Polymorphonuclear Cell Infiltration Research Articles

Topical diosmetin attenuates nociception and inflammation in a ultraviolet B radiation-induced sunburn model in mice.

Burns are a global health problem and can be caused by several factors, including ultraviolet (UV) radiation. Exposure to UVB radiation can cause sunburn and a consequent inflammatory response characterised by pain, oedema, inflammatory cell infiltration, and erythema. Pharmacological treatments available to treat burns and the pain caused by them include nonsteroidal anti-inflammatory drugs (NSAIDs), opioids, antimicrobials and glucocorticoids, which are associated with adverse effects. Therefore, the search for new therapeutic alternatives is needed. Diosmetin, an aglycone of the flavonoid diosmin, has antinociceptive, antioxidant and anti-inflammatory properties. Thus, we evaluated the antinociceptive and anti-inflammatory effects of topical diosmetin (0.01, 0.1 and 1%) in a UVB radiation-induced sunburn model in mice. The right hind paw of the anaesthetised mice was exposed only once to UVB radiation (0.75J/cm2) and immediately treated with diosmetin once a day for 5days. The diosmetin antinociceptive effect was evaluated by mechanical allodynia and pain affective-motivational behaviour, while its anti-inflammatory activity was assessed by measuring paw oedema and polymorphonuclear cell infiltration. Mice exposed to UVB radiation presented mechanical allodynia, increased pain affective-motivational behaviour, paw oedema and polymorphonuclear cell infiltration into the paw tissue. Topical Pemulen® TR2 1% diosmetin reduced the mechanical allodynia, the pain affective-motivational behaviour, the paw oedema and the number of polymorphonuclear cells in the mice's paw tissue similar to that presented by Pemulen® TR2 0.1% dexamethasone. These findings indicate that diosmetin has therapeutic potential and may be a promising strategy for treating patients experiencing inflammatory pain, especially those associated with sunburn.

Role of kinin receptors in skin pigmentation

Previous studies have shown that all kinin system is constitutively expressed in the normal and inflamed skin, with a potential role in both physiological and pathological processes. However, the understanding regarding the involvement of the kinin system in skin pigmentation and pigmentation disorders remains incomplete. In this context, the present study was designed to determine the role of kinins in the Monobenzone (MBZ)-induced vitiligo-like model. Our findings showed that MBZ induces higher local skin depigmentation in kinin receptors knockout mice (KOB1R, KOB2R and KOB1B2R) than in wild type (WT). Remarkably, lower levels of melanin content and reduced ROS generation were detected in KOB1R and KOB2R mice treated with MBZ. In addition, both KOB1R and KOB2R show increased dermal cell infiltrate in vitiligo-like skin, when compared to WT-MBZ. Additionally, lack of B1R was associated with greater skin accumulation of IL-4, IL-6, and IL-17 by MBZ, while KOB1B2R presented lower levels of TNF and IL-1. Of note, the absence of both kinin B1 and B2 receptors demonstrates a protective effect by preventing the increase in polymorphonuclear and mononuclear cell infiltrations, as well as inflammatory cytokine levels induced by MBZ. In addition, in vitro assays confirm that B1R and B2R agonists increase intracellular melanin synthesis, while bradykinin significantly enhanced extracellular melanin levels and proliferation of B16F10 cells. Our findings highlight that the lack of kinin receptors caused more severe depigmentation in the skin, as well as genetic deletion of both B1/B2 receptors seems to be linked with changes in levels of constitutive melanin levels, suggesting the involvement of kinin system in crucial skin pigmentation pathways.

Abstract 5366: ATR promotes extracellular matrix stiffness mediated cytoskeleton remodeling and immunosuppression

