Intense Inflammatory Infiltrate Research Articles

Green formulation and characterization of Camellia azalea synthesized copper nanoparticles and its application in the treatment of ovalbumin-induced allergic conjunctivitis

Camellia azalea can treat many stubborn and malignant diseases, according to traditional Chinese medicine. In addition, it has been indicated that copper nanoparticles containing the plant have many therapeutic effects. Recently, metallic nanoparticles have been used due to the antioxidant and anti-inflammatory potentials. Present research discloses the CuNPs biosynthesizing capability of the leaves of pharmacologically important Camellia azalea. For determining the physicochemical properties of the recent nanoparticles, FE-SEM, FT-IR, TEM and UV-Vis were used. This study therefore assessed the antiallergic potential of copper nanoparticles in ovalbumin-induced allergic conjunctivitis and attempted to predict its action mode. ELISA test was applied to measure the serum ovalbumin-specific IgG and IgE and total IgE. Clinical scores for allergic conjunctivitis induced by intraperitoneal ovalbumin sensitization and topical conjunctival challenge in Dunkin-Hartley guinea pigs were determined after a week’s daily treatment with 10 mg kg−1 chlorpheniramine, 30 mg kg−1 prednisolone, 15 and 80 µg kg−1 copper nanoparticles, or 10 mL kg−1 PBS. The 15 and 60 mg kg−1 copper nanoparticles treatment resulted in a significantly (p < 0.01) low clinical score of allergic conjunctivitis. Ovalbumin-specific IgG and IgE as well as total serum IgE also decreased significantly (p < 0.01). The conjunctival tissue in copper nanoparticles treated guinea pigs had mild mononuclear infiltration compared to the PBS-treated ones, which had intense conjunctival tissue inflammatory infiltration.

New Histoprognostic Factors to Consider for the Staging of Colon Cancers: Tumor Deposits, Invasive Tumor Infiltration and High-Grade Budding

Colorectal cancer is a major public health issue due to its high incidence and mortality. It is, therefore, essential to identify histological markers for prognostic purposes and to optimize the therapeutic management of patients. The main objective of our study was to analyze the impact of new histoprognostic factors, such as tumor deposits, budding, poorly differentiated clusters, mode of infiltration, the intensity of inflammatory infiltrate and the type of tumor stroma, on the survival of patients with colon cancer. Two hundred and twenty-nine resected colon cancers were fully histologically reviewed, and survival and recurrence data were collected. Survival was analyzed using Kaplan-Meier curves. A univariate and multivariate Cox model was constructed to identify prognostic factors for overall survival and recurrence-free survival. The median overall survival of the patients was 60.2 months and the median recurrence-free survival was 46.9 months. Overall survival and recurrence-free survival were significantly worse in the presence of isolated tumor deposits (log rank = 0.003 and 0.001, respectively) and for an infiltrative type of tumor invasion (log rank = 0.008 and 0.02, respectively). High-grade budding was associated with a poor prognosis, with no significant difference. We did not find a significant prognostic impact of the presence of poorly differentiated clusters, the intensity of the inflammatory infiltrate or the stromal type. In conclusion, the analysis of these recent histoprognostic factors, such as tumor deposits, mode of infiltration, and budding, could be integrated into the results of pathological reports of colon cancers. Thus, the therapeutic management of patients could be adjusted by providing more aggressive treatments in the presence of some of these factors.

Environmental enteric dysfunction and small intestinal histomorphology of stunted children in Bangladesh.

