Fractions of a chloroform extract of ajenjo leaves (<i>Artemisia mendozana</i> DC. var. <i>mendozana</i>) inhibit the proliferation,viability and clonogenicity of B16-F0 melanoma cells

The ajenjo, Artemisia mendozana DC. var. mendozana (Asteraceae), grows in the Andean foothills of Mendozaand San Juan, Argentina, and is used as a medicinal plant for its antispasmodic and antifungal properties. The aim of thiswork was to obtain fractions of a chloroform extract of ajenjo leaves and to evaluate the in vitro effects on proliferation,viability and clonogenicity of B16-F0 melanoma cells. Using a silica gel chromatography column, 120 fractions werecollected and grouped according to the chromatographic profile in 9 main fractions (F1–F9). Their major compoundsidentified were: terpenes (F1), terpenes and sesquiterpene lactones (F2–F3), sesquiterpenes (F4–F6) and phenols andsesquiterpenes (F7-9). B16-F0 cells were incubated for 72 h with DMSO (vehicle) or 0.1 mg/ml F1–F9. At 72 h ofculture, F1 decreased both the growing index (GI) and cell viability. F2 and F3 both decreased GI and only F3decreased clonogenic activity. F4 and F5 both decreased GI. Only F5 decreased cell viability and F4 decreasedclonogenicity. Consequently, fractions F6–F8 did not affect any of the cell parameters assayed, while F9 decreased cellviability and inhibited clonogenicity.     

The effect of natural products combination on MCF-7 cells exceeds tamoxifen therapeutic dose effects <i>in vitro</i>

Cancer remains to be one of the most severe sicknesses globally. Cases have kept rising over the years. Breast cancer (BC), which is among the leading types of cancers and predominantly affects women, is the second leading cause of cancer mortality. Researchers have developed interventions over the years; however, the BC survival rate has not improved since the 1980s. This has created the need for novel drug interventions that would manage and treat BC more effectively. This study focused on using a combination of natural product extracts such as phytoestrogen (Ziziphus jujube) and Tannin nanoparticles (NP99) together, which we have referred to as (Z.NP99) and tamoxifen (Tam) as one of the leading BC drugs since the 70s, in treating BC. The effectiveness of Tam if used alone in the treatment and if combined with Z.NP99 was evaluated using MCF-7 cells in vitro. The findings showed that the combination treatment of Z.NP99 affected the proliferation and viability of MCF-7 cells more than Tam at a 10 μg/mL dose. Moreover, Z.NP99 with Tam stimulated the maximum reduction of MCF-7 proliferation and viability in a time-dependent manner. Furthermore, Tam and Z.NP99 augmented the DNA fragmentation percentage combined with the upregulation of the apoptotic genes. Additionally, the results showed that the apoptotic impact of Z.NP99 and Tam on MCF-7 cells may be intermediated by down-regulating some genes such as Claudin-1 followed by down-regulating mRNA expression of MMP-9, VEGF, and BCL-2 genes of treated cells. Combining Tam with Z.NP99 considerably enhanced the effectiveness of conventional therapy. As a result, this study suggested that the Z.NP99 was ideal for developing effective natural treatments that would improve BC outcomes.     

Agmatine pretreatment protects retinal ganglion cells (RGC-5 cell line) from oxidative stress <i>in vitro</i>

Agmatine, 2-(4-aminobutyl)guanidine, has been reported to have neuroprotective effects against various neuronal damages. In this study it was investigated whether agmatine pretreatment rescues the retinal ganglion cells from oxidative injury in vitro. After differentiation of transformed rat retinal ganglion cells (RGC-5 cell line) with staurosporine, agmatine (0.0 to 100.0 μM) pretreatment was performed for 2 hours. Subsequently, they were exposed to hydrogen peroxide (0.0 to 2.5 mM) as an oxidative stress. Cell viability was monitored for up to 48 hours with the lactate dehydrogenase (LDH) assay and apoptosis was examined by the terminal deoxynucleotide transferase-mediated terminal uridine deoxynucleotidyl transferase nick end-labeling (TUNEL) method. As a result, differentiated RGC-5 cells were found to have decreased viability after addition of hydrogen peroxide in a dose-dependent manner. This hydrogen peroxide induced cytotoxicity caused apoptosis characterized by DNA fragmentation. Agmatine pretreatment not only increased cell viability but also attenuated DNA fragmentation. In conclusion, agmatine pretreatment demonstrated neuroprotective effects against oxidative stress induced by hydrogen peroxide in differentiated RGC-5 cells in vitro. This suggests a novel therapeutic strategy rescuing retinal ganglion cells from death caused by oxidative injury     

<i>In vitro</i> study of emodin-induced nephrotoxicity in human renal glomerular endothelial cells on a microfluidic chip

Emodin is an effective component of rhubarb with positive pharmacological effects on human health. However, it isalso toxic to different cells or tissues to varying degrees. The effects of emodin on glomerular endothelial cells (GECs) remain tobe tested, and the documented works were always performed in vitro and hardly reflect the real physiological situation. Tostudy the effects of emodin on GECs in a biomimetic environment, we utilized a microfluidic chip to assess thephysiological reaction of human renal glomerular endothelial cells to various concentrations of emodin in this work. Theresults showed that emodin caused cytotoxicity, impaired glomerular filtration barrier integrity to macromolecules, andincreased barrier permeability in a dose-dependent manner. With the increase in emodin concentration, the concentrationof the pro-inflammatory cytokine tumor necrosis factor-α, interleukin (IL)-6, transforming growth factor-β1, andmonocyte chemoattractant protein (MCP-1) increased while the production of inflammatory cytokine IL-6 first increasedand then decreased with the increase in emodin concentration. Our findings shed new light on emodin-inducednephrotoxicity and provide insights for the application of microfluidic chip devices to reveal drug-cell interactions.     

