Presence Of Inflammatory Infiltrate Research Articles

Assessment of 11 β-Hydroxysteroid Dehydrogenase Type I Activity in Patients with Psoriasis Vulgaris

Abstract Psoriasis is a chronic relapsing dermatological disease that affects around 1–3% of the world’s population and the prevalence has been increasing globally over the past years. A complex multifactorial etiology is suggested to be the underlying mechanism, involving both genetic and environmental factors that can initiate psoriatic inflammation, including mild trauma, sunburn, or chemical irritants. The interaction between these factors can disrupt the epidermal homeostasis, which triggers a viscous inflammatory circle that leads to epidermal hyperplasia and psoriasis. This inflammatory cascade is the outcome of interaction between epidermal DCs, T lymphocytes and keratinocytes. However, the initial stimulus for DCs stimulation and T cells differentiation is still unclear. In psoriasis, there is a hormonal disturbance in the form of defective GC secretion associated with HPA axis dysfunction. Recently, accumulating evidence established the presence of cutaneous steroidogenesis. The most important of this pathway is the final step controlled by 11βHSD. This enzyme by its 2 types can control local levels of activated GCs in the skin. The balance between 11βHSD1 and 11βHSD2 enzymes, which convert active cortisol/inactive cortisone, is the key to maintain skin barrier function, hence a healthy skin. This case-control study was conducted at Ain Shams University hospitals. The study was conducted on thirty-one patients with dermatologist-confirmed diagnosis of chronic plaque psoriasis and thirty age matched healthy controls. Punch biopsies were taken from lesional and non lesional sites in patients and from controls for assessment of the enzyme by ELISA, besides histopathological and immunohistochemical assessment. Our study showed that 11βHSD1 activity was significantly decreased in psoriasis patients in comparison to healthy volunteers. Besides, 11βHSD1 enzyme level was found to be decreased in lesional than non lesional skin in psoriasis patients. Further histopathological assessment revealed that the lower enzyme levels are associated with greater epidermal acanthosis and higher degree of immunostaining by anti-FGF-2, which indicates the major role of 11βHSD1 in controlling psoriatic inflammation. This was supported by the positive correlation between the non lesional enzyme level and the longest period of remission that we detected in our patients. Surprisingly, there was no significant correlation between the lesional or non lesional enzyme levels in patients with PASI score. Yet, the calculation of PASI score depends on several external factors that should be considered. Interestingly, PSS was not correlated with 11βHSD1 level in psoriasis patients but was negatively correlated with the enzyme level only in healthy controls. This supports the role of stress in modifying enzyme activity, however, in psoriasis epidermal proliferation and the presence of inflammatory infiltrate definitely affects the enzyme level. Better understanding of the complex symphony between all the players of psoriasis pathogenesis will reveal the role of 11βHSD1 and factors affecting its level in psoriasis vulgaris.

#2989 Superposition of atypical hemolytic syndrome and tubulointerstitial nephritis, a case report

Abstract Background and Aims Hemolytic uremic syndrome is a heterogeneous group of diseases that have in common the development of thrombotic microangiopathy with diffuse endothelial damage, resulting in the classic triad composed of microangiopathic hemolytic anemia, thrombocytopenia and acute kidney injury, on the other hand, tubulointerstitial nephritis is characterized by the presence of inflammatory infiltrates and edema and of the interstitial compartment that causes deterioration of renal function. Method The following is the case of a patient with acute kidney injury of etiology composed of thrombotic microangiopathy and tubulointerstitial nephritis. Results 27-year-old female patient without significant history, began 3 weeks prior to admission with fever, chills, asthenia, nausea and vomiting of food content, approximately 5 episodes per day for 3 days, presenting intermittent episodes of jaundice of the mucous membranes and conjunctivae, with progressive increase of the symptoms to the point of preventing daily activities, she was admitted for study, detecting hemolytic anemia, thrombocytopenia and severe acute kidney injury, so she was referred to our center was decided for evaluation by Nephrology and Hematology. In our institution, urinary sediment with dysmorphic erythrocytes was documented, peripheral blood smear without the presence of schistocytes, positive Coombs 4 crosses, renal ultrasound with data compatible with acute kidney injury, and supportive management with hemodialysis was initiated. Due to suspicion of autoimmune glomerulopathy, an autoantibody panel was indicated with negative antinuclear antibodies, anti-double-stranded DNA, anti-Sm, anti-B2 glycoproteins, and lupus anticoagulant, with normal complement values, stool culture, and negative stool Shiga protein assay, angiotomography of the abdomen where the integrity of renal circulation is verified, so renal biopsy with immunohistochemical study is performed, finding data compatible with thrombotic microangiopathy and acute tubulointerstitial nephritis. The patient is maintained with supportive care, steroids and hemodialysis as required with progressive recovery of renal function until renal replacement therapy is weaned, she is discharged with a steroid regimen and clinical nephrology and hematology follow-up. Conclusion Acute kidney injury is a complex and multifactorial entity, which requires a comprehensive approach in order to identify the best therapeutic option and to offer a better outcome in the renal function of these patients. The case of our patient presented a sudden deterioration of renal injury composed of two entities with different pathophysiology, demonstrating that, although the approach to these two entities converge in certain etiologies, it is imperative that the clinician does not forget the wide variety of factors that contribute to the development of renal injury.

