Leukocyte Migration Inhibition Research Articles

Estimation of the phenolic content and in vivo evaluation of the anti-inflammatory and anti-ulcer activity of Rhus pentaphylla leaf and fruit extracts from Morocco

The present study aimed to estimate the phenolic compounds and evaluate the anti-inflammatory and anti-ulcer activity of Rhus pentaphylla leaf and fruit extracts in rodents. Phytochemical identification of the extracts was performed using HPLC technique. The anti-inflammatory property of the extracts was studied using three animal models of inflammation: xylene-induced ear swelling in Swiss albino mice, paw edoema and carrageenan-induced air pocketing in Sprague Dawley rats. The antiulcer activity of the extracts was evaluated through HCl/ethanol-induced acute gastric lesions in rats. HPLC results revealed the presence of epicatechin, caffeic acid and rutin in the leaf extract, while quercetin and kaempferol were identified in the fruit extract. Both leaf and fruit extracts demonstrated significant anti-inflammatory potential. Additionally, the extracts inhibited leukocyte migration and exhibited a strong gastroprotective effect. These remarkable in vivo pharmacological activities of Rhus pentaphylla extracts, linked to their bioactive compounds, demonstrate their potential for possible therapeutic applications.

Potential Role and Mechanism of Mulberry Extract in Immune Modulation: Focus on Chemical Compositions, Mechanistic Insights, and Extraction Techniques.

Mulberry is a rapidly growing plant that thrives in diverse climatic, topographical, and soil types, spanning temperature and temperate countries. Mulberry plants are valued as functional foods for their abundant chemical composition, serving as a significant reservoir of bioactive compounds like proteins, polysaccharides, phenolics, and flavonoids. Moreover, these compounds displayed potent antioxidant activity by scavenging free radicals, inhibiting reactive oxygen species generation, and restoring elevated nitric oxide production induced by LPS stimulation through the downregulation of inducible NO synthase expression. Active components like oxyresveratrol found in Morus demonstrated anti-inflammatory effects by inhibiting leukocyte migration through the MEK/ERK signaling pathway. Gallic and chlorogenic acids in mulberry leaves (ML) powder-modulated TNF, IL-6, and IRS1 proteins, improving various inflammatory conditions by immune system modulation. As we delve deeper into understanding its anti-inflammatory potential and how it works therapeutically, it is crucial to refine the extraction process to enhance the effectiveness of its bioactive elements. Recent advancements in extraction techniques, such as solid-liquid extraction, pressurized liquid extraction, superficial fluid extraction, microwave-assisted extraction, and ultrasonic-assisted extraction, are being explored. Among the extraction methods tested, including Soxhlet extraction, maceration, and ultrasound-assisted extraction (UAE), UAE demonstrated superior efficiency in extracting bioactive compounds from mulberry leaves. Overall, this comprehensive review sheds light on the potential of mulberry as a natural immunomodulatory agent and provides insights into its mechanisms of action for future research and therapeutic applications.

Inhibition of leukocyte migration after ischemic stroke by VE-cadherin mutation in a mouse model leads to reduced infarct volumes and improved motor skills.

To distinguish between the genuine cellular impact of the ischemic cascade by leukocytes and unspecific effects of edema and humoral components, two knock-in mouse lines were utilized. Mouse lines Y731F and Y685F possess point mutations in VE-cadherin, which lead to a selective inhibition of transendothelial leukocyte migration or impaired vascular permeability. Ischemic stroke was induced by a model of middle cerebral artery occlusion. Analysis contained structural outcomes (infarct volume and extent of brain edema), functional outcomes (survival analysis, rotarod test, and neuroscore), and the extent and spatial distribution of leukocyte migration (heatmaps and fluorescence-activated cell sorting (FACS) analysis). Inhibition of transendothelial leukocyte migration as in Y731F mice leads to smaller infarct volumes (52.33±4719 vs. 70.43±6483mm3 , p=.0252) and improved motor skills (rotarod test: 85.52±13.24s vs. 43.06±15.32s, p=.0285). An impaired vascular permeability as in Y685F mice showed no effect on structural or functional outcomes. Both VE-cadherin mutations did not influence the total immune cell count or spatial distribution in ischemic brain parenchyma. Selective inhibition of transendothelial leukocyte migration by VE-cadherin mutation after ischemic stroke in a mouse model leads to smaller infarct volumes and improved motor skills.