There is lack of information on the histological characteristics of the intestinal mucosa in Bangladeshi children. Collection of intestinal biopsy samples and assessment of the histomorphological features is considered to be the traditional gold standard for diagnosis of environmental enteric dysfunction (EED). The purpose of the study was to evaluate the intestinal histological characteristics of stunted children aged between 12-18 months with possible EED. 110 children with chronic malnutrition (52 stunted with length-for-age Z score, LAZ<-2 and 58 at risk of stunting with LAZ <-1 to -2) from the Bangladesh Environmental Enteric Dysfunction (BEED) study protocol who underwent upper gastrointestinal (GI) endoscopy were selected for this study. To explore the association of EED with childhood stunting, upper GI endoscopy was done and the biopsy specimens were studied for histopathology. Villous height and crypt depth were measured and the presence and intensity of inflammatory infiltrates in the lamina propria was investigated. Bivariate analysis was performed to examine the relationship between stunting and histologic morphology. More than 90% children irrespective of nutritional status were diagnosed to have chronic non-specific duodenitis on histopathology. Half of the children from both groups had villous atrophy as well as crypt hyperplasia and lymphocytic infiltration was present in more than 90% children, irrespective of groups. However, no statistically significant difference was observed when compared between the groups. The prevalence of chronic non-specific duodenitis in Bangladeshi children, irrespective of nutritional status, was high. A significant number of these children had abnormal findings in intestinal histomorphology. Trial registration number: ClinicalTrials.gov ID: NCT02812615 Date of first registration: 24/06/2016. https://clinicaltrials.gov/ct2/results?cond=NCT02812615&term=&cntry=&state=&city=&dist.

Inmixed and Bellicose-Adenosquamous Carcinoma-Uterine Cervix

Adenosquamous carcinoma of uterine cervix is a preponderantly high grade neoplasm configured of admixed glandular articulations and squamous epithelium wherein glassy cell carcinoma emerges as an exceptional, aggressive variant. Adenosquamous carcinoma is posited to arise from sub-columnar reserve cells confined to endocervical basal epithelium. Cytological smears depict papillary articulations admixed with bland columnar epithelial cells and high grade squamous intraepithelial lesion. The biphasic tumour is composed of well defined, malignant components of glandular articulations and poorly differentiated squamous epithelium. Glassy cell carcinoma demonstrates solid cell nests comprised of pleomorphic, polygonal tumour cells pervaded with glassy, eosinophilic cytoplasm, enlarged, eosinophilic nuclei, prominent nucleoli and distinct cytoplasmic membrane. Neoplastic cells appear enmeshed within an intense inflammatory cell infiltrate with significant eosinophils. Glassy cell carcinoma appears immune reactive to MUC1, MUC2, epithelial membrane antigen (EMA), CAM5.2 or p63 with focal immune reactivity to carcinoembryonic antigen (CEA). Adenosquamous carcinoma requires segregation from neoplasms such as adenocarcinoma with coexisting squamous intraepithelial lesion (SIL), adenoid basal carcinoma, cervical extension of endometrial adenocarcinoma, and squamous cell carcinoma with focal mucin droplets, large cell non keratinizing squamous cell carcinoma or lymphoepithelioma-like carcinoma. Tumefaction may be appropriately treated by radical abdominal hysterectomy followed by adjuvant chemotherapy and radiation therapy.

DOES GALLIC ACID HAVE A POTENTIAL REMEDIAL EFFECT IN EXPERIMENTAL CORROSIVE BURN INJURY TO THE ESOPHAGUS?

Gallic acid, acting as an antioxidant, anti-precipitant and cytoprotective agent, was used as a possible remedial natural component for treating experimentally induced esophageal burn. Wistar rats (n=24) were divided into three groups. Control group was given 1 mL 0.9% NaCl. Experimental esophageal burn was induced with 1 mL 40% NaOH application to the esophagus in groups 2 and 3. Gallic acid® (20 mg/kg) was administered to the treated group via oral gavage for 10 days. Removed tissues were fixed and paraffin blocks were prepared. Histopathological examination was performed after the sections had been stained with hematoxylin-eosin. Tumor necrosis factor alpha and caspase-3 antibodies were used on immunohistochemical analysis. In the esophageal burn group, necrosis, degeneration and numerous apoptotic cells, as well as intense inflammatory cell infiltration and fibrosis in the muscle layer were observed under light microscope. In the treated group, remodeling of epithelial cells with marked reduction in the connective tissue collagen content was observed, as well as marked reduction in the volume of collagen and abundance of inflammatory cells in blood vessels. Gallic acid treatment may help heal esophageal burns and prevent complications.