Case Report: Role of comprehensive rehabilitation In gaining recovery in a rare case of Polymyositis

Polymyositis (PM) is an inflammatory myopathy, a group of illnesses characterized by the presence of inflammatory infiltrates in striated muscle. Proximal muscular weakness is the most common clinical symptom of polymyositis. The exact cause of polymyositis is uncertain, however current research points to an autoimmune condition. We report a rare case of 27-year-old female that presented with the complaints of weakness in upper and lower limbs bilaterally for 2 years which was gradually progressive in nature along with difficulty in swallowing. So after physiotherapy assessment the patient was advised to undergo blood investigations which suggested an increased creatinine phosphokinase enzyme level along with reduced vitamin B12. To make a confirmatory diagnosis the patient underwent electromyography which suggested the presence of myopathy. Although polymyositis is more common in people aged 50 to 70 years this case suggests that it can have an earlier presentation. So along with medical management, a structured physiotherapy protocol was planned. Early diagnosis and management is key to recovery and better quality of life.

PSIX-9 Activation of Multiple Inflammatory Signaling Pathways May Contribute to the Pathogenesis of Idiopathic Cystitis in Felines

Abstract Feline idiopathic cystitis (FIC) is a common urinary disorder in domestic cats. Clinical signs include straining or failing to urinate, frequent urination, and discolored urine. Inflammation is a key characteristic in the pathogenesis of cystitis. We assessed circulating inflammatory markers in a retrospective study in cats housed in the Hill’s colony diagnosed with cystitis (n = 4; 15.2 to 18.6yr) and in control cats (n = 8; 5.9 to 16.5 yr). Blood samples were collected at end-of-life immediately before necropsy after they had lived their full lives. End-of-life pathology reports indicated inflammation and presence of inflammatory infiltrates in the bladder with some cats also showing renal inflammation. RNA was extracted from blood collected in PAXgene tubes, and gene expression investigated using NanoString nCounter platform. Analysis was performed using the nSolver software. There was a significant increase in IRAK4, a kinase that activates the NFκB pro-inflammatory signaling pathway, in FIC when compared with controls (1.56, P < 0.05). Small molecule inhibitors of IRAK4 are being developed as targets of inflammation. Signaling molecules in the NFκB pathway were also significantly increased including Rel A (p65 subunit; 1.3 fold), NFκB1 (p100 subunit; 1.3 fold), IKBKB (1.29 fold), and TANK (1.46 fold) in FIC cats (all P < 0.05). Another signaling pathway contributing to pro-inflammatory signaling is the JAK/STAT pathway. There was a significant increase in JAK2 (1.65) and STAT3 (1.48). Blockade of JAK2/STAT3 pathway can attenuate cytokine synthesis and reduce inflammation. STAT6 was also significantly increased in FIC cats (1.35). Activation of STAT6, by IL-4 and IL-13, can lead to systemic inflammation. Activation of the complement immune pathway was also seen in FIC. While the complement system is important for an innate immune response, its sustained activation leads to chronic pathological inflammation. There was a significant increase in complement C1s, an initiator of classical complement activation pathway, in FIC cats (1.43; P < 0.05). A late component, complement C7, was also upregulated in FIC (2.01 fold; P < 0.05). C7 binds to the C5b-6 complex which eventually becomes part of the terminal complement complex. Increased abundance of C7 and eventual terminal complement complex are associated with increased inflammation and may be important in the progression of FIC. There was also an increase in PECAM1 in FIC cats (1.87; P < 0.05). Signaling through PECAM1 leads to the activation of neutrophils and monocytes. DUSP1 was also elevated in FIC (2.24; P < 0.05). Increased abundance of DUSP1 is hypothesized to be a predictor of chronic subclinical inflammation in humans with cardiovascular disease. Our results identify activation of pro-inflammatory signaling pathways that may be potential targets for alleviating inflammation in cystitis. Pathogenesis of FIC is likely multi-factorial, and nutritional intervention to attenuate the activation of inflammatory pathways may be an important strategy in the management of cystitis.

Human Periapical Odontogenic Granulomas: Aspects of Microvessel Density (MVD), Heterogeneity of Blood Vessels and Mast Cells Density (MCD).

