Functional annotation of proteins in Catharanthus roseus shoot cultures under biogenic zinc nanotreatment.

Nano-interactions are well known for their positive as well as negative impacts on the morphological and physiological systems of plants. Keeping in mind, the conformational changes in plant proteins as one of the key mechanisms for stress adaptation responses, the current project was designed to explore the effect of glutathione-capped and uncapped zinc nano-entities on Catharanthus roseus shoot cultures. Zinc nanotreatment (0.05 µg/mL) significantly induced ester production in C. roseus shoots as detected by Gas Chromatography-Mass spectrometry. These nanotreated shoots were further subjected to peptide-centric nano-LC-MS/MS analysis. Mass spectrometry followed by a Heat map revealed a significant effect of zinc nanoparticles on 59 distinct classes of proteins as compared to control. Proteins involved in regulating stress scavenging, transport, and secondary metabolite biosynthesis were robustly altered under capped zinc nanotreatment. UniProt database identified majority of the localization of the abundantly altered protein in cell membranes and chloroplasts. STRING and Cytoscape analysis assessed inter and intra coordination of triosephosphate isomerase with other identified proteins and highlighted its role in the regulation of protein abundance under applied stress. This study highlights the understanding of complex underlying mechanisms and regulatory networks involved in proteomic alterations and interactions within the plant system to cope with the nano-effect.

Functionalization of ampicillin and gentamicin with biogenic copper nanoparticles (CuNPs) remodel antimicrobial and cytotoxic outcome against MDR clinical isolates.

The present study reports the functionalization of antibiotic-conjugated Alternanthera pungens and Trichodesma indicum copper nanoparticles (CuNPs). Initially, antibiotic profiling of multi-drug resistant (MDR) clinical isolates against five antibiotics was verified and then gentamicin and ampicillin conjugates of CuNPs were prepared. Biosynthesized nanostructures were characterized through UV-visible spectroscopy, Fourier-transformed infrared spectroscopy, X-ray diffraction and scanning electron microscope. Biogenic synthesized CuNPs displayed highest antibacterial activity (24.0-31.3 mm inhibition zones) when capped with gentamicin as compared to the ampicillin-conjugated NPs which showed resistance against most of the bacterial species. A. pungens-derived conjugates of gentamicin (CuAp-GNT) along with the vehicle revealed 4.86 ± 0.20% and 4.25 ± 2.96% hemolytic potential and highest MDA production in S. typhimurium (3.18 ± 1.52 µg/mL and 6.31 ± 3.49 µg/mL) and K. pneumoniae (2.99 ± 0.90 µg/mL and 4.06 ± 1.20 µg/mL). Similarly, CuAp-GNT also showed highest DNA protection ability by displaying 1342.99 ± 11.87 band intensity. All-inclusive, CuAp showed more promising effects when conjugated with gentamicin indicating that capping of gentamicin with the active components of the plant-based copper nanostructures increases the antibacterial capacity of the drug. Hence, conjugation of antibiotics with bio-based sources offers great potential for identifying potent drug leads.

Biological screening of three selected folklore medicinal plants from Pakistan.

This study investigates the pharmacological potential of Adiantum incisum, Alternanthera pungens and Trichodesma indicum. Biological activities of plant extracts (aqueous, methanolic and n-hexane extracts of whole plants) were screened by antitumor potato disc assay (10000, 1000, 100, 10 ppm doses), antifungal tube dilution assay (50, 25, 12.5, 6.25mg/ml) and antioxidant DPPH/reducing power assays (250, 200, 150, 100, 50µg/ml). Significant amount of alkaloids (230.83±30.20mg/g) in Adiantum incisum with lowest amount of phenolics in Alternanthera pungens (43.45 ±14.22µg/mg) were detected. Significant antitumor potential (p<0.05) was revealed by Trichodesma indicum n-hexane extract (85% tumor inhibition; IC50 <10ppm). Moderate to significant antifungal activity (50.73%-78.3%) was shown against Aspergillus niger by Adiantum incisum extracts. Hexane extract of Trichodesma indicum revealed significant antifungal activity (98.9% inhibition) against Mucor specie. Methanolic extracts of all plants displayed significant DPPH radical scavenging potential (96.72%-60.33%; IC50 <50µg/ml) and Ferric power reducing ability with absorbance values (0.164-0.942) very close to standard ascorbic acid. Present study supports the extensive use of these plants in folk medicine and also promotes elaborative in-vivo investigations, isolation of pure therapeutic compounds and formulation of plant-based drugs.

