Novel benzofurane-pyrazole derivatives with anti-inflammatory, cyclooxygenase inhibitory and cytotoxicity evaluation

Novel benzofurane-pyrazolone hybrids have been synthesized for evaluating their anti-inflammatory and cytotoxic properties. 4-(2-chloroacetyl)-1,5-dimethyl-2-phenyl-1,2-dihydro-3H-pyrazol-3-one were reacted with α-hydroxy aldehyde or α-hydroxy ketone derivatives to obtain nine novel pyrazolone derivatives. Structures were successfully elucidated by 1H NMR, 13C NMR, IR and HRMS. Enzyme inhibitory activity was measured on cyclooxygenases (COXs) as considered to address anti-inflammatory activity. Compound 2 showed the highest activity on both COX-1 and COX-2 subtypes with 12.0 μM and 8.0 μM IC50, respectively. This activity was found close to indomethacin COX-2 inhibition measured as 7.4 μM IC50. Rest of the compounds (1, 3–9) showed 10.4–28.1 μM IC50 on COX-2 and 17.0–35.6 μM IC50 on COX-1 (Compound 1 has no activity on COX-1). Tested compounds (1–9) showed activity on NO production. Only compound was the 4, which showed a low inhibition on IL-6 levels. Cell viability was up to 60% at 100 μM for all compounds (1–9) on RAW 264.7 and NIH3T3 cell lines, thus compounds were reported to be noncytotoxic.

Biological Evaluation and Molecular Docking Studies of Novel 1,3,4-Oxadiazole Derivatives of 4,6-Dimethyl-2-sulfanylpyridine-3-carboxamide

To date, chronic inflammation is involved in most main human pathologies such as cancer, and autoimmune, cardiovascular or neurodegenerative disorders. Studies suggest that different prostanoids, especially prostaglandin E2, and their own synthase (cyclooxygenase enzyme-COX) can promote tumor growth by activating signaling pathways which control cell proliferation, migration, apoptosis, and angiogenesis. Non-steroidal anti-inflammatory drugs (NSAIDs) are used, alongside corticosteroids, to treat inflammatory symptoms particularly in all chronic diseases. However, their toxicity from COX inhibition and the suppression of physiologically important prostaglandins limits their use. Therefore, in continuation of our efforts in the development of potent, safe, non-toxic chemopreventive compounds, we report herein the design, synthesis, biological evaluation of new series of Schiff base-type hybrid compounds containing differently substituted N-acyl hydrazone moieties, 1,3,4-oxadiazole ring, and 4,6-dimethylpyridine core. The anti-COX-1/COX-2, antioxidant and anticancer activities were studied. Schiff base 13, containing 2-bromobenzylidene residue inhibited the activity of both isoenzymes, COX-1 and COX-2 at a lower concentration than standard drugs, and its COX-2/COX-1 selectivity ratio was similar to meloxicam. Furthermore, the results of cytotoxicity assay indicated that all of the tested compounds exhibited potent anti-cancer activity against A549, MCF-7, LoVo, and LoVo/Dx cell lines, compared with piroxicam and meloxicam. Moreover, our experimental study was supported by density functional theory (DFT) and molecular docking to describe the binding mode of new structures to cyclooxygenase.

First Report of Phytophthora parsiana Causing Crown and Root Rot on Guayule in the United States