Abstract The stiffening of the extracellular matrix (ECM), a phenomenon that often promotes metastasis and immunosuppression in many cancers, is a process that is not yet fully understood. Our study reveals that ATR, a molecule primarily recognized for its role in maintaining the genome, plays a crucial role in ECM stiffness-induced immune evasion and metastasis in triple negative breast cancer (TNBC) through DNA repair independent manner. In response to ECM stiffness, ATR activates SUN2, a component of the linker of nucleoskeleton and cytoskeleton (LINC) complex. This activation enhances the interaction of SUN2 with outer nuclear membrane proteins KASH1 and KASH2. The interaction between SUN2 and KASH1/2 facilitates the nuclear localization of β-catenin, which in turn promotes epithelial mesenchymal transition (EMT) and reduces the expression of E-cadherin. E-cadherin is essential for attracting CD103+ immune cells that perform anti-tumor immunity. Interestingly, inhibiting ATR reduces EMT in tumor cells, leading to an increase in CD103+ dendritic cell infiltration while decreasing neutrophil and polymorphonuclear myeloid-derived suppressor cell (PMN-MDSC) infiltration. Further, we found that ATR levels are inversely correlated with immunotherapy response rates in patients. The alteration in immune cell composition sensitizes tumors to immunotherapy, thereby suppressing tumor growth and metastasis. In conclusion, the role of ATR in remodeling the immune environment could be crucial for optimizing the clinical use of ATR inhibitors. This makes ATR a promising target for overcoming immunotherapy resistance. We suggest early or prolonged treatment with an ATR inhibitor to potentially reduce metastasis rates and improve the prognosis of TNBC patients. Furthermore, we suggest using ATR levels as a marker to predict response rates and prognosis in TNBC patients. For patients with higher ATR expression levels, combination treatment with ATR inhibitors and immunotherapy may yield better outcomes. Enhanced anti-tumor immunity could increase the chances for patients to eliminate cancer cells and help end breast cancer. Finally, our research suggests the combined treatment of ATR inhibition and immunotherapy for patients without necessarily applying DNA damage stimulation. This approach could potentially reduce toxicity while still benefiting patients. Citation Format: Xinyi Tu, Zhenkun Lou, Robert Mutter. ATR promotes extracellular matrix stiffness mediated cytoskeleton remodeling and immunosuppression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5366.

Intratumoral injection of interferon gamma promotes the efficacy of anti-PD1 treatment in colorectal cancer

Immune checkpoint inhibitors (ICIs) offer new options for the treatment of patients with solid cancers worldwide. The majority of colorectal cancers (CRC) are proficient in mismatch-repair (pMMR) genes, harboring fewer tumor antigens and are insensitive to ICIs. These tumors are often found to be immune-deserted. We hypothesized that forcing immune cell infiltration into the tumor microenvironment followed by immune ignition by PD1 blockade may initiate a positive immune cycle that can boost antitumor immunity. Bioinformatics using a public database suggested that IFNγ was a key indicator of immune status and prognosis in CRC. Intratumoral administration of IFNγ increased immune cells infiltration into the tumor, but induced PD-L1 expression. A combined treatment strategy using IFNγ and anti-PD-1 antibody significantly increased T cell killing of tumor cells in vitro and showed synergistic inhibition of tumor growth in a mouse model of CRC. CyTOF found drastic changes in the immune microenvironment upon combined immunotherapy. Treatment with IFNγ and anti-PD1 antibody in CT26 tumors significantly increased infiltration of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs). IFNγ had a more pronounced effect in decreasing intratumoral M2-like macrophages, while PD1 blockade increased the population of CD8+Ly6C + T cells in the tumor microenvironment, creating a more pro-inflammatory microenvironment. Additionally, PD1 induced increased expression of lymphocyte activating 3 (LAG3) in a significant fraction of CD8+ T cells and Treg cells, indicating potential drug resistance and feedback mechanisms. In conclusion, our work provides preclinical data for the Combined immunotherapy of CRC using intratumoral delivery of IFNγ and systemic anti-PD1 monoclonoal antibody.

Hypomethylation at H19DMR in penile squamous cell carcinoma is not related to HPV infection

ABSTRACT Penile squamous cell carcinoma (SCC) is a rare and aggressive tumour mainly related to lifestyle behaviour and human papillomavirus (HPV) infection. Environmentally induced loss of imprinting (LOI) at the H19 differentially methylated region (H19DMR) is associated with many cancers in the early events of tumorigenesis and may be involved in the pathogenesis of penile SCC. We sought to evaluate the DNA methylation pattern at H19DMR and its association with HPV infection in men with penile SCC by bisulfite sequencing (bis-seq). We observed an average methylation of 32.2% ± 11.6% at the H19DMR of penile SCC and did not observe an association between the p16INK4a+ (p = 0.59) and high-risk HPV+ (p = 0.338) markers with methylation level. The average methylation did not change according to HPV positive for p16INK4a+ or hrHPV+ (35.4% ± 10%) and negative for both markers (32.4% ± 10.1%) groups. As the region analysed has a binding site for the CTCF protein, the hypomethylation at the surrounding CpG sites might alter its insulator function. In addition, there was a positive correlation between intense polymorphonuclear cell infiltration and hypomethylation at H19DMR (p = 0.035). Here, we report that hypomethylation at H19DMR in penile SCC might contribute to tumour progression and aggressiveness regardless of HPV infection.

