Fruits Of Piper Nigrum Research Articles

Exploring multitarget potential of Piper nigrum fruit constituents for Alzheimer's disease: An AI-driven strategy

BackgroundAlzheimer's disease (AD) is a neurodegenerative disease that leads to development of cognition and memory dysfunctions. Currently, there is no known cure for AD, although limited medications are approved for the management of disease condition. Various plant-based leads give new hope for considering phytoconstituents as anti-AD drugs. The Piper nigrum L. fruit extract was reported to have anti-Alzheimer's activity. It creates an interest in the finding of active moieties that may be accountable for anti-AD activity. Hypothesis/PurposeIdentification of multitarget directed ligands isolated from Piper nigrum fruits through AI based studies. Study designThe phytochemical analysis of alkaloid fraction was carried out by LCMS, followed by the evaluation of constituents through in silico studies. MethodsThe fruits methanolic extract was prepared by cold maceration technique. The chemical profiling of the alkaloidal fraction was carried out using LCMS/MS analysis. The obtained compound's target hit genes were identified through network pharmacology studies using String, Metascape, and Cytoscape tools. Further, docking studies and MD simulations were carried out using AutoDock4 and Desmond-Maestro software. Then, electrochemical properties of hit compound P4 were determined using Gaussview6 software. ResultsFrom LCMS/MS analysis data, 29 compounds were considered based on compound intensity and accuracy (>95 %). Then, 41 common gene targets were identified from AD genes and compound-targeted genes. The 41 common genes in the PPI network suggested that AChE and BACE1 were the most abundant proteins. Further, docking studies revealed the hit compound P4 binding interaction and energies when compared to other 28 ligands. The molecular dynamics studies showed that P4-AChE and P4-BACE1 complexes were stable, and there were no RMSD and RMSF fluctuations were observed up to 100 ns. Further, PCA and MM-GBSA analysis data supported that complexes (P4-AChE and P4-BACE1) were stable. The DFT and surface properties indicated that compound P4 was ideal candidate for AD treatment and must be considered for further biological activity studies. ConclusionThe study identified compound P4 (dehydropipernonaline) from alkaloidal fraction of Piper nigrum fruits, suggesting it may be hit candidate for AD treatment.

Green Synthesis of Copper Oxide Nanoparticles from Waste Solar Panels Using Piper nigrum Fruit Extract and Their Antibacterial Activity

To promote environmental development and sustain resource circularity, recycling metals from electronic waste is essential. Electronic waste is a significant secondary source of metals, with its production increasing rapidly and most remaining unrecycled. In solar panels, copper is the second-most-valuable metal after silver. We propose an innovative method to recycle copper from waste solar panels and convert it into copper oxide nanoparticles (CuONPs) using a green synthesis method. Synthesizing CuONPs is advantageous due to their large surface area compared to bulk material. Nitric acid, a strong oxidizing agent, was used to leach copper from solid copper wires in waste solar panels. A green synthesis method, following a bottom-up approach, was employed using Piper nigrum fruit extract to synthesize CuONPs. The synthesized nanoparticles were characterized using various qualitative and quantitative techniques. Spectroscopic analysis confirmed the formation of CuONPs, and transmission electron microscopy revealed that the nanoparticles were spherical with sizes up to 60 nm. Biomolecules from the Piper nigrum extract were detected on the surface of the crystalline CuONPs. These nanoparticles exhibited antibacterial activity against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus using a well-diffusion method, showing a larger zone of inhibition for E. coli compared to S. aureus. This research demonstrates the complete recovery of copper from waste solar panels and its conversion into useful CuONPs, which have potential medicinal applications.

Biosynthesis, Antibacterial and Antifungal Activities of Ag Nanoparticles Prepared Using Piper nigrum Fruit Extract

