Polymer Capped Silver Nanoparticles from Ziziphus nummularia Leaves Extract: Potent Antibacterial and Antioxidant Activity

Generally, synthesis and encapsulation process improve therapeutic value of nano encapsulated drugs. The silver nanoparticles (AgNPs) biosynthesized from Ziziphus nummularia leaves and encapsulated with polyvinyl pyrrolidone (PVP) polymer as antibacterial agents, due to its high bioavailability, better encapsulation and less toxic properties. The nanoparticles (AgNPs) biosynthesized from Ziziphus nummularia leaves and capped with polyvinyl pyrrolidone (PVP) polymer, The acquired AgNPs and polymeric functionalized AgNPs were fully characterised by the UV- Visible spectroscopy , Transmission electron microscopy (TEM), X-Ray diffraction pattern (XRD) and Fourier transform infrared spectroscopy (FTIR).The crystalline Ag NPs and Polymer Functionalized AgNPs have a face-centered cubic structure with an average size of 9.20 nm, according to X-ray Diffraction spectroscopy. Fourier Transform Infrared spectroscopy revealed that biomolecules such as proteins are incapable of reducing metal ions and the formation of an encapsulating layer in terms of metal ions. High-Resolution transmission electron microscopy revealed that Polymer functionalized AgNPs ranged in size of 10 nm. AgNPs and Polymer functionalized AgNPs showed effective antimicrobial and antioxidant activity. The biosynthesized monodisperse silver nanoparticles and encapsulated silver nanoparticles demonstrated better antimicrobial and antioxidant activity which can be used in various biomedical applications.

A Multi-Pronged Approach to Assessing Inflammatory Inhibitory Activity of a New Cyclic Alkaloid Compound from Root Bark of Ziziphus Nummularia (Aubrev.)

[(16-methoxy-10-(3-methyl-butyl)-2-oxa-6, 9, 12-triaza-tricyclo [13.3.1.03, 7] nonadeca-1(18), 13, 15 (19), 16-tetraene-8, 11-Dione], a putative cyclic alkaloid compound (IC) isolated from the root bark of Ziziphus nummularia, showed potential anti-inflammatory potential. Nitric oxide (NO), prostaglandin-E2 (PGE2), and tumour necrosis factor-alpha (TNF- α) levels were measured in vitro to assess IC's potential. ADME simulations and molecular docking of IC by TNF- α receptor were also performed. The in vivo potentials of IC and ethanolic extract (EE) were investigated by assessing carrageenan-induced paw oedema and arachidonic acid/xylene-induced ear oedema. TNF-α inhibition was higher in IC than in others, with a maximal percent inhibition of 88.00 percent at 50.11 µM. IC generated hydrogen bonds with ASP 45 and GLN 47, according to in silico research. Carrageenan, xylene, and arachidonic acid-induced oedema were all significantly reduced by IC. As a result, IC may have clinical potential in the future treatment of inflammation.

Ziziphus nummularia Attenuates the Malignant Phenotype of Human Pancreatic Cancer Cells: Role of ROS

Pancreatic cancer (PC) is the fourth leading cause of all cancer-related deaths. Despite major improvements in treating PC, low survival rate remains a major challenge, indicating the need for alternative approaches, including herbal medicine. Among medicinal plants is Ziziphus nummularia (family Rhamnaceae), which is a thorny shrub rich in bioactive molecules. Leaves of Ziziphus nummularia have been used to treat many pathological conditions, including cancer. However, their effects on human PC are still unknown. Here, we show that the treatment of human pancreatic ductal adenocarcinoma cells (Capan-2) with Ziziphus nummularia ethanolic extract (ZNE) (100–300 μg/mL) attenuated cell proliferation in a time- and concentration-dependent manner. Pretreatment with N-acetylcysteine, an ROS scavenger, attenuated the anti-proliferative effect of ZNE. In addition, ZNE significantly decreased the migratory and invasive capacity of Capan-2 with a concomitant downregulation of integrin α2 and increased cell–cell aggregation. In addition, ZNE inhibited in ovo angiogenesis as well as reduced VEGF and nitric oxide levels. Furthermore, ZNE downregulated the ERK1/2 and NF-κB signaling pathways, which are known to drive tumorigenic and metastatic events. Taken together, our results suggest that ZNE can attenuate the malignant phenotype of Capan-2 by inhibiting hallmarks of PC. Our data also provide evidence for the potential anticancer effect of Ziziphus nummularia, which may represent a new resource of novel anticancer compounds, especially ones that can be utilized for the management of PC.

