Induce Granuloma Formation Research Articles

Safety and Immunogenicity of Recombinant Bacille Calmette-Guérin Strain VPM1002 and Its Derivatives in a Goat Model.

A more effective vaccine against tuberculosis than Bacille Calmette-Guérin (BCG) is urgently needed. BCG derived recombinant VPM1002 has been found to be more efficacious and safer than the parental strain in mice models. Newer candidates, such as VPM1002 Δpdx1 (PDX) and VPM1002 ΔnuoG (NUOG), were generated to further improve the safety profile or efficacy of the vaccine. Herein, we assessed the safety and immunogenicity of VPM1002 and its derivatives, PDX and NUOG, in juvenile goats. Vaccination did not affect the goats' health in regards to clinical/hematological features. However, all three tested vaccine candidates and BCG induced granulomas at the site of injection, with some of the nodules developing ulcerations approximately one month post-vaccination. Viable vaccine strains were cultured from the injection site wounds in a few NUOG- and PDX- vaccinated animals. At necropsy (127 days post-vaccination), BCG, VPM1002, and NUOG, but not PDX, still persisted at the injection granulomas. All strains, apart from NUOG, induced granuloma formation only in the lymph nodes draining the injection site. In one animal, the administered BCG strain was recovered from the mediastinal lymph nodes. Interferon gamma (IFN-γ) release assay showed that VPM1002 and NUOG induced a strong antigen-specific response comparable to that elicited by BCG, while the response to PDX was delayed. Flow cytometry analysis of IFN-γ production by CD4+, CD8+, and γδ T cells showed that CD4+ T cells of VPM1002- and NUOG-vaccinated goats produced more IFN-γ compared to BCG-vaccinated and mock-treated animals. In summary, the subcutaneous application of VPM1002 and NUOG induced anti-tuberculous immunity, while exhibiting a comparable safety profile to BCG in goats.

High prevalence of Ichthyophonus sp. infections in Northeast Atlantic mackerel (Scomber scombrus).

High prevalence of Ichthyophonus sp. infections in Northeast Atlantic mackerel (Scomber scombrus).

AN UNUSUAL CASE REPORT OF SARCOIDOSIS IN AN HIV PATIENT

SESSION TITLE: Fellows Diffuse Lung Disease Posters SESSION TYPE: Fellow Case Report Posters PRESENTED ON: October 18-21, 2020 INTRODUCTION: Coexistence of sarcoidosis in HIV patient is a rare phenomenon and has been attributed to the impairment of immune system that may interfere with granuloma formation.(1) We present an unusual case report of sarcoidosis in HIV patient. CASE PRESENTATION: 58-year-old female with history of HIV on highly active antiretroviral therapy (HAART) (CD4 count: 746 and viral load <20/ml) presented with incidental basilar pulmonary nodules during work up for abdominal pain. Subsequent CT chest showed multiple nodular densities in all lung fields with hilar and mediastinal lymphadenopathy. She is lifetime nonsmoker and had no respiratory symptoms. PET-CT showed avid mediastinal lymph nodes and bone lesions. Bronchoscopy with EBUS guided TBNA was inconclusive with negative fungal and mycobacterial cultures. Thereafter, IR guided bone biopsy was pursued which revealed non-caseating ” naked ” granulomas. Diagnosis of sarcoidosis was established after excluding other granulomatous inflammation, infection, and malignancy. No treatment was initiated as the patient was asymptomatic. DISCUSSION: The introduction of HAART has led to sustained increase in CD4+ T lymphocyte cells, consequently leading to the development of so called “immune restoration disease”. Sarcoidosis is an immune mediated systemic disease resulting from exaggerated cellular immune response to a variety of antigens or self-antigens, in which T lymphocyte triggering, proliferation, and activation is sustained by the CD4+ type 1 (TH1) lymphocytes. TH1 cells produce IFN-?, IL-2, and TNF leading to attraction and activation of mononuclear phagocytes that induce granuloma formation. These immunological features are not clearly exhibited in coexistent sarcoidosis/HIV infection. Before HAART therapy, reports indicated a prevalence of lymphocytic alveolitis due to an increase in CD8 cells, with a very low CD4/CD8 ratio. In a retrospective cohort study, it was concluded that the development of sarcoidosis appears to depend on preservation or restoration of peripheral CD4+ lymphocyte count exceeding 200 cells/µl compared to alternative specific etiologies of granulomatous inflammation.(2) It is worth noting that in all reported clinical cases, sarcoidosis developed several months after HAART therapy compared to acute onset of other typical immune reconstitution syndromes.(1) The need to initiate steroid therapy, as in other immune reconstitution granulomatous processes, remains controversial and is based on patient’s symptoms. CONCLUSIONS: Our case highlights the importance of considering “immune restoration disease” including sarcoidosis and other granulomatous inflammation/infection in HIV patient on HAART therapy with abnormal chest imaging in addition to slow growing lymphoproliferative disorder. Reference #1: Sarcoidosis and HIV infection: a case report and a review of the literature, Postgrad Med J. 2003;79:535–38 Reference #2: Sarcoidosis Following HIV Infection, Chest 2003 Sep; 124 (3), 929–935 DISCLOSURES: No relevant relationships by Moises Cossio, source=Web Response No relevant relationships by Domingo Franco-Palacios, source=Web Response No relevant relationships by Kushagra Gupta, source=Web Response No relevant relationships by Shahzad Hussain, source=Web Response No relevant relationships by Venkateswara Kollipara, source=Web Response No relevant relationships by Toribiong Uchel, source=Web Response