Periapical odontogenic granulomas are among the most encountered pathology that involve the alveolar bone, with severe consequences such as bone resorption, the presence of inflammatory infiltrate and the formation of abnormal vascularization. The present study aimed to quantify the existence of the microvessel density (MVD), mast cell density (MCD) and heterogeneity of the encountered blood vessels. A total of 37 patients diagnosed with odontogenic periapical granulomas were included, and the gender distribution, age and localization of the pathological lesions was assessed. After the surgical removal of the periapical odontogenic granuloma, the collected tissue was fixed in 10% buffered formalin. Primary processing, morphological analysis and immunohistochemical staining was performed in order to characterize the altered tissue. The results outlined the presence of a high number of mast cells, especially in the area of the inflamed tissue; the high heterogeneity of the blood vessels; and increased MVD with positive CD34. The conclusions of the study focus on the key role of the mast cells and their implication in the initiation and development of the angiogenesis process, triggering the inflammatory response of the host. Nevertheless, periapical odontogenic granulomas develop as an inflammatory response to the interaction between the host's immune system and microbial invasion.

Role of transduodenal endoscopic ultrasound-guided fine-needle aspiration/biopsy (EUS-FNA/FNB) for diagnosis of retroperitoneal fibrosis (Ormond's disease).

Retroperitoneal fibrosis (RPF), often referred to as Ormond's disease when it is of idiopathic origin, is a rare disease characterized by the presence of inflammatory infiltrates and periaortic masses in the retroperitoneum. For a definite diagnosis, a biopsy and subsequent pathological examination is required. Currently accepted methods for retroperitoneal biopsy include open, laparoscopic, or CT-guided approaches. However, transduodenal endoscopic ultrasound-guided fine-needle aspiration/biopsy (EUS-FNA/FNB) for diagnosis of RPF has attracted only little attention in the literature. We report two male patient cases who presented with leukocytosis, elevated C-reactive protein, and a suspicious retroperitoneal mass of unknown origin on computed tomography. One patient also reported left lower quadrant pain, whereas the other patient suffered from back pain and weight loss. In both patients, idiopathic RPF was successfully diagnosed by using transduodenal EUS-FNA/FNB with 22- and 20-gauge aspiration needles. Histopathology revealed dense lymphocytic infiltrates and fibrosis. The procedures lasted approximately 25 and 20 minutes, respectively, and in both patients no serious adverse events occurred. Treatment included steroid therapy and administration of Azathioprine. We demonstrate that using EUS-FNA/FNB to diagnose RPF is a feasible, fast, and safe method, which should always be considered as a first-line diagnostic modality. Hence, this case report emphasizes that gastrointestinal endoscopists are likely to play an important role in the setting of suspected RPF.

Pathological changes in the nervous structures of the aortic wall tissue adjacent to an unstable atherosclerotic plaque

To study of nerve structures in the aortic wall in atherosclerosis using a complex of immunohistochemical markers. The objects of the study were excised fragments of the wall of the thoracic and abdominal aorta along with visually determined unstable atherosclerotic plaques. To study nerve structures on paraffin sections, immunohistochemical reactions were performed for the PGP 9.5 protein, tyrosine hydroxylase, and synaptophysin. It has been established that pronounced pathological changes are observed in the nervous structures of the aortic wall near unstable atherosclerotic plaques. Reactive, dystrophic, and severe degenerative changes in neurocytes, nerve fibers, and glial cells are described in the elements of the nervous apparatus of the adventitia (microganglia, nerve trunks, and nerve plexuses). It was found that only sympathetic neurons and their postganglionic fibers remain in the intramural ganglia, while the structures of the parasympathetic nervous apparatus undergo degeneration. Destruction of perivascular nerve plexuses and vasa vasorum in the adventitia, as well as degeneration of varicose axons of the main terminal synaptic plexus at the border of adventitia and superficial smooth muscle layer of the media were demonstrated. It is assumed that the presence of inflammatory infiltrates in the adventitia and intima, denervation and death of vasa vasorum can serve as factors determining the development of the atherosclerotic process.