Distinct morpho-physiological and biochemical features of arid and hyper-arid ecotypes of Ziziphus nummularia under drought suggest its higher tolerance compared to semi-arid ecotype

Tree species in the arid and semi-arid regions use various strategies to combat drought stress. Ziziphus nummularia, native to the Thar Desert in India, is highly drought-tolerant. To identify the most drought-tolerant ecotype of Z. nummularia, one ecotype each from semi-arid (Godhra, annual rainfall > 750 mm), arid (Bikaner, 250–350 mm) and hyper-arid (Jaisalmer, < 150 mm) regions was selected along with two other Ziziphus species; Z. mauritiana and Z. rotundifolia, and screened for parameters contributing to drought tolerance. Among these, Z. nummularia (Jaisalmer) (CIAHZN-J) was the most drought-tolerant. The tolerance nature of CIAHZN-J was associated with increased membrane stability, root length and number, length of hairs and thorns, root dry/fresh weight ratio, seed germination (at −0.5 MPa), proline content (31 fold), catalase and sugar content (2 to 3 fold). Apart from these characteristics, it also exhibited the longest duration to reach highest cumulative drought stress rating, maintained higher relative water content for a longer period of time with reduced leaf size, leaf rolling and falling of older leaves, and displayed sustained shoot growth during drought stress. To determine drought tolerance in Ziziphus, we developed a morphological symptom-based screening technique in this study. Additionally, transcriptome profiling of CIAHZN-J in response to drought revealed the up-regulation of genes involved in sugar metabolism and transport, ABA biosynthesis, osmo-regulation, ROS homeostasis and maintaining water potential. Expression profiles and semi-quantitative reverse transcription PCR results further correlated with the physiological and biochemical mechanisms. In conclusion, CIAHZN-J is an excellent genetic stock for the identification of drought-responsive genes and can also be deployed in crop improvement programmes for drought tolerance.

Assessing Genetic Variability and Population Structure of Ziziphus Nummularia as Characterized by Phenotypic Traits and SSR Markers with Climatic Distribution

Genus of Ziziphus (family Rhamnaceae), approximately 170 species of prodigious economic and ecological importance. Studies on the genetic diversity within Pakistani Ziziphus species are limited, and to date, no single report on the application of DNA barcoding attempt for identification of Ziziphus species is available in the literature. Therefore, the current study was designed to biogeographic distribution, to assess diversity based on phenotypic traits, SSRs markers, and relationships among Ziziphus nummularia (Burm.f.) Wight & Arn. genotypes. For the first time we use ArcGIS modeling, and their cartographic function for the identification of diversity in climatic variables such as temperature, mean diurnal, humidity, precipitation, and precipitation seasonality and the using of these techniques to determine the important variable which is responsible for the recent distribution of the Ziziphus species. A total of 11 phenotypic traits were noted and have significant phenotypic variation among the traits. The study has used 40 simple sequence repeats (SSR) markers for gaining insights into the genetic diversity within 180 genotypes of Z. nummularia. Successful amplification was achieved with 27 SSRs and was applied for understanding the population structure and relationships among the genotypes. A total of 120 alleles were amplified from Z. nummularia genotypes collected from three districts of the Malakand division, alleles per locus ranged from 2 to 6, averaging 4.4286. whereas polymorphism information content (PIC) from 0.332 to 0.794 in locus JSSR-490 and JSSR-97, within mean value was 0.671 per locus, expected heterozygosity (He), was 0.575, observed heterozygosity (Ho), 0.6618 and average gene diversity 0.494. Flow estimates (6.415) indicated frequent gene flow within Z. nummularia genotypes. Analysis of molecular variance (AMOVA) revealed high genetic variability (80%). The genetic relationship inferred from the neighbor-joining (NJ) phylogeny separate genotypes into three clusters and Bayesian model-based STRUCTURE analyses and PCoA analysis resolved all genotypes and indicted that the KP, populations, Swat and Buner, represent excesses of two different migration routes, with one designated from Swat and Dir (L). The overall results indicated the prevalence of genetic variability and relationships among Z. nummularia across geographical boundaries has retained unique alleles and this may facilitate the development of agronomically desirable and genetically improved Ziziphus cultivars and collections that can help achieve efficient conservation.