Schistosoma mansoni FES Tyrosine Kinase Involvement in the Mammalian Schistosomiasis Outcome and Miracidia Infection Capability in Biomphalaria glabrata.

Schistosomiasis is a neglected tropical disease (NTD) caused by helminthes from the Schistosoma genus. This NTD can cause systemic symptoms induced by the deposition of parasite eggs in the host liver, promoting severe complications. Functional studies to increase knowledge about parasite biology are required for the identification of new drug targets, because the treatment is solely based on praziquantel administration, a drug in which the mechanism of action is still unknown. Protein kinases are important for cellular adaptation and maintenance of many organisms homeostasis and, thus, are considered good drug targets for many pathologies. Accordingly, those proteins are also important for Schistosoma mansoni, as the parasite relies on specific environmental signals to develop into its different stages. However, the specific roles of protein kinases in S. mansoni biology are not well understood. This work aims at investigating the tyrosine-protein kinase FES (Feline Sarcoma) functions in the maintenance of S. mansoni life cycle, especially in the establishment of mammalian and invertebrate hosts’ infection. In this regard, the verification of Smfes expression among S. mansoni stages showed that Smfes is more expressed in infective free-living stages: miracidia and cercariae. Schistosomula exposed to SmFES-dsRNA in vitro presented a reduction in movement and size and increased mortality. Mice infected with Smfes-knocked-down schistosomula exhibited a striking reduction in the area of liver granuloma and an increased rate of immature eggs in the intestine. Female adult worms recovered from mice presented a reduced size and changes in the ovary and vitellarium; and males exhibited damage in the gynecophoral canal. Subsequently, miracidia hatched from eggs exposed to SmFES-dsRNA presented changes in its capability to infect and to sense the snail mucus. In addition, the SmFES RNAi effect was stable from miracidia to cercariae. The establishment of infection with those cercariae reproduced the same alterations observed for the knocked-down schistosomula infection. Our findings show that SmFES tyrosine kinase (1) is important in schistosomula development and survival; (2) has a role in adult worms pairing and, consequently, female maturation; (3) might be essential for egg antigen expression, thus responsible for inducing granuloma formation and immunomodulation; and (4) is essential for miracidia infection capability. In addition, this is the first time that a gene is kept knocked down during three different S. mansoni life stages and that a tyrosine kinase is implicated in the parasite reproduction and infection establishment in the mammalian host. Accordingly, SmFES should be explored as an alternative to support schistosomiasis treatment and morbidity control.

Anti-Inflammatory Effect of Ethanolic Extract of Carica Papaya Leaves and Indomethacin in Cotton Pellet Induced Granuloma in Animal Model

Background: Anti-inflammatory effect can be exerted by Carica papaya.&#x0D; Objective: The purpose of the present study was to see the anti-inflammatory effect of ethanolic extract of Carica papaya leaves and indomethacin in carrageenan induced rat paw edema animal model.&#x0D; Methodology: This was an animal study carried out in the Department of Pharmacology at Dhaka Medical College, Dhaka, Bangladesh during the period from July 2014 to June 2015 for a period of one (01) year. The leaves of Carica Papaya collected from Botanical garden, Mirpur, Dhaka, Bangladesh. The animals were divided into four groups. The animals were divided into four groups. In all the animals granuloma were induced by implantation of autoclaved cotton pellet on the 1st day and they treated as follows: Group I were served as control that was received normal saline. Group II were received ethanolic extract of Carica Papaya leaves. Group III were received ethanolic extract of Carica Papaya leaves. Group IV were received indomethacin. On the 15th day animals were anesthetized, implanted pellets were dissected out, dried at hit air oven and the final weights were measured.&#x0D; Result: The experiment was carried out on 48 Long Evan Norwegian rats. Rats were divided in 4 groups of six animals each. The mean increase of weight of cotton pellet was 48.06±1.02, 24.48±0.36, 22.42±0.56 and 18.02±0.43 in group I, II, III and IV respectively. The inhibition of granuloma formation was 49.06%, 53.35% and 62.51% in group II, III and IV respectively.&#x0D; Conclusion: In conclusion the ethanolic extract of Carica Papaya leaves has effects on cotton pellet induced granuloma formation in experimental rat.&#x0D; Journal of Current and Advance Medical Research 2019;6(1):2-5

Evaluation of Anti-Inflammatory Activities of a Triterpene β-Elemonic Acid in Frankincense In Vivo and In Vitro.