Nanomaterials are promising alternatives to antibiotics with the potential to handle the menace of emerging multidrug resistance in pathogenic microbes. We report a simple biosynthetic approach to prepare Ag nanoparticles (NPs) with strong antibacterial and antifungal actions by using Piper nigrum fruit extract as a bioreducing agent. The biosynthesized material was characterized by SEM, AFM, TEM, HRTEM, XRD and UV–Visible absorption spectroscopy. SEM, AFM and TEM analyses provided evidence that the average diameter of the Ag NPs was 18[Formula: see text]nm, while XRD and HRTEM confirmed the FCC structure of Ag NPs. The biosynthesized Ag NPs demonstrated extremely strong antibacterial activity against Bacillus oceanisediminis and Escherichia coli, and showed extremely strong antifungal action against Schizosaccharomyces pombe, inhibiting their growth at very low concentrations of 3[Formula: see text][Formula: see text]g/mL, 4[Formula: see text][Formula: see text]g/mL and 3[Formula: see text][Formula: see text]g/mL, respectively. The molecular origin of their antibacterial and antifungal activity was studied using various molecular and biochemical assays. The results of these assays revealed a major role for reactive oxygen species (ROS) in the antifungal and antibacterial actions of the biosynthesized Ag NPs. Furthermore, morphological changes in cells and cell wall damage upon Ag NP treatment were observed by scanning electron microscopy. In addition, significant enhancement in protein leakage and fragmentation of DNA was also observed upon treatment of S. pombe, B. oceanisediminis and E. coli with Ag NPs. In conclusion, the observed extremely strong antibacterial and antifungal activities of biosynthesized Ag NPs are due to the oxidative stress triggered by ROS, causing cell wall damage and subsequent protein leakage and DNA fragmentation.

Evaluation of antimicrobial activity of a herbal mixture

Due to adverse effects of using allopathic medicines, the use and search for pharmaceuticals made from plants has increased recently. A mixture of herbal extracts known as “Chukku Kashaya” is widely used as home remedy in Kerala for respiratory disorders. Additionally, it relieves cold and fever symptoms. The present investigation focused on evaluating the antimicrobial activity of methanolic extracts obtained by soxhlet extraction of leaves of Ocimum tenuiflorum and Plectranthus amboinicus and fruits of Piper nigrum and rhizome of Zingiber officinalae and their combination in equal ratios and their activity was determined using agar well diffusion method against Staphylococcus aureus (gram positive), Klebsiella pneumoniae (gram negative), and Pseudomonas aeruginosae (gram negative). Streptomycin was used as positive control. Zingiber officinalae showed maximum zone of inhibition against all the bacteria and herbal mixture showed moderate zone of inhibition. The mixture was more active than Ocimum tenuiflorum , Piper nigrum and Plectranthus amboinicus. Staphylococcus aureus was shown to be the most sensitive to all the extracts and herbal mixtures, whereas Klebsiella pneumoniae showed the least activity. Though Z.officinalae has higher antimicrobial activity, there is limited literature support to prove its activity against fever, cold and other symptoms of respiratory tract infections. Z.officinalae is mainly used for GI related disorders. This herbal blend is more efficient because it includes of O.tenuiflorum : having antipyretic, analgesic and voice improving activity ; P.amboinicus: providing relief from cough, asthma, bronchitis and other symptoms of respiratory tract infections; P. nigrum which can improve appetite and digestion .

Exploring the Potential Mechanism of Action of Piperine against Candida albicans and Targeting Its Virulence Factors.

Plant-derived compounds have proven to be a source of inspiration for new drugs. In this study, piperine isolated from the fruits of Piper nigrum showed anti-Candida activity. Furthermore, the mechanisms of action of piperine and its impact on virulence factors in Candida albicans, which have not been comprehensively understood, were also assessed. Initially, piperine suppressed the hyphal transition in both liquid and solid media, hindered biofilm formation, and resulted in observable cell distortions in scanning electron microscope (SEM) samples, for both fluconazole-sensitive and fluconazole-resistant C. albicans strains. Additionally, the morphogenetic switches triggered by piperine were found to rely on the activity of mutant C. albicans strains. Secondly, piperine treatment increased cell membrane permeability and disrupted mitochondrial membrane potential, as evidenced by propidium iodine and Rhodamine 123 staining, respectively. Moreover, it induced the accumulation of intracellular reactive oxygen species in C. albicans. Synergy was obtained between the piperine and the fluconazole against the fluconazole-sensitive strain. Interestingly, there were no hemolytic effects of piperine, and it resulted in reduced cytotoxicity on fibroblast cells at low concentrations. The results suggest that piperine could have a dual mode of action inhibiting virulence factors and modulating cellular processes, leading to cell death in C. albicans.

