Perivascular Inflammatory Infiltrates Research Articles

PS-B01-7: VASCULAR SMOOTH MUSCLE CELL NOX5 REGULATES FIBROBLAST PHENOTYPIC SWITCH AND CARDIOVASCULAR FIBROSIS

Objective: We previously observed that mice expressing human Nox5 (hNox5) in VSMC exhibit vascular dysfunction and cardio-renal fibrosis, where mechanisms are unknown. Here we postulated that VSMC-Nox5 promotes fibroblast phenotypic changes leading to myofibroblast differentiation and pro-fibrotic responses. Design and Methods: Fibroblasts were cultured from wildtype (WT) and hNOX5 mice (Nox5 + SM22 +). Mice (20 weeks old) were infused with Ang II (600 ng/Kg/day) for 28 days and renal fibrosis/inflammation studied. Markers of myofibroblasts, pro-fibrotic and inflammatory phenotypes were assessed by qPCR and immunoblotting. Results: Fibroblasts from Nox5 + SM22 + mice exhibited increased mRNA of pro-fibrotic markers, such as Col1A1 (2-ddC:1.74 ± 0.16 vs. WT 0.67 ± 0.11), Col3A1 (2-ddC:1.74 ± 0.18 vs. WT 0.96 ± 0.24) and TIMP3 (2-ddC:2.65 ± 0.25 vs. WT 0.38 ± 0.07), p < 0.05. Myofibroblast phenotype (SMC) markers mRNA, aSMA (2-ddC:1.54 ± 0.05 vs. WT 0.78 ± 0.17) and Myocd (2-ddC:1.36 ± 0.17 vs. WT 0.39 ± 0.22) were also increased, p < 0.05. mRNA expression of CD36 (2-ddC:1.37 ± 0.07 vs. WT 86 ± 0.24), TNFa (2-ddC:1.32 ± 0.2 vs. WT 0.71 ± 0.17) and TNFR1 (2-ddC:1.26 ± 0.04 vs. WT 1.02 ± 0.10) were increased, while CD68 expression was decreased (2-ddC:0.82 ± 0.11 vs. WT 1.36 ± 0.18) in fibroblasts from Nox5+SM22+ mice (p < 0.05). In Nox5 mice fibroblasts, ROS production and TGFb protein expression (AU:1.8 ± 0.05 vs. WT 1.4 ± 0.06), as well as TGFbR2 gene expression (2-ddC:2.04 ± 0.17 vs. WT 0.57 ± 0.12), were increased (p < 0.05). Kidneys from Ang II-infused Nox5+SM22+ mice exhibited significant perivascular fibrosis and inflammatory cell infiltration compared to WT. Kidney expression of vimentin (AU:1.01 ± 0.05 vs WT 0.85 ± 0.03) and aSMA (AU:0.44 ± 0.03 vs WT 0.33 ± 0.01), markers of myofibroblast differentiation, were increased in Nox5 mice (p < 0.05). Gene expression of DNMT3a and TET2, DNA methylation regulatory enzymes, were also increased in fibroblasts from Nox5+SM22+ mice, p < 0.05. Conclusion: VSMC-Nox5 regulates fibroblast phenotypic switch leading to fibrosis; a process that may involve increased production of ROS and TGFb, and activation of DNA methylation pathways.

Nevus Lipomatosus Cutaneous Superficialis Masquerading as Giant LipomaA Rare Case Report

Nevus Lipomatosus Cutaneous Superficialis (NLCS) is an idiopathic, rare skin hamartoma. It occurs due to ectopic deposition of adipocytes in dermis. There are two main clinical forms of NLCS- classic and solitary. Both have different clinical presentations but histologically they show similar morphology characterised by bundles of mature adipocytes in the superficial and deep dermis separated by bundles of collagen. The hallmark of this lesion is absence of connection between subcutaneous fat and adipocyte clusters in dermis.Though NLCS is benign developmental disorder and remains static for a long time, it can be cosmetically unpleasant if grows to a large size. Herein, authors report a case of a 21-year-old male with a solitary painless pedunculated lump in right thigh for 10 years that clinically presented as giant lipoma due to its soft consistency, cut surface of the mass was yellow and fibrofatty. There was no connection of these adipocytes seen with the subcutaneous fat. Focal collections of adipocytes were also seen around the blood vessels. Sparse perivascular mononuclear inflammatory infiltrate also noted. Final diagnosis of NLCS was made.

Nephrotoxicity induced by different diameters of sphere gold nanoparticles with special emphasis on the nephroprotective role of quercetin.