The purpose of this research was to extract and separate the compounds from frankincense, and then evaluate their anti-inflammatory effects. The isolated compound was a representative tetracyclic triterpenes of glycine structure according to 1H-NMR and 13C-NMR spectra, which is β-elemonic acid (β-EA). We determined the content of six different localities of frankincense; the average content of β-EA was 41.96 mg/g. The toxic effects of β-EA administration (400, 200, 100 mg/kg) for four weeks in Kunming (KM) mice were observed. Compared with the control group, the body weight of mice, the visceral coefficients and serum indicators in the β-EA groups showed no systematic variations. The anti-inflammatory effects of β-EA were evaluated in LPS-induced RAW264.7 cells, xylene-induced induced ear inflammation in mice, carrageenin-induced paw edema in mice, and cotton pellet induced granuloma formation in rats. β-EA inhibited overproduction of tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6), monocyte chemotactic protein 1 (MCP-1), soluble TNF receptor 1 (sTNF R1), Eotaxin-2, Interleukin 10 (IL-10) and granulocyte colony-stimulating factor (GCSF) in the RAW264.7 cells. Intragastric administration with β-EA (300, 200, and 100 mg/kg in mice, and 210, 140, and 70 mg/kg in rats) all produced distinct anti-inflammatory effects in vivo in a dose-dependent manner. Following treatment with β-EA (300 mg/kg, i.g.), the NO level in mice ears and PGE2 in mice paws both decreased (p < 0.01). In conclusion, our study indicates that β-EA could be a potential anti-inflammatory agent for the treatment of inflammatory diseases.

MmpL8MAB controls Mycobacterium abscessus virulence and production of a previously unknown glycolipid family

Mycobacterium abscessus is a peculiar rapid-growing Mycobacterium (RGM) capable of surviving within eukaryotic cells thanks to an arsenal of virulence genes also found in slow-growing mycobacteria (SGM), such as Mycobacterium tuberculosis A screen based on the intracellular survival in amoebae and macrophages (MΦ) of an M. abscessus transposon mutant library revealed the important role of MAB_0855, a yet uncharacterized Mycobacterial membrane protein Large (MmpL). Large-scale comparisons with SGM and RGM genomes uncovered MmpL12 proteins as putative orthologs of MAB_0855 and a locus-scale synteny between the MAB_0855 and Mycobacterium chelonae mmpL8 loci. A KO mutant of the MAB_0855 gene, designated herein as mmpL8MAB , had impaired adhesion to MΦ and displayed a decreased intracellular viability. Despite retaining the ability to block phagosomal acidification, like the WT strain, the mmpL8MAB mutant was delayed in damaging the phagosomal membrane and in making contact with the cytosol. Virulence attenuation of the mutant was confirmed in vivo by impaired zebrafish killing and a diminished propensity to induce granuloma formation. The previously shown role of MmpL in lipid transport prompted us to investigate the potential lipid substrates of MmpL8MAB Systematic lipid analysis revealed that MmpL8MAB was required for the proper expression of a glycolipid entity, a glycosyl diacylated nonadecyl diol (GDND) alcohol comprising different combinations of oleic and stearic acids. This study shows the importance of MmpL8MAB in modifying interactions between the bacteria and phagocytic cells and in the production of a previously unknown glycolipid family.

