Bradykinin (BK) is an important mediator in several inflammatory and vascular diseases that acts in part via induction of cyclooxygenase-2 (COX-2). The mechanisms involved in BK-mediated COX-2 induction are unclear.Here we characterized the transcriptional mechanisms involved in human pulmonary artery smooth muscle cells. BK stimulated the activity of a transiently transfected 966-bp (؊917 to ؉ 49) COX-2 promoter luciferase reporter construct. There was no reduction in BK-induced luciferase activity in cells transfected with COX-2 promoter constructs of 674, 407, 239, and 135 bp or constructs with mutated CCAAT/enhancer-binding proteinor NF-B-binding sites. In contrast luciferase activity was reduced in cells transfected with a 407-bp COX-2 promoter fragment containing a mutated cAMP response element (CRE)-binding site, suggesting that the CRE binding site is critical. Electrophoretic mobility shift assays using oligonucleotides specific for the CREbinding region of the COX-2 promoter and consensus oligonucleotides showed strong specific binding. Furthermore BK increased consensus cAMP-responsive luciferase reporter (p6CRE/luc)-mediated luciferase expression. CRE activation occurred by BK inducing cytosolic phospholipase A 2 -mediated arachidonic acid release and rapid prostaglandin E 2 (PGE 2 ) production, thereby increasing cAMP. Indomethacin inhibited BKinduced PGE 2 production, cAMP accumulation, and CRE/luc reporter and COX-2 promoter luciferase activity. Exogenous PGE 2 and EP2 (ONO-AE1 259) and EP4 (ONO-AE1 329) PGE 2 receptor agonists mimicked the effect of BK. Collectively these studies indicate that COX-2 induction by BK in human pulmonary artery smooth muscle cells is mediated by the CRE through a novel autocrine loop involving endogenous PGE 2 .

Methicillin-resistant Staphylococcus aureus (MRSA) is an important human pathogen that causes various infections. The increasing resistance of MRSA to different antibiotics is widely spreading; therefore, plant extracts may be novel therapeutic alternatives. The phytochemical profiling of Cupressus macrocarpa Hartw. ex Gordon leaves in vitro, and in vivo, antimicrobial potential of its extracts against MRSA clinical isolates were explored. A phytochemical tentative identification of 49 compounds was performed in the leaves using LC-ESI-MS/MS; in addition, isolation, and structure elucidation of hesperidin and eriocitrin were achieved for the first time. The diethyl ether extract (DEEL) exhibited the best antibacterial effect with MIC values ranging from 2 to 8 µg/mL, which significantly reduced the growth and efflux activity in 48.78% and 29.26% of isolates, respectively. qRT-PCR showed a significant down expression of norA and norB genes, which significantly affected the bacterial cell morphology and had a non-significant effect on membrane depolarization (using flow cytometry). In a rat model, four groups were wounded and treated with normal saline or DEEL, or infected with MRSA, or infected and treated with DEEL. The regeneration of the epidermis, maturation of granulation tissue, and reduction of inflammatory cell infiltration were observed after treatment with DEEL. Thus, C. macrocarpa leaves may be a promising source for new antimicrobials against MRSA.