Although, gold nanoparticles (GNPs) are attracting more and more attention due to their ease of synthesis, modification, and great potential value in biomedical applications, exhibited harmful effects on human health and other living species. Quercetin (Qur) clarifies diverse pharmacological effects, especially anti-inflammatory, antiapoptotic, and antioxidant ones. This study aimed to evaluate the probable nephrotoxicity induced by different diameters of sphere GNPs, as well as the nephroprotective role of Qur. A total of 54 healthy mature male albino rats were grouped and treated with or without sphere GNPs; 10, 20, and 50 nm and Qur (200 mg/kg b.wt.). The effects of GNPs and Qur were estimated through the collection of blood and kidney samples from euthanized rats and performed biochemical, histopathological, and immunohistochemical investigations. In comparison between different diameters of GNPs, the 10 nm GNPs revealed more significant elevations in all renal function parameters: creatinine, urea, blood urea nitrogen, and uric acid followed by 20 nm then 50 nm. Pre-cotreatment with Qur decreased all renal functional values. Histopathologically, 10 nm revealed the most potent renal pathological changes represented in the renal cortex with cloudy swelling of renal tubules, hypercellularity of some glomeruli, severe congestion of renal blood vessels, focal inter tubular edema, and vascular endotheliosis (degeneration of endothelium). In addition, the renal medulla revealed perivascular inflammatory cellular infiltration, perivascular fibrosis, intra tubular glycogen deposition, and casts deposition of mainly cellular casts. On the other hand, the Qur treatment ameliorated most of these pathological changes. The size of GNPs is pivotal in their pathological effect on renal tissues where the small-sized GNPs; 10 nm have more potent cytotoxic, inflammatory, and apoptotic effects rather than the larger ones. Otherwise, Qur clarified a significant mitigating role against the nephrotoxicity of the GNPs.

Lichen planus with IgA vasculitis: a rare association

<p>Lichen planus is an inflammatory skin disorder which has been shown to be associated with thyroid disorders, hepatitis B, hepatitis C. IgA vasculitis is a form of cutaneous small vessel vasculitis (CSVV) characterized by palpable purpura, abdominal pain and joint pain usually seen in children. We report a case of 45-year-old female patient came with violaceous itchy lesions which were distributed over trunk and extremities of 1 month duration. Complete blood count (CBC) and serum biochemistry tests were normal. Histopathology of violaceous lesions showed changes of lichen planus. Purpuric lesions showed mild perivascular and transmural inflammatory infiltrate, composed mainly of neutrophils and few lymphocytes in the superficial dermis. Direct immunofluorescence was positive for IgA (granular pattern along with vessel walls of small vessels). Based on clinical, histopathological findings, a diagnosis of lichen planus with IgA vasculitis was made. The patient was treated with 30 mg prednisolone, gradually tapered over 6 weeks and anti-histamines. Satisfactory response was observed at follow-up at 6 weeks. Our case highlights the rare association of lichen planus with IgA vasculitis. Thorough evaluation of the patient helped in reaching correct diagnosis. </p>

Fingolimod attenuates ovalbumin-induced airway inflammation via inhibiting MAPK/ERK signaling in mice.

The current study was designed to investigate the potential anti-inflammatory and antioxidant effects of fingolimod against Ovalbumin (Ova)-induced allergic airway inflammation compared to dexamethasone. Fingolimod was given (0.5 mg/kg/day, p.o.) for sensitized mice 1 hbefore Ova challenge from Days 19 to 24. Fingolimod significantly inhibited Ova-induced elevation of inflammatory cells and eosinophils numbers in bronchoalveolar lavage fluid(BALF) and reduced concentrations of immunoglobulin E in serum and of sphingosine-1-phosphate, interleukin(IL)-4, and IL-13 in BALF. Fingolimod inhibited microvascular leakage and edema as reflected by the decreased lung/body weight index. These findings were supported by histopathological examination results showing that fingolimod substantially decreased perivascular edema and inflammatory cell infiltration. Fingolimod also attenuated Ova-induced oxidative stress as evidenced by decreased malondialdehyde concentration along with increasing concentrations of reduced glutathione and superoxide dismutase in lung tissues. Fingolimod also significantly decreased monocyte chemoattractant protein-1(MCP-1), p-ERK,and p-P38 in lung tissues of Ova-challenged mice. In conclusion, the current study demonstrated the anti-inflammatory and antioxidant effects of fingolimod in allergic airway inflammation that might be associated with the downregulation of mitogen activated kinases signaling to decrease T helper 2 cytokine secretion (IL-4 and IL-13) and MCP-1 expression, along with the inhibition of oxidative stress.

Shiitake Flagellate Dermatitis

A man in his early 70s presented with a 3-day history of intensely pruritic erythematous papules and papular vesicular eruption arranged in a flagellate pattern on his neck, trunk, (Figure 1), and extremities with no general symptoms. On questioning, he had eaten undercooked shiitake mushrooms 2 days before the onset of the cutaneous lesions. Laboratory investigation was unremarkable. A skin biopsy revealed spongiosis, marked papillary dermal edema, and a dermal perivascular inflammatory infiltrate mainly composed of lymphocytes and eosinophils.