Abstract B019: Tumor-derived PTHrP molds an immunosuppressive microenvironment

Abstract Purpose[JP1] : Molecular phenotyping of PDAC [PJ2] has revealed an aggressive squamous/basal/quasi-mesenchymal[JP3] subtype, clinically presenting as highly metastatic and harboring genetic amplifications of KRAS. Our previous work indicated that a squamous identity gene, Parathyroid Hormone Like Hormone (PTHLH; encoding for the PTHrP protein), is frequently co-amplified with KRAS and skews PDAC towards a squamous identity. Further, PTHrP is secreted by tumor cells and promotes tumor growth and metastasis through autocrine signaling. However, pathological analysis of PTHrP-null tumors indicated that the tumor microenvironment (TME) was re-organized upon loss of PTHrP. Thus, we hypothesized that PTHrP is a tumor-derived factor that shapes the TME through its systemic secretion and mobilization of immunosuppressive myeloid cells. Methods: In vivo experiments used KrasG12D; p53R172H; Pdx1-Cre; YFP (KPCY) and KPCY-PthlhloxP/WT (KPCY-Pthlhhet) for spontaneous tumor formation in immune competent mice. Orthotopic implantation experiments used Pthlh overexpressing KPCY tumor cells (herein KPCY-PthlhOE) or empty vector control cells (KPCY-PthlhEV). Tumor immunophenotyping was done using flow cytometry. Cytokine expression was performed using qPCR and ELISA. Results: In vivo investigation of tumors from KPCY-Pthlhhet mice revealed less MDSC and greater CD8+ T cell infiltration than KPCY controls, demonstrating the necessity of PTHrP for immunosuppression[PJ4]. Orthotopic implantation of KPCY-PthlhOE cell lines reduced overall TME immune cell recruitment, while simultaneously increasing the proportion of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC) infiltration. A concomitant decrease in the frequency of T cells in the KPCY-PthlhOE tumors was also observed, showing the sufficiency of PTHrP to organize an immunosuppressive TME[PJ5]. Additionally, a reduction of PD-1+ and CD44high CD8+ T cells revealed they were less activated. Interestingly, recombinant human PTHrP was sufficient to induce MDSC mobilization in vivo.[JP6] To interrogate the mechanism of MDSC trafficking into the TME, cytokine production by was analyzed in vitro. KPCY-PthlhOE exhibited a pronounced increase and decrease of Cxcl1 and Cxcl10 expression, respectively, relative to the KPCY-EV control. Conclusions: The work herein describes a new mechanism for the mobilization and infiltration of immune suppressive MDSC in the tumor microenvironment, initiated by PTHrP. The PTHrP-dependent immunosuppressive TME excludes and inhibits effector immune cells, such as CD8+ T cells. This phenomenon has been described in squamous-like PDAC but may also apply to other squamous adenocarcinomas. [PJ7] Further elucidation of the mechanism by which PTHrP is the genesis of the immune suppressive TME may spur the development of novel treatments for these more clinically severe cancer subtypes. Citation Format: Calvin P. Johnson, Boyang Ma, Jason Pitarresi. Tumor-derived PTHrP molds an immunosuppressive microenvironment [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Pancreatic Cancer; 2023 Sep 27-30; Boston, Massachusetts. Philadelphia (PA): AACR; Cancer Res 2024;84(2 Suppl):Abstract nr B019.

Combination of radiotherapy and Anlotinib enhances benefit from immunotherapy to liver metastasis and abscopal tumor from lung cancer

Many studies have shown that liver metastasis can weaken the efficacy of immunotherapy. Immunotherapy combined with radiotherapy or anti-angiogenic therapy has been proven to have synergistic anti-tumor effects. So we devote to explore whether the combination of the three therapies can exert effective anti-tumor effects on liver metastasis. The clinical information of 118 patients with liver metastasis were collected to compare the intrahepatic progression-free survival between immunotherapy and immunotherapy combined with other treatments. We used Lewis lung cancer (LLC) cell to establish a mouse liver metastasis tumor model and record tumor burden and survival. Tumor-infiltrating immune cells detected by flow cytometry. RNA sequencing was performed and the proportion of immune cells were analyzed by TIMER2.0 database. Compared with immunotherapy group, the combination therapy group showed a trend for longer median intrahepatic progression-free survival. Radiotherapy combined with PD-1 inhibitor and Anlotinib can inhibit liver metastasis and subcutaneous tumor growth and prolong the survival compared with other groups in vivo. Compared with the anti-PD-1 treatment group, triple therapy can increase CD4+T, CD8+T, and IFN-γ+CD8+T cells and decrease infiltration of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC) in tumors. PPAR signaling pathway were significantly activated and CD8+T and dendritic cells (DC) were increased in the triple therapy group compared to the PD-1 inhibitor combined with Anlotinib group. Radiotherapy combined with PD-1 inhibitor and Anlotinib can effectively exert anti-tumor efficacy and reshape the tumor immune microenvironment by increasing the infiltration of anti-tumor immune cells and reducing the infiltration of immunosuppressive immune cells.