Abstract-Increased levels of inflammatory cytokines contribute to the pathophysiology of pulmonary hypertension.Prostacyclin (PGI 2 ) analogues, which relax pulmonary vessels mainly through cAMP elevation, have a major therapeutic role. In this study, we show that prolonged incubation with bradykinin (BK), interleukin-1␤ (IL-1␤), and transforming growth factor-␤ 1 (TGF-␤ 1 ) markedly impairs cAMP accumulation in human pulmonary artery smooth muscle cells in response to short-term incubation with prostaglandin E 2 (PGE 2 ) and the PGI 2 analogues iloprost and carbaprostacyclin. A similar reduction in cAMP accumulation in response to a direct adenylyl cyclase activator, forskolin, suggested that the effect was attributable to downregulation of adenylyl cyclase. Reverse transcriptase-polymerase chain reaction studies showed downregulation of adenylyl cyclase isoforms 1, 2, and 4. The effect of IL-1␤, BK, and TGF-␤ 1 on cAMP levels was abrogated by the selective COX-2 inhibitor NS398. Furthermore, it was mimicked by prolonged incubation with the COX-2 product PGE 2 and PGI 2 analogues or the COX substrate arachidonic acid, suggesting that it was mediated by endogenous prostanoids produced by COX-2. Consistent with this, IL-1␤, BK, and TGF-␤ 1 all induced COX-2 and PGE 2 release. These results show that BK, IL-1␤, and TGF-␤ 1 downregulate adenylyl cyclase in human pulmonary artery smooth muscle cells via COX-2 induction and prostanoid release. This suggests a novel mechanism whereby mediators and cytokines produced in pulmonary hypertension may impair the therapeutic effects of prostacyclin analogues such as iloprost and carbaprostacyclin. Key Words: interleukin-1␤ Ⅲ transforming growth factor-␤ 1 Ⅲ bradykinin Ⅲ cAMP Ⅲ adenylyl cyclase P rostacyclin (PGI 2 ) analogues are effective treatment for pulmonary hypertension. 1 Long-term treatment with intravenous PGI 2 improved survival rates and reduced vascular resistance in primary and secondary pulmonary hypertension. 2 PGI 2 analogues act mainly through cAMP, a relaxant second messenger, 3 by binding to adenylyl cyclase-coupled prostacyclin receptors. 4 -6 PGI 2 analogues also regulate pulmonary artery remodeling. 7 Administration of PGI 2 analogues may compensate for defective PGI 2 production in pulmonary hypertension. Pulmonary vascular tone and remodeling is controlled by the balance between vasoconstrictor and vasodilator mediators. 8 In pulmonary hypertension, there is an imbalance with excess thromboxane A 2 and reduced dilator PGI 2 production 9 and pulmonary artery PGI 2 synthase expression. 10 Endothelin-1 (ET-1) plays an important role in pulmonary hypertension. ET-1 levels are elevated in patients with pulmonary hypertension. ET-1 has a growth regulatory effect on pulmonary smooth muscle cells, partly via K ϩ channels. 11 Inflammatory cytokines and mediators contribute to the increased pulmonary resistance and remodeling in pulmonary hypertension, 12 including interleukin-1 (IL-1), IL-6, ET-1, and prostanoids. 13,14 IL-1␤ is interesting because its elevate...

We have previously shown that interleukin (IL)-1beta, transforming growth factor (TGF)-beta1, or bradykinin (BK) impair cAMP generation in response to prostacyclin analogs in human pulmonary artery smooth muscle (PASM), suggesting that inflammation can impair the effects of prostacyclin analogs on PASM in pulmonary hypertension. Here we explored the biochemical mechanisms involved. We found that IL-1beta, BK, and TGF-beta1 reduced adenylyl cyclase isoform 1, 2, and 4 mRNA, increased Galphai protein levels, and reduced prostacyclin receptor (IP receptor) mRNA expression. In contrast, Galphas protein levels were unchanged. Protein kinase A (PKA) (H-89, KT-2750, PKIm) and p38 mitogen-activated protein (MAP) kinase (SB-202190) inhibitors attenuated these effects, but protein kinase C (bisindolylmaleide) or phosphoinositol 3-kinase (LY-294002) inhibitors did not. Fluorescent kemptide assay and Western blotting confirmed that PKA and p38 MAP kinase were activated by IL-1beta, BK, and TGF-beta1. These studies suggest that IL-1beta, BK, and TGF-beta1 impair IP receptor-mediated cAMP accumulation by multiple effects on different components of the signaling pathway and that these effects are PKA and p38 MAP kinase dependent.

Schizophrenia remains one of the most chronic and highly disabling mental disorders. Lumateperone is a recent FDA-approved atypical antipsychotic drug for the treatment of schizophrenia. However, the internal FDA pathologist raised concerns regarding pigment deposition associated with degeneration in different tissue in animal studies with lumateperone treatment. The adrenal gland may be implicated in lumateperone side effects, and quercetin may have the ability to fulfill this treatment gap. To prove this hypothesis, 40 male guinea pigs were used and divided into four groups; control, quercetin-treated, lumateperone-treated, and quercetin/lumateperone cotreated orally for 28 consecutive days. Behavioral forced swim (FST) and open field (OF) tests were done at the end of treatment. Retro-orbital blood samples were taken to assess hormones: adrenocorticotropic hormone (ACTH), cortisol, dehydroepiandrosterone acetate (DHEA), and aldosterone, along with an assessment of oxidative stress parameters: malondialdehyde (MDA), glutathione (GSH), and superoxide dismutase (SOD). Adrenal glands were extracted for histopathological assessment with H&E, Mallory trichome staining, immunostaining, and electron microscopy studies. Lumateperone-treated group showed a significant reduction in the activity in FST and OF with histopathological deterioration in adrenal secretory function and structure and increased expression of interleukin-6 (IL-6), CASPASE-3, collagen deposition, and decreased proliferating cell nuclear antigen (PCNA). Cytoplasmic vacuolation, pyknosis of the nuclei, increase in the lysosome, lipofuscin pigment, and cellular infiltration with diminishing in the number of secretory granules could all be observed in lumateperone-treated group. Coadministration of quercetin and lumateperone showed improvement of the previously deteriorated parameters. Quercetin had a prophylactic effect against lumateperone depressive-like effect on animal behavior and its possible adrenal damage.Graphical AbstractConceptual framework for the proposed mechanism of action of coadministration of quercetin and lumateperone.