Obstacles in Diagnosing and Treating a Case of Primary Cutaneous CD8-Positive Aggressive Epidermotropic T-Cell Lymphoma; A Case Report

Cutaneous T-cell lymphomas (CTCL) have a wide spectrum of clinical, histological, immunophenotypic features and genetic alterations which make their diagnosis challenging. Due to the rarity and poor characterization of Primary cutaneous CD8-positive aggressive epidermotropic T-cell lymphoma ((PCAECyTCL), it remains a provisional entity in WHO classification (2018) (1). The constant immunohistochemical finding in PCEACyTCL is CD8 +/ CD4 - expression that is non-specific. So, it needs a consensus on the correlation of clinical & lab for accurate diagnosis (2).In addition to the aggressive presentation of the disease and the associated burden of infection make treatment decisions very challenging. Herein, we are presenting our experience in treating an aggressive case of PCEACyTCL, and how we dealt with the disease and its complications. a 58-year-old male, with a history of Psoriasis, started with him 30 years and he was on topical treatment for it. Referred to us after 3 months journey in the Dermatological department with rapidly progressive maculopapular lesions & tumors, showing central ulcerations, necrosis, and complicated by purulent bacterial infection involving his face, scalp, neck, all 4 limbs, whole trunk, perineum with the oral cavity and oropharynx. There was also generalized lymphadenopathy (Figure 1). His lab results were within the normal range with no evidence of any abnormal infiltration by lymphocytes in peripheral blood and Bone marrow examination. Punch skin biopsy showed that the dermis was the seat of dense localized upper & mid dermal wedge-shaped angiocentric & perivascular inflammatory cell infiltrate composed mainly of activated lymphocytes mixed with histiocytes, plasma cells & neutrophils with atypical, transformed lymphocytes. The infiltrate is encroaching upon the epidermis with degeneration of the basal cell layer & epidermotropism. The Immunohistochemistry revealed diffuse positivity of CD3, weak positivity of CD4, and strong positivity of CD8 with negativity for CD7, CD20, CD30, and ALK in the atypical, transformed T-lymphocytes (Figure 2). The patient was diagnosed to have pcAECyTCL. The staging was done using Contrast CT of the chest, abdomen, and pelvis and showed generalized lymphadenopathy with pulmonary nodules, so he was staged (IVB). After mild improvement of the patient's general condition by anti-biotics, painkillers, and supportive treatment, he started CHOP/Etoposide (doxorubicin 50 mg/m2, cyclophosphamide 750 mg/m2, and vincristine 1.4 mg/m2 by intravenous infusion on day 1, etoposide 100 mg/m2 intravenously days 1-3) to be repeated/3 weeks, with G-CSF as our patient showed aggressive myelosuppression after cycles. There was a very promising improvement in his skin lesions (Figure 3), oral ulcers, and general condition. After 3 cycles of chemotherapy, a PET scan showed: Non metabolically active lymphadenopathy. Metabolically active diffuse skin lesions forming irregular skin thickening over the trunk & limbs (SUV max 5). So, he continued further 3 cycles with preparation for an ASCT. Unfortunately, before stem cell transplantation, the skin lesions progressed again with the same aggression. So, he was given 1 cycle of GEMOX (Gemcitabine1 gm/m2 and Oxaliplatin 100 mg/m2 to be repeated/14 days) then he got COVID-19 infection and is still under treatment for it. In our patient, psoriasis can be considered a susceptible predisposing factor for pcAECyTCL according to the proposed theory by Cai, Fleming and Yan, and others (3,4). One of the obvious problems of pcAECyTCL is the exhibition of T-helper-1 cytokine profile, this makes many treatment options inappropriate as Interferons (5). Theoretically, multi-agent chemotherapies appear to be better options, but they carry the risk of toxicity and myelosuppression, and their response is unsatisfactory till now. There are emerging novel therapies but still not applied to this type of lymphoma due to its rarity (6, 7, 8). Bastidas et al. was suggesting that Ruxolitinib may play a future role in its treatment, but this is still in need of future research (9). The CHOP/etoposide protocol was promising in our patient, but it didn't give a durable response. The plan after improvement from COVID-19 infection is to give a targeted therapy as Brentuximab if available or an aggressive protocol as Hyper-CVAD followed by ASCT hoping for CR, longer OS, and better quality of life. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Abstract 14566: The Role of Eosinophils in Pulmonary Hypertension

Introduction: Pulmonary hypertension (PH) is a severe life-threatening disease with insidious onset and high mortality, featured by pulmonary vascular remodeling and perivascular inflammatory cell infiltration. However, the precise immune mechanisms underlying PH remain unclear. Although an enhanced Th2 immune response in PH has been reported for years, the role of eosinophils (EOS), one of the Th2 effector cells, in PH is seldom reported. Investigating the roles and molecular mechanisms of EOS may present a novel paradigm of PH development. Hypothesis: EOS protect against PH through lipid metabolites. Methods: EOS infiltration and chemotaxis were investigated in peripheral blood and lung tissues from PH patients and sugen/hypoxia-induced PH mice. ΔdblGATA mice and anti-IL5 antibody (TRFK5)-treated mice were applied to investigate the role of EOS in PH development. Flow cytometry-based assays were used to quantify infiltration of different immune cells. The culture supernatants and lysates of EOS were collected to explore the mechanisms in co-culture cell experiments. Results: EOS accumulated in lung tissues from PH patients and mice, and the infiltration of EOS into pulmonary tissues during PH may be promoted by fibroblast-derived CCL11. Furthermore, EOS deficiency increased neutrophil (p=0.0038) and monocyte/macrophage (p=0.0344) infiltration and promoted pulmonary arterial smooth muscle cell proliferation, leading to aggravated PH in mice. Mechanistically, arachidonate 15-lipoxygenase (ALOX15), which catalyzes the production of pro-resolving lipid mediators, is specifically expressed by EOS. Conclusions: EOS play a beneficial role during PH development via ALOX15 and its downstream lipid mediators, suppressing perivascular inflammation and maintaining smooth muscle cell homeostasis.

