Isolation and Characterization of Phenylalanine Ammonia Lyase (PAL) Genes in Ferula pseudalliacea: Insights into the Phenylpropanoid Pathway.

Phenylalanine ammonia lyase (PAL) is a key enzyme regulating the biosynthesis of the compounds of the phenylpropanoid pathway. This study aimed to isolate and characterize PAL genes from Ferula pseudalliacea Rech.f. (Apiales: Apiaceae) to better understand the regulation of metabolite production. Three PAL gene isoforms (FpPAL1-3) were identified and cloned using the 3'-RACE technique and confirmed by sequencing. Bioinformatics analysis revealed important structural features, such as phosphorylation sites, physicochemical properties, and evolutionary relationships. Expression analysis by qPCR demonstrated the differential transcription profiles of each FpPAL isoform across roots, stems, leaves, flowers, and seeds. FpPAL1 showed the highest expression in stems, FpPAL2 in roots and flowers, and FpPAL3 in flowers. The presence of three isoforms of PAL in F. pseudalliacea, along with the diversity of PAL genes and their tissue-specific expression profiles, suggests that complex modes of regulation exist for phenylpropanoid biosynthesis in this important medicinal plant. The predicted interaction network revealed associations with key metabolic pathways, emphasizing the multifaceted roles of these PAL genes. In silico biochemical analyses revealed the hydrophilicity of the FpPAL isozyme; however, further analysis of substrate specificity and enzyme kinetics can clarify the specific role of each FpPAL isozyme. These comprehensive results increase the understanding of PAL genes in F. pseudalliacea, helping to characterize their contributions to secondary metabolite biosynthesis.

Peptidic Compound as DNA Binding Agent: In Silico Fragment-based Design, Machine Learning, Molecular Modeling, Synthesis, and DNA Binding Evaluation.

BACKGROUND: Cancer remains a global burden, with increasing mortality rates. Current cancer treatments involve controlling the transcription of malignant DNA genes, either directly or indirectly. DNA exhibits various structural forms, including the G-quadruplex (G4), a secondary structure in guanine-rich regions. G4 plays a crucial role in cellular processes by regulating gene expression and telomerase function. Researchers have recently identified G4-stabilizing binding agents as promising anti-cancer compounds. Additionally, peptides have emerged as effective anticancer pharmaceuticals due to their ability to form multiple hydrogen bonds, electrostatic interactions, and van der Waals forces. These properties enable peptides to bind to specific areas of DNA chains selectively. However, despite these advancements, designing G4-binding peptides remains challenging due to a lack of comprehensive information. OBJECTIVE: In our present study, we employed an in silico fragment-based approach to design G4- binding peptides. This innovative method combines machine learning classification, molecular docking, and dynamics simulation. METHODS: AutoDock Vina and Gromacs performed molecular docking and MD simulation, respectively. The machine learning algorithm was implemented by Scikit-learn. Peptide synthesis was performed using the SPPS method. The DNA binding affinity was measured by applying spectrophotometric titration. RESULTS: As a result of this approach, we identified a high-scoring peptide (p10; sequence: YWRWR). The association constant (Ka) between p10 and the ctDNA double helix chain was 4.45 × 105 M-1. Molecular modeling studies revealed that p10 could form a stable complex with the G4 surface. CONCLUSION: The obtained Ka value of 4.45 × 105 M-1 indicates favorable interactions. Our findings highlight the role of machine learning and molecular modeling approaches in designing new G4-binding peptides. Further research in this field could lead to targeted treatments that exploit the unique properties of G4 structures.

Alpha-Pinene and Tannic Acid Inhibit Trichomonas vaginalis Protozoan Cells by Inducing Apoptosis.