Depletion of macrophages deteriorates bisphosphonate-related osteonecrosis of the jaw-like lesions in mice.

Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is a potentially intractable disease with no definitive pathophysiology and no treatment and prevention strategies. This study aimed to investigate whether time-selective depletion of macrophages worsens BRONJ-like lesions in mice. A murine model of high-prevalence BRONJ-like lesions in combination with zoledronate/chemotherapeutic drug administration and tooth extraction was created according to the methods of our previous studies. Daily intra-oral submucosal administration of clodronate-loaded liposomes, which temporarily depletes systemic macrophages, was performed immediately after tooth extraction. Spleens, femora, tibiae, and maxillae were dissected 2weeks after extraction to evaluate BRONJ-like lesions and systemic conditions by micro-computed tomography analysis, histomorphometric and immunofluorescent analyses, and serum chemistry with ELISA. Depletion of macrophages significantly decreased the numbers of local and systemic macrophages and osteoclasts on the bone surface, which markedly worsened osseous healing, with increased necrotic bone and empty lacunae in the existing alveolar bone and newly formed bone in the extraction sockets, and soft tissue healing, with decreased collagen production and increased infiltration of polymorphonuclear cells. Interestingly, the depletion of macrophages significantly shifted macrophage polarization to M1 macrophages through an increase in F4/80+CD38+ M1 macrophages and a decrease in F4/80+CD163+ M2 macrophages, with decreases in the total number of F4/80+ macrophages. These data demonstrated that severe inhibition of osteoclasts in bone tissue and polarization shifting of macrophages in soft tissue are essential factors associated with BRONJ.

A new histone deacetylase inhibitor remodels the tumor microenvironment by deletion of polymorphonuclear myeloid-derived suppressor cells and sensitizes prostate cancer to immunotherapy

BackgroundProstate cancer (PCa) is the most common malignancy diagnosed in men. Immune checkpoint blockade (ICB) alone showed disappointing results in PCa. It is partly due to the formation of immunosuppressive tumor microenvironment (TME) could not be reversed effectively by ICB alone.MethodsWe used PCa cell lines to evaluate the combined effects of CN133 and anti-PD-1 in the subcutaneous and osseous PCa mice models, as well as the underlying mechanisms.ResultsWe found that CN133 could reduce the infiltration of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs), and CN133 combination with anti-PD-1 could augment antitumor effects in the subcutaneous PCa of allograft models. However, anti-PD-1 combination with CN133 failed to elicit an anti-tumor response to the bone metastatic PCa mice. Mechanistically, CN133 could inhibit the infiltration of PMN-MDSCs in the TME of soft tissues by downregulation gene expression of PMN-MDSC recruitment but not change the gene expression involved in PMN-MDSC activation in the CN133 and anti-PD-1 co-treatment group relative to the anti-PD-1 alone in the bone metastatic mice model.ConclusionsTaken together, our work firstly demonstrated that combination of CN133 with anti-PD-1 therapy may increase the therapeutic efficacy to PCa by reactivation of the positive immune microenvironment in the TME of soft tissue PCa.

Efficacy of Transdermal Delivery Nano Ethosomal Gel from Ashitaba Leaves on In-vivo Burn Wound Healing in Albino Rats

This study aimed to evaluate the in-vivo burn wound healing process in albino rats treated using transdermal delivery nano ethosomal gel from ashitaba leaves. Ethosomal vesicles were formulated using soy lecithin, cholesterol, ethanol, water, and ashitaba leaf nanoparticles using the cold method. A total of 25 male rats were randomly divided into 5 groups, i.e., (C-) treated without nanoparticle extract, (C+) treated using 1% Silver Sulfadiazine®, treatment group treated nano ethosomal gel from ashitaba leaves with the respective doses were (T1) 1%, (T2) 2,5%, and (T3) 5%. Therapy was initiated on day 1 or after being induced by a burn wound for 14 days, twice a day. On the 15th day, a termination was carried out to take skin tissue from burn scars, then histopathology preparations were made with routine staining. Microscopic observations with an optical microscope on collagen, polymorphonuclear cell infiltration, angiogenesis, and re-epithelialization. The T3 group that was treated using 5% nano ethosomal gel showed the best burn wound healing, this may be caused by compounds in ashitaba leaf nanoparticles which have antioxidant, anti-inflammatory, and antibacterial effects, thus the use of transdermal delivery therapy of 5% ashitaba leaf nano ethosomal gel was effective for wound treatment burn on rat skin.

