise in the expression of genes linked to cell surface receptors. We conclude that a comparison of microglia transcriptomic activity during brain development and post-stroke adult brain might provide us with new clues about how neurodegeneration occurs in the adult brain. This information could very useful to develop drugs to slow down or limit the post-stroke pathology and improve clinical outcome.

Breast MRI-the Importance of Type II and III Dynamic Curves Evaluation and Framing in BI-RADS 4C and 5 Score.

BACKGROUND: Breast Magnetic Resonance Imaging (MRI) offers the highest sensitivity in detecting breast cancer among existing clinical and imaging techniques, making it a crucial component of breast imaging protocols. This study aims to investigate MRI importance in correlation with previous imaging discordant procedures performed as echography and/or mammography to evaluate characteristics and framing in high-risk BI-RADS 4C or 5 categories based on morphological features and kinetic curves of masses found in the breasts of patients from our database. METHODS: A retrospective study with related statistical analysis was performed on a group of 33 cases, selected from a total of 488 patients who underwent breast MRI examinations at SPAD Imaging International S.R.L. Craiova, between 01.01.2021 and 31.12.2023, aged between 33 and 75 years. In all patients, MRI images parameters were analysed. RESULTS: In 33 patients, 23 had a single lesion and 10 had multiple lesions, 9 of them in the ipsilateral breast and, as a particularity, one of them, located in the contralateral breast. In 21 of the total patients with multiple or single lesions they had type III curves, which were classified in the BI-RADS 5 category, considering both criteria-morphology and type of curve, where the other previous techniques had not mentioned an increased risk, hence revealing that the situation in a percentage of 63.63 in the case of MRI investigation proved to be clearly superior. CONCLUSION: Combining both kinetic and morphologic criteria can enhance the diagnostic accuracy of MRI in breast lesion evaluation.

Clinical and Morphological Study of Cervical Squamous Intraepithelial Lesions.

Squamous intraepithelial lesions (SILs) are cancer precursors targeted by secondary prevention of cervical cancer programs that are sometimes difficult to grade accurately. Mena is an actin regulatory protein involved in membrane protrusion, cell motility, in tumor invasion and metastasis. We studied retrospectively 68 cases of patients diagnosed with squamous intraepithelial lesions that received expedited treatment (treatment without colposcopic biopsy). We analyzed demographic, behavioral data, obstetrical and medical history, from the patients' medical charts and we studied the cervical fragments or cones harvested after the excisional procedure. Our study failed to identify a correlation between SILs and risk factors such as low socioeconomic status, combined oral contraceptive use, intrauterine device use, parity, gravity, except for the tobacco smoking habit that proved to be related to the cervical lesions' development. Mena was expressed in most of the analyzed SILs and its expression was correlated with lesions' grade in terms of both area and intensity, suggesting that Mena stains especially abnormal cells and that its expression intensity correlates with the risk of malignant transformation. Further studies are needed to validate Mena as an early stage of cervical carcinogenesis marker.

The influence of SARS-CoV-2 on the immune system elements and on the placental structure. Clinical, histological and immunohistochemical study.

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.

Treatment of skin defects with PRP enriched with hyaluronic acid - histological aspects in rat model.

Tissue healing is a complex, dynamic process, characterized by the replacement of devitalized and absent cell and tissue structures. This can be obtained by different methods, these being found in the "reconstructive scale", which although it is very rich does not offer a universally valid solution for closing skin wounds. In plastic surgery, platelet-rich plasma (PRP) has proven effective in the treatment of skin graft donor areas, burn wounds, skin grafts, tendons, or varicose ulcers. Also, hyaluronic acid (HA) has found its utility in different areas of medicine, other than the esthetics field, with satisfactory results after its use in various lesions. The aim of our study was to find a method of healing wounds with skin defect that shortens the time of complete epithelialization compared to native healing, which is accessible to any patient both by its simplicity and by the lowest possible costs. So, we decided to test a preparation consisting of PRP and granular HA in this type of wounds on a group of 30 Wistar rats. Corroborating the macroscopic data with the microscopic ones, an important similarity can be observed between the healing of the adjuvant-treated lesion at 14 days postoperatively and the healing of the lesion left to natural healing at 21 days, thus shortening the healing period by seven days.