PSUN275 Autoimmune Dysglycemia Syndrome (ADS) Associated with Multiple Rheumatological Conditions

Abstract Background Autoimmune dysglycemia syndrome is rare entity that can be induced by drugs or associated with rheumatic diseases. This syndrome includes the insulin autoimmune syndrome caused by anti-insulin antibodies; and type B insulin resistance syndrome caused by anti-insulin receptor antibodies. Patients may present with hyperglycemia, hypoglycemia, or both. Clinical case A 41-year-old Peruvian woman; pulmonary tuberculosis at age 14. In 2008 she presented polyarthritis, elevated ESR and CRP, she received methotrexate and deflazacort for 3 months, presenting remission and suspending medication. In 2009, she was admitted to the emergency room with seizures that resolved after intravenous administration of 33% dextrose. She was hospitalized for study, during her stay she presented episodes of fasting hypoglycemia and postprandial hyperglycemia. She reported hyporexia, weight loss, alopecia, and joint pain. On examination: she presented malar and soft palate erythema, mild pretibial edema. Laboratory tests: fasting blood glucose: 52-69mg/dl, albumin 1,6g/L; fasting insulin: 69.3uIU/mL, C-peptide: 3ng/mL; Anti-insulin antibodies: 4,7 (NV < 0,5), proteinuria 4,2 g/24h, ANA 1/2560 mottled pattern. Anti-cardiolipin, Anti-Sm and Anti-DNA (-). Hemoglobin: 10.6, Leukocytes: 3,160, lymphocytes: 1,540, platelets: 166,000. Renal biopsy showed membranous glomerulonephritis. The determination of antibodies against the insulin receptor is not available in Peru. With the diagnosis of autoimmune dysglycemia associated with SLE, she received prednisone 30mg/d, azathioprine 125mg/d, hydroxychloroquine 400mg/d; fractionated normocaloric diet, NPH + crystalline insulin on a correction scale, she was discharged with a progressive decrease in NPH insulin and metformin 850mg bid. Her glucose levels stabilized, control of proteinuria and joint symptoms was achieved with azathioprine, hydroxychloroquine, prednisone 10mg. The patient remained asymptomatic with rheumatological treatment, she suspended metformin and endocrinological follow-up in 2011. In 2013 she presented recurrent episodes of Raynaud's phenomenon and symptoms of proximal muscle weakness, crusted ulcers on the pads of the fingers of both hands, malar rash and exanthema in heliotrope. Laboratory tests: CPK: 8347U/L. Electromyogram: Spontaneous activity with polyphasic short small motor unit potential and early recruitment. Muscle biopsy showed perifascicular atrophy, perivascular and perimysial inflammatory infiltrate with CD4+ T lymphocytes and B lymphocytes. Glucose, insulin, C3, and C4 levels were normal, and anti-insulin antibodies (-). The diagnosis of dermatomyositis was established. She received prednisone 60mg, hydroxychloroquine, methotrexate 10mg, and nifedipine. In August 2020, telemedicine care in endocrinology and rheumatology concluded that there were no episodes of hypoglycemia, glucose: 86 mg/dl, insulin: 12uIU/ml; anti-insulin antibodies (-); CPK levels 136IU/L, C3, C4 normal, no proteinuria and anti ds-DNA (-). Conclusion Autoimmune conditions could cause dysglycemia mainly through different autoantibodies which may cause hyperglycemia and in rare cases hypoglycemia. This case report emphasizes the importance of long-term follow-up in patients with autoimmune conditions including SLE and ADS. Many of these patients are prompt to develope a new autoimmune disease over time. Presentation: Sunday, June 12, 2022 12:30 p.m. - 2:30 p.m.

S2774 Sweet Syndrome as One of the Manifesting Symptoms of Inflammatory Bowel Disease