Antinociceptive Efficacy of 15-Deoxy-Δ12,14-Prostaglandin J2 Therapy in Response to Experimentally Induced Temporomandibular Joint Arthritis: A Systematic Review of Studies in Rats

The aim of the present systematic review was to assess the antinociceptive efficacy of 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) therapy in rats with experimentally induced temporomandibular joint (TMJ) arthritis. The focused question was “Is 15d-PGJ2 therapy effective in the management of TMJ nociception?” Indexed databases were searched without time and language restrictions up to and including September 2023 using different key words. Original studies were included. Risk of Bias (RoB) was assessed using the SYRCLE tool. Six studies performed in male Wistar rats with experimentally induced TMJ arthritis were included. The observation or follow-up period ranged between 45 min and 14 days. Four studies reported that 15d-PGJ2 therapy retards the production of proinflammatory cytokines in TMJ tissues. Four studies reported that 15d-PGJ2 therapy inhibits leukocyte migration and plasma extravasation in TMJ tissues. In one study, the expression of decay-accelerating factor in TMJ tissues increased after 15d-PGJ2 therapy. One study showed that 15d-PGJ2 inhibits nociception in a dose-dependent manner via the activation of peripheral kappa/delta opioid receptors. Prior sample-size-estimation (SSE) was performed in none of the studies and all studies had a high RoB. Due to a high RoB, methodological variations, and the absence of prior SSE within the included studies, it is demanding to derive an absolute verdict regarding the antinociceptive efficacy of 15d-PGJ2 therapy in response to experimentally induced TMJ arthritis.

Specific Immunological Investigation in the Management of Patients with Spine and Joint Pathology

Abstract Objective: The immunological investigation is used for the improvement of the quality of the treatment of various diseases. The aim of the study was to estimate the significance of specific immunological methods in the management of inflammatory diseases. Materials and Methods: Determination of the level of autoimmune lymphocytotoxic and granulocytotoxic antibodies (ALA and AGA) and reaction of inhibition of leukocyte migration were used for investigation of persons with orthopedic, metabolic, and lung pathology. Results: The study showed increased serum levels of ALA and AGA, decreased nonspecific LIF production and increased level of LIF in the presence of own serum, increased leukocyte migration to synovial membrane, bone, cartilage, Candida lusitaniae, Staphylococcus epidermidis, Staphylococcus aureus in persons with periprosthetic infection.. Decreased leukocyte migration to the antigens of S. aureus, Streptococcus pyogenes, Escherichia coli, and Pseudomonas aeruginosa and connective tissue antigens was determined in persons with inflammatory diseases of bones. Increased ALA, reduced LIF, and leukocyte migration enhancement to synovial membrane were revealed in impaired glucose metabolism. The persons with anemia were characterized by increased ALA, sensitization to bone and cartilage, Staphylococcus, and humoral sensitization to E. coli. Increased serum levels of ALA, AGA, decreased level of nonspecific LIF production and sensitization to connective tissue, Streptococcus and E. coli were determined in persons with impaired cholesterol level. Chronic obstructive pulmonary disease patients were characterized by sensitivity to S. aureus, P. aeruginosa, and humoral sensitization to the lung tissue. Conclusions: Inflammatory diseases are characterized by specific signs of immunological status and require definition of individual tactics of the treatment.

Індикативні особливості тканинної та мікробної сенсибілізації в патогенезі генералізованого пародонтиту, афілійованого з ревматоїдним артритом