Guayule (Parthenium argentatum A. Gray) is a perennial shrub plant (approximately 50 cm in height) cultivated in the southwestern United States. It produces natural low-allergenic latex, resins and high-energy biofuel feedstock. During August 2021, a crown and root rot disease was observed on 2-year-old plants of direct-seeded guayule cultivar ‘Az 2’ in research plots located in Pinal county, Arizona, where a record 36 cm of rainfall fell during monsoon season. Symptoms included yellowing of leaves, wilting, and plant death. Average disease incidence was 16%. Isolation from necrotic crown and root tissues on 10% clarified V8-PARP (Jeffers and Martin 1986) yielded Phytophthora-like colonies. Three isolates were subcultured on V8 agar and chlamydospores and hyphal swellings were abundant in 2-week-old cultures. All three isolates produced abundant noncaducous and nonpapillate sporangia ranging from 33 to 54 μm × 20 to 39 μm (average 45.5 × 28.5 μm, n = 20) in soil water extract solution. Isolates did not produce oospores after 2 weeks on carrot agar at 20°C in the dark. Isolates had optimum vegetative growth at 30 oC and grew well at 35 oC. There was no growth at 5 and 40 oC. Genomic DNA was extracted from the mycelia of three isolates using DNeasy Plant Pro Kit (Qiagen Inc., Valencia, CA) according to the manufacturer’s instructions. The internal transcribed spacer (ITS) region of rDNA, mitochondrially encoded cytochrome c oxidase 1 (cox 1) gene, and beta-tubulin (β-tub) gene were amplified with primers ITS1/ITS4 (White et al., 1990), COXF4N/COXR4N and TUBUF2/TUBUR1 (Kroon et al., 2004) and the resulting 3 amplicons were sequenced (GenBank Accession No. OK438221, OK484426, and OK484427). A BLASTn search of 811-bp amplicon (OK438221) revealed 99% match (762/766) with ITS sequences MG865562 which was Phytophthora parsiana Ex-type CPHST BL 47 from Iran. BLAST analysis of the 867-bp amplicon (OK484427) showed 99% identity (866/867) with the COX 1 sequence of P. parsiana (KC733455) from Virginia. BLAST analysis of the 941-bp amplicon (OK484426) showed 99% identity (928/938) with the β-tub sequence of P. parsiana (AY659746). To fulfill Koch’s postulates, pathogenicity tests were conducted twice on 2-week-old ‘Az 2’ guayule seedlings grown in 10 plants per 1.9-liter pot filled with a steam-disinfested potting mix. Pots were placed in a plastic container and watered three times a week by flooding, to create waterlogged conditions. Plants were maintained in a greenhouse with 12 h day/12 h night (15-28 oC) and fertilized weekly with a 20-20-20 fertilizer at 1mg/ml. Fifty plants in 5 pots were challenged with a P. parsiana isolate by drenching each pot with 50 ml of a 1×105 zoospore/ml suspension. Fifty plants in 5 pots, serving as a control, received each 50 ml of distilled water. Symptoms of wilting, root rot, and plant death were observed 1 week afterward in inoculated plants, whereas control plants remained asymptomatic. P. parsiana was reisolated from necrotic roots of inoculated plants but not from control plants. To our knowledge, this is the first report of crown and root rot in guayule caused by P. parsiana in Arizona. P. parsiana is a species known for causing root rot on woody plants such as pistachio in California (Fichtner et al., 2016) and Iran (Mostowfizadeh-Ghalamfarsa et al., 2008). Arizona is home of desert woody guayule plant. P. parsiana may represent a significant barrier to commercialization of guayule for rubber in low desert areas of Arizona. The origin, distribution, and virulence of the pathogen on Arizona guayule is currently unknown. Disease resistance evaluation may help identify resistance in guayule germplasm that are useful in breeding for resistant cultivars.

10 reasons, why to use naproxen – in the light of new scientific reports

Naproxen is a non-steroidal anti-inflammatory drug with a wide range of use, used in patients with various pain syndromes in the musculoskeletal system. It has anti-inflammatory, analgesic and antipyretic properties through non-selective and reversible inhibition of COX-1 and COX-2 isoforms. In Poland, it is registered in the treatment of rheumatic diseases and acute inflammations of the musculoskeletal system. Some naproxen preparations are also approved for the treatment of migraine and fever. The paper presents the characteristic features of the substance.

Modulation of Enzyme-Catalyzed Synthesis of Prostaglandins by Components Contained in Kidney Microsomal Preparations

In the kidney, prostaglandins formed by cyclooxygenase 1 and 2 (COX-1 and COX-2) play an important role in regulating renal blood flow. In the present study, we report our observations regarding a unique modulatory effect of renal microsomal preparation on COX-1/2-mediated formation of major prostaglandin (PG) products in vitro. We found that microsomes prepared from pig and rat kidneys had a dual stimulatory–inhibitory effect on the formation of certain PG products catalyzed by COX-1 and COX-2. At lower concentrations, kidney microsomes stimulated the formation of certain PG products, whereas at higher concentrations, their presence inhibited the formation. Presence of kidney microsomes consistently increased the Km values of the COX-1/2-mediated reactions, while the Vmax might be increased or decreased depending on stimulation or inhibition observed. Experimental evidence was presented to show that a protein component present in the pig kidney microsomes was primarily responsible for the activation of the enzyme-catalyzed arachidonic acid metabolism leading to the formation of certain PG products.