Micropropagation of <i>Ilex dumosa</i> (Aquifoliaceae) from nodal segments in a tissue culture system

Micropropagation of Ilex dumosa var. dumosa R. (“yerba señorita”) from nodal segments containing one axillary bud was investigated. Shoot regeneration from explants of six-year-old plants was readily achieved in ¹/₄ strength Murashige and Skoog medium (¹/₄ MS) plus 30 gr·L^-1 sucrose and supplemented with 4.4 µM BA. Further multiplication and elongation of the regenerated shoots were obtained by subculture in a fresh medium of similar composition with 1.5 gr·L^-1 sucrose. Rooting induction from shoots were achieved in two steps: 1) 7 days in ¹/₄ MS (30 gr·L^-1 sucrose, 0.25 % Phytagel^®) with 7.3 µM IBA and 2) 21 days in the same medium without IBA and 20 µM of cadaverine added. Regenerated plants were successfully transferred to soil. This micropropagation schedule can be implemented in breeding programs of Ilex dumosa.     

Gene expression of 49 kDa apyrase,cytoskeletal proteins,ATPase, ADPase and amino acid contents of <i>Pisum sativum</i> (L.) cells germinated in E<i>uryops arabicus</i> (Steud. ex Jaub. & Spach) water extract

The present research reports of quick and marked changes induced by plant extract of Euryops arabicus in thegene expression of 49-kDa apyrases, cytoskeletal proteins, ATPases, ADPase and amount of amino acid of pea (Pisumsativum L. var. Alaska). Pellets of cytoskeletals proteins (27000 xg) were probed with anti-apyrase antibody, biotinylatedanti-rat, actin and alpha and beta-tubulin for Western blotting. ATPase and ADPase activities were determined basedon the hydrolytic efficacy of adenine triphosphate and adenine diphosphate. By 72 hours, the abundance of apyrases,cytoskeletal proteins and amount of amino acid in pellets of 27000 xg of germinated pea seeds in E. arabicus extractswere sharply increased than those sown in distilled water. All the samples exhibited that the stems had more amountfrom apyrases, cytoskeletal proteins, amino acids and ATPase and ADPase activities than primary leaves and primaryroots that were germinated either on E. arabicus water extract or in distilled water. Based on the enzyme’s capability tohydrolyse nucleotide triphosphate and nucleotide diphosphate as well as the direct association between expression of49-kDa apyrase and cytoskeletal proteins, E. arabicus water extract had an important effect on plant germinations.     

Identification of <i>PtGai</i> (a DELLA protein) in trifoliate orange and expression patterns in response to drought stress

Gibberellins (GAs) are an important hormone in regulating plant growth and development, and DELLA protein is an essential negative regulator of GA signal transduction. The aim of the study was to clone a GA-inhibiting protein DELLA from trifoliate orange (Poncirus trifoliata L. Raf.) and to analyze the bioinformations and expression patterns of the protein gene in tissues and in response to drought stress. A DELLA protein was isolated from trifoliate orange and named as PtGai (Genebank number: MZ170959). The PtGai protein had 1731 bp open reading frames, along with 576 amino acid codes, and also grouped with sweet orange (XM_006430552.4). The PtGai protein sequence was 65% homology with the sequences of DELLA proteins in other plant families. PtGai protein existed in the nucleus based on the prediction of subcellular localization. PtGai protein could be expressed in roots, stems, and leaves, along with the highest expression in stems. PtGai was upregulated by drought stress in leaves and roots, along with the decrease of root total GA concentration and the inhibition of shoot and root biomass production. It indicated the characteristics of PtGai protein and the roles of PtGai in GA synthesis and plant growth.     

Role of foliar spray of plant growth regulators in improving photosynthetic pigments and metabolites in <i>Plantago ovata</i> (Psyllium) under salt stress–A field appraisal

Salinity is one of the major abiotic factors that limit the growth and productivity of plants. Foliar application of plant growth regulators (PGRs) may help plants ameliorate the negative impacts of salinity. Thus, a field experiment was conducted at the Botanical Garden University of Balochistan, Quetta, to explore the potential role of PGRs, i.e., moringa leaf extract (MLE; 10%), proline (PRO; 1 µM), salicylic acid (SA; 250 µM), and thiourea (TU; 10 mM) in ameliorating the impacts of salinity (120 mM) on Plantago ovata, an important medicinal plant. Salinity hampered plant photosynthetic pigments and metabolites but elevated oxidative parameters. However, foliar application of PGRs enhanced photosynthetic pigments, including Chl b (21.11%), carotenoids (57.87%) except Chl a, activated the defense mechanisms by restoring and enhancing the metabolites, i.e., soluble sugars (49.68%), soluble phenolics (33.34%), and proline (31.47%), significantly under salinity stress. Furthermore, foliar supplementation of PGRs under salt stress led to a decrease of about 43.02% and 43.27% in hydrogen peroxide and malondialdehyde content, respectively. Thus, PGRs can be recommended for improved photosynthetic efficiency and metabolite content that can help to get better yield under salt stress, with the best and most effective treatments being those of PRO and MLE to predominately ameliorate the harsh impacts of salinity.