216 ARRHYTHMOGENIC CARDIOMYOPATHY: IS THERE EVIDENCE OF AN INFLAMMATORY MYOCARDIAL DISEASE?

Abstract Background and aim Arrhythmogenic cardiomyopathy (ACM) is a heredo-familiar primary heart muscle disease, mainly due to mutations of desmosomal genes and characterized by progressive myocardial atrophy with fibro-fatty replacement, causing electrical instability at risk of sudden cardiac death (SCD). Since the first anatomopathological descriptions, the hypothesis that myocardial inflammation could have an important role in the onset and evolution of ACM has been advanced. With this work we aim to describe the prevalence of inflammatory cells in ACM and their characterization through histological and immunohistochemical (IHC) analysis in endomyocardial biopsy (EMB) and heart specimens coming from either SCD or heart transplantation (HTx). Material and methods We analysed EMB and whole heart specimens (from juvenile SCD registry – below 40 years of age at the time of death – and HTx) with a diagnosis of ACM, focusing on the presence of inflammatory infiltrates. For the whole hearts, in a subgroup of cases the inflammatory infiltrate was evaluated on 9 myocardial sections by applying a semi-quantitative score according to the number of sections involved (0 = none; 1 = 1 section involved; 2 = 2-5 sections; 3 = >5 sections) and an IHC panel for myocarditis on 1 right ventricular and 1 left ventricular section (CD45, CD3, CD8, CD4, CD68, CD20, CD138 and vimentin- as a control for fibroblast). Results Among the 63 analysed EMB (41 M, mean age 45 years, range 18-71 years), 26 cases (41%) showed foci of inflammation with CD3+ > 7/mm2. In all cases viral genome (DNA and RNA virus) was absent. A total of 106 heart specimens with a diagnosis of ACM were enrolled, including 73 juvenile SCD (49 M, mean age 26 years) and 33 HTx (18 M, mean age 45 years). Inflammation was reported in 95% of SCD an d 67% of HTx. The inflammatory infiltrate was focal or multifocal in all, except one case of juvenile SCD with diffuse myocarditis in the postero-lateral wall of the left ventricle. In the 24 reviewed cases, the inflammation was focal with a mean inflammatory score of 1.75 ±0.9 in the HTx group and 1.5±0.9 in the SD group (p = NS). By IHC we showed that: a) there is no significant difference in the inflammatory score between SD and HTx; b) the inflammation is mostly near-scarring (13 cases); c) inflammatory cells consist prevalently of cytotoxic T lymphocytes CD3+CD8+ (with CD4+<CD8+ both in SD and HTx, p < 0.05). Conclusions Our pathological data confirm that ACM is an inflammatory cardiomyopathy with a higher prevalence of inflammation in SCD cases. The inflammatory infiltrate is mainly composed of cytotoxic T lymphocytes with a variable presence of macrophages. These data, combined with the recent experimental studies demonstrating the role of immune mediators in the evolution of the disease, support the potential application of immunosuppressive therapy in ACM.

Synovial fluid and radiographic evaluation of joints from dogs with visceral leishmaniasis

BackgroundPolyarthritis has been associated with canine visceral leishmaniasis (CanVL), and co-infection with Ehrlichia canis is common and may alter clinical manifestations.MethodsA total of 89 dogs presenting CanVL were subdivided into two groups: (1) G1, consisting of 46 dogs seronegative to Ehrlichia spp., and (ii) G2, consisting of 43 dogs seropositive to Ehrlichia spp. Eight joints (carpal, tarsal, stifles and elbows) from each dog were evaluated by radiography and synovial fluid (SF) cytologic analysis.ResultsOverall, 74 of the 89 (83.1%) dogs presented joint abnormalities suggestive of osteoarthritis by radiography (G1: 40/46 [86.9%]; G2: 34/43 [79.0%]), with no statistically significant between-group difference. All dogs with abnormal joint X-ray images presented radiographic lesions bilaterally, independent of the characteristics of the lesion. Soft tissue swelling around the joint and joint space narrowing were more commonly observed in G1 than in G2 dogs. There was no significant between-group difference in terms of other radiographic abnormalities suggestive of osteoarthritis (evident trabecular pattern, subchondral bone sclerosis, osteolysis, osteolytic–proliferative lesions or bone proliferation). SF from 174/315 (55.2%) and 152/307 (49.5%) joints from G1 and G2 dogs, respectively, presented an inflammatory infiltrate, but there was no significant association between the presence of inflammatory infiltrate and group. There was also no statistical difference between groups in either of the evaluated joints in terms of the percentage of neutrophils or mononuclear cells. Leishmania spp. amastigotes were found in 69/315 (21.9%) joints from G1 dogs and in 100/307 (32.5%) joints from G2 dogs (Fisherʼs exact test, P = 0.002, odds ratio = 0.5, 95% confidence interval = 0.4–0.8). The neutrophilic infiltrate was significantly higher in joints with amastigote forms in both G1 (Mann–Whitney U-test, U(18) = 817, Z = -3.76, P = 0.0001) and G2 dogs (Mann–Whitney U-test, U(18) = 6543, Z = − 5.06, P < 0.0001).ConclusionsA high prevalence of arthritis in dogs with CanVL was found, and all dogs presented involvement in multiple joints. Although no difference was observed between groups in terms of the number of dogs with polyarthritis and the presence of an inflammatory infiltrate in SF, Leishmania spp. amastigotes were found more frequently in joints from G2 dogs. Further studies evaluating SF in dogs co-infected with L. infantum and E. canis should be performed to evaluate this finding.Graphical