Cyclooxygenase Enzyme and Lipid Peroxidation Inhibitory Terpenoids and Steroidal compounds as Major Constituents in Cleome viscosa Leaves

Bioassay guided study of Cleome viscosa Linn. (Cleomaceae) leaves led to the isolation of a new cembrenoid diterpene (1) and three known compounds (2-4) from the hexane extract. The chemical structures of these compounds were elucidated by spectroscopic methods such as NMR (1D and 2D), HRMS and IR and identified and afforded compound 1, malabaric acid (2), stigmast-4-en-3-one (3) and stigmast-4-ene-3,6-dione (4). This is the first report of compounds 1 and 2 from C. viscosa Linn. Isolates were evaluated for anti-inflammatory activity using in vitro cyclooxygenase enzyme (COX-1 and -2) inhibitory assays. The novel cembrenoid diterpene (1) exhibited IC50 values of 8.4 μM for COX-1 enzyme and 45.2 μM for COX-2 enzyme, respectively. Similarly, malabaric acid (2) exhibited IC50 values of 11.5 μM for COX-1 enzyme and 46.9 μM for COX-2 enzyme, respectively. Their inhibitory activities were in par with non-steroidal anti-inflammatory drugs aspirin, ibuprofen and naproxen. Sterols 3 and 4 gave IC50 values of 62.6 and 67.9 μM, respectively for COX-1 enzyme while indicating weak COX-2 enzyme inhibition. Lipid peroxidation inhibitory (LPO) and MTT assays were used to determine antioxidant activity of these compounds. Compounds 1-4 showed LPO inhibition with IC50 values between 82 and 100 µM and moderate antioxidant activity in the MTT assay. Biological activities reported for these compounds are for the first time and it support anecdotal medicinal claims of C. viscosa Linn. leaves.

Platelet ACKR3/CXCR7 Favors Anti-Platelet Lipids over an Atherothrombotic Lipidome and Regulates Thrombo-inflammation

Platelet ACKR3/CXCR7 surface expression is enhanced and influences prognosis in coronary artery disease-(CAD) patients, who exhibit a distinct atherothrombotic platelet lipidome. Current investigation validates the potential of ACKR3/CXCR7 in regulating thrombo-inflammatory response, through its impact on the platelet lipidome. CAD patients-(n=230) with enhanced platelet-ACKR3/CXCR7 expression exhibited reduced aggregation. Pharmacological CXCR7-agonist-(VUF11207) significantly reduced pro-thrombotic platelet response in blood from ACS patients-(n=11) ex vivo. CXCR7-agonist administration reduced thrombotic functions and thrombo-inflammatory platelet-leukocyte interactions post myocardial infarction-(MI) and arterial injury in vivo. ACKR3/CXCR7-ligation did not affect surface availability of GPIbα, GPV, GPVI, GPIX, αv-integrin, β3-integrin, coagulation profile-(APTT, PT), bleeding time, plasma-dependent thrombin generation-(thrombinoscopy) or clot formation-(thromboelastography), but counteracted activation-induced phosphatidylserine exposure and procoagulant platelet-assisted thrombin generation. Targeted-(micro-UHPLC-ESI-QTrap-MS/MS) and untargeted-(UHPLC-ESI-QTOF-MS/MS) lipidomics analysis revealed that ACKR3/CXCR7-ligation favored generation of anti-thrombotic lipids-(dihomo-γ-linolenic acid-DGLA, 12-hydroxyeicosatrienoic acid-12-HETrE) over cyclooxygenase-COX-1-(thromboxane-TxA2), or 12-lipoxygenase-LOX-(12-HETE) metabolized pro-thrombotic, and phospholipase derived atherogenic-(lysophosphatidylcholine-LPC) lipids, in healthy subjects and CAD patients, contrary to anti-platelet therapy. Through 12-HETrE, ACKR3/CXCR7-ligation coordinated with Gαs-coupled prostacyclin receptor-(IP) to trigger cAMP-PKA mediated platelet inhibition. ACKR3/CXCR7-ligation reduced generation of lipid agonists-(arachidonic acid-AA,TxA2), lipid signaling intermediates-(lyophosphatidylinositol-LPI, diacylglycerol-DG), which affected calcium mobilization, intracellular signaling, consequently platelet interaction with physiological matrices and thrombo-inflammatory secretion-(IL1β,IFN-γ,TGF-β,IL-8), emphasizing its functional dichotomy from pro-thrombotic CXCR4. Moreover, CXCR7-agonist regulated heparin-induced thrombocytopenia-(HIT)-sera/IgG-induced platelet and neutrophil activation, heparin induced platelet aggregation-(HIPA), generation of COX-1-(TxA2), 12-LOX-(12-HETE) derived thrombo-inflammatory lipids, platelet-neutrophil aggregate formation, and thrombo-inflammatory secretion (sCD40L, IL-1β, IFN-γ, TNF-α, sP-selectin, IL-8, tissue factor-TF) ex vivo. Therefore, ACKR3/CXCR7 may offer a novel therapeutic strategy in acute/chronic thrombo-inflammation exaggerated cardiovascular pathologies, and CAD.