P160 Paracoccidioidomycosis severe: Bizarre presentation in the face

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM IntroductionParacoccidioidomycosis is a systemic mycosis caused by the dimorphic fungus Paracoccidioides brasiliensis. The disease is restricted to Latin America. It is the principal systemic mycosis in Brazil, with higher incidences in the southern, southeastern and central regions.ObjectivesIt's to report a case of paracoccioidomycosis with a bizarre presentation on the face.Case ReportA 54-year-old male farmer from Açailândia, Maranhão, with a history of an extensive ulcerated lesion that started in the right wing of the nose. After 6 months of evolution, the lesion had already spread reaching the nasal and oral mucosa, evolving with acute respiratory failure, requiring tracheostomy (Fig.1). He was referred to the Cancer Hospital for a biopsy, which was referred for pathological examination, showing an intense inflammatory infiltrate consisting of lymphocytes, histiocytes, neutrophilis with giant, multinucleated cells that phagocytize refrigerant bodies, compatible with paracoccidioidomycosis. Material sent for direct examination and culture, showed positive for Paracoccidioides sp. After PCR amplification of the genetic material and sequencing, P. brasiliensis was identified. Chest tomography showed sparse ground-glass attenuation, but affecting both lungs and radiodense striations in the lower lobes bilaterally. Treatment was established with liposomal amphotericin B, with a good therapeutic response (Fig. 2). The patient was discharged from the hospital using itraconazole and still upper outpatient follow-up, with good healing of the lesions.ConclusionParacoccidioidomycosis can get a severe presentation, distancing the physician from the diagnosis, showing that in all these situations, simple tests, such as the direct examination of lesional material or sputum can elucidate the diagnosis. We need to think about it in endemic area.

IMMUNOHISTOCHEMICAL EXPRESSION OF METALLOPROTEINASES AND TFN-Α BY MACROPHAGES AND LANGERHANS CELLS IN RADICULAR CYSTS

Objectives This research aimed to evaluate the relationship between Langerhans cells, macrophages, matrix metalloproteinases (MMPs-9, -13), and tumor necrosis factor (TNF)-α in the lining epithelium of cystic lesions to elucidate the cyst expansion. Study Design A sectional and retrospective study was performed with 20 radicular cysts (RCs) and 20 residual radicular cysts (RRCs) by anti-CD68, anti-CD1a, anti-MMP-9, anti-MMP-13, and anti-TNF-α antibodies. Results The immunoexpression of MMP-13 and CD68 was significantly higher in RCs when compared to RRCs (P = .011 and P = .012, respectively). An intense inflammatory infiltrate was significantly correlated with CD68 immunoexpression in RCs (P = .025). Macrophages demonstrated a significant positive correlation with MMP-13 (P = .015). A moderate correlation was observed between MMP-9 and MMP-13 (P = .01). TNF-α expression was more common in RCs (P = .001). Langerhans cells were more frequently expressed in atrophic epithelium (P = .041) and significantly correlated with TNF-α (P = .014) in RCs. Conclusions Macrophages induce a greater release of TNF-α. Higher immunoexpression of MMP-13 and MMP-9 is observed in the early stages of RCs compared to RRCs. The microorganisms are the main factors triggering a proinflammatory immune response and greater cystic expansion in the early stages of these cysts. This research aimed to evaluate the relationship between Langerhans cells, macrophages, matrix metalloproteinases (MMPs-9, -13), and tumor necrosis factor (TNF)-α in the lining epithelium of cystic lesions to elucidate the cyst expansion. A sectional and retrospective study was performed with 20 radicular cysts (RCs) and 20 residual radicular cysts (RRCs) by anti-CD68, anti-CD1a, anti-MMP-9, anti-MMP-13, and anti-TNF-α antibodies. The immunoexpression of MMP-13 and CD68 was significantly higher in RCs when compared to RRCs (P = .011 and P = .012, respectively). An intense inflammatory infiltrate was significantly correlated with CD68 immunoexpression in RCs (P = .025). Macrophages demonstrated a significant positive correlation with MMP-13 (P = .015). A moderate correlation was observed between MMP-9 and MMP-13 (P = .01). TNF-α expression was more common in RCs (P = .001). Langerhans cells were more frequently expressed in atrophic epithelium (P = .041) and significantly correlated with TNF-α (P = .014) in RCs. Macrophages induce a greater release of TNF-α. Higher immunoexpression of MMP-13 and MMP-9 is observed in the early stages of RCs compared to RRCs. The microorganisms are the main factors triggering a proinflammatory immune response and greater cystic expansion in the early stages of these cysts.