Relevance. A lot of research has been devoted to the pathogenesis of generalised periodontitis, but very often researchers do not take into account the fact that generalised periodontitis often occurs against the background of internal diseases of the body, which cause not only special manifestations of the course of generalised periodontitis, but also can affect the results of its prevention and treatment. In this sense, rheumatoid arthritis, which is accompanied by chronic systemic connective tissue damage, is noteworthy. Purpose: to determine the place and integrative features of tissue and microbial sensitisation in the pathogenesis of generalised periodontitis associated with rheumatoid arthritis. Material and methods of the study. Immunological studies were performed in 335 patients divided into 3 groups: Group 1 - the main group - consisted of 136 patients with GP in the setting of RA. Group 2 - control group - 71 patients with GP diseases without signs of RA. Group 3 - comparative - 128 people without GP disease, who had musculoskeletal diseases not related to RA and practically healthy people. The periodontal condition of patients was assessed in two groups using the following parameters: Silness-Loe index (PI), bleeding index (BOP), probing pocket depth (PPD), recession level (REC), clinical attachment level (CAL) and tooth loss. To determine microbial and tissue sensitisation, the leukocyte migration inhibition reaction was used with microbial antigens of streptococci and staphylococci, and tissue antigens (bone and synovial). All the data obtained were processed by the mathematical method of variation statistics. Results of the study. In the majority of patients with GP affiliated with RA, a pronounced sensitisation to streptococcal antigen was detected, which in the chronic course of GP was 70.3±4.3%. The acute course in this group was accompanied by a high level of sensitisation in a slightly lower percentage of patients in this group and amounted to 62.5±7.0%. In the control group, the frequency of sensitised individuals to streptococcal antigen was significantly lower and amounted to 37.5±8.9 in the chronic course and 44.0±6.1 in the acute course. In practically healthy people without pathology of internal organs, musculoskeletal system and without GP disease, this indicator was only 26.0±8.0. Conclusion. The presence of microbial and tissue sensitisation in patients with GP and RA indicates that similar and cross antigens of streptococcus and staphylococcus microorganisms with connective tissue are a potential trigger for the development of both GP and RA, which is the reason to consider GP and RA as comorbid diseases with similar pathogenetic mechanisms of development. Key words: generalised periodontitis, periodontal condition, rheumatoid arthritis, sensitisation, antigen.

CORTICOSTEROIDS USAGE IN THE INTENSIVE CARE UNIT: SPECIFIC RECOMMENDATIONS BASED ON RECENT EVIDENCE

Corticosteroids are a class of drugs that mimic the effects of cortisol, a hormone naturally produced by the adrenal glands. They exert a wide range of effects on the body's immune, metabolic, and inflammatory responses. In the context of critically ill patients in the intensive care unit (ICU), corticosteroids are frequently utilised due to their potent anti-inflammatory and immunosuppressive properties. In the ICU setting, corticosteroids are commonly employed to manage various conditions such as severe sepsis, acute respiratory distress syndrome (ARDS), exacerbations of chronic obstructive pulmonary disease (COPD), and adrenal insufficiency. Their mechanism of action involves suppression of pro-inflammatory cytokines, inhibition of leukocyte migration, and stabilisation of cell membranes, among other effects. The choice of corticosteroid, dosing regimen, and duration of therapy in the ICU depends on the specific clinical condition being treated and individual patient factors. For instance, corticosteroids such as hydrocortisone are often administered to attenuate the systemic inflammatory response and improve hemodynamic stability in the management of septic shock. Dosing may vary but commonly involves an initial bolus followed by continuous infusion or intermittent dosing. In ARDS, corticosteroids may reduce lung inflammation and prevent further tissue damage. Methylprednisolone is a commonly utilised corticosteroid in this context, with dosing typically initiated at a high dose and then tapered gradually based on clinical response. For patients with exacerbations of COPD, corticosteroids help to reduce airway inflammation and improve lung function. Oral or intravenous corticosteroids such as prednisone or methylprednisolone are often prescribed for a short duration during exacerbations. In cases of adrenal insufficiency, corticosteroid replacement therapy is essential to restore physiological cortisol levels and prevent adrenal crisis. Hydrocortisone is the corticosteroid of choice in this scenario, with dosing adjusted based on the degree of adrenal dysfunction and stress level. Despite their efficacy, corticosteroids are associated with a range of potential adverse effects, including immunosuppression, hyperglycemia, fluid retention, electrolyte abnormalities, and increased risk of infection. Therefore, carefully monitoring patients receiving corticosteroid therapy in the ICU is paramount, with adjustments to minimise risks while optimising therapeutic benefits. In summary, corticosteroids play a crucial role in managing critically ill patients in the ICU, offering potent anti-inflammatory and immunomodulatory effects. However, their use requires careful consideration of the underlying condition, patient characteristics, and potential adverse effects, with dosing and duration tailored to individual needs.