Genetic variation is predominantly structured by geography rather than host in feather mites (Acariformes: Sarcoptiformes) associated with tanagers (Aves: Thraupidae) in Brazil

Feather mites are the most common ectosymbionts on birds. These obligatory symbionts are mainly transmitted during their host’s parental care, which creates high host specificity. Due to this intimate relationship, it is thought that their geographic distribution is restricted by their host distribution, or that a host species harbors the same mite composition across its whole range. However, our knowledge regarding the geographic distribution of feather mites remains scarce, with only a few studies indicating disconnections between mite and host distributions, especially in widespread hosts. Here, we investigate the feather mites distribution on four tanager species, three widespread – Thraupis sayaca (L.), T. palmarum (Wied), and Stilpnia cayana (L.) from Northern and Southern Brazil; and the Amazonian T. episcopus (L.). Feather mites were identified using the molecular barcode marker COX-1 using K2P genetic distances. We found a strong genetic structure between Northern and Southern populations of tanagers of more than 10%, even among conspecific hosts. Therefore, the mite distribution on Brazilian tanagers is predominantly shaped by geography rather than by host species. These features in turn reflect historical horizontal transmissions among the hosts, suggesting a high potential for frequent host switches in these symbionts.

Meloxicam in pain syndrome treatment of comorbid diseases patients

The issue nonsteroidal anti-inflammatory drugs (NSAIDs) use safety is associated with a high frequency of adverse events (AEs) from the gastrointestinal tract and cardiovascular risks. Patients with lower back pain (LBP) and osteoarthritis (OA), as a rule, have comorbid diseases, such as arterial hypertension (AH), coronary heart disease (CHD), gastrointestinal tract (GIT) diseases, which significantly complicates the appointment of NSAIDs. The main guideline in NSAIDs appointment is the selective ability to inhibit cyclooxygenase-1 and -2 (COX). The ratio of the activity of NSAIDs when blocking COX-1/COX-2 allows us to judge their potential toxicity. And, then higher the selectivity of NSAIDs, then lower its toxicity. For example, the ratio of COX-1/COX-2 in meloxicam is 0.33, diclofenac – 2.2, tenoxicam – 15, piroxicam – 33, indomethacin – 107. To the predominantly selective COX-2 NSAIDs include meloxicam, which has little effect on the GIT, the lowest relative risk (RR) of complications from the cardiovascular system (CVS). The therapeutic efficacy of meloxicam is comparable to piroxicam and diclofenac. A number of studies have shown the high efficacy of meloxicam, both with per oral (p/o) administration (7.5–15 mg/d), and with intramuscular (i/m) administration (1.5 ml), and when injected into trigger zones. Both with p/o and the injectable form of meloxicam has minimal GIT AEs and absence local reaction in the injection area. The drug can be recommended both as a combination therapy and prescribed in monotherapy.