Antibacterial evaluation of silver nanoparticles synthesized from lychee peel: individual versus antibiotic conjugated effects.

This paper describes the extracellular synthesis of silver nanoparticles from waste part of lychee fruit (peel) and their conjugation with selected antibiotics (amoxicillin, cefixim, and streptomycin). FTIR studies revealed the reduction of metallic silver and stabilization of silver nanoparticles and their conjugates due to the presence of CO (carboxyl), OH (hydroxyl) and CH (alkanes) groups. The size of conjugated nanoparticles varied ranging from 3 to 10 nm as shown by XRD. TEM image revealed the spherical shape of biosynthesized silver nanoparticles. Conjugates of amoxicillin and cefixim showed highest antibacterial activity (147.43 and 107.95%, respectively) against Gram-negative bacteria i.e. Alcaligenes faecalis in comparison with their control counterparts. The highest reduction in MIC was noted against Gram-positive strains i.e. Enterococcus faecium (75%) and Microbacterium oxydans (75%) for amoxicillin conjugates. Anova two factor followed by two-tailed t test showed non-significant results both in case of cell leakage and protein estimation between nanoparticles and conjugates of amoxicillin, cefixime and streptomycin. In case of MDA release, non-significant difference among the test samples against the selected strains. Our study found green-synthesized silver nanoparticles as effective antibacterial bullet against both Gram positive and Gram negative bacteria, but they showed a more promising effect on conjugation with selected antibiotics against Gram negative type.

Physiological and biochemical mechanisms of allelopathy mediated by the allelochemical extracts of Phytolacca latbenia (Moq.) H. Walter.

In allelopathy, one plant suppresses the growth and development of other plant/plants by negatively affecting a variety of physiological and biochemical reactions. We checked the effects of methanolic extracts (allelochemical extracts) of Phytolacca latbenia (Moq.) H. Walter on antioxidant enzyme activities such as peroxidases (PODs), super oxide dismutases (SODs) and catalase (CAT) and on total protein contents (TPC), cellular injury (CI), and malondialdehyde (MDA) in the germinating seeds of Brassica napus L. (dicot) and Triticum aestivum L. (monocot). Both the crude methanolic extract root (CMER) and crude methanolic extract aerial (CMEA) of P. latbenia at 10000 ppm significantly reduced the POD activity in both the test seeds. The activity of SODs was significantly decreased by both CMER and CMEA in B. napus germinating seeds. A linear increase in the activity of CAT, CI, and MDA contents was found in both the test seeds with the increasing concentrations of CMEA and CMER, while TPC of the germinating seeds was found decreased. It is inferred that both the CMEA and CMER inhibited/delayed the seed germination, reduced the seedling growth by affecting a variety of biochemical and physiological attributes, and also caused cellular membrane injury in the germinating seeds of both the monocot and dicot seeds.

Green solvent-based extraction of three Fabaceae species: A potential antioxidant, anti-diabetic, and anti-leishmanial agents.