Anti-inflammatory and antioxidant properties of Bertholletia excelsa(H.B.K) bark extract

Bertholletia excelsais native to the Amazon Rainforest and is popularly known as the Brazil nut. It has socioeconomic importance due its nuts being a great export product. There are fewstudies in the literature regarding the biotechnological potential of its bark, although it is used in folk medicine. The aim of this study was to determine the chemical constituents, anti-inflammatory and antioxidant properties of B. excelsabark extract(BEB). Twelve substances were identified by LC/MS/MS, and cytotoxicity tests were carried out, as well as analyses of nitric oxide production and elimination of free radicals. BEB caused cytoprotection against oxidative stress in macrophages, increased HMOX-1 expression, overcame the antioxidant effects of GPx-1 and reduced its expression and was able to inhibit leukocyte migration in use peritonitis. BEB efficiently attenuated oxidative stress due to its antioxidant and anti-inflammatory properties and, as such, can be used as a safe and effective source of a natural herbal medicine.

Metal Hypersensitivity Following Total Ankle Arthroplasty: Case Series and Literature Review

Metal hypersensitivity following total ankle arthroplasty (TAA) is an underreported complication that can manifest as dermal, systematic, and orthopaedic-specific symptoms. It is critical to recognize that metal hypersensitivity is a diagnosis of exclusion; only after all other potential sources of failure, such as loosening or infection are ruled out, can this diagnosis be considered. Aside from imaging and common laboratory analysis, skin patch testing, leukocyte migration inhibition test (LMIT), and lymphocyte transformation test (LTT) are the testing options available for metal allergy. With regards to management of metal hypersensitivity, nonoperative modalities involving topical dermatological management are generally preferred. Severe cases of metal allergy may necessitate operative management consisting of explantation of the implant and either revision TAA with a custom hypoallergenic implant or conversion to an ankle fusion. We present 3 cases of presumed metal hypersensitivity following TAA. In all 3 cases, other forms of TAA failure were carefully ruled out. One patient underwent explantation and conversion to hypoallergenic implant, 1 patient underwent explantation and ankle arthrodesis with hypoallergenic hardware, and 1 patient elected for conservative care. In patients who underwent explantation and conversion to hypoallergenic hardware, no further symptoms associated with metal allergy were noted. Additional research is necessary to improve diagnostic accuracy of metal allergy and make treatment options more effective and accessible.Level of Evidence: Retrospective case series, IV.

α1-adrenoceptor ligands inhibit chemokine receptor heteromerization partners of α1B/D-adrenoceptors via interference with heteromer formation

We reported previously that α1-adrenoceptor (α1-AR) ligands inhibit chemokine receptor (CR) heteromerization partners of α1B/D-AR. The underlying mechanisms are unknown and in vivo evidence for such effects is missing. Utilizing CCR2 and α1B-AR as prototypical partners, we observed in recombinant systems and THP-1 cells that α1B-AR enhanced whereas its absence inhibited Gαi signaling of CCR2. Phenylephrine and phentolamine reduced the CCR2:α1B-AR heteromerization propensity and inhibited Gαi signaling of CCR2. Phenylephrine cross-recruited β-arrestin-2 to CCR2, and reduced expression of α1B/D-AR, CR partners (CCR1/2, CXCR4) and corresponding heteromers. Phentolamine reduced CR:α1B/D-AR heteromers without affecting β-arrestin-2 recruitment or receptor expression. Phenylephrine/phentolamine prevented leukocyte infiltration mediated via CR heteromerization partners in a murine air pouch model. Our findings document that α1-AR ligands inhibit leukocyte migration mediated by CR heteromerization partners in vivo and suggest interference with α1B-AR:CR heteromerization as a mechanism by which CR partners are inhibited. These findings provide new insights into the pharmacology of GPCR heteromers and indicate that an agonist and antagonist at one GPCR can act as antagonists at heteromerization partners of their target receptors.

The anti-inflammatory and analgesic activities of 2Br-Crebanine and Stephanine from Stephania yunnanenses H. S.Lo.