Background: Trichomoniasis is one of the most common sexually transmitted infections worldwide. The growing concern of drug resistance of this infection has cautioned the need for new drug development. We evaluated the potential antiproliferative and apoptotic effect of α-pinene and tannic acid (TA) on Trichomonas vaginalis cells. In addition, the cytotoxicity of agents on Vero cells was investigated. Methods: Trichomonas cells were axenically cultured in TYI-S-33 medium. In vitro antiproliferative activity of α-pinene, TA, and metronidazole was investigated against Trichomonas cells. The assays were carried out in triplicate using microtiter plate and trypan blue staining method. Annexin V/PI staining with flow cytometry was used to evaluate apoptosis induction. In addition, the cytotoxic effect was measured by MTT assay. Results: α-Pinene and TA exhibited significant inhibition of the Trichomonas cells and the lowest IC50 values were 22.9 µg/ml and 140 µg/ml at 48 hours' incubation, respectively. The CC50 was found at 116 µg/ml for α-pinene and 473 µg/ml for TA, after 48 hours of treatment. The flow cytometry study demonstrated that the natural compounds induced apoptosis in Trichomonas cells. After 24 hours of treatment, the induction of apoptosis was 5.2% - 36.6% at concentrations of 3.9 - 62.5 µg/ml for α-pinene and TA induced-apoptosis was 6.1% - 53.8% at concentrations of 125-2000 µg/ml. Conclusion: Although the results show the antiproliferative and apoptotic effect of α-pinene and TA on Trichomonas cells, in vivo studies are needed to further clarify the effects of these compounds.

Botanical description, phytochemical constituents, ethnobotany, traditional medicinal use, and pharmacological activities of Stachys lavandulifolia Vahl.

Stachys lavandulifolia Vahl known as "mountain tea", is a perennial flowering plant belonging to the Lamiaceae family and is widespread in Iran, Armenia, Azerbaijan, Iraq, Turkey and Turkmenistan. S. lavandulifolia is widely used in traditional medicine for its analgesic, anti-inflammatory and anxiolytic properties. This plant has different chemical compounds classes including terpenoids, iridoids, flavonoids and phenylethanoids that have been isolated from the aerial parts of it. This review covers the plant botany, traditional medicinal uses and chemical composition of S. lavandulifolia, along with its biological and pharmacological activities including clinical trial data. The information of this review article was obtained from different scientific databases such as Google scholar, Science Direct, Hindawi, SID, Scopus, PubMed, and ACS as well as traditional Persian books. Pharmacological and clinical studies, especially Anxiolytic activity and anti-inflammatory on the plant are relatively low, so these studies are suggested in the future. Also, phytochemical investigation on root of the plant is necessary.

Chimgin from Ferula haussknechtii as AChE inhibitor and confirmation of the absolute configuration.

Alzheimer's disease (AD) is the most common cause of dementia worldwide and is classified as a neurodegenerative disorder. From a drug design perspective, natural products (NPs) are more drug-like and are highly compatible with biological systems compared to most synthetic libraries. NPs provide a more efficient and cost-effective approach to new drug discovery. However, the complexity of NPs makes their identification a challenging task. Chimgin, a bicyclic monoterpene with three chiral centers, exhibits a wide range of biological activity. Despite this, the exact structure of chimgin has remained unclear until now. In this study, we quantified the amount of chimgin in Ferula haussknechtii using analytical Reversed-phase high-pressure liquid chromatography equipped with photodiode array detector (RP-HPLC-PDA). Furthermore, we determined the absolute configuration of chimgin through electronic circular dichroism (ECD) spectroscopy and time-dependent density functional theory (TDDFT) calculations. Finally, we evaluated its inhibitory effect on AChE through in vitro and in silico studies. The extraction process yielded an output of 2.82 ± 0.10% with an exact amount of 0.62 ± 0.04 mg of chimgin per 100 g of plant. Based on the results of ECD and TDDFT calculation, the absolute configuration of chimgin was determined to be 1S, 2S, 4S. Chimgin exhibited an inhibitory effect on AChE with an IC50 of 37.43 µM and its mechanism of action was found to be competitive. HighlightsChimgin was isolated from the roots of Ferula haussknechtii.The amount of chimgin in the plant was determined by RP-HPLC-PDA.Its absolute configuration of chimgin was determined using ECD.In vitro acetylcholinesterase activity of the chimgin was evaluated.The docking and molecular dynamic simulation of chimgin was done.Communicated by Ramaswamy H. Sarma.

Ferulago bernardii as a New Source of α-Pinene Binds to ctDNA: In†Silico and in†Vitro Studies.