Effectivity test of ethanol extract of black pepper (Piper nigrum) against mortality of Aedes aegypti and its effect on midgut morphological changes

Aedes aegypti is the vector of Dengue Hemorrhagic Fever (DHF). Dengue vector management usually depends on synthetic larvicides. However, it can lead to insect resistance and leave chemical pollutant residues. Therefore natural larvicides are needed for their environmentally friendly and do not induce insect resistance. Piper nigrum fruit is expected to contain secondary metabolites beneficial as larvicides to Aedes aegypti. This study aims to test the effectivity of the ethanol extract of Piper nigrum fruit on the mortality of Aedes aegypti larvae and determine the effect of extract exposure on larval midgut morphological changes. The study used a completely randomized design with five experimental groups and four repetitions as follows: K (aquades + 0% extract), P1 (0.25% extract), P2 (0.50% extract), P3 (0.75% extract), P4 (1% extract). Data analysis used one-way ANOVA and continued to the LSD Post hoc test. The extract's effectiveness was analyzed using the probit test and LC50 confirmation. Changes in the morphology of larval midgut were observed descriptively and presented in the form of tables and figures. The results showed that the 1% treatment resulted in the highest larval mortality of 93% and the lowest in the 0.25% treatment of 43%. The LC50 value was 2,666 based on the probit analysis. The ethanol extract of Piper nigrum fruit affected midgut morphological changes of Aedes aegypti larvae characterized by peripheral membrane damage, microvilli thinning, epithelial cell swelling and vacuolation, and basement membrane damage.

Green synthesis of Piper nigrum copper-based nanoparticles: in silico study and ADMET analysis to assess their antioxidant, antibacterial, and cytotoxic effects.

Nanobiotechnology is a popular branch of science that is gaining interest among scientists and researchers as it allows for the green manufacturing of nanoparticles by employing plants as reducing agents. This method is safe, cheap, reproducible, and eco-friendly. In this study, the therapeutic property of Piper nigrum fruit was mixed with the antibacterial activity of metallic copper to produce copper nanoparticles. The synthesis of copper nanoparticles was indicated by a color change from brown to blue. Physical characterization of Piper nigrum copper nanoparticles (PN-CuNPs) was performed using UV-vis spectroscopy, FT-IR, SEM, EDX, XRD, and Zeta analyzer. PN-CuNPs exhibited potential antioxidant, antibacterial, and cytotoxic activities. PN-CuNPs have shown concentration-dependent, enhanced free radical scavenging activity, reaching maximum values of 92%, 90%, and 86% with DPPH, H2O2, and PMA tests, respectively. The antibacterial zone of inhibition of PN-CuNPs was the highest against Staphylococcus aureus (23mm) and the lowest against Escherichia coli (10mm). PN-CuNPs showed 80% in vitro cytotoxicity against MCF-7 breast cancer cell lines. Furthermore, more than 50 components of Piper nigrum extract were selected and subjected to in silico molecular docking using the C-Docker protocol in the binding pockets of glutathione reductase, E. coli DNA gyrase topoisomerase II, and epidermal growth factor receptor (EGFR) tyrosine to discover their druggability. Pipercyclobutanamide A (26), pipernigramide F (32), and pipernigramide G (33) scored the highest Gibbs free energy at 50.489, 51.9306, and 58.615kcal/mol, respectively. The ADMET/TOPKAT analysis confirmed the favorable pharmacokinetics, pharmacodynamics, and toxicity profiles of the three promising compounds. The present in silico analysis helps us to understand the possible mechanisms behind the antioxidant, antibacterial, and cytotoxic activities of CuNPs and recommends them as implicit inhibitors of selected proteins.

New natural pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) and iNOS inhibitors identified from Penicillium polonicum through in vitro and in vivo studies.