Oxidative stress status assessment of rats' brains injury following subacute exposure to K-oximes

Oxidative stress status and morphological injuries in the brain of Wistar rats induced by repeated application of selected acetylcholinesterase reactivators – asoxime, obidoxime, K027, K048, K074, and K075 were evaluated. Each oxime in a dose of 0.1 of LD50/kg im was given 2x/week for 4 weeks. Markers of lipid peroxidation (malondialdehyde, MDA), and protein oxidation (advanced oxidation protein products, AOPP), as well as the activity of antioxidant enzymes (catalase, CAT, superoxide dismutase, SOD, glutathione reductase, GR, and glutathione peroxidase, GPx), were estimated in the brain tissue homogenates on day 35 of the study. Brain alterations were carefully quantified by semiquantitative grading scales – brain damage score (BDS). Oxidative stress parameters, MDA and AOPP were significantly highest in the asoxime-, obidoxime- and K075-treated groups (p < 0.001). The activity of SOD and CAT was significantly elevated in the obidoxime-, K048-, and K075-treated groups (p < 0.001). Besides, GR was markedly decreased in the obidoxime- and K074-treated groups (p < 0.01), while treatment with K048, K074 and K075 induced extremely high elevation in GPx levels (p < 0.001). In the same groups of rats, brain alterations associated with polymorphonuclear cell infiltrate were significantly more severe than those observed in animals receiving only asoxime or K027 (p < 0.001). The presented results confirmed that treatment with different oximes significantly improved the oxidative status and attenuated signs of inflammation in rats’ brains. Presented results, together with our previously published data can help to predict likely adverse systemic toxic effects, and target organ systems, which are crucial for establishing risk categories, as well as in dose selection of K-oximes as drug candidates.

THE COMPLEX EFFECT OF VITAMIN D AND ALPHA TOCOPHEROL ACETATE ON THE LUNG MICROCIRCULATION IN THE EXPERIMENTAL INDUCTION OF SYSTEMIC SCLEROSIS

Abstract. The aim of the study was to investigate the effects of vitamin D3 (vitamin D) and alpha-tocopherol acetate (vitamin E) on the microcirculatory vessels of the lungs in an experimental model of systemic sclerosis (SSc).&#x0D; Methods. We divided the experimental animals into four separate groups: an intact group (IG) consisting of 15 animals, a control group (CG) of 20 animals, experimental group 1 (EG1) and experimental group 2 (EG2) consisting of of 25 animals each. Randomization of groups was carefully performed using the weight variance minimization method. To induce SSc in laboratory animals of EG1, subcutaneous injections of 0.5 ml of 5% sodium hypochlorite (NaClO) were administered 3 times a week for 6 weeks in a row. The CG of rats received an isotonic solution following the same scheme, while the IG was not subjected to any injections. In addition to NaClO, the EG2 received intramuscular injections of vit E (10 mg/100 g) and vit D (1000 IU/100 g) for three weeks during the second half of the experiment.&#x0D; After 8 weeks, the animals were euthanized, and lung tissue samples were obtained for histological and electron microscopic analysis. The histological preparations were examined using light microscopy and photographed using a Leica DME light microscope, a DCM 900 digital microscope camera, and a Nicon Coolpix P5100 camera. Meanwhile, the electron microscopy was performed to visualize the ultrastructural characteristics of the lung tissue by use of "PEM-125K" (Selmi, Ukraine) with subsequent photography at magnifications from 2000 to 20000 times.&#x0D; All animal procedures were carried out in strict compliance with bioethical principles. The Ethics Commission of the IFNMU granted approval for the study under document number 117/20 on November 19, 2020.&#x0D; Results. The histological analysis of the lung specimens of CG animals showed a normal morphological structure. In the EG1, the lungs exhibited abnormalities in the structure of the hemocapillaries, including narrowing of the vessel lumen due to thickening of the vascular wall, edema, and infiltration of polymorphonuclear cells. However, in the EG2, there was an improvement in the histological features, with a reduction in edematous processes and a decrease in the intensity of polymorphonuclear cell infiltration.&#x0D; The electron microscopic examination of the lung samples from EG1 animals revealed the presence of platelet adhesion and aggregation, as well as aggregates of erythrocytes within the hemocapillary lumen. Swelling of endothelial cells and narrowing of the hemocapillary lumen were also observed, along with uneven thickening of the basement membrane. In contrast, the combination of vitamins D and E in EG2 animals resulted in improved rheological properties of blood and ultrastructural characteristics of the hemomicrocirculatory elements. The lumen of the hemocapillaries contained single erythrocytes and inactive platelets. The endothelial cells’ membraines showed minimal alterations, with clear contours and no micropinocytotic vesicles or cytoplasmic growths. The nuclei of the endothelial cells had a fine-grained matrix with evenly distributed chromatin granules.&#x0D; Conclusions. This study proves that the combined use of vitamins D and E has a positive effect on the state of the microcirculatory vessels in the lungs of experimentally induced systemic sclerosis.