Iatrogenic Postvaccinal Injection Site Granulomas in Cattle

Background: The Brazilian government has established disease eradication and control programs to protect livestock from pathogens that affect animal health and compromise animal protein production and food safety and quality. Vaccination campaigns against foot-and-mouth disease and other infectious diseases in cattle except brucellosis can be carried out bynon-veterinarian employees. However, vaccination errors can result in the formation of granulomas at injection sites that can affect animal welfare and production. The present study aimed to report two cases of granulomas at injection sites due to the inadvertent administration of vaccines containing saponins and mineral oil as adjuvants.Cases: Case 1. The history was that a 2-year-old Devon steer was down for 7 days and was vaccinated 20 days ago with a vaccine containing saponins and mineral oil as adjuvants. Case 2. A 7-month-old Holstein heifer was examinated due to a 40-day history of ataxia, forelimb paresis progressing to tetraparesis, and vaccination with a vaccine containing saponins and mineral oil as adjuvants 60 days ago. These two animals were admitted in the Veterinary Hospital from the University of Passo Fundo (UPF) with a clinical history of incoordination and permanent decubitus after vaccination. The disease had a similar clinical course in both animals. Clinical signs included the presence of a palpable cervical mass at the site of vaccination, forelimb paresis that progressed to tetraparesis, and decubitus scars. Treatment included intravenousadministration of anti-inflammatory steroids, antibiotic therapy, daily cleansing of the pressure sores, all four limbs were massaged, animals underwent several physiotherapy sessions, slings were used to mantain recumbent patients in a standing position and supportive therapy consisted of fluid therapy and oral supplementation. Animals remained hospitalizedfor approximately 40 days. Pacients experienced temporary improvement during treatment, and would walk with an uncoordinated gait. The clinical picture worsened after treatment was discontinued, necessitating euthanasia. At necropsy, gross lesions were similar in both animals. In case 1, the trapezius was firm and pale and had multiple granulomas whichextended into the cervical vertebral column at C3-C4 and invaded the spinal canal compressing the spinal cord. In case 2, there was extensive damage to the trapezius by granulomatous inflammation; numerous nodular granulomas exuded milky contents. These lesions extended deep into the muscle fibers and infiltrated the vertebral column at C5-C6, with involvement of the medullary canal and spinal cord compression. Microscopically there was severe, diffuse pyogranulomatous myositis. Each pyogranuloma had a central clear space. Nodal architecture was effaced by these inflammatory nodules.Discussion: Some drugs and vaccines contain irritating adjuvants and induce granuloma formation at the inoculation site. In both cattle, vaccines were injected intramuscularly in the cranial third of the neck in close proximity to the cervical vertebrae and surrounding tissues. These vaccines possibly induced an exuberant inflammatory reaction at the inoculationsite. An exacerbated inflammatory response following the administration of adjuvanted vaccines by improperly trained personnel caused substantial tissue damage at the injection site. Severe, locally extensive lesions were found at necropsy affecting adjacent structures including skeletal muscles and spinal cord. The clinical signs of ataxia and forelimb paresis thatprogressed to tetraparesis were due to the marked pyogranulomatous inflammation in C3-C4 in case 1 and in C5-C6 in case 2. The present study reinforces the importance of good farming practices and properly trained personnel working at farms.Keywords: bovine, vaccine, infection site, granuloma, iatrogenic.

Retinal Pigment Epithelial Cells Control Early Mycobacterium tuberculosis Infection via Interferon Signaling.

Mycobacterium tuberculosis (Mtb) bacilli have been found in retinal pigment epithelial (RPE) cells from uveitis patients without signs of systemic tuberculosis (TB) infection. RPE cells are important for ocular immune privilege and uveitis development. To address a potential role for Mtb-infected RPE cells in the development of uveitis, we delineated the response to Mtb infection in human RPE cells and primary human macrophages, the main target cell of Mtb. Primary human RPE cells, the human RPE cell line ARPE-19, and monocyte-derived proinflammatory M1 and anti-inflammatory M2 macrophages were infected with DsRed-expressing Mtb strain H37Rv. Infection rates and clearance were addressed along with RNA sequencing analysis, a confirmation analysis by dual-color reverse-transcriptase multiplex ligation-dependent probe amplification (dcRT-MLPA) and cytokine secretion. RPE cells robustly controlled intracellular outgrowth of Mtb early after infection. The response in RPE cells to control Mtb survival was dominated by interferon (IFN) signaling and further characterized by prominent regulation of cell death/survival-associated genes and low-level production of Th1-associated cytokines. In contrast, macrophages engaged a plethora of responses including IFN signaling and communication between innate and adaptive immune cells to induce granuloma formation. Together, our data demonstrate that RPE cells display a strong response to Mtb infection that appears, however, incomplete in comparison to the macrophage response to Mtb. The RPE response might reflect a balance between mechanisms aimed at Mtb eradication and mechanisms that limit retinal inflammation.

Mycobacterial cord factor enhances migration of neutrophil-like HL-60 cells by prolonging AKT phosphorylation.