Introduction: Sweet's syndrome (SS), a subtype of neutrophilic dermatosis (ND), is an uncommon skin disorder characterized by the abrupt appearance of painful erythematous skin lesions, fever, and neutrophilia. Diagnosis of SS is made via biopsy showing a dense neutrophilic infiltrate of the dermis and epidermal sparing. SS is associated with hematologic malignancies or drug reactions and less commonly with inflammatory bowel disease (IBD). Treatment of SS, and other NDs, mirrors treatment of the underlying condition and is typically steroid responsive. Identification is important as SS is an extra systemic manifestation of underlying disease and indicates a poor prognosis. We present an 18 year old male with SS as a manifesting symptom of IBD. Case Description/Methods: An 18 year old male presented with 1 month of abdominal pain, oral ulcerations, hematochezia, and a left lower extremity lesion. He had a diffusely tender abdomen, multiple ulcers bilaterally on his buccal mucosa, cheilitis, and a 1 cm ulcerated lesion on his left leg. He was febrile and blood tests revealed anemia, thrombocytosis, neutrophilia, elevated inflammatory markers, and a positive C-ANCA. His leg and oral lesions were biopsied as well as a cratered esophageal ulcer found during endoscopy. Sigmoidoscopy demonstrated a circular ulceration of the anal mucosa and severe patchy inflammation in the descending and sigmoid colon. He was started on methylprednisolone for empiric treatment of presumed IBD. The skin biopsy showed ND with an interstitial and perivascular inflammatory infiltrate with epidermal necrosis without vasculitis most consistent with SS. He was diagnosed with Crohn’s Disease and has seen improvement of his skin lesions, oral lesions, and abdominal symptoms after a prolonged steroid taper. (Figure) Discussion: While NDs are well described in association with IBD, SS is not a subtype of ND that commonly occurs with less than 100 cases described. Biopsy is the definitive diagnostic study and should be done promptly as ND’s differential includes drug-reaction, malignancy, and the skin lesions may appear similar to infectious lesions. The two former conditions will respond to treatment though may relapse if not identified, and the latter will be exacerbated by immunosuppression. For Physicians treating IBD, knowledge of the appearance of ND can ensure prompt biopsy and initiation, or withdrawal, of appropriate therapies.Figure 1.: Top Row: Mucocutaneous findings in the Mouth and Left Lower Extremity Bottom Row: Perivascular mixed inflammatory infiltrate primarily composed of neutrophils with a few eosinophils present with no evidence of vasculitis. The infiltrate is primarily limited to the dermis.

The role of calcium-sensitive receptor in ovalbumin-induced airway inflammation and hyperresponsiveness in juvenile mice with asthma.

The role of the calcium-sensitive receptor (CaSR) was assessed in a juvenile mouse model of asthma induced by ovalbumin (OVA). The experiment was divided into normal control, OVA, and OVA +2.5/5mg/kg NPS2143 (a CaSR antagonist) groups. OVA induction was performed in all groups except the normal control, followed by assessing airway hyperresponsiveness (AHR) and lung pathological changes. Serum OVA-specific IgE and IgG1 were detected with an enzyme-linked immunosorbent assay (ELISA), and inflammatory cells were counted in bronchoalveolar lavage fluid (BALF). Real-time quantitative polymerase chain reaction, ELISA, and western blotting were performed to detect gene and protein expression. NPS2143 improved the OVA-induced AHR in mice, and AHR was higher in the OVA +2.5mg/kg NPS2143 group than in the OVA +5mg/kg NPS2143 group. Furthermore, NPS2143 reduced the production of OVA-specific IgE and IgG1 in serum and the number of eosinophils and lymphocytes in BALF in OVA mice with reduced CaSR expression in lung tissues. Besides, OVA-induced mice exhibited peribronchial and perivascular inflammatory cell infiltration, which was accompanied by severe goblet cell hyperplasia/hyperplasia and airway mucus hypersecretion. Furthermore, these mice exhibited increased levels of Interleukin (IL)-5, IL-13, MCP-1, and eotaxin, which were alleviated by NPS2143. The 5mg/kg NPS2143 showed more effective than the 2.5mg/kg treatment. CaSR expression was elevated in the lung tissues of OVA-induced asthmatic juvenile mice, whereas the CaSR antagonist NPS2143 reduced AHR and attenuated the inflammatory response in OVA-induced juvenile mice, possibly exerting therapeutic effects on childhood asthma.

Abstract 127: Nox5 Expression In A Vascular Smooth Muscle Cell-specific Manner Induces Fibroblast To Myofibroblast Differentiation