Feeding habits and habitat use of barking deer (Muntiacus vaginalis) in Himalayan foothills, Pakistan

Northern red muntjac (Muntiacus vaginalis; “barking deer”) is a shy and small-sized cervid mammal, limited to the outer Himalayan foothill forests in Pakistan. Habitat characteristics were measured by locating direct and indirect signs. To quantify habitat utilization of barking deer, 80 field surveys were conducted in the study area along transects. 1200 Quadrats at 50 m intervals were deployed along these transect lines to determine microhabitat factors associated with seasonal distribution. The food composition of the barking deer was determined through fecal droppings analysis by micro-histological technique. Forty-five fecal samples of barking deer were collected from the study area (Murree-Kotli Sattian-Kahuta National Pak); summer (28) and winter (17). The micro-histological analysis revealed that more plant species are available in its habitat during the summer season (27) as compared to winter (19). Due to browsing nature barking deer mostly feed on trees in both seasons. While shrubs are slightly higher in winters. In summer barking deer consumed 10 Trees, 6 Shrubs, 5 Herbs, and 6 kinds of grass species. Dominant tree species were Phyllanthus emblica and Acacia modesta. Dominant shrub species were Ziziphus nummularia and Justicia adhatoda. In winter barking deer consumed 8 Trees, 7 Shrubs, 3 Herbs, and 1 Grass. Dominant tree species were Bauhinia variegata and Acacia modesta while shrubs included Ziziphus nummularia and Carissa opaca.

Evaluation of Cholinesterase Inhibitory Potential of Different Genotypes of Ziziphus nummularia, Their HPLC-UV, and Molecular Docking Analysis

Ziziphus nummularia is an important source of valuable phytoconstituents, which are widely used in traditional medicine system of Indo-Pak sub-continent. In this study we investigated the distribution of phenolic compounds in the fruit pericarps of six different genotypes (ZNP01-06) of Z. nummularia growing in the unexplored hilly areas of Pakistan. The methanolic extracts of these genotypes were screened for total phenolic content (TPC), total flavonoid content (TFC), antioxidant, and cholinesterase inhibitory potentials. The observed biological potentials were explained in terms of the outcome of molecular docking and HPLC analyses. Among them, genotype ZNP02 displayed high TPC (88.50 ± 1.23 μg/mL) and showed potent scavenging activity against DPPH (67.03 ± 1.04 μg/mL) and ABTS (65.3 ± 1.74 μg/mL) in comparison to ascorbic acid (68.7 ± 0.47 μg/mL). Moreover, genotypes ZNP01, ZNP02, and ZNP04 displayed potent inhibition against acetyl and butyryl cholinesterases (AChE and BChE) with IC50 values of 21.2, 20.5, and 23.7 μg/mL (AChE) and 22.7, 24.4, and 33.1 μg/mL (BChE), respectively. Furthermore, the individual compounds in the most potent species ZNP01 responsible for potent enzyme inhibition (identified through HPLC-UV analysis), were computed via docking simulation software to the enzyme structures. Among these compounds rutin exhibited significant binding affinity with value of −9.20 kcal/mol. The differences amongst the phytochemical compositions of the selected genotypes highlighted the genotypic variations in them. Based on our results it was concluded that the selected plant can be used as remedy of oxidative stress and neurodegenerative diseases. However, further studies are needed to isolate responsible compounds and test the observed potential in vivo, along with toxicological evaluations in animal models.