Ferulago bernardii Tomk & M. Pimen belongs to Apiaceae family. Various species of the Ferulago genus have antioxidant, anticholinesterase, cytotoxic, and antiproliferative effects. In this study, the essential oil of F. bernardii was extracted using the Clevenger apparatus. The essential oil compounds were identified using GC-MS/FID. The interaction between the essential oil and DNA strands was evaluated through spectrophotometric titration. The molecular mechanism of the interaction between the main components of the essential oil and different DNA strands was assessed using molecular dynamics simulation. Based on the results, 92.03±1.20 % of the essential oil consisted of α-pinene. Therefore, the essential oil could serve as a suitable source of α-pinene. α-pinene is a monoterpene hydrocarbon that has various effects, including anti-inflammatory, antioxidant, antimicrobial, and antitumor properties. The binding constant of the essential oil to DNA strands (Ka ) was determined to be 5.40±0.47×10-3  M-1 . Molecular dynamics simulation demonstrated that α-pinene could interact with AT and CG rich DNA strands and indirectly stabilize G-Quadruplex. Given the different applications for α-pinene and its high percentage in the essential oil, it is suggested that researchers pay more attention to F. bernardii in the pharmaceutical, cosmetic, and food industries.

In vitro Antibacterial Activity and Wound Healing Effects of Achillea millefolium Essential Oil in Rat.

Objectives: In this study we aimed to evaluate the in vitro antibacterial activity and wound healing properties of Achillea millefolium essential oil (AMEO) in full-thickness wound model in rat. The antibacterial activity of AMEO was evaluated against Staphylococcus aureus and Pseudomonas aeruginosa using the broth dilution method. Methods: The 2 cm × 2 cm full-thickness excisional wounds were created on the back of animals. Topical therapy was applied twice a day using 1%, 2%, and 3% w/w AMEO ointments, and the measurement of the wounds area was carried out every 3 days, after that the wound closure percentage was calculated in these days. Hydroxyproline content and histopathological evaluation of wound tissue samples were carried out on day 7 and 14 post wounding. Eucerin was used for the treatment of vehicle control group and negative control group received no treatment. Results: Our results revealed the bacteriostatic activity of AMEO against S. aureus and P. aeruginosa. Wound healing activity evaluation of AMEO showed the significant increase (p < 0.05) in the wound closure percentages in rats treated with AMEO 1% and 2% comparing to those of non-treatment group. In addition, hydroxyproline contents of tissue significantly (p < 0.01) increased in AMEO 1% and 2% comparing to non-treatment group. Histopathological evaluations of wound tissue samples on day 7 and 14 demonstrated higher accumulation of collagen fibers, reduction of edema and inflammation and also formation of tissue appendages in 1% and 2% AMEO treated groups in comparison with non-treatment group. Conclusion: The results of this study indicated that AMEO has the potential to be used as a safe and effective wound healing agent.

Antibacterial and Anti-Glucosyltransferase Activity of Verbascum speciosum Against Cariogenic Streptococci.

Objectives: Dental caries is a prevalent chronic human infection worldwide and several plants have shown anticariogenic properties through antibacterial activity against oral pathogens. The present study aimed to assess anticariogenic activity of Verbascum speciosum, in search of novel agents for the prevention and treatment of dental caries. Methods: Hydro-alcoholic extracts from flowers and total aerial parts of the plant were prepared by maceration. Antibacterial activity of the extracts against Streptococcus mutans (ATCC 35668) and Streptococcus sobrinus (ATCC 27607) was investigated by agar diffusion and microdilution techniques. Inhibitory concentration-fifty values of the flowers' extract against Streptococcus mutans glucosyltransferase enzymes were determined. The total flavonoid content of the extracts was determined using an aluminum chloride reaction. Results: Verbascum speciosum flowers' extract showed significantly higher flavonoid content and antibacterial activity; with minimum inhibitory concentrations of 100 and 200 µg/mL for Streptococcus mutans and Streptococcus sobrinus, respectively. The extract inhibited the synthesis of glucan by cell-associated and extracellular glucosyltransferase enzymes in a dose-dependent manner with higher activity against the extracellular enzyme. Conclusion: This study indicated effective anticariogenic activity of Verbascum speciosum flowers extract. This extract can be considered as an alternative to current anticaries therapies or an additive to dental care products.

Combined Antimicrobial Activity of Extracts from Quercus infectoria Galls and Scrophularia striata Aerial Parts for an Anticariogenic Herbal Mouthwash.