Mice expressing human Nox5 (hNox5) in VSMC exhibit cardfiovascular fibrosis, where mechanisms are unknown. We postulated that VSMC-Nox5 promotes fibroblast to myofibroblast differentiation that contributes to fibrosis. Fibroblasts were cultured from wildtype (WT) and hNOX5 mice. Mice (20 weeks old) were infused with Ang II (600 ng/Kg/day) for 28 days and renal fibrosis/inflammation studied. Markers of myofibroblasts (αSMA), and pro-fibrotic and inflammatory signaling molecules were assessed by qPCR and immunoblotting. Inflammatory infiltrate was assessed by FACS. Fibroblasts from Nox5 mice exhibited increased mRNA of markers of myofibroblast αSMA (2 -ddC :1.54±0.05 vs. WT 0.78±0.17) and Myocd (2 -ddC :1.36±0.17 vs. WT 0.39±0.22), as well as, pro-fibrotic markers, Col1A1 (2 -ddC :1.74±0.16 vs. WT 0.67±0.11), Col3A1 (2 -ddC :1.74±0.18 vs. WT 0.96±0.24) and TIMP3 (2 -ddC :2.65±0.25 vs. WT 0.38±0.07), p<0.05. mRNA expression of CD36 (2 -ddC :1.37±0.07 vs. WT 86±0.24), TNFα (2 -ddC :1.32±0.2 vs. WT 0.71±0.17) and TNFR1 (2 -ddC :1.26±0.04 vs. WT 1.02±0.10) were increased, while CD68 expression was decreased (2 -ddC :0.82±0.11 vs. WT 1.36±0.18) in fibroblasts from Nox5 mice (p<0.05). In Nox5 mice fibroblasts, ROS production and TGFβ protein expression (AU:1.8±0.05 vs. WT 1.4±0.06), as well as TGFβR2 gene expression (2 -ddC :2.04±0.17 vs. WT 0.57±0.12), were increased (p<0.05). mRNA of DNMT3a and TET2, DNA methylation regulatory enzymes, were also increased in fibroblasts from Nox5 mice, p<0.05. Kidneys from Ang II-infused Nox5 mice exhibited significant perivascular fibrosis and inflammatory cell infiltration compared to WT, as well as increased protein expression of TGFβ (AU: 3.59±0.8 vs. WT 1.54±0.2) and IL-11 (AU: 0.64±0.08 vs. WT 0.39±0.04), p<0.05; where levels of macrophage F4/80+ cells (%:24±2 vs WT 18±1, p<0.05) and levels of cytotoxic CD8+ T cells were increased, p<0.05. Kidney expression of vimentin (AU:1.01±0.05 vs WT 0.85±0.03) and αSMA (AU:0.44±0.03 vs WT 0.33±0.01), were increased in Nox5 mice (p<0.05). Nox5 regulates fibrosis by inducing fibroblast-to-myofibroblast differentiation, possibly through increased ROS. These processes may be important in Nox5-assocated cardiovascular-renal fibrosis.

32573 Pyoderma gangrenosum: Ulcerative to pustular variety

Pyoderma gangrenosum (PG) is a chronic, rare, noninfectious neutrophilic dermatosis of unknown etiology. Four clinical variants of PG have been described: ulcerative, bullous, vegetative, and pustular. Pustular pyoderma gangrenosum is an uncommon variant where the evolution from pustules to ulcers may occur. Although most PG patients present only a single clinical variant, the coexistence of different varieties has been reported in the literature. A 50-year-old woman presented with a 2-year history of multiple painful pustules on her trunk and four extremities. The patient reports having started with an ulcer with dark reddish undermined edges on her right leg. The pustules progressed to hemorrhagic crusts and some of them to ulcers. The dermatosis occurred in periods of exacerbation and remission. The patient presented a positive pathergy sign. Her medical history included diabetes mellitus type 2, arterial hypertension, and deep venous thrombosis. A biopsied specimen from the pustule showed an intraepidermal unilocular pustule containing abundant polymorphonuclear cells, red blood cells, and plasma cells. The dermis exhibits a perivascular mixed inflammatory infiltrate composed of neutrophils, lymphocytes, and some eosinophils. Immunofluorescence was negative. Laboratory examination showed C-reactive protein 31 mg/dL and sedimentation velocity 38 mg/dL. Treatment based on prednisone was given with a good response. This is a rare case of pustular PG that showed the evolution from ulcerative to pustular PG. It is important to recognize to make an early diagnosis and prevent the progression of pustular PG.

Misdiagnosis of neutrophilic erythema of infancy as leukocytoclastic vasculitis: A potential diagnostic pitfall in an infantile inflammatory dermatosis.

The authors declare no conflict of interest. The data that support the findings of this study are openly available in NFEI at https://pubmed.ncbi.nlm.nih.gov/. F IGURE 1 Annular erythematous plaques in an infant. Erythematous annular plaque on the right thigh (A) and progression after 3 weeks (B) F IGURE 2 Histopathology of annular erythematous plaque. Punch biopsy specimen from the right leg shows perivascular and interstitial dermal inflammatory infiltrate (A, ×100) primarily composed of neutrophils (B, ×400). Perivascular and interstitial inflammatory infiltrate with primarily neutrophilic inflammation with occasional admixed eosinophils (C, ×400). Leukocytoclasis is present (D, ×400) and definitive vessel wall damage is not observed (A–D)

Multiple skin-colored facial papules in a child

Multiple skin-colored facial papules in a child

Wenyang Huazhuo Tongluo formula alleviates pulmonary vascular injury and downregulates HIF-1α in bleomycin-induced systemic sclerosis mouse model