The Fabaceae is renowned for its diverse range of chemical compounds with significant biological activities, making it a valuable subject for pharmacological studies. The chemical composition and biological activities of three Fabaceae species were investigated using methanol separately and in combination with dimethyl sulfoxide (DMSO) and glycerol for extraction. The results revealed the highest phenolic (49.59 ± 0.38 mg gallic acid equivalent/g), flavonoid (29.16 ± 0.39 mg rutin equivalent/g), and alkaloid (14.23 ± 0.54 mg atropine equivalent/g) contents in the Caesalpinia decapetala methanol extracts. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity and DNA protection activity were the highest (0.88 ± 0.43 µg/mL IC50 and 2149.26 band intensity) in Albizia julibrissin methanol extracts. The α-amylase activity was highest in all methanol extracts (<15 µg/mL IC50 values), while the α-glucosidase inhibition potential was highest (<1 µg/mL IC50 value) in the methanol-glycerol and methanol-DMSO extracts. Pearson coefficient analysis showed a strong positive correlation between the DPPH and α-amylase assays and phytochemicals. Anti-leishmanial activity was observed in decreasing order: A. julibrissin (74.75 %) > C. decapetala (70.86 %) > Indigofera atropurpurea (65.34 %). Gas chromatography-mass spectrometry revealed 33 volatile compounds and, aamong these (Z)-9-octadecenamide was detected in the highest concentration ranging from 21.85 to 38.61 %. Only the methanol extracts of the examined species could be assessed for in vivo studies for immediate applications.

Evaluation of potential inhibitory effects on acetylcholinesterase, pancreatic lipase, and cancer cell lines using raw leaves extracts of three fabaceae species.

The present study examined the biological potential and phytochemicals of Sophora mollis, Mucuna pruriens, and Indigofera atropurpurea methanolic leaf extracts. In vitro anti-acetylcholinesterase and anti-lipase assays were performed using different concentrations of plant extracts, and the IC50 values were determined. The cytotoxic potential of the selected plant extracts was assessed against HeLa, PC3, and 3T3 cell lines using an MTT assay. S. mollis leaf extract displayed the highest inhibition percentage (114.60% ± 19.95 at 1000 µg/mL) for the anti-acetylcholinesterase activity with a prominent IC50 value of 75.9 µg/mL. The anti-lipase potential was highest with the M. pruriens leaf extract (355.5 µg/mL IC50), followed by the S. mollis extract (862.7 µg/mL IC50). Among the cell lines tested, the cytotoxic potential of the I. atropurpurea extract (91.1 ppm IC50) against the PC3 cell line was promising. High-performance liquid chromatography revealed gallic acid, chlorogenic acid, caffeic acid, vanillic acid, rutin trihydrate, and quercetin dihydrate in varying concentrations in all plant species. The concentration of chlorogenic acid (69.09 ppm) was highest in M. pruriens, and the caffeic acid concentration (45.20 ppm) was higher in S. mollis. This paper reports the presence of bioactive therapeutic compounds in selected species of the Fabaceae family that could be micro-propagated, isolated, and utilized in pharmaceutical industries.

DAT and PRX1 gene expression modulates vincristine production in Catharanthus roseus L. propagates using Cu, Fe and Zn nano structures.

Underlying mechanism of nanostructures upon monoterpene induction in Catharanthus roseus has not been explored yet. In the current study, Copper, Iron and Zinc nanoparticles were biosynthesized by Eriobotrya japonica seed extract and capped with reduced glutathione. Biosynthesized nanoparticles and their capped analogues were characterized by UV-visible spectrophotometer, FTIR, XRD and SEM. Selected concentration of nanostructures were used in plant tissue culture media which instigated the production of alkaloids, tannins and flavonoids without significantly affecting the growth index of propagated calli and shoots cultures of C. roseus. Accelerated vincristine production was noticed in propagated calli and shoots under copper and zinc nanostress (1645-1865 µg/ml respectively) with the least effect by iron nanostructure. Highest concentration of calcium was recorded in in vitro shoots under capped (3.42 mg/ml ± 7.16) and uncapped (4.41 mg/ml ± 20.44) Zn nanoparticles compared to control (2.82 mg/ml ± 13.41). Real time PCR depicts nano-zinc mediated increased expression of DAT and PRX1 genes of TIA pathway. Significant correlation among PRX1/DAT gene expression with vincristine production and calcium accumulation in the presence of nanostress validate by PCA. This study paved way the opportunities of metal biogenic nanomaterials as an ideal drug modulator in plant tissue culture studies.

Cancer and Disease Diagnosis - Biosensor as Potential Diagnostic Tool for Biomarker Detection.