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Background: Osteoporosis is a major health problem. Teriparatide is a recombinant parathyroid hormone used as antiosteoporotic therapy. Lepidium sativum (LS) is widely used as a traditional herbal therapy for hypertension, diabetes and renal disorders. The LS seeds are widely known as a good traditional alternative medication for fracture healing.Objectives: Current research focused on evaluation of the Lepidium sativum versus teriparatide effect on glucocorticoidinduced osteoporosis Materials and Methods: 60 adult male guinea pigs were randomly divided into six equal groups: control groupI (distilled water); LS treated group (300 mg/kg suspended in distilled water orally by gastric tube), teriparatide treated group (4 mcg/ kg subcutaneously twice weekly), glucocorticoid treated group (3.5 mg/kg subcutaneously), teriparatide and glucocorticoids treated group and glucocorticoids and lepidium sativum treated group as pervious groups. At the end of the study, animals were anaesthetized and sacrificed. Femur bones of each animal were excised for histological and immunohistochemical studies (caspase-3 and osteoponotin). Results: Glucocorticoids induced bone resorption manifested as resorption cavities, thickened periosteum associated with decreased and irregular cortical and trabecular bone thickness. Marked reduced irregular collagen fibers were detected by trichrome staining. Immunohistochemically, this group showed positive immunoreactivity for caspase-3 in osteocytes and decrease in osteopontin deposits in bone matrix. Moreover, there was significant increase in number of osteoclasts associated with decrease in number of osteoblasts. Significant decrease in serum calcium level and increase in serum alkaline phosphatase were detected. Administration of either teriparatide or Lipidium sativum with glucocorticoids improved biochemical, histological and morphometric bone changes. They reduced osteocytes apoptosis and osteoclasts increase. Lipidium sativum was more effective improving changes induced by glucocorticoids. Conclusions: Glucocorticoids induced bone resorption. Despite the high cost of teriparatide, it did not achieve the desired protective effect. LS is cheap, available and its protective effect is promising with no adverse effects.

Background: Remdesivir is a novel broad spectrum antiviral drug previously used to treat Ebola. It is a pro-drug nucleoside with antiviral activity that is opposed to SARS-CoV-2 and coronavirus. Aim: Current research was planned to evaluate and compare the potential ameliorative impact of the hematopoietic-stem-cell mobilized by the granulocyte colony-stimulating factor (G-CSF) versus BM-MSC on the effect of novel antiviral remdesivir on the kidney. Materials and Methods: Rats divided into four groups: control group, Remdesivir treated group (20 mg/kg/day IV on the first day followed by 10 mg/kg/day for 6 days), Remdesivir + BM-MSCs group (3x10⁶ cells/ml of PKH26 labelled MSC) and Remedesivir+ Filgrastim group (70 μg/kg/day/5 days). At the end of the experiment, animals were anaesthetized and sacrificed. Both animal kidneys were excised for histological, immunohistochemistry, and electron microscopy studies. Biochemical and morphometric assessments had been performed. Results: Remdesivir caused distortion and degeneration of both the glomeruli and the renal tubules associated with Bowman's space widening. It greatly increased the deposition of collagen and enhanced the expression of caspase 3, IL-6, and TGF-β1. Ultrastructure changes were observed in the form of thickening of glomerular basement membrane, dilated basal plasma membrane infoldings of tubular epithelium and mitochondrial degeneration. Biochemically, decreased antioxidant enzymes, reduced glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) with increased serum urea and creatinine were also recorded. Both BM-MSCs and G-CSF improved histological structure and function of the kidney. Conclusion:Prescribing drugs such as remdesivir should be carried out with severe care. BM-MSCs and G-CSF are an efficient and ideal option to protect patients from irreversible kidney damage.