Overexpression of pro-inflammatory cytokines and iNOS have been found to be concomitant with several chronic inflammatory diseases and hence targeting their inhibition would be a useful therapy for inflammation. In view of this, study on discovery of natural pro-inflammatory cytokines inhibitory lead molecules from Penicillium polonicum, an endophytic fungus isolated from the fresh fruits of Piper nigrum was performed. When the culture broth extract of P. polonicum (EEPP) was subjected to LPS-induced cytokines expression (ELISA in RAW 264.7 cells), it exhibited inhibition of TNF-α, IL-6 and IL-1β and this encouraged us to do chemical investigation on EEPP to explore the bioactive components. Four compounds isolated and characterised as 3,5-di-tert-butyl-4-hydroxy-phenyl propionic acid (1), 2,4-di-tert-butyl phenol (2), indole 3-carboxylic acid (3) and tyrosol (4) were tested for their effect on the production of TNF-α, IL-1β and IL-6 in RAW 264.7 cells (ELISA). All the compounds exhibited a highly significant (P<0.0001) inhibition effect, particularly against IL-1β (IC50: 4-0.91μM, 1-2.81μM, 3-4.38μM, and 2-5.54μM). Tyrosol (4) was most active with IC50 values of 0.91, 2.67 and 4.60μM against IL-1β, IL-6 and TNF-α, respectively. On observing the potential activity of the compounds, two compositions C1 and C2 were prepared by mixing equimolar concentrations of compounds 1, 2, 3 & 4 (C1) and compounds 1, 2, 3, 4 & piperine (C2) in equal ratio. A synergistic effect was observed with C1 exhibiting potential suppression of IL-6 secretion (IC50 1.91μM) and C2 against IL-1β (IC50 5.98μM). Also, the individual compounds and C1 were effective in controlling iNOS expressions in RAW 264.7 cells (RTPCR). Further, the in vivo performance of the compounds and compositions were studied under two in vivo inflammatory models (LPS-induced endotoxaemia and carrageenan-induced paw oedema). Compounds 1, 2, 3, 4, C1 and C2 at 50mg/kg oral dose showed a significant control over the LPS-stimulated TNF-α, IL-1β and IL-6 levels in plasma. C1, C2 and 1 exhibited>50% pan-cytokine inhibition effect. Under the carrageenan-induced anti-inflammatory model, a significant reduction in the paw oedema measured in terms of the difference in the paw thickness was observed. Further, attenuation of pro-inflammatory cytokines levels following ELISA and RT-PCR experiments in the paw tissue homogenate was in agreement with paw thickness results. All compounds and C1 decreased the iNOS gene expression levels, and also the MPO activity and NO production in the paw tissue homogenate with tyrosol (4) as the most active molecule. Further, the mechanism of action was explored by testing the effect of the compounds on the expression of inflammatory markers using western blot analysis (in vitro). They were found to regulate the expression of pro-form and matured-form of IL-1β by inhibiting NFκB. Also, the compounds reduced the translocation of the NF-κB subunit p65 to the nucleus. Thus, compounds 3,5-di-tert-butyl-4-hydroxy-phenyl propionic acid (1), 2,4-di-tert-butyl phenol (2), indole 3-carboxylic acid (3) and tyrosol (4) are reported as new natural multiple pro-inflammatory cytokines inhibitory leads. The interesting results of C1 might lay a footing for the development of a new anti-inflammatory composition.

Biosynthesized Ag–ZnO nanohybrids exhibit strong antibacterial activity by inducing oxidative stress

We report facile biosynthesis of Ag–ZnO nanohybrids consisting of Ag nanoparticles decorated ZnO nanobullets prepared by decorating wet chemically synthesized ZnO nanobullets with Ag nanoparticles through bioreduction of Ag + ions with aqueous extract of Piper nigrum fruits. The prepared nanomaterials were well characterized by FESEM, TEM, HRTEM, EDX, XRD, XPS, PL and UV–vis spectroscopy. FESEM and TEM analyses on the nanohybrids revealed ∼18 nm Ag nanoparticles decorating ZnO nanobullets with average size ∼48 nm. XRD results revealed hexagonal wurtzite ZnO with 22.4 nm crystallite size and FCC Ag with 18.7 nm crystalline size. Ag–ZnO nanohybrids exhibited strong antibacterial action against Escherichia coli, Bacillus oceanisediminis and Pseudomonas entomophila and efficiently inhibited their growth at 100 μg/mL, 50 μg/mL and 125 μg/mL, respectively. The molecular basis of antibacterial action of Ag–ZnO nanohybrids against E. coli was investigated using different biochemical and molecular assays. Addition of antioxidant histidine suppressed the antibacterial action of Ag–ZnO nanohybrids towards E. coli due to its ROS scavenging action. Bradford assay results showed enhanced protein leakage from Ag–ZnO nanohybrids treated E. coli, while TBARS assay results confirmed lipid peroxidation triggered by ROS. SEM on Ag–ZnO nanohybrids treated E. coli confirmed significant damage to the cell wall leading to morphology change. The antibacterial activity of Ag–ZnO nanohybrids against E. coli is mainly due to the ROS-induced oxidative stress, which caused enhanced lipid peroxidation, cell wall damage leading to significant protein leakage and DNA fragmentation.