Objectives: Dental caries is one of the most prevalent human diseases worldwide. The disease initiates with bacterial adherence to the tooth surface followed by the formation of dental plaques. Mutans streptococci and Candida albicans are principal oral microorganisms involved in the initiation and development of dental caries. Phytochemicals have been shown to possess promising antimicrobial properties against a wide range of microorganisms and can be used for the prevention and treatment of dental caries. Herein, we reviewed literature on plants that are traditionally used for their antimicrobial properties or possess promising anticariogenic activity. We selected aerial parts of Scrophularia striata (S. striata) and galls of Quercus infectoria (Q. infectoria) and investigated their antimicrobial activity against cariogenic microorganisms. Methods: Water soluble fractions were obtained from hydroalcoholic extracts of S. striata and Q. infectoria and their antimicrobial activity against Streptococcus mutans (S. mutans), Streptococcus sobrinus (S. sobrinus), and Candida albicans (C. albicans) was evaluated separately and in combination. The extracts were then used for preparing an herbal mouthwash whose stability and tannic acid content were evaluated over 60 days. Results: Q. infectoria gall extract possesses efficient antimicrobial activity that was synergistically enhanced in the presence of S. striata extract. Mouthwash prepared using these extracts showed desirable organoleptic characteristics, antimicrobial activity, and stability. Conclusion: Extracts of S. striata and Q. infectoria galls can be used together for preparing dental products with effective anticariogenic properties. Our study highlights the importance of extensive pharmacological investigations when using herbal products alone or in combination with other chemical substances.

Radioprotective effect of Malva sylvestris L. against radiation-induced liver, kidney and intestine damages in rat: A histopathological study.

Background: Ionizing radiation (IR) is widely used in the treatment of cancer in radiotherapy. One of the main concerns of patients with gastrointestinal cancers undergoing radiotherapy is the harmful side effects of IR on normal tissues. The liver, kidney, and duodenum are usually exposed to high doses of radiation in the treatment of some cancers in abdominal region radiotherapy. We aimed to assess the radioprotective effects of Malva sylvestris L. against IR damages to the abdominal region. Materials and methods: This current study was conducted on 45 rats divided randomly into nine groups of five: A) negative control group, B) sham group, C) irradiation group, D) mallow treatment-1(200gr/kg), E) mallow treatment-2(400gr/kg), F) mallow treatment-3(600gr/kg), G) mallow treatment-4(200gr/kg) plus irradiation, H) mallow treatment-5(400gr/kg) plus irradiation, I) mallow treatment-6(600gr/kg) plus irradiation. Irradiation was performed with a 6Gy x-ray. Histopathological evaluations were performed 10 days after irradiation. Results: The histopathological examination results confirmed that preventive therapy with the effective dose of mallow reduced the liver, kidney, and intestine damage induced by radiation. The dose of 400 mg/kg was more effective than other selected dose in improving the damage caused by irradiation in the studied tissues. Conclusion: This study concludes that Malva sylvestris L. contributed to significant improvements in radiation-induced histological parameters of the liver and kidney and, to a lesser extent, in the intestine. These results collectively indicate that mallow is an effective radioprotective agent.

In silico molecular modeling, neuro-behavioral profile, and toxicity assessment of the essential oil of Ferula gummosa Boiss. as an anti-seizure agent.