Trehalose 6,6'-dimycolate (TDM), or cord factor, is a crucial stimulus of immune responses during Mycobacterium tuberculosis infection. Although TDM has immuno-stimulatory properties, including adjuvant activity and the ability to induce granuloma formation, the mechanisms underlying these remain unknown. We hypothesized that TDM stimulates transendothelial migration of neutrophils, which are the first immune cells to infiltrate the tissue upon infection. In this study, it was shown that TDM enhances N-formylmethionyl-leucyl-phenylalanine (fMLP)-induced chemotaxis and transendothelial movement by prolonging AKT phosphorylation in human neutrophils. TDM induced expression of macrophage-inducible C-type lectin, a receptor for TDM, and induced secretion of pro-inflammatory cytokines and chemokines in differentiated HL-60 cells. In 2- and 3-D neutrophil migration assays, TDM-stimulated neutrophils showed increased fMLP-induced chemotaxis and transendothelial migration. Interestingly, following fMLP stimulation of TDM-activated neutrophils, AKT, a crucial kinase for neutrophil polarization and chemotaxis, showed prolonged phosphorylation at serine 473. Taken together, these data suggest that TDM modulates transendothelial migration of neutrophils upon mycobacterial infection through prolonged AKT phosphorylation. AKT may therefore be a promising therapeutic target for enhancing immune responses to mycobacterial infection.

Anti-inflammatory and Analgesic Effects of Polygonum orientale L. Extracts.

Background and Purpose: Polygonum orientale L. (family: Polygonaceae), named Hongcao in China, is a Traditional Chinese Medicinal and has long been used for rheumatic arthralgia and rheumatoid arthritis. However, no pharmacological and mechanism study to confirm these clinic effects have been published. In this investigation, the anti-inflammatory, analgesic effects and representative active ingredient compounds of P. orientale have been studied.Methods: Dried small pieces of the stems and leaves of P. orientale were decocted with water and partitioned successively to obtain ethyl acetate and ethyl ether extract of P. orientale (POEa and POEe). Chemical compositions of them were analyzed by UPLC-Q-Exactive HRMS. Anti-inflammatory and analgesic effects of POEa and POEe were evaluated using xylene induced ear edema, carrageenan induced paw edema, Freunds’ complete adjuvant induced arthritis, and formaldehyde induced pain in rat. Their mechanisms of anti-inflammatory and analgesic effects were also studied via assays of TNF-α, IL-1β, IL-6, and PGE2 in serum.Results: UPLC-Q-Exactive HRMS analysis showed that POEa and POEe mainly contained flavonoids including orientin, isoorientin, vitexin, luteolin, and quercetin. Furthermore, anti-inflammatory effects of POEa and POEe were evident in xylene induced ear edema. The paw edema in Freund’s complete adjuvant and carrageenan were significantly (P < 0.05, 0.01) inhibited by POEa (5, 7.5 g/kg). POEe (7.5 g/kg) was significantly (P < 0.05, 0.01) inhibited Freunds’ complete adjuvant induced paw edema and cotton pellet induced granuloma formation. Similarly, POEe significantly (P < 0.05, 0.01) inhibited the pain sensation in acetic acid induced writhing test. POEa (5, 7.5 g/kg) significantly (P < 0.05, 0.01) inhibited formaldehyde induced pain in both phases. POEa (7.5 g/kg) markedly (P < 0.05) prolonged the latency period of hot plate test after 30 and 60 min. The concentrations of TNF-α, IL-1β, IL-6, and PGE2 were significantly (P < 0.01) decreased by POEa (3.75, 5 g/kg).Conclusion: POEa and POEe have anti-inflammatory and analgesic effects, which was mainly relevant to the presence of flavonoids, including orientin, isoorientin, vitexin, luteolin, and quercetin. The mechanism of anti-inflammatory and analgesic effects of POEa may be to decrease the concentrations of TNF-α, IL-1β, IL-6, and PGE2 in serum.

Matrix Metalloproteinase-9 (MMP-9) Level in Tuberculosis Exposed and Infected Children

Background: Lung Tuberculosis (TB) is a chronic infectious disease caused by Mycobacterium tuberculosis (M. Tb). Excessive body's immune response to M. Tb will contribute to the host tissue damage. M. Tb infection stimulates gene expression and MMP-9 secretion from monocyte which is infected by M. Tb. M. Tb molecular mechanisms inducing granuloma formation which is the ESAT-6 protein that stimulates the production of matrix metalloproteinase-9 (MMP-9) from epithelial cells near the infected macrophages. Objective: To evaluate the levels of Matrix Metalloproteinase-9 (MMP-9) in children who lived in the same house with a tuberculosis person. Methods: A cross sectional study was conducted at the Lung Health Community Center (BBKPM) Makassar, from May to July 2015. The study population was children aged 3 months to 18 years. Results: Levels of MMP-9 were examined in 55 samples of the study, 29 samples infected with TB (52.7%), and 26 samples exposed to TB (47.3%). There is no significant difference in the levels of MMP-9 and genders, age, nutritional status, status of BCG, as well as the levels of MMP-9 in the group of exposed and infected to TB (p>0.05). Conclusion: There was no significant differences between the levels of MMP-9 in the group of exposed and infected to TB.