Ethnopharmacological relevance: Crebanine (Cre) and Stephanine (Step) are isoquinoline aporphine-type alkaloids that are extracted from Stephania yunnanenses H. S. Lo. Plants of the Stephania genus are often used for treatment of stomach pain, abdominal pain, and rheumatoid arthritis. Both Cre and Step exhibit strong activities but are also associated with a certain level of toxicity, 10,11-dibrominecrebanine (2Br-Cre) is a bromine-modified derivative of Cre that we prepared and tested in order to reduce toxicity and enhance efficacy. Aim of this study: To investigate the anti-inflammatory and analgesic effects of 2Br-Cre and Step based on previous research findings and explore the specific biological mechanisms involved. Materials and methods: The anti-inflammatory and analgesic effects of 2Br-Cre and Step were investigated using a range of experimental models, including xylene-induced ear edema, carrageenan-induced pleurisy, carrageenan-induced paw edema, the hot-plate test, the naloxone antagonism test and the acetic acid writhing test. A model of chronic constriction injury (CCI) of the sciatic nerve was also established to investigate therapeutic effects. A RAW264.7 cell model was established using lipopolysaccharide (LPS) to estimate the effects of these compounds on cytokines levels. Results: 2Br-Cre and step significantly inhibited ear edema, paw edema and presented anti-inflammatory activity in the pleurisy model by inhibiting leukocyte migration and nitric oxide (NO) production, and by reducing the levels of PGE2. 2Br-Cre and Step significantly increased the pain threshold of mice subjected to heat stimulation; the effect was blocked by naloxone, thus suggesting that the analgesic effects of 2Br-Cre and Step were mediated by opioid receptors. 2Br-Cre and Step inhibited the frequency of writhing and prolonged the latency of writhing, and reduced the abnormal increase in the levels of BDNF in the serum and brain, thus alleviating the pain caused by CCI. In addition, 2Br-Cre and Step significantly inhibited the production of several inflammatory cytokines (IL-6, IL-1β and TNF-α) by LPS-induced RAW264.7 macrophages (p < .01). Conclusion: 2Br-Cre and Step exerted remarkable anti-inflammatory and analgesic effects. As a structural modification of Cre, 2Br-Cre retains the anti-inflammatory and analgesic activity of Cre but with better efficacy. Consequently, 2Br-Cre should be investigated further as a lead compound for analgesia.

Fuzapladib reduces postsurgical inflammation in the intestinal muscularis externa.

Postoperative ileus (POI) is a surgical complication that induces emesis and anorexia. Fuzapladib (FUZ), an inhibitor of leukocyte-function-associated antigen type 1 (LFA-1) activation, a leukocyte adhesion molecule, exerts anti-inflammatory effects by inhibiting leukocyte migration into the inflammatory site. In this study, we examined the prophylactic impact of FUZ on POI in a mouse model. POI model mice were generated by intestinal manipulation, and the effect of FUZ on intestinal transit and the infiltration of inflammatory cells into the ileal muscularis externa was assessed. The increased number of macrophages was significantly suppressed by FUZ, whereas the infiltration of neutrophils into the ileal muscularis externa was not sufficiently inhibited in the POI model mice. Additionally, FUZ did not ameliorate delayed gastrointestinal transit in POI model mice. In conclusion, our results suggest that FUZ does not improve delayed gastrointestinal transit but partially inhibits inflammation in the ileal muscularis externa in POI model mice. FUZ may be a potential anti-inflammatory agent for the management of post-surgical inflammation.

Antinociceptive and anti-inflammatory properties of aqueous extract obtained from Serjania marginata Casar leaves

Ethnopharmacological relevanceSerjania marginata Casar (Sapindaceae Family) Leaves are popularly used against abdominal pain.Antiulcer properties of S. marginata were scientifically described, however rare studies showed the antinociceptive effects of this plant. Aim of studyIn this study, we investigated the antinociceptive and anti-inflammatory effects of aqueous extract obtained from Serjania marginata leaves (AESM) in nociception/inflammation models. Material and methodsAESM was analyzed in FIA-ESI-IT-MS and Mass spectrometer LTQ XL. AESM oral administration (p.o.) (30, 100 and 300 mg/kg), dexamethasone subcutaneous injection (1 mg/kg, s.c.) and morphine (5 mg/kg, s.c.) were tested against the acetic acid-induced nociception, carrageenan-induced acute inflammatory paw edema/hyperalgesia, formalin-induced nociception and carrageenan-induced pleurisy in Swiss mice. ResultsFlavonoids rutin was detected in the phytochemical analysis of this extract. Oral treatment of AESM 300 mg/kg significantly reduced the number of acetic acid-induced abdominal writhing. AESM (100 and 300 mg/kg) significantly inhibited formalin-induced nociception, mechanical hyperalgesia and paw edema in carrageenan-model. Furthermore, AESM significantly inhibited leukocyte migration and protein exudation in the carrageenan-induced pleurisy test. ConclusionThis study confirms the antinociceptive, and anti-inflammatory activity of AESM, which may explain, in part, the popular use of this plant as a natural antinociceptive agent. This pharmacological action can be caused by flavonoids such as rutin and other compounds present in AESM.