E-Cadherin Modulation and Inter-Cellular Trafficking in Tubular Gastric Adenocarcinoma: A High-Resolution Microscopy Pilot Study.

Despite the numerous advances in tumor molecular biology and chemotherapy options, gastric adenocarcinoma is still the most frequent form of gastric cancer. One of the core proteins that regulates inter-cellular adhesion, E-cadherin plays important roles in tumorigenesis as well as in tumor progression; however, the exact expression changes and modulation that occur in gastric cancer are not yet fully understood. In an attempt to estimate if the synthesis/degradation balance matches the final membrane expression of this adhesion molecule in cancer tissue, we assessed the proportion of E-cadherin that is found in the Golgi vesicles as well as in the lysosomal pathway We utilized archived tissue fragments from 18 patients with well and poorly differentiated intestinal types of gastric cancer and 5 samples of normal gastric mucosa, by using high-magnification multispectral microscopy and high-resolution fluorescence deconvolution microscopy. Our data showed that E-cadherin is not only expressed in the membrane, but also in the cytoplasm of normal and tumor gastric epithelia. E-cadherin colocalization with the Golgian vesicles seemed to be increasing with less differentiated tumors, while co-localization with the lysosomal system decreased in tumor tissue; however, the membrane expression of the adhesion molecule clearly dropped from well to poorly differentiated tumors. Thus E-cadherin seems to be more abundantly synthetized than eliminated via lysosomes/exosomes in less differentiated tumors, suggesting that post-translational modifications, such as cleavage, conformational inactivation, or exocytosis, are responsible for the net drop of E-cadherin at the level of the membrane in more anaplastic tumors. This behavior is in perfect accordance with the concept of partial epithelial-to-mesenchymal transition (P-EMT), when the E-cadherin expression of tumor cells is in fact not downregulated but redistributed away from the membrane in recycling vesicles. Moreover, our high-resolution deconvolution microscopy study showed for the first time, at the tissue level, the presence of Lysosome-associated membrane glycoprotein 1 (LAMP1)-positive exosomes/multivesicular bodies being trafficked across the membranes of tumor epithelial cells. Altogether, a myriad of putative modulatory pathways is available as a treatment turning point, even if we are to only consider the metabolism of membrane E-cadherin regulation. Future super-resolution microscopy studies are needed to clarify the extent of lysosome/exosome exchange between tumor cells and with the surrounding stroma, in histopathology samples or even in vivo.

Subtle Immunoreactivity Differences in the Fractal Patterns of Membrane E-Cadherin in Gastric Adenocarcinoma.

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.

Accuracy of Endoscopic Ultrasonography for Gastric Cancer Staging.

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.

Gastric Cancer Angiogenesis Assessment by Dynamic Contrast Harmonic Imaging Endoscopic Ultrasound (CHI-EUS) and Immunohistochemical Analysis-A Feasibility Study.

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.

Descriptive Analysis of Patients with Gastric Tumors Referred to the Largest Emergency Hospital in Oltenia Region-Romania, Between 2015-2020.

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.

Clinical and pathological aspects of condyloma acuminatum - review of literature and case presentation.

Condyloma acuminatum (CA) is a pathology caused by the human papillomavirus (HPV). It is manifested by the appearance of warts in the vulvar, pubic, and anorectal regions, but can occur in other areas. It is a common disease that can be prevented by using measures such as condoms or vaccine. Topical, local, pharmacological, surgical, and excisional therapy options are available for this pathology. Macroscopically, it appears as a vegetative tumor, with a single implantation base that branches towards the periphery, with a cauliflower appearance. CA is defined microscopically by acanthosis, parakeratosis, papillomatosis and koilocytosis. Immunohistochemical studies can detect the presence of various HPV strains or viral antigens and can emphasize certain specific characteristics; e.g., in the case presented in this study, we observed that the tumor had a fulminant evolution due to a strong vascular base identified with anti-cluster of differentiation (CD) 34 antibody, by the existence of epithelial cells with a high degree of cell proliferation, as evidenced by the anti-Ki67 antibody, the inactivation of the tumor suppressor gene and the appearance of immunolabeling for the anti-p53 antibody, by the strong immunoreactivity for p63 which reveals the existence of cells with dysplastic and neoplastic transformation potential, but also by detecting the immunolabeling for p16INK4a that is associated with the existence of HPV. Also, the tumor was immunoreactive for cytokeratin (CK) AE1∕AE3, partially immunoreactive for CK5∕6 in the basal layer and negative for CK7, which demonstrates the squamous epithelial origin of the described tumor. Subepithelial cells of the inflammatory system have been identified, such as macrophages immunolabeled with anti-CD68 antibody, T-lymphocytes immunolabeled with anti-CD3 antibody and rare B-lymphocytes immunolabeled with anti-CD20 antibody, which demonstrates the strong cellular response to remove the virus from the structure. Surgical and excisional treatment was helpful for the patient, because she was able to resume normal sexual activity and defecation, and on the other hand, microscopic studies showed the potential for malignant transformation of CA.