ORAL ERYTHEMA MULTIFORME—A CASE REPORT

An acute inflammatory condition that affects the skin and mucosa, erythema multiforme is caused by a cell-mediated immune response. This paper reports the case of a patient diagnosed with erythema multiforme. A 29-year-old male patient reported pain in the mouth and throat lesions. During intraoral examination the presence of erosive and ulcerated lesions was noted, with erythematous coloration, disseminated, diffuse limits, irregular format, hemorrhagic and ulcerated surface in the buccal mucosa, soft palate, and lateral border of the tongue. Incisional biopsy was performed. Histopathology revealed a parakeratinized epithelium with hyperplasia, acanthosis, hydropic degeneration, focal exocytosis, and ulcers covered with fibrinous exudate. The lamina propria of fibrous connective tissue was permeated by intense mononuclear inflammatory infiltrate. The conclusion was a chronic nonspecific inflammatory process with perivascular disposition, confirming the diagnosis of erythema multiforme. Total remission was obtained with prednisolone 20 mg for 30 days without relapse for 3 years.

THE ROLE OF ORAL LESIONS IN THE DIAGNOSIS OF BEHÇET'S SYNDROME

Behçet's syndrome is an inflammatory disorder known as a singular model of vasculitis. A 47-year-old male patient was referred to the stomatologist by a rheumatologist after hospitalization for thrombosis of the iliac vein and inferior vena cava and high C-reactive protein and erythrocyte sedimentation rate values. The patient reported recurrent folliculitis and oral and scrotum ulcerations. Intraoral examination revealed ulcerations dispersed through the mucosa. An excisional biopsy was performed in a right buccal mucosa lesion. Histopathologic analysis demonstrated nonkeratinized stratified squamous epithelium with areas of exocytosis and ulceration. Connective tissue presented an intense mixed inflammatory infiltrate, congested vessels, hemorrhage, and vasculitis. HLA-B genotyping identified the presence of HLA-B15. The histopathologic report associated with the other findings enabled the rheumatologist to diagnose the condition. The patient is on daily continuous oral use of colchicine 0.5 mg and prednisolone 20 mg, in addition to weekly subcutaneous administration of 0.1 mL of methotrexate 25 mg/mL and has been without recurrence for 8 months.

GLANDULAR CHEILITIS: CASE REPORT

Glandular cheilitis is a rare chronic inflammatory disease affecting the minor salivary glands, predominantly in the lower lip. An 18-year-old Black woman presented with macrocheilia with a soft consistency in the lips. There were yellowish crusts in the vermilion border that, when removed, revealed multiple punctate red, painful papules in lip mucosa. After ruling out associated systemic disease and infections, an incisional biopsy was performed in the lower lip. Histopathologic findings consisted of enlarged salivary ducts with accumulation of material suggestive of mucus, intense inflammatory mononuclear infiltrate, and areas of ductal metaplasia and acinar degeneration in the minor salivary glands. A final diagnosis of glandular cheilitis was made. Topical clobetasol propionate was prescribed 3 times per day and progressive improvement was observed over 30 days. The patient did not present recurrence in 6 months of follow-up.

Influence of supplement administration of omega-3 on the subcutaneous tissue response of endodontic sealers in Wistar rats.