Hepatic and blood alterations in Lithobates catesbeianus tadpoles exposed to sulfamethoxazole and oxytetracycline

In this study the effects of environmentally realistic concentrations of the antibiotics sulfamethoxazole (SMX) and oxytetracyclyne (OTC) on Lithobates catesbeianus tadpoles were evaluated, through the analyzes of the frequencies of micronucleus and nuclear abnormalities in erythrocytes, alterations in leucocytes, liver histopathology, and changes in hepatic esterase activities and oxidative stress biomarkers. The animals were exposed for 16 days at concentrations of 0 (control), 20, 90 and 460 ng L−1. No significant difference was found in the frequencies of micronucleus and nuclear abnormalities. The two highest concentrations of SMX and all concentrations of OTC caused a significant increase in the number of lymphocytes. A significant decrease in the number of neutrophils compared to the control group was observed for all concentrations tested of both antibiotics. Also, decrease in the activity of glutathione S-transferase and high histopathological severity scores, indicating liver damage, were found in tadpoles exposed to the two highest concentrations of SMX and all concentrations of OTC. The main changes in the liver histopathology were the presence of inflammatory infiltrate, melanomacrophages, vascular congestion, blood cells and eosinophils. Esterase activities were unchanged. Indeed, the two highest concentrations of OTC caused a reduction in the activities of superoxide dismutase and glucose 6-phosphate dehydrogenase, while the highest concentration inhibited the activity of glutathione peroxidase and increased protein carbonyl levels. These results evidences that environmentally realistic concentrations of SMX and OTC in aquatic environments are capable to significantly disrupt tadpoles’ physiology, possibly affecting negatively their survival rate in natural environments.

Abstract 3319: Purified poloxamer 188 mitigates radiation-induced lung toxicity

Abstract The dose-limiting factor in radiation therapy (RT) is damage to healthy tissues, which presents as radiation-induced lung toxicity (RILT). Acute RILT, pneumonitis, is an early inflammatory-driven toxicity that develops within weeks to months following RT. Purified poloxamer 188 (PP188) is a triblock copolymer that associates with and protects cell membranes from oxidative insult, minimizing secondary inflammatory damage. This study aimed to investigate the effects of PP188 to mitigate the onset of acute radiation pneumonitis in a rat model of RILT, and to test the effect of PP188 on the efficacy of RT on tumor growth of a syngeneic CT26.WT murine colon carcinoma in Balb/c mice. To evaluate the efficacy of PP188 to mitigate RILT in healthy tissues, Sprague Dawley rats were given 200mg/kg PP188 or vehicle via IV 1hr prior to a single dose of partial volume (6mm) 20Gy X-irradiation to the right lung, and then were followed for 6 weeks. To validate PP188 treatment does not interfere with RT efficacy to reduce tumor volume, Balb/c mice were implanted with CT26.WT murine colon adenocarcinoma in the high axilla space, given 5Gy SARRP irradiation, and tumor volume was monitored. Rats were evaluated for histopathological changes indicative of acute RILT, including leakage of proteins into alveolar space, thickening of alveolar septa, and alteration of bronchial epithelium and capillaries; for presence of inflammatory infiltrate (Hanker-Yates Peroxidase Leukocyte kit); and for immunohistochemical expression of alpha-smooth muscle actin (SMA). Without PP188, significant inflammatory infiltrate was observed in all lungs bilaterally (p&amp;lt;0.001) contributing to significant bronchiole epithelium thickening (p&amp;lt;0.001) and alveolar septa thickening (p&amp;lt;0.001). Within the thickened alveoli there was significant SMA expression (p&amp;lt;0.001) indicative of cells undergoing a myofibroblast transition that was not observed in the PP188+RT animals or controls. Morphological assessment demonstrated that PP188 significantly attenuated pathological changes in the functional subunits of the lung, alveoli, as well as the vasculature including bronchiole epithelium and small capillaries, with no significant differences from control animals. Finally, it was shown that PP188 does not abrogate radiation-induced tumor volume reduction. The unique mechanism of PP188 provides radioprotection to healthy tissue without interfering with tumor control, and has the potential to mitigate radiation-induced toxicities in the majority of cancers treated with RT. Citation Format: Sarah G. Dennis-Little, Wilfred Ngwa, Marikki Laiho, Graham Warren, Michael J. Yost. Purified poloxamer 188 mitigates radiation-induced lung toxicity [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 3319.