Pulmonary fibrosis in a mouse model of sarcoid granulomatosis induced by booster challenge withPropionibacterium acnes

Pulmonary fibrosis (PF) associated with chronic sarcoidosis remains poorly understood, and no experimental model is currently available for this condition. Previous studies have shown that Propionibacterium acnes (PA) was associated with sarcoidosis and induced granuloma formation in mice. Here, we investigated whether repeated challenge with PA induces persistent inflammation leading to sarcoidosis followed by PF in mice. Specifically, C57BL/6 mice were inoculated intraperitoneally and subjected to intratracheal challenge with PA, and then were booster-challenged with either PA or phosphate-buffered saline on day 28. Inflammation, granulomata, and features of fibrosis were evaluated every 7 days until day 70. Complete remission of lung granulomata was apparent on day 42 in the sarcoid-remission group. However, granulomata was present from days 21 to 70 in mice that received PA boosting. Inflammatory cell counts and Th1 cytokine levels in lung lavage fluids were elevated up to day 70. Furthermore, fibrotic changes in the lungs were observed around granulomatous and peribronchovascular regions after PA boosting. Taken together, these findings suggest that development of PF following sarcoidosis may result from continuous PA infection and inflammation. Repeated boosting with PA to induce PF might be a useful model for future studies of sarcoidosis-associated PF.

Anti-inflammatory, analgesic and anti-pyretic activities of standardized root extract of Jasminum sambac

Ethnopharmacological relevanceThe plant Jasminum sambac L. (Oleaceae) is cultivated throughout India. The leaves and roots of the plant are used traditionally in the treatment of inflammation, fever and pain. The leaves of the plant have been reported to posses significant anti-inflammatory and analgesic activities. Objective: To scientifically validate anti-inflammatory, analgesic and anti-pyretic activities of roots from Jasminum sambac. Materials and methodsEthanol root extract of Jasminum sambac (EJS) was standardized using HPTLC and was subjected to acute oral toxicity study. Further, analgesic activity of EJS at 100, 200 and 400mg/kg, p.o. was evaluated using writhing test on Swiss albino mice and tail-flick test on Charles Foster albino rats. Anti-inflammatory activity of EJS was assessed by carrageenan-induced rat paw edema, cotton pellet-induced granuloma and Freund׳s adjuvant-induced arthritis models, while antipyretic activity was evaluated using Brewer׳s yeast induced pyrexia. In addition, biochemical parameters such as alkaline phosphatase (ALP), aspartate transaminase (AST), alanine transaminase (ALT), lipid peroxidation (LPO), superoxide dismutase (SOD) and catalase (CAT) in blood serum and edematous tissue of rats exposed to acute (carrageenan) and granulomatous tissue in sub-chronic (cotton pellet granuloma) inflammation models were also evaluated. ResultsPhytochemical analysis of EJS revealed the presence of flavonoids, phenols, saponins, tannins and carbohydrates in major quantities, while the quantity of hesperidin in EJS (using HPTLC) was found to be 4.25%w/w. EJS at 400mg/kg, p.o. reduced writhing count up to 49.21%, whereas in tail-flick test, EJS in a dose dependent manner increased latency in flicking tail. EJS at 400mg/kg, p.o. showed significant anti-inflammatory activity after 2nd, 3rd, 4th and 6thh of treatment in carrageenan-induced edema, while a 33.58% inhibition in cotton pellet induced granuloma formation was observed at same dose level. EJS significantly (p<0.001) inhibited adjuvant-induced arthritis and also showed significant antipyretic activity. Further, a significant reversal in alterations of all the biochemical parameters (except ALP) in tissues was also observed. ConclusionsThe study confirms the anti-inflammatory, analgesic and antipyretic activity of EJS which may be attributed to the presence of various phytoconstituents quantified especially hesperidin which have already been reported for its significant role in the treatment of inflammation and associated problems.

Multiwall Carbon Nanotubes Directly Promote Fibroblast-Myofibroblast and Epithelial-Mesenchymal Transitions through the Activation of the TGF-β/Smad Signaling Pathway

A number of studies have demonstrated that MWCNTs induce granuloma formation and fibrotic responses in vivo, and it has been recently reported that MWCNT-induced macrophage activation and subsequent TGF-β secretion contribute to pulmonary fibrotic responses. However, their direct effects against alveolar type-II epithelial cells and fibroblasts and the corresponding underlying mechanisms remain largely unaddressed. Here, MWCNTs are reported to be able to directly promote fibroblast-to-myofibroblast conversion and the epithelial-mesenchymal transition (EMT) through the activation of the TGF-β/Smad signaling pathway. Both of the cell transitions may play important roles in MWCNT-induced pulmonary fibrosis. Firstly, in-vivo and in-vitro data show that long MWCNTs can directly interact with fibroblasts and epithelial cells, and some of them may be uptaken into fibroblasts and epithelial cells by endocytosis. Secondly, long MWCNTs can directly activate fibroblasts and increase both the basal and TGF-β1-induced expression of the fibroblast-specific protein-1, α-smooth muscle actin, and collagen III. Finally, MWCNTs can induce the EMT through the activation of TGF-β/Smad2 signaling in alveolar type-II epithelial cells, from which some fibroblasts involved in pulmonary fibrosis are thought to originate. These observations suggest that the activation of the TGF-β/Smad2 signaling plays a critical role in the process of the fibroblast-to-myofibroblast transition and the EMT induced by MWCNTs.