ETHNOPHARMACOLOGICAL RELEVANCE: Ferula gummosa Boiss., known in Persian as "Baridje," belongs to the Apiaceae family. All parts of this plant, especially the root, contain galbanum. Galbanum, the oleo-gum resin of F. gummosa, is one of the essential traditional herbal medicines in Iran, which is used as a tonic for epilepsy and chorea, memory enhancement, gastrointestinal diseases, and wound healing. AIM OF THE STUDY: We investigated the toxicity, anticonvulsant effects, and molecular modeling of the essential oil (EO) distilled from the oleo-gum resin of F. gummosa. MATERIALS AND METHODS: Gas chromatography-mass spectrometry was used to identify the EO components. The cytotoxicity of EO on HepG2 cell lines was assessed by the MTT method. Male mice were arranged as follows: negative control groups (sunflower oil (10 ml/kg, i.p.) or saline (10 ml/kg, p.o.)), EO groups (0.5, 1, 1.5, and 2.5 ml/kg, p.o.), and positive control groups (ethosuximide (150 mg/kg, p.o.) or diazepam (1.0 or 2 mg/kg, i.p.)). The motor coordination and neurotoxicity of EO were studied using the rota-rod test. Open-field, novel object recognition, and passive avoidance learning tests were used to investigate the effect of EO on locomotor activity and memory function. An acute pentylenetetrazole-induced seizure model was utilized to evaluate the anticonvulsant properties of the EO. The interaction of the EO main components with the GABAA receptor was investigated by coarse-grained molecular dynamics simulations. RESULTS: ß-pinene, sabinene, α-pinene, and ρ-cymene were the main components of EO. The IC50 of the EO at 24, 48, and 72 h was found to be 59.90, 12.96, and 3.93 µl/ml, respectively. No adverse effects were observed in memory, motor coordination, and locomotor activity in mice treated with EO. Administration of EO (1, 1.5, and 2.5 ml/kg) improved survival rates in mice receiving pentylenetetrazole (PTZ; to induce an epileptic seizure). Sabinene was able to bind to the binding site of benzodiazepines at the GABAA receptor. CONCLUSIONS: Acute treatment with the EO of F. gummosa caused antiepileptic effects and could effectively increase the survival rate in PTZ-treated mice with no significant toxicity.

In vitro antitrichomonal activity of Satureja khuzestanica and main essential oil components carvacrol, thymol, and eugenol.

INTRODUCTION: Human trichomoniasis is a widespread sexually transmitted disease and the concern of drug resistance in the parasite is growing. Hence, this study was performed to evaluate in vitro antitrichomonal activity of Satureja khuzestanica, carvacrol, thymol, eugenol, and phytochemical evaluation of the S. khuzestanica oil. METHODOLOGY: Extracts and essential oil of S. khuzestanica, and the components were prepared. Then, susceptibility testing was performed using the microtiter plate method and Trichomonas vaginalis isolates. The minimum lethal concentration (MLC) of the agents was determined in comparison with metronidazole. Also, the essential oil was investigated by gas chromatography-mass spectrometry and gas chromatography-flame ionization detector. RESULTS: After 48 hours of incubation, carvacrol and thymol were the most effective antitrichomonal agents with MLC of 100 µg/mL, followed by the essential oil and hexanic extract (MLC = 200 µg/mL), then eugenol and methanolic extract (MLC = 400 µg/mL), in comparison with the metronidazole MLC of 6.8 µg/mL. Overall, 33 identified compounds accounted for 98.72% of the total essential oil composition with carvacrol, thymol, and p-cymene being the major constituents. CONCLUSIONS: The results suggested the potency of S. khuzestanica and its bioactive ingredients against T. vaginalis. Thus, further in vivo studies are required to evaluate the efficacies of the agents.

Isolation and purification of terpenoid compounds from Ferula haussknechtii and evaluation of their antibacterial effects.

The roots of F. haussknechtii are used by local people in order to treat wounds and urinary infections. Ferula species are rich in bioactive compounds with biological effects. In line with our previous studies about screening antibacterial natural products, five terpenoid derivatives were purified from Ferula haussknechtii. The separation and purification were performed by column chromatography. Their structures were determined by 1 D and 2 D NMR as hawraman 8-p-hydroxybenzoyl-tovarol (1), ferutinin (2), lancerotriol 6-(p-hydroxybenzoate) (3), chimganin (4), and chimgin (5). Then, the antibacterial effects of the purified compounds were evaluated by measuring their MIC values against Gram-positive and Gram-negative bacteria. The results showed that compound (1) had the most antibacterial effect on Bacillus cereus (MIC = 16 µg/mL). The antibacterial effects of F. haussknechtii compounds are in line with their local application and it is suggested that further studies should be conducted to determine their mechanism of action.

Efficacy of a standardized herbal product from Pistacia atlantica subsp. Kurdica in type 2 diabetic patients with hyperlipidemia: A triple-blind randomized clinical trial.