A New Lignan (Polonilignan) and Inhibitors of Nitric Oxide Production from Penicillium polonicum, an Endophytic Fungi of Piper nigrum.

Inhibiting nitric oxide (NO) or its production is found to be of therapeutic benefit. To discover natural small molecule inhibitors of NO production, a bioassay- and LC/MS-guided chemical investigation was done on the metabolites of endophytic fungus isolated from fresh Piper nigrum fruits. The isolated pure strain was identified as Penicillium polonicum by 16S rDNA sequence comparison. The culture broth extract of P. polonicum (EEPP) exhibited a significant reduction of NO production (Griess method) in LPS-stimulated RAW 264.7 cells (P<0.0001). To understand the chemical constituents of bioactive EEPP, column chromatography and p-TLC studies were carried out, which yielded eight pure compounds. They were characterised as botryosphaeridione (1), 3-(3,5-di-tert-butyl-4-hydroxy)phenylpropionic acid (2), variabilone (3), 2,4-di-tert-butylphenol (4), indole-3-carboxylic acid (5), tyrosol (6), ethyl ferulate (7) and a new lignan (8) based on the spectral analysis. The structure elucidation of the new lignan, named polonilignan (8), was based on HR-MS, 1 H- & 13 C-NMR, H-H COSY, HSQC and HMBC spectra. Compounds 2, 4, 5 and 6 showed a significant decrease (P<0.0001) in the production of NO in LPS-induced RAW 264.7 cells. Tyrosol (6) and indole-3-carboxylic acid (5) controlled nitrite release with IC50 values of 22.84 and 55.01 μM, respectively. This is the first report of (i) P. polonicum as an endophytic fungus of pepper fruits, (ii) isolation of compounds 1-8 except 6 from P. polonicum culture broth extract and (iii) NO inhibition effect of 2, 4, 5 and 6.

Evaluation of Pharmacokinetic Profile of Cirpusins®, Extract of Cyperus rotundus in Presence of Bioavailability Enhancer, BioPerine®

Obesity, a condition accompanying abnormal or excessive accumulation of fat in the body has become a major public health concern globally. Various herbs hailed from traditional systems of medicine like Ayurveda have gained importance in recent years to combat obesity. One such herb Cyperus rotundus (nutgrass), is known for its beneficial effects in obesity, in Ayurveda. Cirpusins® a patented extract from the dried rhizomes of Cyperus rotundus (standardized for minimum of 6% total identified stilbenes - Piceatannol, Scirpusin A &amp; Scirpusin B) and BioPerine®, a bioavailability enhancer, obtained from the dried fruits of Piper nigrum (standardized to minimum 95% piperine). In this study, we examined the pharmacokinetic profile and bioavailability of Cirpusins® in Sprague-Dawley rats at two different doses (45 and 90 mg/kg) in the presence and absence of BioPerine® respectively. The results signifies that addition of BioPerine® (0.45 mg/kg) with Cirpusins® (45 mg/kg) enhances the bioavailability by reducing the effective dose of Cirpusins® (90 mg/kg) to half.

Synergistic effect of Trikatuk, a traditional Thai formulation, on antioxidant and alpha-glucosidase inhibitory activities

Oxidative stress and hyperglycemia are known to be responsible for several diseases, including diabetes. To prevent these diseases, efforts are ongoing to identify novel antioxidants with hypoglycemic effects. Trikatuk is a traditional Thai formulation composed of three herbs in equal quantities: the fruits of Piper nigrum and Piper retrofractum and the rhizomes of Zingiber officinale. This formulation has been reported to have antioxidant activity and the three individual herbs have been traditionally used for managing diabetes. Therefore, the present study aimed to investigate the phytochemical contents, antioxidant activity (DPPH and ABTS radical scavenging assays), and alpha-glucosidase inhibitory activity of Trikatuk. As Trikatuk is a combination of herbs, the synergistic effect and two different preparation methods from powder or extract mixtures were also explored. The results revealed that Trikatuk, particularly prepared from a mixture of extracts, exhibited high contents of piperine, total phenolics and total flavonoids, as well as significant antioxidant and alpha-glucosidase inhibitory activities. Trikatuk also exhibited additive or synergistic effects in all experiments. Overall, Trikatuk is potentially an alternative traditional medicine and dietary supplement offering antioxidant activity and diabetes management.