Inhibition of foreign body giant cell formation by 4- hexylresorcinol through suppression of diacylglycerol kinase delta gene expression

Grafted macromolecules often induce granuloma formation with foreign body giant cell (FBGC) infiltration, and this is the main reason for graft failure. Diacylglycerol kinase (DAGK) is an important intracellular mediator of FBGC formation in macrophages. In this study, 4-hexylresorcinol (4HR) inhibited DAGKδ in a macrophage cell line (RAW264.7 cells). As a result of DAGK-δ inhibition by 4HR, FBGC formation was significantly inhibited in RAW264.7 cells. Silk fibroin is a well-known natural macromolecule, and when it is grafted into bone defects, it results in granuloma formation with massive FBGC formation. 4HR-incorporating silk graft materials displayed significant reduction of granuloma formation and increases in the extent of new bone formation in a rabbit calvarial defect model. In conclusion, 4HR could inhibit foreign body reaction via a DAGK-mediated pathway.

Evaluation of anti‐inflammatory effect of analogs of L‐carnitine in a model of chronic inflammation in vivo (1056.1)

In this project we developed a model of chronic inflammation induced by subcutaneous implantation of cotton tassel to induce granuloma formation and evaluate drug efficacy of 3 analogs of L‐carnitine. Chronic granulomatous inflammation is a specific type of chronic inflammatory reaction, characterized by the accumulation of modified macrophages (epitheloid cells). Wistar male rats with granuloma were treated with the three analogs at a dose of 2mg/kg. Different serum interleukins were determined and histological analysis of thymus, spleen and granuloma was made. The analysis shows a reduced level of pro‐inflammatory interleukins (Il‐6 and TNF‐α) in animals treated with analogs having a greater effect the analog 2 and 3. Histological analysis showed the presence of epithelioid cells and giant cells in animals treated with L‐carnitine and analog 1, whereas animals treated with analogue 2 and 3 showed leukocyte infiltration. With regard the thymus, only was seen an increase in medulla area with the analog 2. While in the spleen was observed a reduction of medulla area with the analogs 2 and 3. In conclusion the analogs had anti‐inflammatory and anti‐granuloma activity, but they had different efficacy.Grant Funding Source: CONACYT

Foreign body granuloma secondary to leuprorelin acetate.