BACKGROUND: Hyperlipidemia is one of the consequences of type 2 diabetes mellitus (T2DM) that puts the patients at the risk of getting cardiovascular disease. Pistacia atlantica subsp. Kurdica is used by local people for the improvement of lipid and glucose indices. This study was carried out to evaluate clinical effects of P. atlantica subsp. Kurdica on hyperlipidemia related to T2DM patients. MATERIALS AND METHODS: In this randomized, triple-blind, and placebo-controlled study, type 2 diabetes patients with hyperlipidemia were randomly allocated to receive either P. atlantica kurdica or placebo capsule for 2 months. 58 Patients were followed up at the beginning and after the end of each month for assessment of lipid profiles, glucose indices, and liver and renal function tests. The herbal capsule was standardized according to volatile and nonvolatile compounds by GC-MS/FID and RP-HPLC-PDA, respectively. The total phenolic and flavonoid contents were also determined with a spectrophotometer. RESULTS: After 1 and 2 months of the intervention period, mean differences of 2HPP, total cholesterol, and LDL-c have been reduced significantly (P < 0.05) between the two groups, but there were not reported any significant statistical changes in FBS, HbA1c, TG, HDL-c, ALT, AST, and Cr. The herbal capsule was standardized according to benzoic acid, ballic acid, rutin, and quercetin standard (6.5, 2.1, 1.4, 0.7 mg, respectively), and also α-pinene as major volatile constituent 28.51%. CONCLUSION: According to results the P. atlantica kurdica is effective in the improvement of lipid profile and glucose indices in line with its local application. CLINICAL TRIAL ID: (IRCT201708109014N178).

An arginine-rich peptide inhibits AChE: template-based design, molecular modeling, synthesis, and biological evaluation.

Life expectancy is growing especially in developed countries. In this regard, aging-associated diseases such as Alzheimer's disease (AD) are more common. Multi interconnected pathological factors involved in AD demand multi-target therapeutics. AChE, as a well-known target in AD, decreases the acetylcholine (ACh) in cholinergic synapse and, besides, increases the rate of amyloid-beta (Aß) aggregation. To block the destructive effects of AChE on cholinergic neurons in AD, we designed a peptidic inhibitor of the peripheral anionic site (PAS). The PAS plays a crucial role to attract and direct the ACh to the enzyme active site and increase the rate of Aß aggregation by changing the folding state. We utilized the template-based approach in combination with molecular docking, molecular dynamic simulation, and data mining to design a peptide library. Scoring was performed according to binding energy and the interaction profile of AChE inhibitors. The best candidate (p8, RMLRTTRY) was synthesized using solid-phase peptide synthesis, purified by RP-HPLC, and identified by ESI-MS. The inhibitory effect of p8 on AChE was 102.2 ± 15.2 µM. The kinetic and molecular modeling studies indicated the mixed inhibition mechanism for p8. The Arg residues in p8 had an essential role in binding to PAS.

Chemical composition and bioactivities of essential oils from different plant parts of Ferula pseudalliacea Rech.f. as an endemic plant from Iran.

The essential oils of leaf, flower, immature and mature fruit from Ferula pseudalliacea (Apiaceae) which grow wildly in Iran as an endemic plant were obtained by hydro-distillation and subsequently analysed by GC-FID and GC-MS. The oils obtained in yields 0.4-4.0% (w/w) and the analysis of GC-MS/FID chromatogram was resulted in the identification of 43-47 compounds, representing 90.9 - 96.7% of the total oils. α-Pinene was the main compound in all samples. The oils exhibited mild activity against Bacillus pumilus, Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus with or more than 15 mm zones of growth inhibition and MIC values of 9-15 mg/ml. Essential oil of immature and mature fruits showed potent antioxidant activity with IC50 values of 35.4 ± 0.4 and 39.1 ± 0.4 µg/mL, respectively. So, this plant as an antibacterial and antioxidant agent can be useful in food and medicine industry. [Formula: see text].

The Promising Effect of Peucedanum chenur Chloroformic Extract on Prevention of Human Colorectal Cancer Progression by Modulating miR-135b, miR-21, and APC Genes.