Clinical and pathological prognostic factors in Merkel cell carcinoma.

e21574 Background: From 25 to 50% of radically resected Merkel Cell Carcinomas (MCCs) develop local or distant recurrence with a 5 year Overall Survival (OS) of 35% and 13%, respectively. Due to the rarity of the disease, there are limited data about clinical/histological risk factors related to patients (pts) prognosis. Methods: We retrospectively evaluated a series of 57 MCCs pts treated with surgical curative intent at two Italian Centres from 2000 to 2020. For 43 of them, archival histological material was retrieved for further analysis. The following clinical and histological data were analyzed and related to time to relapse (TTR), OS and cancer specific survival (CSS): gender, age, Charlson Comorbidity Index value, immunodepression, surgical margins, re-excision and/or radiotherapy (RT) in case of positive margins, stage, perineural/lymphovascular infiltration, presence of inflammatory infiltrate (tumoral infiltrating lymphocyte (TILs) (&gt; 1 positive cell per high-power field) and its composition CD3+, CD4+ T and CD8+ T cells), positivity for MCC-polyomavirus (MCCpV), Ki67 index and PDL1 expression CPS. Results: Pts were mainly female (52%), median age was 75 years (49-99), 70% were immunocompetent and 51% presented T1 stage. All pts received surgery as main treatment approach; in case of positive margins, pts received re-excision or RT, when feasible. During a median follow-up of 37 months (1- 188), 40% had a locoregional or distant recurrence. Median TTR of relapsed pts was 3 months (1-88); median OS and CSS in the whole population was 117 months (1-188) and not reached, respectively. At univariate analysis, only male gender was associated with a higher risk of relapse (HR 1.6; 1.07-2.55; p 0.02). Older age (HR 1.09; 1.04-1.14, p &lt; 0.001), T stage ≥2 (HR 4.79, 1.99-11.53, p &lt; 0.001) and positive margins after re-excision (HR 17.49, 1.09-279.74, p 0.043) were associated with shorter OS and negatively related to CSS. Re-excision and/or RT in cases of positive margins at diagnosis improved OS (HR 0.154, 95% CI 0.028-0.851, p = 0.032) and CSS (HR 0.15; 0.02-0.85, p 0.03). MCCpV positivity (HR 0.23, 0.08-0.62, p 0.004), TILs (HR 0.30, 0.11-0.80, p 0.017), a moderate or high content of CD3+ T cells (HR 0.33, 0.119-0.968, p 0.04), CD8+ T cells (HR 0.35, 0.12-0.99, p 0.04), and PDL1 CPS &gt; 1 (HR 0.26, 0.08-0.82, p 0.02) resulted in longer OS. The impact of MCCpV positivity (HR 0.27; 0.08-0.88 p 0.03) and high TILs (HR 0.31, 0.09-0.99, p 0.04) was also confirmed for CSS. Conclusions: In primarily surgically treated MCCs, female gender, younger age, low T stage, positivity for MCCpV and tumoral immune cells infiltration were associated with better prognosis, as well as an aggressive management comprising re-excision and/or RT in case of positive margins.

Inflammatory myopathies: an update for neurologists.

Idiopathic inflammatory myopathies (IIM) are a heterogenous group of treatable myopathies. Patients present mainly to the rheumatologist and neurologists, complaining of acute or subacute onset of proximal weakness. Extramuscular manifestations may occur, including involvement of the lungs, skin, and joints. Classically, the diagnosis used to be made based on the creatine kinase level increase, abnormalities in electroneuromyography and presence of inflammatory infiltrates in the muscle biopsy. Recently, the importance of autoantibodies has increased, and now they may be identified in more than half of IIM patients. The continuous clinicoseropathological improvement in IIM knowledge has changed the way we see these patients and how we classify them. In the past, only polymyositis, dermatomyositis and inclusion body myopathy were described. Currently, immune-mediated necrotizing myopathy, overlap myositis and antisynthetase syndrome have been considered the most common forms of IIM in clinical practice, increasing the spectrum of classification. Patients previously considered to have polymyositis, in fact have these other forms of seropositive IIM. In this article, we reviewed the new concepts of classification, a practical way to make the diagnosis and how to plan the treatment of patients suffering from IIM.

Hibiscus (Hibiscus sabdariffa L.) supplementation increases butyrate synthesis and reduces inflammatory cells, attenuating the formation of aberrant crypt foci in BALB/c mice induced to pre-neoplastic lesions.

The development of colorectal cancer involves some morphological changes, and in the initial stage, pre-neoplastic lesions called aberrant crypt foci (ACF) appear. Thus, an intervention with sources of bioactive compounds such as Hibiscus sabdariffa L., rich in phenolic compounds and anthocyanins, could attenuate the risk of developing these lesions due to its antioxidant, anti-inflammatory and anti-proliferative properties. Therefore, the aim of this study was to evaluate the effects of 5% and 10% supplementation of dehydrated H. sabdariffa calyces (DHSC) during the development of 1,2-dimethylhydrazine-induced preneoplastic lesions in male BALB/c mice. The characterization of DHSC was carried out. The in vivo experiment lasted 12 weeks, and the animals were randomly divided into 3 experimental groups: the control group (CON) and the supplemented groups with 5% DHSC and 10% DHSC. The activities of liver enzymes catalase and superoxide dismutase were determined. In addition, ACF, short chain fatty acids (SCFA), presence of inflammatory infiltrates, goblet cells and leukocytes in the colonic mucosa were quantified. There was a significant reduction in ACF and the presence of inflammatory infiltrates in the colon of animals in groups 5DHSC and 10DHSC. In addition, the 10DHSC group showed an increase in the activity of the catalse enzyme, in the production of butyrate and in the presence of NK cells in the colon, in addition to more hypertrophied goblet cells. Based on these findings, it is suggested that DHSC supplementation may be recommended to attenuate cellular responses in the early stage of preneoplastic lesions.