Analgesic and anti-inflammatory potential of ethanolic extract from Serjania erecta leaves

Ethnopharmacological relevanceThe infusion of Serjania erecta Radlk (Sapindaceae) (popular name “cipó-cinco-folhas”) leaves is used in popular medicine to treat back pain. The anti-inflammatory, anti-hyperalgesic and anti-nociceptive properties of the ethanolic extract from S. erecta leaves (EESE) has not been yet completely clarified. Aim of the studyThe present study investigated the anti-hyperalgesic, anti-nociceptive and anti-inflammatory properties of EESE in experimental models in mice. Material and methodsEESE was fractionated by chromatographic techniques and the compound was identified by nuclear magnetic resonance (NMR), infrared (IR) spectrum, ultraviolet (UV) methods. Mice received a single dose of EESE by oral route (30, 100, and 300 mg/kg, p.o.) and were submitted to nociception induced by formalin, pleurisy induced by carrageenan and peritonitis induced by zymosan models. Mice also received EESE (30 and 100 mg/kg, p.o.) for 22 days in Complete Freund Adjuvant (CFA) model and another group received EESE for 7 days (30 and 100 mg/kg, p.o.) in pleurisy induced by Bacillus Calmette-Guerin (BCG). The cytotoxicity (MTT), phagocytic and chemotactic inhibitory activities of EESE were performed in in vitro assays. ResultsThe fractionation of EESE led to the identification of kaempferol-3-O-α-L-rhamnopyranoside. The oral administration of all doses of EESE decreased the nociceptive response induced by formalin. EESE significantly inhibited leukocyte migration in carrageenan-induced pleurisy and zymosan peritonitis models. The daily administration of EESE during for 7 days inhibited the leukocyte migration and the mycobacteria growth of pleural material obtained from animals which received BCG. EESE significantly reduced edema, cold allodynia and mechanical hyperalgesia responses induced by CFA. EESE did not induce cytotoxicity, and also decreased the leukocyte phagocytic activity, as well as, neutrophil chemotaxis. ConclusionsEESE showed analgesic and anti-inflammatory properties in acute and persistent experimental models in mice. EESE also reduced in vitro leukocyte chemotaxis and phagocytic activity without inducing cytotoxicity. The continuous oral treatment with EESE was effective against hyperalgesia and inflammation and these results could explain the popular use of S. erecta as an analgesic natural agent.

The Essential Oil of Hyptis crenata Inhibits the Increase in Secretion of Inflammatory Mediators.

Hyptis crenata is a plant of great ethnopharmacological importance widely distributed in South American countries. In Northeast Brazil, teas or infusions of its aerial parts are used in folk medicine to treat several acute and chronic inflammatory diseases. In a previous work we have demonstrated that the essential oil of H. crenata (EOHc) has an antiedematogenic effect. The aim of this work was to evaluate the effect of EOHc on cytokines secretion and cellular infiltration. Peritonitis and paw edema models induced by carrageenan were used to determine leucocyte count, myeloperoxidase (MPO) activity, nitrite, and cytokines secretion. EOHc (10-300 mg/kg) significantly inhibited leucocyte migration and reduced the neutrophil count (control: 1.46 × 103 ± 0.031 × 103/mL) of the total leucocytes population in extracellular exudate (control: 2.14 × 103 ± 0.149 × 103/mL) by 15.00%, 43.29%, 65.52%, and 72.83% for the doses of 10, 30, 100, and 300 mg/kg EOHc, respectively (EC50: 24.15 mg/kg). EOHc (100 mg/kg) inhibited the increase in myeloperoxidase activity and completely blocked the increase in nitrite concentration induced by carrageenan. EOHc markedly reduced the pro-inflammatory cytokines (IL-6, MCP-1, IFN-γ, TNF-α, and IL-12p70) and increased IL-10, an anti-inflammatory cytokine (compared to control group, p &lt; 0.05). This study demonstrates that EOHc has a long-lasting anti-inflammatory effect mediated through interference on MPO activity, and nitrite, and cytokines secretion. This effect, coupled with low EOHc toxicity, as far as results obtained in mice could be translated to humans, suggests that EOHc has great potentiality as a therapeutic agent.