Analysis of cancer biomarkers has enormous promise for advancing our molecular understanding of illness and facilitating more precise and timely diagnosis and follow-up care. MicroRNA, exosomes, ctDNA, CTCs, and proteins are only some of the circulating biomarkers that can be detected by liquid biopsy instead of the more intrusive and time-consuming process of doing a tissue biopsy. As the cancer diagnosis bio-markers reveal ultra-low levels in the early stages of the disease, highly sensitive approaches are urgently required. Researchers have taken an interest in a optical biosensor for detecting cancer biomarkers as a potential tool for early disease diagnosis. These techniques have the potential to aid in the development of effective treatments, ultimately leading to a higher rate of patient survival. This review briefly discuss the i) understanding of cancer and biomarkers for early diagonosis purpose ii) Molecular methods and ii) biosensor-based diagnostics. The reseach primary focus on advancement in biosensor design using various concepts ie., Electrochemical, Chemiluminescence and Colorimetric, Surface plasmons (SP), Surface plasmon resonance (SPR), localized surface plasmon resonance (LSPR), Fluorescence, Fiber-based sensors, Terahertz based biosensors, and Surface enhanced Raman spectroscopy (SERS). As a result of the local electric field amplification around plasmonic (usually gold and silver) nanostructures, surface-enhanced Raman spectroscopy (SERS) has emerged as a rapid, selective, and sensitive alternative to conventional laboratory analytical methods, making significant strides in a number of biosensing applications but still under developing stage to be used as diagnostic tool in clinical research.

Litchi chinensis inspired nanoformulations: a synergy guided approach for unraveling promising cytotoxic attributes of metal and nonmetal conjugates.

In present study, diverse Litchi chinensis-mediated nanostructures in combination with 5-fluorouracil drug were fabricated viz. Au, Se, Ag, Ag-Se, Ag-Au, 5-FU Ag-Se and 5-FU Ag-Au with subsequent characterization and scrutinization of their anticarcinogenic capabilities. UV-Visible spectroscopic analysis confirmed the state transition for each precursor salt. XRD and transmission electron microscopy analysis revealed spherical/quasispherical nanostructures with monoclinic crystalline organization ranged between 18 nm and 38 nm. FTIR analysis revealed fabricated nanoparticles to be capped with various phytoconstituents. DLS and Zeta potential analysis of unloaded and drug-loaded bielemental nanoparticles (BNPs) showed comparatively large hydrodynamic particle size distribution and sufficient stability of nanoparticles. BNPs showed promising lethality concentrations for brine shrimp (LC50 < 2 µg/ml) and antitumor (LC50 < 10 µg/ml) assessments. These findings were in positive correlation with the antioxidant inhibitory concentrations IC50 (74.2-180.1 µg/ml) of the tested entities. Ag-Se and Ag-Au were loaded with 5-FU (loading efficiency of 47% ± 1.14 and 25% ± 0.32, respectively) in light of their promising cytotoxic actions. All nanostructures showed profound hemocompatibility with maximum hemolytic activity as low as 2.4%. Highly significant difference (P < 0.01) was observed in antineoplastic potentials of unloaded and 5-FU loaded BNPs against HepG2 and HT144, with most substantial IC50 for 5-FU Ag-Au (8.95 ± 2.86 µg/ml). 5-FU Ag-Au was identified as a significant inducer of DNA fragmentation with maximum relative tail moment (HepG2: 3.45 ± 0.21) among all treatments.

Determination of ROS Scavenging, Antibacterial and Antifungal Potential of Methanolic Extract of Otostegia limbata (Benth.) Boiss.