617 Indocyanine Green: Harnessing Novel Methods to Identify Burn Wound Healing Potential

IntroductionObjective determination of burn wound healing potential remains elusive and significantly impacts decision making for surgery, the extent of tissue excised intraoperatively and the use of donor site-sparing alternative tissue therapies. Indocyanine green angiography (ICGA) has promise as an adjunct to evaluate healing potential, but feasibility has limited adoption in clinical practice. Delayed fluorescence imaging of indocyanine green (ICG), in a method called second-window ICG (SWIG), is a new technique used intraoperatively to guide tumor resection via increased peritumoral endothelial permeability. The objective of this study is to examine ICGA and SWIG fluorescence in burns requiring excision and grafting, and to correlate SWIG fluorescence to microscopic localization of inflamed and necrotic tissue.MethodsDeep partial thickness, indeterminate depth or full thickness burns were identified in adult patients scheduled for excision and grafting. 24 hours prior to surgery, baseline bright light and fluorescence images were obtained before the administration of up to 5 mg/kg ICG intravenously. ICGA was performed within 5 minutes of infusion initiation. On the day of surgery, bright light and SWIG fluorescence images were obtained before and after burn excision. The excised tissue was imaged ex-vivo to determine the presence of fluorescence in the tissue compared to that remaining within the wound bed. Excised tissue was processed for histologic analysis of cellular architecture, viability, inflammation and necrosis. Macroscopic ICGA and SWIG fluorescence images were compared to the associated microscopic tissue sections to determine the presence of inflammatory infiltrate, localization of non-viable tissue, and co-localization of ICG fluorescence.ResultsICGA imaging performed preoperatively demonstrated variable fluorescence throughout the burns without a clear cutoff value to delineate deep partial versus full thickness burns. SWIG imaging revealed a speckled fluorescence pattern prior to burn excision that became diffuse after excision suggesting a potential utility of SWIG to intraoperatively identify excision completion. ICGA and SWIG fluorescence demonstrated an inverse relationship, and SWIG fluorescence was associated with non-viable tissue.ConclusionsICGA imaging alone was unreliable to delineate the need for surgical intervention. SWIG imaging of burn injuries may represent a valuable tool to guide the extent of excision intraoperatively and reduce unnecessary excision of viable tissue. Further studies are needed to understand SWIG fluorescence at the inflammation-necrosis border and how ICGA fluorescence along with SWIG can synergistically improve detection of healing potential in burn patients.

Topographical distribution and histological characterization of stromal fibrosis in invasive breast carcinoma

Carcinoma breast is associated with various changes in organ and at the tissue level. The desmoplastic response in a tumor due to tumor hypoxia may result in dense or sparse, homogenous or focally hetrogenous fibrosis of the breast tissue. Peritumoral and stromal fibrosis has been assessed as an marker of intratumoral hypoxia and is being studied as histological surrogate prognostic parameter in breast cancer. We studied the topographical distribution and histological characterization of fibrosis and correlated it with various other histopathological parameters. 50 patients of carcinoma breast presenting to our institute were included in the study. We obtained a statistically significant correlation of presence of fibrosis with presence of lymphovascular invasion (p value =0.043), while we could not obtain any significant difference with age, menopause, tumor size, tumor grade, pathological stage, lymph node involvement, presence of necrosis, adipocytic invasion, duration of tumor, laterality, presence of inflammatory infiltrate and with ER, PR, HER2Neu status. This is a limited study in a small number of patients focussing on various stromal changes in breast cancer. This study has further characterized the heterogeneous nature of fibrosis in breast carcinoma using special staining which may facilitate the physician in making patient diagnosis, prognostication and outcome.

The Neuropathology of Autoimmune Ataxias.