Dear Editor: Leuprorelin acetate is a synthetic analogue of the naturally occurring luteinizing hormone releasing hormone (LHRH), and is produced in 1- and 3-month (3.75 mg and 11.25 mg, respectively) depot preparations, typically administered intramuscularly or subcutaneously, in the upper arm or abdomen1. To date, several cases of leuprorelin acetate granuloma have been reported in the Japanese and European literature1,2, though not in Korean dermatologic journals. Herein, we report a case of a foreign body granuloma, resulting from leuprorelin acetate injections. A 44-year-old woman presented to our clinic, with three pruritic nodules on the left upper arm. Four months prior, the patient had been treated for endometriosis with leuprolide (LEUPLIN® DPS injection 11.25 mg; Takeda, Osaka, Japan), via two subcutaneous injections in the left upper arm. Three months later, three discrete nodules developed on and near the injection sites. Clinical examination revealed three, 2 to 3 cm-sized, firm, erythematous nodules on the left upper arm (Fig. 1). A 3-mm punch biopsy was performed, under the differential diagnosis of bacterial, fungal, or mycobacterial infection, traumatic panniculitis, or injection site granuloma. Pathologic findings revealed a marked degeneration of adipose tissue, and an inflammatory cell infiltrate, mainly composed of lymphohistiocytes. Foreign body giant cells with translucent intracytoplasmic vacuoles were also noted (Fig. 2), and Ziehl-Neelsen, Gram, and periodic acid-Schiff stains were all negative. The patient was treated with intralesional triamcinolone injections (2.5 mg/ml). On follow-up 1 month later, the size of the lesion was noted to have decreased. Fig. 1 Three, pruritic, 2~3-cm, erythematous nodules at the site of a previous leuprorelin injection on the left upper arm. Fig. 2 Histopathology of skin lesions. (A) An acanthotic epidermis with a marked degeneration of adipocytes, and an intense inflammatory cell infiltrate, composed of lymphocytes and neutrophils. (B) Foreign-body-type giant cells with translucent intracytoplasmic ... LHRH analogues are widely used to treat sex-hormone-responsive diseases2. Currently, two LHRH analogues are available: leuprorelin acetate (LEUPLIN®; Takeda), and goserelin acetate (Zoladex®; AstraZeneca, Cheshire, U.K.)3. In these depot preparations, LHRH is coupled to biodegradable microcapsules, composed of lactic/glycolic acid polymers1. As LHRH has lipolytic activity in vitro, the degenerated lipid granules may induce foreign body reactions4. Moreover, lactic/glycolic acid polymers remain in tissues for several months, likely further contributing to granulomatous inflammation1. Though leuprorelin acetate granulomas can in some cases be painful, most are asymptomatic. Firm, subcutaneous nodules or sterile abscesses can develop1. Histopathologic analysis of such lesions reveal multi-nucleated giant cells, containing round, translucent microspheres and granulomas, with secondary foreign body reactions4. In the case described here, leuprorelin acetate was twice injected subcutaneously in the upper arm, with erythematous nodules developing at the injection site, three months later. The histologic features were identical to those of previously reported leuprorelin acetate granulomas. Leuprorelin acetate granulomas are rare in Western countries, though many cases have been reported in Japan, with this discrepancy explained by the difference in injection methods3. In Western countries, leuprorelin acetate is administered intramuscularly, while in Korea as well as in Japan, it is generally administered subcutaneously3. In one study by Shiota et al.3, the incidence rate of leuprorelin acetate granuloma was 4.2% (5 out of 118 cases), with all cases associated with 11.25 mg leuprorelin acetate depot injections. All five patients with granulomas were transitioned to treatment with goserelin acetate, with recurrence only occurring in one individual. Based on these findings, goserelin acetate would seem to have a lower risk of inducing granuloma formation, than leuprorelin acetate. Therefore, switching the patient to intramuscular goserelin acetate is recommended.

Aluminum enhances inflammation and decreases mucosal healing in experimental colitis in mice.

The increasing incidence of inflammatory bowel diseases (IBDs) in developing countries has highlighted the critical role of environmental pollutants as causative factors in their pathophysiology. Despite its ubiquity and immune toxicity, the impact of aluminum in the gut is not known. This study aimed to evaluate the effects of environmentally relevant intoxication with aluminum in murine models of colitis and to explore the underlying mechanisms. Oral administration of aluminum worsened intestinal inflammation in mice with 2,4,6-trinitrobenzene sulfonic acid- and dextran sodium sulfate-induced colitis and chronic colitis in interleukin 10-negative (IL10−/−) mice. Aluminum increased the intensity and duration of macroscopic and histologic inflammation, colonic myeloperoxidase activity, inflammatory cytokines expression, and decreased the epithelial cell renewal compared with control animals. Under basal conditions, aluminum impaired intestinal barrier function. In vitro, aluminum induced granuloma formation and synergized with lipopolysaccharide to stimulate inflammatory cytokines expression by epithelial cells. Deleterious effects of aluminum on intestinal inflammation and mucosal repair strongly suggest that aluminum might be an environmental IBD risk factor.

Evaluation of antinociceptive and anti-inflammatory activities of extract and fractions of Eugenia jambolana root bark and isolation of phytoconstituents

Eugenia jambolana Lam., Myrtaceae, is a widely distributed and traditionally well known plant in India. The root bark of the plant was extracted with ethanol and then successively fractionated into petroleum ether fraction, chloroform fraction, n-butanol fraction and methanol fraction. The extract and fractions of the plant material were evaluated for the antinociceptive activity by acetic acid-induced writhing test and formalin-induced nociception test, and anti-inflammatory activity was screened by carrageenan-induced rat paw edema, cotton pellet induced granuloma formation and adjuvant induced arthritis in rat models. The test materials showed the antinociceptive and anti-inflammatory effect in dose dependent manner and the petroleum ether fraction was found to be most potent among the test materials. At 400mg/kg b.w., p.o. dose petroleum ether fraction significantly inhibited 54.28% writhing response and 73.77% formalin induced nociception in mice. The fraction with same dose showed significant 79.31% inhibition of carrageenan-induced rat paw edema, 57.78% anti-proliferative effect and 77.93% inhibition of adjuvant induced arthritis. The bioactive petroleum ether fraction was then subjected to column chromatography which led to isolate three compounds, namely, β-sitosterol, stigmasterol and lupeol. These compounds were characterized and identified by IR, 1H NMR, 13C NMR and mass spectroscopy.