BackgroundVascular damage, autoimmune abnormalities, and fibrosis are the three pathological features of systemic sclerosis (SSc).However, pulmonary vascular damage is the main factor affecting the progression and prognosis of SSc. The main purpose of this study was to explore the molecular mechanism of Wenyang Huazhuo Tongluo Formula in alleviating pulmonary vascular injury in bleomycin-induced SSc mouse model.MethodsMasson staining and H&E staining were used to analyze the degree of pulmonary vascular fibrosis and the infiltration of leukocyte cells in lung tissue ofbleomycin-induced SSc mouse models treated with saline (BLM group), Wenyang Huazhuo Tongluo Formula (WYHZTL group) and HIF-1α inhibitor KC7F2 (KC7F2 group). Blood vessel exudation was determined by analyzing the cell number and albumin concentration in bronchoalveolar lavage fluid using a cell counter and ELISA assay, respectively. The degree of vascular injury was assessed by measuring the expression levels of vWF, E-selectin, ICAM-1, VCAM-1, VE-cadherin and claudin-5 in serum and pulmonary vascular endothelial cells using ELISA and immunofluorescence staining. Finally, the effect of Wenyang Huazhuo Tongluo Formula on the expression of HIF-1α was detected using immunofluorescence staining.ResultsWenyang Huazhuo Tongluo Formula and KC7F2 significantly inhibited bleomycin-induced pulmonary vascular fibrosis and the level of perivascular inflammatory cell infiltration. The number of cells and the concentration of albumin were significantly reduced in the bronchoalveolar lavage fluid of the WYHZTL group and KC7F2 group compared with the BLM group. In addition, treatment with Wenyang Huazhuo Tongluo Formula and KC7F2 significantly downregulated the expression levels of vWF, E-selectin, ICAM-1, VCAM-1 and HIF-1α, but upregulated the expression of VE-cadherin and claudin-5 in serum and pulmonary vascular endothelial cells, compared with treatment with saline.ConclusionsThis study reveals that Wenyang Huazhuo Tongluo Formula plays a new role in the treatment of SSc by alleviating pulmonary vascular damage. Furthermore, we found that Wenyang Huazhuo Tongluo Formula alleviates pulmonary vascular injury and inhibits HIF-1α expression.

Bullous systemic lupus erythematosus with nephrotic syndrome in an adolescent girl: a case report

Bullous systemic lupus erythematosus is an uncommon autoimmune subepidermal vesiculobullous disease occurring in less than 5% of patients with systemic lupus erythematosus. The diagnosis of bullous systemic lupus erythematosus is based on clinical picture, histopathological findings, direct immunofluorescence and other immunologic tests. Importantly, establishing the diagnosis of bullous systemic lupus erythematosus requires the occurrence of systemic lupus erythematosus according to the American College of Rheumatology criteria. We report a case of bullous systemic lupus erythematosus in a 16-year-old Vietnamese girl, who demonstrated an acquired vesiculobullous eruption associated with nephrotic syndrome and fulfilled the European League Against Rheumatism/ American College of Rheumatology 2019 classification criteria of systemic lupus erythematosus. Clinical examination showed numerous tense, clear fluid-filled vesicles and bullae affecting the whole body with the predilection for the face, neck, chest, abdomen and bilateral inner thigh areas. Erosions were observed on the tongue and the buccal mucosa. Histopathologic examination showed subepidermal vesicle containing abundant neutrophils and perivascular mixed inflammatory cell infiltration. Microabscesses at tips of dermal papillae and the features of a leukocytoclastic vasculitis were not seen. Direct immunofluorescence of perilesional skin biopsies demonstrated linear deposition of IgG, IgA, IgM and C3 in the basement membrane zone. Linear immunoglobulin deposition displayed a u-serrated pattern with the predominant staining intensity of IgG. Serologic tests showed positive antinuclear antibody. Complement components assay revealed low levels of C3 (66.5 mg/dL) and C4 (3,9mg/dL). Proteinuria level was increased to 3.5g/24 hours. The final diagnosis of bullous systemic lupus erythematosus with nephrotic syndrome was made. The patient was treated with a high-dose oral glucocorticoid scheme and showed significant clinical improvement.

NOX5 EXPRESSION IN A VASCULAR SMOOTH MUSCLE CELL-SPECIFIC MANNER INDUCES FIBROBLAST PHENOTYPIC SWITCH AND PRO-FIBROTIC EFFECTS