A biosynthetic pathway for the selective sulfonation of steroidal metabolites by human gut bacteria.

Members of the human gut microbiome enzymatically process many bioactive molecules in the gastrointestinal tract. Most gut bacterial modifications characterized so far are hydrolytic or reductive in nature. Here we report that abundant human gut bacteria from the phylum Bacteroidetes perform conjugative modifications by selectively sulfonating steroidal metabolites. While sulfonation is a ubiquitous biochemical modification, this activity has not yet been characterized in gut microbes. Using genetic and biochemical approaches, we identify a widespread biosynthetic gene cluster that encodes both a sulfotransferase (BtSULT, BT0416) and enzymes that synthesize the sulfonate donor adenosine 3'-phosphate-5'-phosphosulfate (PAPS), including an APS kinase (CysC, BT0413) and an ATP sulfurylase (CysD and CysN, BT0414-BT0415). BtSULT selectively sulfonates steroidal metabolites with a flat A/B ring fusion, including cholesterol. Germ-free mice monocolonized with Bacteroides thetaiotaomicron ΔBT0416 exhibited reduced gastrointestinal levels of cholesterol sulfate (Ch-S) compared with wild-type B. thetaiotaomicron-colonized mice. The presence of BtSULT and BtSULT homologues in bacteria inhibited leucocyte migration in vitro and in vivo, and abundances of cluster genes were significantly reduced in patients with inflammatory bowel disease. Together, these data provide a mechanism by which gut bacteria sulfonate steroidal metabolites and suggest that these compounds can modulate immune cell trafficking in the host.

Novel alantolactone derivative AL-04 exhibits potential anti-inflammatory activity via modulation of iNOS, COX-2 and NF-κB

Natural compounds and their synthesized analogues continue to be valuable sources in the discovery and development of novel anti-inflammatory agents. AL-04 is a thiol analogue derived from a natural sesquiterpene alantolactone, that demonstrated potential anti-inflammatory activity in vitro in comparison to its parent compound. However, the anti-inflammatory mechanism of action of AL-04 has not been elucidated. In this context, we investigated the signaling pathway that primarily mediate the anti-inflammatory activity of AL-04 and its effect on principal inflammatory mediators including iNOS, COX-2 and ROS. Furthermore, the anti-inflammatory activity was investigated in vivo in carrageenan induced paw oedema model in addition to the exploration of anti-nociceptive activity and acute toxicity. The results suggested that treatment with AL-04 significantly decreased the LPS-induced upregulation of pro-inflammatory cytokines and mediators in addition to the downregulated transcription of TNF-α and IL-6 in RAW 264.7 cell line. Furthermore, mRNA and the protein expression of COX-2 and iNOS were also significantly attenuated with AL-04 at a concentration of 10 µM. Western blot studies further suggested that AL-04 downregulated LPS-stimulated NF-κB p65 expression. In addition to this the anti-inflammatory activity of AL-04 was demonstrated in carrageenan induced paw oedema model with significant inhibition of oedema in a dose-dependent manner. The anti-inflammatory activity of AL-04 was further demonstrated in balb/c mice by inhibition of leukocyte migration and vascular permeability. Besides, AL-04 also inhibited thermally and chemically induced pain in tail-flick and acetic acid induced writing assays respectively in balb/c mice suggesting the analgesic potential of the compound. Acute toxicity studies further suggested the appreciable safety of AL-04 at high dose of 2000 mg/kg with no indications of toxicity or changes in biochemical and haematological parameters. Overall, the study insinuates the anti-inflammatory potential of AL-04 and paves way for further exploration of the compound as a safer therapeutic anti-inflammatory agent.

The anti-inflamatory effect of Andira anthelmia lectin in rats involves inhibition of the prostanoid pathway, TNF-α and lectin domain.