MICROSCOPIC CHANGES IN RAT’S GINGIVA UNDER CONDITIONS OF EXPERIMENTAL PERIODONTITIS AND THE USE OF MESENCHYMAL STEM CELLS

MICROSCOPIC CHANGES IN RAT’S GINGIVA UNDER CONDITIONS OF EXPERIMENTAL PERIODONTITIS AND THE USE OF MESENCHYMAL STEM CELLS

FDG PET/CT in Aseptic Splenic Abscess

HomeRadiologyRecently Published PreviousNext Reviews and CommentaryFree AccessImages in RadiologyFDG PET/CT in Aseptic Splenic AbscessWeiting Liu, Xiaohong Ou Weiting Liu, Xiaohong Ou Author AffiliationsFrom the Department of Nuclear Medicine, West China Hospital of Sichuan University, No. 37 Guoxue Alley, Chengdu 610041, People’s Republic of China.Address correspondence to X.O. (email: [email protected]).Weiting LiuXiaohong Ou Published Online:Mar 28 2023https://doi.org/10.1148/radiol.222955MoreSectionsPDF ToolsImage ViewerAdd to favoritesCiteTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinked In A 32-year-old woman presented with fever and arthralgia for 1 month. She had a past medical history of rheumatoid arthritis with 2 years of glucocorticoid treatment. At physical examination, splenomegaly was found. Her laboratory data showed leukocytosis (15.86 × 109/L; reference range, 3.5–9.5 × 109/L) and a high erythrocyte sedimentation rate. Serial blood cultures were negative. Intravenous antibiotics and corticosteroid therapy did not relieve her symptoms. Fluorodeoxyglucose (FDG)-PET/CT examination was performed, showing abnormal standardized uptake values in the spleen (Figure). Primary splenic lymphoma was suspected, and the patient underwent laparoscopic splenectomy. At pathologic examination (Figure), the diagnosis of aseptic splenic abscesses as a complication of rheumatoid arthritis was made. Corticosteroids and supportive treatment were then initiated, and her clinical condition gradually improved.Images in a 32-year-old woman presenting with fever and arthralgia for 1 month. She had a past medical history of rheumatoid arthritis with 2 years of glucocorticoid treatment. A fluorodeoxyglucose (FDG)-PET examination was performed. (A) Maximum intensity projection whole body image shows intense diffuse splenic uptake and moderate bone marrow uptake. (B) Axial-fused PET/CT abdominal image shows an enlarged spleen with heterogeneously increased FDG standardized maximum uptake value (standardized maximum uptake value,13.94; liver standardized maximum uptake value, 3.76). At pathologic examination, macroscopic examination of the spleen showed many nonconfluent white nodules (not shown). (C) With use of hematoxylin and eosin, a photomicrograph with 400× magnification shows numerous areas of necrosis surrounded by intense polymorphonuclear cell infiltration (arrows) and fibroblastic proliferation at the periphery of the splenic nodules.Download as PowerPointOpen in Image Viewer Splenic abscesses generally occur in patients with immune dysfunction (1), presenting as solitary or multifocal lesions. Aseptic abscesses are most commonly associated with nonspecific inflammatory bowel disease and other autoimmune diseases (spondyloarthropathy, rheumatoid arthritis, lupus erythematosus, and nodular arteritis), or autoinflammatory (synovitis, acne, pustulosis, hyperostosis, and osteitis) and neoplastic ones (myeloma, monoclonal gammopathy) (2). Histopathologic examination helps to exclude other conditions. The cornerstones of aseptic abscesses are mature polymorphonuclear neutrophils in the center of the splenic nodules, with histiocytes and single giant cells present at the periphery (2). It is unclear whether glucocorticoids could be a factor that promotes the formation of aseptic splenic abscesses. The presence of disseminated sterile splenic microabscesses with intense FDG uptake related to rheumatoid arthritis is rare.Disclosures of conflicts of interest: W.L. No relevant relationships. X.O. No relevant relationships.