Piper nigrum Fruit Extract as an Antibiotic Resistance Reversal Agent in MDR Bacteria

Antibiotic resistance development and spread in clinical pathogens is an immense threat that has already outpaced the discovery and development of novel and more effective antibiotic drugs. Recently the focus has been shifted to medicinal plants as novel therapeutic options for reversing antibiotic resistance by targeting different resistance mechanisms. Piper nigrum is a plant that has the potential to reverse antibiotic resistance and increase the efficacy of the current drugs. In the present study, seven different antibiotics, clindamycin, gentamicin, levofloxacin, amikacin, tigecycline, imipenem, and tetracycline, were used against antibiotic-resistant Staphylococcus aureus and Salmonella typhi. Antibiotic resistance reversal analysis was tested by the AST disc method. Increased zones of inhibition of S. aureus by four antibiotics, clindamycin (9 mm), gentamicin (7 mm), levofloxacin (9 mm), and amikacin (9 mm) were recorded after using P. nigrum extract. In addition, the use of P. nigrum extract also increased the zone of inhibition of S. typhi with amikacin (11 mm), gentamicin (10 mm), tigecycline (9 mm), levofloxacin (11 mm), and imipenem (10 mm). This study suggests that P. nigrum extracts can be used as natural antibiotic resistance reversal agents that increase the effectiveness of current antibiotics and can reverse antibiotic resistance.

Deciphering the impact and mechanism of Trikatu, a spices-based formulation on alcoholic liver disease employing network pharmacology analysis and in vivo validation.

Trikatu Churna (TC) comprising Zingiber officinale rhizome, Piper longum, and Piper nigrum fruit, is effective in treating liver diseases and has high nutraceutical values. However, the efficacy of TC in treating alcoholic liver disease (ALD) and its mechanism remain largely unknown. This study evaluated the hepatoprotective effects of different doses of TC as well as to identify the bioactive components and determine their mechanism of action against ethanol-induced ALD. A compound-target network analysis model of TC was established to identify its potential bioactive compounds and pathways that might regulate its hepatoprotective effects. Further, in-vivo studies were performed to validate the potential of TC (200 mg/kg and 400 mg/kg b.w.) in the treatment and management of ALD. The study revealed that both the dosages of TC demonstrate significant (p > 0.0001) hepatoprotective effects by improving body weight, total bilirubin, serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), serum alkaline phosphate (ALP), total cholesterol, total protein, globulin, albumin, and liver morphology. The High-performance thin-layer chromatography (HPTLC) fingerprinting of TC showed the presence of piperine. Network pharmacology identifies the role of TC in regulating various signaling processes including Advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE), Hypoxia-inducible factors (HIF-1), Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-Kappa B), and Phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling to exert its anti-inflammatory, antioxidant and anti-apoptotic role in managing ALD. Based on the bioinformatics analysis, some of the key targets of TC were found to be Prostaglandin-Endoperoxide Synthase 2 (PTGS2) or Cyclooxygenase-2 (COX-2), Sirtuin 1 (SRT1), and caspase-3. These effects may serve as a novel therapeutic option for the treatment of ALD. These preclinical validation studies for the ethnopharmacological potential of TC in ALD treatment further paved the way for researchers to perform next-level translational and clinical studies. Further, in-depth experimental studies for the validation of these bioinformatics-based results will give a clearer picture of mechanisms.

The terminal enzymatic step in piperine biosynthesis is co-localized with the product piperine in specialized cells of black pepper (Piper nigrum L.).