To investigate the effect and mechanisms of Andira anthelmia lectin in rat models of acute inflammation. AAL anti-inflammatory activity was evaluated in Wistar rat models of paw edema and peritonitis. AAL (0.01-1mg/kg i.v.) was injected 30min before stimulation with carrageenan and with initial and late phase inflammatory mediators into the animals paw or peritoneum for evaluation of cell migration (optical and intravital microscopy), paw edema (plethysmometry and histopathology); hyperalgesia (analgesimetry). AAL inhibited leukocyte migration induced by carrageenan, mainly neutrophils to the peritoneal fluid, decreasing leukocyte adhesion. In the peritoneal fluid, AAL reduced the gene expression of TNF-α and cyclooxygenase, as well the levels of PGE2. AAL inhibited the paw edema induced by carrageenan, serotonin, histamine, TNF-α, PLA2 and PGE2, but not by L-arginine. In this model, AAL also inhibited mechanical hypernociception induced by TNF-α, PGE2, db-cAMP and capsaicin, and the activity of myeloperoxidase in the paw tissues. AAL presents anti-inflammatory effect in acute models of rat inflammation involving the participation of prostaglandins, TNF-α and lectin domain.

Dynamics of T-lymphocyte lymphokine production from piglets with bronchopneumonia in response to non -specific mitogens &lt;em&gt;in vitro&lt;/em&gt;

The aim of the study was to evaluate the production of lymphokines by piglets' T-lymphocytes during the disease of acute nonspecific bronchopneumonia, in response to nonspecific mitogens in vitro. Blood from 3.5-month–old piglets of a large white breed was used as a model for the study (group 1 - patients with clinical signs of lung disease - underwent antibiotic and vitamin therapy in appropriate doses, group 2 - clinically healthy - intact were under observation). The functional activity of T-lymphocytes was evaluated in vitro, the level of lymphokine production in the reaction of inhibition of leukocyte migration (RTML) with nonspecific mitogens of T-lifmocytes - concanavalin A (Con A) and phytohemagglutinin (PHA) was taken into account before treatment and 7, 14 and 21 days after its initiation. In the first days of the disease, it was found that lymphocytes isolated from sick animals react differently to the introduction of concanavalin A and phytohemagglutinin cells into the culture – the reaction in response to PHA is more pronounced. The percentage of inhibition of leukocyte migration in response to the introduction of PHA and KonA was the lowest for the entire study period on the 7th day of treatment, which characterized the highest production of lymphokines in response to both mitogens. From the 14th day, the production of lymphokines in both cases was not as active. This is due to the fact that at the beginning of the inflammatory process, T-lymphocytes actively produced lymphokines that inhibit the migration of leukocytes, which should contribute to the formation of a focus of inflammation and limit secondary alterations. After antibiotic treatment and relief of the inflammatory process, chemotactic stimuli acquire the opposite orientation, this contributes to the dispersion of immunocompetent cells from the focus of inflammation and is necessary for the adequate course of proliferative processes in the lung tissue.

Evaluation of the in vivo and in vitro anti-inflammatory activity of a new hybrid NSAID tetrahydropyran derivative.

Evaluate the anti-inflammatory activity in vivo and invitro of cis-(±)-acetate of 4-chloro-6-(naphtalene-1-yl)-tetrahydro-2H-pyran-2-yl) methyl 2-(2-(2,6-diclorofenylamine) phenyl (LS19). Male Swiss mice were analyzed in the paw edema, ear edema, and air pouch tests, and in vitro COX inhibition, cytotoxicity evaluation, and cytokine and nitric oxide determination tests. The compound showed effect on the carrageenan- and bradykinin-induced paw edema and capsaicin-induced ear edema tests. In addition, the compound was able to inhibit leukocyte migration to decrease the levels of the pro-inflammatory cytokines tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) and to increase the levels of the anti-inflammatory cytokine IL-10. The compound was also able to reduce levels of TNF-α, IL-6, and nitric oxide in the RAW 264.7 cell line and to inhibit COX activity. LS19 did not induce any significant changes in the viability of RAW 264.7 cells, demonstrating safety for these cell lines. The compound LS19 did not reduce the production of gastric mucus and induced a smaller increase in the extent of gastric lesions than that developed by the administration of diclofenac. In summary, the new compound proved to be safer and it had additional mechanisms compared to diclofenac.