Histomorphometric evaluation of skin wounds in rats submitted to biomodulatory therapies – research protocol

INTRODUCTION: The use of biomodulatory therapies in order to help tissue repair has been increasingly common in different areas of health. OBJECTIVE: This study aims to comparatively evaluate the effects of 660 nm laser photobiomodulation, ozone therapy, and ozonated oil on repair through histomorphometric analysis in skin wounds in rats. Forty Wistar rats will be divided into 4 groups of 10 animals each, Control Group (GC), Laser Group (LG), Ozone Gas Group (OGG), and Ozonated Oil Group (OOG). MATERIALS AND METHODS: Standard skin wounds will be made on the back of the animals, and the different experimental groups will be treated with the biomodulatory therapies described for three consecutive days. Five and ten days after surgery, five rats from each group will be euthanized. Skin fragments, including the wound area, will be removed for histological processing and subsequent staining of histological sections with Hematoxylin-eosin and Sirius red. Micrographs of the histological sections will be obtained and ten standard images will be captured for quantitative evaluation of the variables collagen area, number of blood vessels, and epithelium thickness. The variables infiltrate of polymorphonuclear and monomorphonuclear inflammatory cells, as well as edema, will be analyzed semiquantitatively. Statistical analysis of the study variables will be performed, with a significance level of p&lt;0.05. CONCLUSION: It is expected to verify which of the biomodulatory therapies used can favor the resolution of tissue repair, in particular, by promoting collagen biosynthesis.

Prime-Pull Immunization of Mice with a BcfA-Adjuvanted Vaccine Elicits Sustained Mucosal Immunity That Prevents SARS-CoV-2 Infection and Pathology.

Vaccines against SARS-CoV-2 that induce mucosal immunity capable of preventing infection and disease remain urgently needed. In this study, we demonstrate the efficacy of Bordetella colonization factor A (BcfA), a novel bacteria-derived protein adjuvant, in SARS-CoV-2 spike-based prime-pull immunizations. We show that i.m. priming of mice with an aluminum hydroxide- and BcfA-adjuvanted spike subunit vaccine, followed by a BcfA-adjuvanted mucosal booster, generated Th17-polarized CD4+ tissue-resident memory T cells and neutralizing Abs. Immunization with this heterologous vaccine prevented weight loss following challenge with mouse-adapted SARS-CoV-2 (MA10) and reduced viral replication in the respiratory tract. Histopathology showed a strong leukocyte and polymorphonuclear cell infiltrate without epithelial damage in mice immunized with BcfA-containing vaccines. Importantly, neutralizing Abs and tissue-resident memory T cells were maintained until 3 mo postbooster. Viral load in the nose of mice challenged with the MA10 virus at this time point was significantly reduced compared with naive challenged mice and mice immunized with an aluminum hydroxide-adjuvanted vaccine. We show that vaccines adjuvanted with alum and BcfA, delivered through a heterologous prime-pull regimen, provide sustained protection against SARS-CoV-2 infection.

Histopathological Evaluation of Nanocurcumin for Mitigation of Radiation- Induced Small Intestine Injury.

In the current study, we aimed to mitigate radiation-induced small intestinal toxicity using post-irradiation treatment with nano-micelle curcumin. Small intestine is one of the most radiosensitive organs within the body. Wholebody exposure to an acute dose of ionizing radiation may lead to severe injuries to this tissue and may even cause death after some weeks. This study aimed to evaluate histopathological changes in the small intestine following whole-body irradiation and treatment with nanocurcumin. Forty male Nordic Medical Research Institute mice were grouped into control, treatment with 100 mg/kg nano-micelle curcumin, whole-body irradiation with cobalt-60 gamma-rays (dose rate of 60 cGy/min and a single dose of 7 Gy), and treatment with 100 mg/kg nano-micelle curcumin 1 day after whole-body irradiation for 4 weeks. Afterward, all mice were sacrificed for histopathological evaluation of their small intestinal tissues. Irradiation led to severe damage to villi, crypts, glands as well as vessels, leading to bleeding. Administration of nano-micelle curcumin after whole-body irradiation showed a statistically significant improvement in radiation toxicity of the duodenum, jejunum and ileum (including a reduction in infiltration of polymorphonuclear cells, villi length shortening, goblet cells injury, Lieberkühn glands injury and bleeding). Although treatment with nano-micelle curcumin showed increased bleeding in the ileum for non-irradiated mice, its administration after irradiation was able to reduce radiation-induced bleeding in the ileum. Treatment with nano-micelle curcumin may be useful for mitigation of radiationinduced gastrointestinal system toxicity via suppression of inflammatory cells' infiltration and protection against villi and crypt shortening.