Piperine (1-piperoyl piperidine) is responsible for the pungent perception of dried black pepper (Piper nigrum) fruits and essentially contributes to the aromatic properties of this spice in combination with a blend of terpenoids. The final step in piperine biosynthesis involves piperine synthase (PS), which catalyzes the reaction of piperoyl CoA and piperidine to the biologically active and pungent amide. Nevertheless, experimental data on the cellular localization of piperine and the complete biosynthetic pathway are missing. Not only co-localization of enzymes and products, but also potential transport of piperamides to the sink organs is a possible alternative. This work, which includes purification of the native enzyme, immunolocalization, laser microdissection, fluorescence microscopy, and electron microscopy combined with liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), provides experimental evidence that piperine and PS are co-localized in specialized cells of the black pepper fruit perisperm. PS accumulates during early stages of fruit development and its level declines before the fruits are fully mature. The product piperine is co-localized to PS and can be monitored at the cellular level by its strong bluish fluorescence. Rising piperine levels during fruit maturation are consistent with the increasing numbers of fluorescent cells within the perisperm. Signal intensities of individual laser-dissected cells when monitored by LC-ESI-MS/MS indicate molar concentrations of this alkaloid. Significant levels of piperine and additional piperamides were also detected in cells distributed in the cortex of black pepper roots. In summary, the data provide comprehensive experimental evidence of and insights into cell-specific biosynthesis and storage of piperidine alkaloids, specific and characteristic for the Piperaceae. By a combination of fluorescence microscopy and LC-MS/MS analysis we localized the major piperidine alkaloids to specific cells of the fruit perisperm and the root cortex. Immunolocalization of native piperine and piperamide synthases shows that enzymes are co-localized with high concentrations of products in these idioblasts.

Preparation and Standardization of Vrihatsamsharkar Churna: A Polyherbal Formulation

In India the traditional system of medicine is in practice from the ancient times for the treatment of several diseases. Due to lack of standardization or purity of herbal drugs some effective drugs are not regularly used for the treatment which is a major drawback for the herbal sector and pharma industries. That is why the standardization techniques for herbal drugs are required to be developed and make a bond with the main science health stream. There are certain preliminary guidelines given by CCRAS (Central Council for Research in Ayurveda and Siddha) for the standardization of these herbal drugs. Standardization of herbal formulations is a prime requirement for the establishment of the good quality of such herbal drugs. The present paper report the preparation and standardization of a poly herbal formulation which contain Syzygium aromaticum (flower buds), Myristica fragrans (seeds), Piper longum (fruit), Zingiber officinale (rhizome), Piper nigrum (fruit). This “Vrihatsamsharkar” churna poly herbal formulation is used in the treatment of dry cough. Following parameters like microscopy, macroscopy, ash value, extractive value, water soluble ash, bulk density, tapped density, Hausner ratio, Carr’s index, moisture content, swelling index and preliminary tests are calculated and discussed in this paper. These are certain important parameters which are necessarily required for the authentication of any herbal drug or any herbal formulation.

The aflatoxin inhibitors capsaicin and piperine from Capsicum chinense and Piper nigrum fruits modulate the antioxidant system in Aspergillus parasiticus

Several aflatoxin inhibitors can modulate the antioxidant system in fungi. In this work, the effect of the ethanolic extract of Capsicum chinense and Piper nigrum fruits, capsaicin, and piperine on the expression of the aflE, aflG, aflH, aflI, aflK, aflL, aflO, aflP, and aflQ genes involved in the aflatoxin biosynthetic pathway in Aspergillus parasiticus were studied by qRT-PCR analysis. As well as, the effect on the expression of fungal antioxidant genes (sod1, catA, and cat2) and enzymatic activity of catalase (CAT) and superoxide dismutase (SOD). Results reveal that the highest (p < 0.05) radial growth inhibition (68 and 86%) and aflatoxins production inhibition (73 and 80%) was observed with capsaicin and piperine respectively, at 300 µg/mL, instead of the ethanolic extract at the same concentration. The qRT-PCR analysis showed that compounds and extracts at 300 µg/mL induced a down-regulation of aflatoxin genes and an up-regulation on the fungal antioxidant genes. CAT activity increased by 23.15, 36.65, 51.40, and 65.50%, in the presence of C. chinense and P. nigrum extract, capsaicin, and piperine exposure, respectively. While SOD activity was not significantly impacted (p > 0.05). In conclusion, the capsaicin and piperine, two antifungal and anti-aflatoxigenic compounds produce an up-regulation of antioxidant defense genes accompanied by an enhancement of catalase enzymatic activity in A. parasiticus.

Design, Development and Evaluation of Anisomeles indica Herbal Cough Syrup

The objective of the present study was to design, development and evaluation antitussive herbal cough syrup because designing of oral herbal formulation is still a challenge in modern pharmaceutics and the syrup formulation presents many technical problems to the industrial pharmacist. Potential antitussive herbs were used for developing the herbal cough syrup. Decoction of plant Anisomeles indica, Leaves of Adhathoda vasika, stems of Glycyrrhiza glabra, fruits of Piper nigrum and plant of Mentha piperita was prepared. One part of decoction was mixed with five parts of simple syrup IP (1:5) to prepare formulation. The formulations were evaluated by morphological characters, physical parameters like PH, Specific gravity, viscosity etc. Herbal cough syrup was also subjected for the accelerated stability testing (AST) for the period of 72hours at accelerated temperature conditions. No marked changes were noticed in all the evaluated parameters during AST. The laboratory scale preparation of herbal Syrup may be used as a stable, liquid dosage form and the work done in stability testing may help in the progress of shelf-life determination studies. The presence study includes design, development and evaluation of Anisomeles indica herbal cough syrup first time.