Natural substances such as omega-3 have been used in the medical field due to their numerous properties and, in particular, modulating effect on the systemic and local inflammatory processes. Thus, this study evaluated the influence of omega-3 supplementation on the subcutaneous tissue response of endodontic sealers in Wistar Rats. Polyethylene tubes were implanted in the subcutaneous tissue of 48 animals (one empty for control and three filled with Sealapex, AH Plus or Endofill). The animals were treated with omega-3 (TO) or water (TW). Treatments started 15 days before implantation until euthanasia. After 5, 15 and 30 days (n=8), animals were euthanized and polyethylene tubes and surrounding tissue were removed and processed for histological analysis. The inflammatory reaction was analysed by Haematoxylin and Eosin stain and immunolabelling for IL-6 and TNF-α. The collagen maturity was analysed by picrosirius red stain and calcium deposition by von Kossa stain and polarized light. Results were statistically analysed (p < .05). Amongst TW sealer groups, Endofill evoked a more intense inflammatory infiltrate compared with AH Plus and control in the 30-day period (p=.009). However, in TO sealer groups, there was no difference amongst the sealers and control in all periods (p >.05). Comparing each sealer as a function of the supplementation with water or omega-3, there are differences for Endofill (p =.001) and Sealapex (p =.005) in the 30-day period, presenting lower inflammatory infiltrate in the animals treated with omega-3. A higher percentage of immature fibres was observed at 15 and 30 days in the TO group, compared with the TW group (p <.05). The deposition of calcium particles was observed only by Sealapex in all periods, despite the supplementation procedure. Omega-3 supplementation influence the tissue reactions of endodontic sealers, modulating inflammation, the immunolabelling of IL-6 and TNF-α, the repair process and it does not interfere with calcium deposition.

Eosinophilic Granulomatous Myocarditis in Two Pigs

Eosinophilic myocarditis is a human condition that has been rarely documented in animals. We now report two unrelated porcine cases of idiopathic eosinophilic granulomatous myocarditis that resembled the human disease and which were associated with sudden death. The most relevant gross finding in both cases was marked cardiomegaly, accompanied by raised, multifocal to coalescent small white nodules (1-2mm) and poorly demarcated multifocal pale areas in the epicardium. Histologically, there were multifocal to coalescent areas of cardiomyocyte loss with replacement by an intense inflammatory infiltrate of eosinophils and epithelioid macrophages, and proliferation of fibrous connective tissue. Immunohistochemistry for porcine circovirus type 2 (PCV2) and Toxoplasma gondii, in-situhybridization and quantitative polymerase chain reaction tests for PCV2 and porcine circovirus type 3 and aerobic bacterial culture on myocardium samples were negative.

Abstract 774: Microdissected gene expression profiling of recurrent nasopharyngeal carcinoma

Abstract Nasopharyngeal cancer (NPC) is an EBV-driven epithelial carcinoma with a recurrence rate of 30%. Biological pathways and predictive markers for recurrence remain understudied. Further, NPC tumors are associated with an intense inflammatory infiltrate, masking gene expression signals from the tumor epithelial compartment. We perform laser-capture microdissection on a preliminary cohort of 12 tumors, including primary tumors from recurrent and non-recurrent cases matched for stage of disease. Smart-3SEQ, a novel 3’ end RNA-Seq technique which allows for the accurate quantification of transcript abundance in FFPE samples, was performed to obtain gene expression libraries. Tumor epithelial and microenvironment regions from were separately microdissected, giving a total of 42 histologically-pure gene expression libraries. Raw reads were mapped to the human hg19 and EBV genomes and gene expression counts were obtained with Subread. Differential gene expression and gene set enrichment was performed in R with DESeq2 and fgsea respectively. We observed that primary NPC tumors that did not recur were significantly enriched for immune processes, including leukocyte mediated immunity, innate immune response and lymphocyte activation (padj &amp;lt; 1E-9 for all). The top differentially expressed genes in this group included mediators of inflammation such as STAT1 (padj = 0.0204). Critically, these gene expression changes were observed within the tumor epithelial compartment. In contrast, primary NPC tumors that eventually recurred were enriched for known oncogenes including BCL2 (padj = 0.000420). These findings suggest that heterogeneity in gene expression between NPC tumors play an important role in treatment response. Further work to define gene expression subtypes of NPC tumors will be helpful for patient stratification and personalized care. Citation Format: Joshua Tay, Wei Keat Teo, Chuan Keng Goh, Bing Cheng Wu, Kwok Seng Loh. Microdissected gene expression profiling of recurrent nasopharyngeal carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 774.