Analysis of Images of Neutrophil Lymphocyte Ratio and Bacteria Density Level of Helicobacter Pylori in Tissue in Gastritis Patients

Gastritis is an inflammatory disease that can last for a long time. Helicobacter pylori infection is known as the main cause of peptic ulcer and the most common cause of gastritis. Helicobacter Pylori infection will cause mononuclear (MN) and polymorphonuclear (PMN) cell infiltration and will increase the production of proinflammatory cytokines in the gastric mucosa compared to non-Helicobacter Pylori gastritis. Helicobacter Pylori with histopathological examination diagnosis. This type of research is descriptive observational with purposive sampling data collection technique at the Anatomical Pathology Laboratory Installation, RSPAL dr. Ramelan Surabaya. The data used are the results of a biopsy of 60 samples that were determined as gastritis with histopathological examination of the identification of Helicobacter Pylori bacteria in gastric tissue classified by the level of bacterial density using The Updated Sydney System, as well as haemogram data for complete blood examination to further calculate the neutrophil lymphocyte ratio (NLR). The data were processed using descriptive statistical tests to determine the average neutrophil lymphocyte ratio (NLR). Based on the statistics of the r-Spearman test, the density of Helicobacter Pylori, (Pν = 0.000) below &lt; 0.005 indicates that there is a strong relationship between the neutrophil lymphocyte ratio (NLR) and the density of Helicobacter Pylori bacteria.

RETRACTED: Centella asiatica extract in carboxymethyl cellulose at its optimal concentration improved wound healing in mice models

Centella asiatica (C. asiatica) has reported to be one of the traditional herbal remedies, whereas poor water solubility leads to lower bioavailability thereby affecting it remedial efficacy. Therefore, we aimed to evaluate its efficacy through increased bioavailability by using high viscosity Carboxymethyl Cellulose (CMC) as solvent on methanol-based extract on wound healing, in vivo. The preparation was applied as 0.0% (control, CMC alone), 0.25. 0.5 and 1% concentrations of extract of C.asiatica. We evaluated the efficiency of preparations on wound healing progression as progression of wound contraction, tissue proliferation and cells deposition, and relative level of gene expression for genes associated with wound healing. The results showed that 0.5% extract in CMC had significantly higher (P < 0.05) wound contraction than control and other concentrations. The level tissue deposition and the infiltration of polymorphonuclear cells in groups treated with 0.5 % concentration preparation were higher than that other treatments and control. Similarly, the relative level of gene expression in 0.5% concentration treated group were statistically significantly higher (P < 0.05) than that of control. It is believed that the lower concentration of the extract would have lessor effect on wound healing, whereas higher concertation would be interfering the optimal inflammatory tissue deposition; and there by negatively affecting wound healing. The results indicated that C.asiatica can be optimally used at 0.5 % of extract in CMC for wound healing as indicated by speeding the progression of wound closure and by increasing the expression of collagen II and III together with reducing the expression of TGFβ1. However, higher concentrations of the crude extract of C.asiatica could paradoxically resulting in undesired effects. It is recommended that further evaluation should be performed on wider scale and the economic feasibility evaluation should be performed.

SerpinB3 administration protects liver against ischemia-reperfusion injury.

We have investigated the change in SerpinB3 during hepatic ischemia and the potential role of its antiprotease activity in cell protection by the administration of wild-type SerpinB3 (SerpinB3-WT) or active loop-deleted recombinant SerpinB3 protein (SerpinB3-D) in a rat model of ischemia (60 min)/reperfusion (60 min) (I/R). A time-dependent increase of SerpinB3, both at transcription and protein level, was found in ischemic livers after 60, 120 and 180 min. SerpinB3-WT decreased polymorphonuclear cell infiltration and serum enzymes and increased ATP when compared with I/R group. These events were not obtained using SerpinB3-D. No significant changes in both liver SerpinB3 mRNA and protein were found in all I/R groups considered. The present data show that the administration of SerpinB3-WT reduced the I/R injury and this effect appears to be dependent on its anti-protease activity.