Clinicopathologic Relevance of Claudin 18.2 Expression in Gastric Cancer: A Meta-Analysis.

An increasing number of tumor markers have been discovered to have potential efficacy as diagnostic and prognostic tools in gastric cancer. We aimed to assess putative correlations between claudin 18.2 expression and pathological or prognosis features in patients with gastric cancer. MEDLINE, Web of Science, EBSCO, and ClinicalTrials.gov were used to search for relevant studies from their inception to 30 October 2020. Finally, a total of six articles were included in this meta-analysis. Review Manager 5 software was applied to examine the heterogeneity among the studies and to calculate the odds ratio with 95% CI by selecting corresponding models, in evaluating the strength of the relationship. Publication bias test was also conducted. No bias and no significant correlations were found between CLDN 18.2 and TNM stages, Lauren classification, HER2, grading, or overall survival. This meta-analysis expounded that the relationship with CLDN 18.2 and pathological features depends on the percentage of staining of tumor cells for which CLDN 18.2 is considered positive. Our pooled outcomes suggest that targeted therapy for CLDN 18.2 could be effective if certain criteria were established.

Nanoparticle-functionalized dressings for the treatment of third-degree skin burns - histopathological and immunohistochemical study.

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.

In-hospital and short-term prognostic factors in acute pulmonary edema: clinical and morphological features.

INTRODUCTION: There are extensive records which have included patients with acute heart failure (AHF), but specific studies about prognosis in acute pulmonary edema (PE) are scarce and have enrolled a small number of patients. The objectives of this study were to evaluate the predictive factors of short-term evolution in patients with PE. PATIENTS, MATERIALS AND METHODS: This was a prospective, two-center survey of 70 consecutive patients admitted for acute cardiogenic PE. The follow-up was performed one month after discharge. The composite endpoint was in-hospital death, and death of any cause or readmission for heart failure (HF) at one month after discharge. Heart and lung tissue analysis was performed postmortem to identify morphological features of PE. RESULTS: In-hospital mortality was 4.2%, another 14.2% died in the first month, and an additional 10% required rehospitalization for HF. The characteristics significantly associated with end-point occurrence were: history of kidney disease, anemia, diabetes mellitus, lack of prior angiotensin-converting enzyme inhibitor÷angiotensin-receptor blocker treatment, lower systolic blood pressure (BP) at admission, lower diastolic BP at admission, creatinine at admission and at discharge, an increase in creatinine during stay, glomerular filtration rate at admission, serum sodium at admission, decrease in serum sodium during hospitalization, N-terminal prohormone of brain natriuretic peptide (NT-proBNP) at discharge, right ventricle fractional area change, left atrium volume index. We created a multiple logistic regression model and identified five prognostic factors: age, diabetes, creatinine, diastolic BP, serum sodium. This model correctly classified 48 (96%) patients without worsening and 13 (65%) patients with worsening, providing an overall accuracy of 87.1%. Necropsy was performed on five patients and fragments of left ventricle myocardium and lung were harvested for histopathological and immunohistochemical studies. The myocardium exhibited fibrosis areas where the myocytes were completely or partially replaced by collagen fibers. Lung tissue analysis revealed some case-to-case differences, but the common finding was alveoli size larger than normal, with the lumen completely or almost completely covered by an eosinophilic liquid. CONCLUSIONS: The factors that best predicted the short-term outcome in PE were age, diabetes, diastolic BP, creatinine, serum sodium.