Objective: Mice expressing human Nox5 (hNox5) in vascular smooth muscle cells (VSMC) exhibit redox-sensitive vascular dysfunction and cardio-renal fibrosis. Mechanisms underlying pro-fibrotic effects are unclear. Here we assessed whether hNox5 in VSMCs promotes fibroblast phenotypic changes leading to myofibroblast differentiation and pro-fibrotic/inflammatory responses. Design and method: Fibroblasts were cultured from wildtype (WT) and mice expressing hNOX5 in VSMC (Nox5+SM22+). Mice (20 weeks old) were infused with Ang II (600 ng/Kg/day) for 28 days and renal fibrosis/inflammation studied. Markers of myofibroblasts, pro-fibrotic and inflammatory phenotypes were assessed by qPCR and immunoblotting. Inflammatory infiltrate was assessed by FACS. Results: Fibroblasts isolated from Nox5+SM22+ mice exhibited increased mRNA expression of pro-fibrotic markers, such as Col1A1 (2-ddCt: 1.74 ± 0.16 vs. WT 0.67 ± 0.11), Col3A1 (2-ddCt: 1.74 ± 0.18 vs. WT 0.96 ± 0.24) and TIMP3 (2-ddCt: 2.65 ± 0.25 vs. WT 0.38 ± 0.07), p < 0.05. Myofibroblast phenotype (SMC) markers mRNA, aSMA (2-ddCt: 1.54 ± 0.05 vs. WT 0.78 ± 0.17) and Myocd (2-ddCt: 1.36 ± 0.17 vs. WT 0.39 ± 0.22) were also increased in fibroblasts from Nox5 mice, p < 0.05. In terms of inflammatory phenotype, we observed that mRNA expression of CD36 (2-ddCt: 1.37 ± 0.07 vs. WT 86 ± 0.24), TNFa (2-ddCt: 1.32 ± 0.2 vs. WT 0.71 ± 0.17) and TNFR1 (2-ddCt: 1.26 ± 0.04 vs. WT 1.02 ± 0.10) were increased, while CD68 expression was decreased (2-ddCt: 0.82 ± 0.11 vs. WT 1.36 ± 0.18) in fibroblasts from Nox5+SM22+ mice (p < 0.05). In Nox5 mice fibroblasts, key regulators of fibrosis such as ROS production and TGFb protein expression (AU: 1.8 ± 0.05 vs. WT 1.4 ± 0.06), as well as TGFbR2 gene expression (2-ddCt: 2.04 ± 0.17 vs. WT 0.57 ± 0.12), were increased (p < 0.05). Kidneys from Ang II-infused Nox5+SM22+ mice exhibited significant perivascular fibrosis and inflammatory cell infiltration compared to WT. Kidney expression of vimentin (AU: 1.01 ± 0.05 vs WT 0.85 ± 0.03) and aSMA (AU: 0.44 ± 0.03 vs WT 0.33 ± 0.01), markers of myofibroblast differentiation, were increased in Nox5 mice (p < 0.05). In kidneys from Nox5+SM22+ mice, increased frequencies of macrophage F4/80 + (%: 24 ± 2 vs WT 18 ± 1, p < 0.05). Conclusions: Our findings suggest that Nox5 in VSMCs influences fibroblast phenotypic switching to myofibroblasts that likely influences cardiovascular and renal fibrosis in hypertension.

Effects of YM155 on the proliferation and apoptosis of pulmonary artery smooth muscle cells in a rat model of high pulmonary blood flow-induced pulmonary arterial hypertension

ABSTRACT Introduction Proliferation and apoptosis of pulmonary artery smooth muscle cells (PASMCs) play an important role in the occurrence and development of pulmonary arterial hypertension (PAH). The purpose of this study was to investigate the effects of survivin inhibitor YM155 on the proliferation and apoptosis of PASMCs in rats with PAH induced by high pulmonary blood flow. Methods Thirty male Sprague-Dawley (SD) rats were randomly divided into control, model, and YM155 intervention groups. A rat model of PAH induced by high pulmonary blood flow was established, and it was confirmed by assessments of right-ventricular pressure (RVP) and right ventricular hypertrophy index (RVHI). Immunohistochemical staining and western blot analysis were used to detect the expression of survivin, and the proliferation and apoptosis of PASMCs. Lastly, the effects of in vivo treatment of YM155 were tested. Results The increased expression of survivin mRNA and protein were observed in the model group, accompanied by pulmonary arteriolar wall thickening, lumen stenosis, and perivascular inflammatory cell infiltration. Elevated expression of survivin and pulmonary vascular remodeling were significantly mitigated after YM155 treatment. Specifically, the YM155 intervention group had a significantly lower PASMC proliferation rate and a higher PASMC apoptotic rate. Conclusion YM155 suppressed PASMC proliferation and promoted PASMC apoptosis by inhibiting survivin expression and thereby reducing pulmonary vascular remodeling in high pulmonary blood flow-induced PAH in vivo.

Patologia da periodontite crônica em ovinos

ABSTRACT: Periodontitis is an inflammatory process of infectious origin affecting the teeth and their supporting structures, causing significant economic losses and reducing animal welfare. Bacteria in the gingival biofilm are one of the main factors in initiating inflammatory lesions. Bacteria act directly on tissues or indirectly through substances that cause tissue damage. Studies on the etiopathogenesis of periodontitis in Brazilian sheep herds are scarce. The present study aimed to characterize histologically periodontal lesions of culled sheep from the Brazilian breed, Santa Inês. Periodontal lesions, such as periodontal pockets containing plant tissue and bacteria, replacement of the periodontal ligament by connective tissue and inflammatory cells, superficial pustules, hydropic epithelial degeneration, and epithelium hyperplasia, were observed. Submucosal changes were characterized by granulation tissue, edema, swelling of the endothelial cells, bacteria, and predominantly perivascular lymphoplasmacytic inflammatory infiltrate. In the alveolar bone, osteoclastic resorption and bone apposition were observed. This study revealed subacute to chronic inflammation, alveolar bone resorption, and cortical bone apposition in ovine periodontitis. Thus, these findings can contribute to the evolution of knowledge about the etiopathogenesis of ovine periodontitis and, possibly, the development of measures to control the disease.