Wide spectrum medicinal significance augments plant utilization as the primary source of significant pharmaceutical agents. In vitro investigation of antioxidant and antimicrobial activity highlights the therapeutic potential of Otostegia limbata. Methanol extract of the plant (MEP) shows considerable dose dependent antioxidant ability at six concentrations (7.81 µg/mL to 250 µg/mL) in 2.2-diphenyl-1-picrylhydrazyl (DPPH) assay, phosphomolybdate assay (PMA) and reducing power assay (RPA). The plant capability to scavenge free radicals in the mixture ranged from 37.89% to 63.50% in a concentration-dependent manner. MEP was active against five tested bacterial strains in the agar-well diffusion method. Staphylococcus aureus, gram-positive bacteria was found to be most susceptible followed by S. epidermidis with 18.80 mm and 17.47 mm mean zone of inhibition. The mean inhibition zone against gram-negative strains Klebsiella pneumonia, Pseudomonas spp. and Escherichia coli were 15.07 mm, 14.73 mm, and 12.17 mm. MEP revealed potential against Alternaria spp. and Aspergillus terreus fungal strains evaluated through agar-tube dilution assay. Aspergillus terreus was more sensitive than Alternaria spp. with an average 78.45% and 68.0% inhibition. These findings can serve as a benchmark for forthcoming scrutiny such as bioactive components discovery and drug development.

Understanding Hyaluronan Receptor (CD44) Interaction, HA-CD44 Activated Potential Targets in Cancer Therapeutics.

Cancer is a complex mechanism involving a series of cellular events. The glycoproteins such as hyaluronan (HA) are a significant element of extracellular matrix (ECM), involve in the onset of cancer developmental process. The pivotal roles of HA in cancer progression depend on dysregulated expression in various cancer. HA, also gain attention due to consideration as a primary ligand of CD44 receptor. The CD44, complex transmembrane receptor protein, due to alternative splicing in the transcription process, various CD44 isoforms predominantly exist. The overexpression of distinct CD44 isoforms (CD44v) standard (CD44s) depends on the tumour type and stage. The receptor proteins, CD44 engage in a variety of biological processes, including cell growth, apoptosis, migration, and angiogenesis. HA-CD44 interaction trigger survival pathways that result in cell proliferation, invasion ultimately complex metastasis. The interaction and binding of ligand-receptor HA-CD44 regulate the downstream cytoskeleton pathways involve in cell survival or cell death. Thus, targeting HA, CD44 (variant and standard) isoform, and HA-CD44 binding consider as an attractive and useful approach towards cancer therapeutics. The use of various inhibitors of HA, hyaluronidases (HYALs), and utilizing targeted Nano-delivery of anticancer agents and antibodies against CD44, peptides gives promising results in vitro and in vivo. However, they are in clinical trials with favourable and unfavourable outcomes, which reflects the need for various modifications in targeting agents and a better understanding of potential targets in tumour progression pathways.

Additive-based stability assessment of biologically designed CuO and GSH-CuO nanospheres and their applicability as Nano-biosensors.

Uncapped and Glutathione capped Cupric oxide nanospheres were synthesized by the interaction of Berberis lycium (Bl) root extract with corresponding salt solution. CuO nanospheres were best optimized by mixing 2% Bl extract solution with 1 mM CuSO4·5H2O (pH 11, 90 °C) Reduced glutathione (0.25 mM) in solution form was added in respective emulsion after 24 h. Synthesis of nanospheres was ensured by distinct surface plasmonic resonance peaks shown by CuO (370-420 nm). Addition of glutathione resulted in sharp blue shift and lowered absorbance values in UV spectra suggesting the decrease in nanoparticles' size and concentration. Average particle sizes as deduced with XRD were found to be 18.52 and 16.57 nm for CuO and GSH-CuO nanospheres respectively. Additive based stability assessment of synthesized nanospheres revealed CuO and GSH-CuO nanospheres to be highly stable in the presence of Catechin hydrate among various tested chemical compounds while ascorbic acid appeared as a strong destabilizing agent. TMB was oxidized by H2O2 in the presence of synthesized enzymes likewise horseradish peroxidase; though exhibited moderate results. Glutathione stabilized cupric oxide nanospheres exhibited the potential to be modulated further into efficient nanozymes as these showed better affinity towards chromogenic substrate TMB (Km value 0.32 mM) and better catalytic efficiency (0.075 mM-1 s-1) compared to uncapped CuO nanomimetics (1.6 mM, 0.033 mM-1 s-1). All of the tested additives served as inhibitors to the peroxidase mimicking potential of CuO and GSH-CuO nanozymes.