PURPOSE: The therapeutic use of herbal medicines for the diseases, including cancer, is increasing due to their lower side effects. The present research evaluated the effect of Peucedanum chenur chloroformic extract (PCCE) on cell proliferation against HCT-116 human colorectal cancer cell line. METHODS: The cytotoxic effect of PCCE was evaluated by MTT assay. The activity of the Wnt/B-catenin pathway was assayed through measuring the expression of miR-135b, miR-21, and APC genes by real-time PCR. The flow cytometry and scratch tests were used to study the cell cycle and cell migration, respectively. Also, the antioxidant activity of PCCE was measured by DPPH and iron-chelating tests. RESULTS: The results showed the downregulation of miR-135b and miR-21 and overexpression of the APC gene. Furthermore, PCCE decreased the free radicals, cell migration, and cell proliferation. The antioxidant activity of PCCE was confirmed by standard tests. CONCLUSION: Altogether, our findings suggest that purified compounds of PCCE could be developed as a potent chemo-preventive drug for the treatment of CRC.

Clinical and phytochemical studies of Plantago major in pressure ulcer treatment: A randomized controlled trial.

BACKGROUND: Plantago major L. is used by local people to improve various wounds and lesions such as pressure ulcer. In this study, the therapeutic effects of P. major topical formulation on the stage 1 pressure ulcer in patients have been investigated. MATERIALS AND METHODS: This randomized triple blind clinical trial study was performed on 130 patients. During the 14 days of study, each of the test and control groups was checked according to check list. Also the topical formulation was standardized by HPLC based on the amount of quercetin. RESULTS: The findings of this study indicated a significant difference in resolution of the damage between the test and control groups. Topical formulation was standardized by HPLC based on the quercetin (1.88 mg/100g) and no side effects associated with this topical formulation was found. CONCLUSION: The results confirmed the traditional use of P. major in resolution of the damage. CLINICAL TRIAL ID: (IRCT201609209014N117).

To be ionized or not to be ionized: the vital role of physicochemical properties of galbanic acid derivatives in AChE assay.

Alzheimer's disease is a progressive neurodegenerative disorder and patients suffer from memory loss, a decline in language skill and impairment in other cognitive functions. In the cholinergic hypothesis, dysfunction of cholinergic neurons especially in the hippocampus and cerebral cortex contributes to cognitive decline in patients. So agents that enhance acetylcholine concentration could improve cognitive function. AChEIs are among the most studied anti-Alzheimer agents. Galbanic acid as a natural compound with a sesquiterpene coumarin scaffold is a weak inhibitor of AChE. In the present contribution, we discussed the impact of carboxylic group ionization on inhibitory effects. We performed in vitro and in silico studies on galbanic acid, methyl and ethyl galbanates as AChE inhibitors. The order of inhibitory effect on AChE was obtained as ethyl galbanate ∼ methyl galbanate > galbanic acid. Our study highlights the important role of the physicochemical properties of natural lead compounds in each specific assay.Communicated by Ramaswamy H. Sarma.

Design, synthesis and biological evaluation of anticholinesterase peptides: Fragment-based vs. template-based peptide design.

The prevalence of Alzheimer's disease (AD) has become a substantial global concern. Approved AChE inhibitors have been used for symptomatic treatment of AD. Binding of amyloid ß (Aß) to the peripheral anionic site of AChE facilitates the formation of Aß plaques. Blocking this proposed protein-protein interaction by inhibition of the peripheral anionic site of AChE, in addition to increasing the level of ACh, reduces the Aß aggregation and might qualify to slow down the progression of disease besides the palliative treatment. Targeting protein-protein interactions consider as one of the most challenging issues in the realm of drug design in which peptides have potentials to excel in. In the present study, we applied two virtual fragment-based and template-based approaches to design peptidic inhibitors of the PAS of AChE. Based on the in silico studies, high scored peptides p2 (WTWYGYWVW) and p10 (NHRMLTRRY) obtained from fragment-based and template-based design respectively. Regarding in vitro results, p2 (IC50 = 16 ± 3.2 µM) and p10 (IC50 = 23.6 ± 4.9 µM) showed significant AChE inhibitory effects. The molecular mechanism of inhibition studied by Lineweaver-Burk plots was mixed inhibition for both peptides. The in vitro results conformed to the in silico results and showed that both peptides occupied the CAS and PAS of AChE. The comparison of two peptide-design approaches revealed that the fragment-based design had more chemical diversity and showed priority to the template-based design. According to the obtained results, peptidic inhibitors of AChE designed by the proposed fragment-based approach might be more efficient in comparison to traditional approaches.