Neuroprotective Effects of Black Pepper Cold-Pressed Oil on Scopolamine-Induced Oxidative Stress and Memory Impairment in Rats.

Oxidative stress is usually associated with many neurodegenerative diseases. In this study, the gas chromatography–mass spectrometry (GC–MS) analysis of cold-pressed oil (CPO) from black pepper (Piper nigrum) fruits was performed and its neuroprotective effects were evaluated for the first time. The analysis of CPO revealed the presence of the lignan sesamin (39.78%), the alkaloid piperine (33.79%), the monoterpene hydrocarbons 3-carene (9.53%) and limonene (6.23%), and the sesquiterpene β-caryophyllene (10.67%). Black pepper hydrodistilled oil (HDO) was also comparatively analyzed by GC–MS to show the impact of oil isolation by two different methodologies on their components and class of compounds identified. HDO analysis revealed 35 compounds (99.64% of the total peak areas) mainly composed of monoterpene hydrocarbons (77.28%), such as limonene (26.50%), sabinene (21.36%), and β-pinene (15.53%), and sesquiterpene hydrocarbons (20.59%) represented mainly by β-caryophyllene (19.12%). Due to the low yield obtained for HDO (0.01% v/w), only CPO was chosen for the evaluation of its neuroprotective potential. Alzheimer-type dementia was induced in rats by scopolamine intraperitoneal injection (1.5 mg/kg/day) for seven days. CPO was administered orally (100 mg/kg) for a week before scopolamine administration and then concomitantly for another week. Donepezil (1 mg/kg, orally) was used as a reference drug. CPO administration significantly improved the rat behaviors as evaluated by the Morris water maze test, evident from prolongation in time spent in the platform quadrant (262.9%, compared to scopolamine) and increasing in the crossing time by 18.18% compared to the control group. The rat behavior tested by passive avoidance, showed prolongation in the step-through latency compared to control. Moreover, CPO significantly (p < 0.05) ameliorated the activities of antioxidant enzymes such as catalase, superoxide dismutase (SOD) and reduced malondialdehyde (MDA) equivalents by 22.48%, 45.41%, and 86.61%, respectively, compared to scopolamine. Furthermore, CPO administration decreased scopolamine-induced elevated acetylcholinesterase levels in rats’ hippocampi by 51.30%. These results were supported by histopathological and in silico molecular docking studies. Black pepper oil may be a potential antioxidant and neuroprotective supplement.

Evaluation of Additive or Synergistic Effect of Piper nigram and Ocimum sanctum Extracts for their Antidepressant Activity

Depression is a state of excessive sensitivity to criticism, fear of rejections, lack of self-interest, loss of pleasure. In the traditional systems of medicine, many plants and formulations have been used to treat depression for thousands of years. In recent times, research on the plants increased globally and so many plants provide the evidence to cure diseases. Ocimum sanctum, popularly known as Tulsi is one of the sacred herbs for Hindus in the Indian subcontinent. It has a versatile role in traditional medicine. The fruits of Piper nigrum are used to make black pepper. This hotly pungent spice is one of the earliest known and most widely used spices in the world today. Wide range of animal tests for antidepressant agents are commonly used. The Forced swim test and Tail suspension test in mice were mostly used. Hence in the present study Forced swim test was used as animal model of depression. In present study immobility time in Forced swim test was significantly decreased by a combination of Piper nigrum fruit extract and Ocimum sanctum extract treated groups compared to control group. The combination of extracts (50 mg/kg each) activity was comparable to standard drug Fluoxetine. Treatment with extracts does not modify the locomotor activity of mice, which indicates that they exert antidepressant effects without modifying significantly locomotor activity. Therefore, the present study confirms the combination of alcoholic extract of Piper nigrum (AEPN) fruit and aqueous extract of Ocimum sanctum (AEOS) possessing additive/synergistic antidepressant activity.