Influence of chlorhexidine digluconate on biocompatibility of cements: Morphological and immunohistochemistry analysis.

In this study, the in vivo biocompatibility was evaluated by using conventional ionomer cements modified with Chlorhexidine (CHX) in different time intervals. In total, 105 male Wistar rats were randomized into seven groups: control, groups M, M10, M18 and groups RL, RL10, RL18 (M-Meron and RL-RivaLuting, and added CHX-10% and CHX-18%, respectively). Histological analyses of inflammatory infiltrate and collagen fibers, and immunohistochemistry of CD68+ for macrophages (MOs) and multinucleated giant cells (MGCs) were performed. Data were analyzed using the Kruskal-Wallis and Dunn (p< .05) tests. Intense inflammatory infiltrate was demonstrated in Group Riva CHX-18% within 7 and 15 days (p< .05), without differences after 30 days. For collagenization, healing of the groups was compatible with that of control in 15 and 30 days (p> .05). Immunomarking of CD68+ was more significant in the groups with higher concentration of CHX. There was significant difference in quantity of MGCs in groups with 18% CHX, Meron (p= .001) in 7 days, and in Riva at 30 days (p= .001). Significant difference was also found in quantities of MOs in Groups Meron and Riva in 7 days (p= .001), and only in Riva at 15 and 30 days (p= .001). The cements with addition of CHX demonstrated biocompatibility with tissues. Riva CHX-18% had the most effect on cells of the inflammatory process but showed satisfactory tissue repair. RESEARCH HIGHLIGHTS: The concentration of 18% chlorhexidine was shown to be biocompatible with tissues; the slow release of chlorhexidine by the cements could significantly prolong its antibacterial effect on the oral medium.

Gingival tissue as a reservoir for human immunodeficiency virus type 1: Preliminary results of a cross-sectional observational study.

Despite combined antiretroviral therapy (cART), total cure of immunodeficiency virus type 1 (HIV-1) infection remains elusive. Chronic periodontitis (CP) is strongly associated with HIV-1 infection. This condition is characterized by an intense inflammatory infiltrate mainly constituted of immune cells which in turn may be a valuable source of HIV-1 reactivation. This study aimed to determine if gingival tissue could act as a reservoir for HIV-1. Twelve patients with HIV-1 and CP and 12 controls (no HIV-1-infection and no CP) were evaluated in a cross-sectional study. RNA viral load and interleukin (IL) levels were determined in blood plasma and saliva. Histological sections of gingival tissue were stained with fluorescent antibodies against p24 antigen and different cellular biomarkers. In six of the 12 patients, HIV RNA load was detected, despite cART; in three of them, expression of viral RNA was also detected in saliva. The levels of IL-2, IL-6, and IL-12 were higher in blood and saliva of patients with HIVand CP than controls. HIV-1 p24 antigen was detected by immunostaining in gingival biopsies of 10 of the 12 patients but in no controls. Immune markers for T cells and antigen-presenting cells were also identified in most patients and some controls. These preliminary data showing the detection of HIV-1 p24 antigen in the gingival biopsies of a significant part of patients with HIV-1 and CP under cART together with the presence of immune cells, plead for the existence of a HIV-1 reservoir in the gingival tissue of this population.

Biocompatibilidade in vivo de cartilagem elástica tratada em soluções alcalinas

ABSTRACT This study verified the in vivo biocompatibility of bovine elastic cartilage decellularized with alkaline solution in relation to the non-decellularized cartilage implanted in rats. Fifty Wistar rats were divided into two groups, with the experimental group (EG) receiving subcutaneous implants of cartilage treated in alkaline solution and the control group (CG) receiving subcutaneous implants of untreated cartilage. In both groups, the implants were removed on days 3, 7, 14, 21 and 42 with the preparation of histological slides stained with Hematoxylin and Eosin for the quantification of inflammatory cells, fibroblasts, angiogenesis, and cartilage degradation. The results showed that EG presented a less intense inflammatory infiltrate and better organization of collagen fibers compared to CG. It was concluded that the alkaline treatment provided better biocompatibility for elastic cartilage when implanted subcutaneously in rats.