Autoimmune-mediated ataxia has been associated with paraneoplastic disease, gluten enteropathy, Hashimoto thyroiditis as well as autoimmune disorders without a known associated disease. There have been relatively few reports describing the neuropathology of these conditions. This review is an attempt to consolidate those reports and determine the ways in which autoimmune ataxias can be neuropathologically differentiated from hereditary or other sporadic ataxias. In most instances, particularly in paraneoplastic forms, the presence of inflammatory infiltrates is a strong indicator of autoimmune disease, but it was not a consistent finding in all reported cases. Therefore, clinical and laboratory findings are important for assessing an autoimmune mechanism. Such factors as rapid rate of clinical progression, presence of known autoantibodies or the presence of a malignant neoplasm or other autoimmune disease processes need to be considered, particularly in cases where inflammatory changes are minimal or absent and the pathology is largely confined to the cerebellum and its connections, where the disease can mimic hereditary or other sporadic ataxias.

An Integrative Approach to Characterize the Early Phases of Dimethylhydrazine-Induced Colorectal Carcinogenesis in the Rat.

This study aimed to characterize an animal model of colorectal cancer (CRC) in the early stages of disease development. Twenty-nine male Wistar rats were divided into two control groups (CTRL1 and CTRL2), receiving EDTA–saline injections and two induced groups (CRC1 and CRC2), receiving 1,2-dimethylhydrazine (DMH) injections for seven consecutive weeks. CRC1 and CTRL1 were euthanized at the 11th week, while CRC2 and CTRL2 were euthanized at the 17th week. DMH treatment decreased microhematocrit values and IL-6, ghrelin, and myostatin serum levels. Histopathological analysis of intestinal sections showed that DMH-treated rats were characterized by moderate to severe epithelial dysplasia. An adenoma was observed in one animal (CRC2 group), and the presence of inflammatory infiltrate at the intestinal level was primarily observed in DMH-treated animals. DMH also induced Ki-67 immunoexpression. The gut microbiota analysis showed a higher abundance of Firmicutes, Clostridia, Clostridiales, Peptostreptococcaceae, Blautia, Romboutsia, and Clostridium sensu stricto in CRC than CTRL rats, whereas Prevotellaceae, Prevotella, Akkermansia, and Lactobacillus levels were more prevalent in CTRL animals. Our results suggest that this model could be helpful to investigate chemoprevention in the early stages of CRC.

Valuation of angiogenesis in bovine xenografts implanted in intracorporal sites of rabbits as models of in-vivo bioreactors

ABSTRACT The aim of this study was to evaluate neovascularization of bovine xenografts implanted in intracorporeal sites of rabbits (bioreactors). 30 rabbits were used, divided into 6 groups, according to the evaluation time (7, 15, 30, 45, and 60 days); each animal received xenogenic implants in 3 different intracorporeal sites (A1 - omentum bag; A2 - intermuscular space of quadriceps femoris; A3 - subperiosteal of ilium bone). Histological assessments graded the presence of angiogenesis, the number of inflammatory cells, newly formed bone tissue, and the presence of giant cells. Histological analyses showed intense angiogenesis in all implanted xenografts. Presence of inflammatory infiltrate and giant cells at the A1 implant site and presence of bone neoformation at the A3 implant site were noted. Degeneration of implants and formation of a fibrous capsule were noted. When comparing the interaction of the site with the days of evaluation, statistical analysis showed a significant difference (p≤0.05) in any time of neovascularization analysis. The vascular endothelial growth factor (VEGF) and inflammatory cells of the omentum in its structure, may have contributed to the greater presence of neovessels and inflammatory cells, a fact that may indicate functionality as a possible bone substitute.

Effect of dexamethasone on experimental enteritis produced by Giardia lamblia in a Meriones unguiculatus model

Our aim was to evaluate the impact of immunosuppression on the development of giardiasis. Thirty-six gerbils (4–6 weeks old) were distributed in four groups containing nine animals each: Control (CT); Control-Infected by Giardia lamblia (CTIn), Immunosuppressed (IS), and Immunosuppressed-Infected by G. lamblia (ISIn). Animals in the IS and ISIn groups received intramuscular dexamethasone solution for 25 days. On the 11th day, the animals in the CTIn and ISIn groups were inoculated with G. lamblia. After 14 days of infection, the 25th day of the experiment, all groups were euthanized. Four hours after euthanasia, the intestinal permeability was evaluated and sections of the duodenum and spleen were harvested for morphometric and histopathological analyses. Immunosuppressed groups showed a significant increase in intestinal permeability compared to control and infected groups. Considering that the infection can become chronic in immunosuppressed groups, we should be alert to the possibilities of chronic inflammatory changes, both locally and systemically, due to the loss of the intestinal barrier. Lesions were observed in the duodenal mucosa of the gerbils of the CTIn group, with reduced villi size, crypt hyperplasia, edema, and the presence of inflammatory infiltrate in the lamina propria. In the ISIn group, we observed no inflammation, long and intact villi, and a significant increase in the area of intestinal mucins, despite the large number of trophozoites identified. Our results suggest that exacerbation of the immune response has a direct relationship with the appearance of lesions during enteritis produced by G. lamblia in the assessed model.