Inflammatory monocytes and microglia play independent roles in inflammatory ictogenesis

BackgroundThe pathogenic contribution of neuroinflammation to ictogenesis and epilepsy may provide a therapeutic target for reduction of seizure burden in patients that are currently underserved by traditional anti-seizure medications. The Theiler's murine encephalomyelitis virus (TMEV) model has provided important insights into the role of inflammation in ictogenesis, but questions remain regarding the relative contribution of microglia and inflammatory monocytes in this model.MethodsFemale C57BL/6 mice were inoculated by intracranial injection of 2 × 105, 5 × 104, 1.25 × 104, or 3.125 × 103 plaque-forming units (PFU) of the Daniel’s strain of TMEV at 4–6 weeks of age. Infiltration of inflammatory monocytes, microglial activation, and cytokine production were measured at 24 h post-infection (hpi). Viral load, hippocampal injury, cognitive performance, and seizure burden were assessed at several timepoints.ResultsThe intensity of inflammatory infiltration and the extent of hippocampal injury induced during TMEV encephalitis scaled with the amount of infectious virus in the initial inoculum. Cognitive performance was preserved in mice inoculated with 1.25 × 104 PFU TMEV relative to 2 × 105 PFU TMEV, but peak viral load at 72 hpi was equivalent between the inocula. CCL2 production in the brain was attenuated by 90% and TNFα and IL6 production was absent in mice inoculated with 1.25 × 104 PFU TMEV. Acute infiltration of inflammatory monocytes was attenuated by more than 80% in mice inoculated with 1.25 × 104 PFU TMEV relative to 2 × 105 PFU TMEV but microglial activation was equivalent between groups. Seizure burden was attenuated and the threshold to kainic acid-induced seizures was higher in mice inoculated with 1.25 × 104 PFU TMEV but low-level behavioral seizures persisted and the EEG exhibited reduced but detectable abnormalities.ConclusionsThe size of the inflammatory monocyte response induced by TMEV scales with the amount of infectious virus in the initial inoculum, despite the development of equivalent peak infectious viral load. In contrast, the microglial response does not scale with the inoculum, as microglial hyper-ramification and increased Iba-1 expression were evident in mice inoculated with either 1.25 × 104 or 2 × 105 PFU TMEV. Inoculation conditions that drive inflammatory monocyte infiltration resulted in robust behavioral seizures and EEG abnormalities, but the low inoculum condition, associated with only microglial activation, drove a more subtle seizure and EEG phenotype.

Perianal squamous cell carcinoma in a Quarter Horse colt

Squamous cell carcinoma (SCC) is a squamous epithelial neoplasm. They are solitary tumors with indolent expansion, are locally invasive, and can be seen in any part of the body. The most commonly affected sites are the head, orbital region, and genitalia of horses. Solar irradiation is one of the main causes, especially in depigmented and hairless areas. SCC is the neoplasm most frequently described in equine species. Several treatments have been described in horses, usually associated with surgical excision with different adjuvant therapies, hoping for a better prognosis and a decrease in cancer-related morbidity. Since the recurrence rates are high, especially in regions that do not have an adequate safety margin after tumor removal, multimodality treatment is essential. The aim of this report was to describe a case of perianal SCC in a Quarter Horse colt clinically treated with the immunomodulatory drug imiquimod. A three-year-old, grey-haired, male Quarter Horse was examined for diagnosis and treatment of an ulcerative lesion in the perianal region. According to the owner's report, the lesion appeared approximately 16 months ago. Based on the medical history and clinical examination, SCC was suspected. To confirm the clinical diagnosis, a biopsy was performed. Histological examination showed infiltrative neoplastic proliferation from the epidermis, consisting of malignant keratinocytes associated with hyperkeratosis, neutrophilic infiltrate, and foci of ulceration. The underlying dermis exhibited an intense chronic inflammatory infiltrate. The findings were compatible with SCC. Due to the location and extension of the lesion, we decided to use immunomodulatory therapy with imiquimod. After 16 weeks of imiquimod application, the lesion completely regressed. The horse was clinically followed for two years, and no recurrence was identified.

Paratesticular liposarcoma: the clinicopathological features of nineteen cases

Objective: To investigate the clinicopathological features and differential diagnoses of paratesticular liposarcoma. Methods: The cases were collected from 2012-2020, from the archives of the Department of Pathology, Peking University Third Hospital, with diagnosis confirmed by histology, immunostaining and FISH tests. Results: Totally 19 patients were enrolled (including 11 in-hospital patients and 8 consultant cases). The patients aged 37-84 years (mean 57 years). The preoperative clinical diagnoses were spermatic cord/inguinal masses (nine patients), scrotal masses (seven patients), and inguinal hernia (three patients). Six lesions recurred after local resection, including one case extending from pelvic liposarcoma. Histologically, there were 10 cases of well-differentiated liposarcoma (WDLPS) and nine cases of dedifferentiated liposarcoma (DDLPS). WDLPSs mostly showed the combined features of lipoma-like, inflammatory and sclerosing subtypes (six patients); the other four WDLPSs had pure lipoma-like subtype features. DDLPSs were low-grade (three patients) or high-grade (six patients), with the morphology resembling myxofibrosarcoma, inflammatory myofibroblastoma, spindle cell sarcoma, pleomorphic undifferentiated sarcoma and pleomorphic liposarcoma. Intense inflammatory cells infiltration was commonly observed in five WDLPSs and two DDLPSs. Ossification was observed in three tumors. Immunohistochemically, the tumors were positive for MDM2 (8/10) and CDK4 (10/10), which were expressed in lipo-differentiating cells, spindle cells in WDLPS, and in dediffferentiated components. S-100 was only expressed by lipocytes (10/10). CD34 expression was positive and diffuse in the stromal cells of WDLPSs and focal or diffuse in dedifferentiated areas (10/10). FISH tests with an MDM2 gene probe were positive (12/12). Conclusions: Paratesticular liposarcoma may be overlooked by both clinicians and pathologists. WDLPS and DDLPS predominate, showing various histologic divergences. The presence of amplification of the 12q14-q15 region (containing the MDM2 and CDK4 genes) is helpful for making the correct diagnosis.

Release of Matrix Metalloproteinases by Macrophages in Radicular Cysts and Residual Radicular Cysts.

Radicular cysts (RCs) and residual radicular cysts (RRCs) are the sequelae of dental caries and that leads to proliferation of epithelial rests of Malassez in periapical tissues. The aim was to evaluate the relationship between Langerhans cells, macrophages, matrix metalloproteinases (MMP-9, MMP-13), and tumor necrosis factor-alpha (TNF-α) in the capsule and lining epithelium of cystic lesions. Twenty RCs and 20 RRCs were submitted to immunohistochemical analysis with anti-CD68, anti-CD1a, anti-MMP-9, anti-MMP-13, and anti-TNF-α antibodies. The Mann-Whitney test and the Spearman correlation test were used for analysis of the data (P<0.05). The immunoexpression of MMP-13 and CD68 was significantly higher in RCs when compared with RRCs (P=0.011 and 0.012, respectively). The presence of an intense inflammatory infiltrate was significantly correlated with the immunoexpression of CD68 in RCs (P=0.025). Expression of CD68 showed a significant positive correlation with MMP-13 (P=0.015). A moderate correlation was observed between MMP-9 and MMP-13 (P=0.010). TNF-α expression was more common in RCs (P=0.001). CD1a was more frequently expressed in atrophic epithelium (P=0.041) and was significantly correlated with TNF-α (P=0.014). Langerhans cells induce a greater release of TNF-α which, in turn, is responsible for the stimulation of M1 macrophages. Higher immunoexpression of MMP-13 and MMP-9 is observed in the early stages of RCs compared with RRCs. Therefore, the toxins of microorganisms present in highly inflamed RCs are the main factors triggering a proinflammatory immune response and greater cystic expansion in the early stages of these lesions.