Biosynthesized Silver Nanoparticles for Cancer Therapy and In Vivo Bioimaging

In the current communication, a simple, environmentally compatible, non-toxic green chemistry process is used for the development of silver nanoparticles (AgZE) by the reaction between silver nitrate (AgNO3) and the ethanolic leaf extract of Zinnia elegans (ZE). The optimization of AgZE is carried out using a series of experiments. Various physico-chemical techniques are utilized to characterize the nanomaterials. The cell viability assay of AgZE in normal cells (CHO, HEK-293T, EA.hy926, and H9c2) shows their biocompatible nature, which is supported by hemolytic assay using mouse RBC. Interestingly, the nanoparticles exhibited cytotoxicity towards different cancer cell lines (U-87, MCF-7, HeLa, PANC-1 and B16F10). The detailed anticancer activity of AgZE on human glioblastoma cell line (U-87) is exhibited through various in vitro assays. In vivo the AgZE illustrates anticancer activity by inhibiting blood vessel formation through CAM assay. Furthermore, the AgZE nanoparticles when intraperitoneally injected in C57BL6/J mice (with and without tumor) exhibit fluorescence properties in the NIR region (excitation: 710nm, emission: 820nm) evidenced by bioimaging studies. The AgZE biodistribution through ICPOES analysis illustrates the presence of silver in different vital organs. Considering all the results, AgZE could be useful as a potential cancer therapeutic agent, as well as an NIR based non-invasive imaging tool in near future.

Different Aspects of Classical Pathway Overactivation in Patients With C3 Glomerulopathy and Immune Complex-Mediated Membranoproliferative Glomerulonephritis

The rare and heterogeneous kidney disorder C3 glomerulopathy (C3G) is characterized by dysregulation of the alternative pathway (AP) of the complement system. C3G is often associated with autoantibodies stabilizing the AP C3 convertase named C3 nephritic factors (C3NeF). The role of classical pathway (CP) convertase stabilization in C3G and related diseases such as immune complex-mediated membranoproliferative glomerulonephritis (IC-MPGN) remains largely unknown. Here, we investigated the CP convertase activity in patients with C3G and IC-MPGN. Using a refined two-step hemolytic assay, we measured the stability of CP convertases directly in the serum of 52 patients and 17 healthy controls. In four patients, CP convertase activity was prolonged compared to healthy controls, i.e. the enzymatic complex was stabilized. In three patients (2 C3G, 1 IC-MPGN) the convertase stabilization was caused by immunoglobulins, indicating the presence of autoantibodies named C4 nephritic factors (C4NeFs). Importantly, the assay also enabled detection of non-immunoglobulin-mediated stabilization of the CP convertase in one patient with C3G. Prolonged CP convertase activity coincided with C3NeF activity in all patients and for up to 70 months of observation. Crucially, experiments with C3-depleted serum showed that C4NeFs stabilized the CP C3 convertase (C4bC2a), that does not contain C3NeF epitopes. All patients with prolonged CP convertase activity showed clear signs of complement activation, i.e. lowered C3 and C5 levels and elevated levels of C3d, C3bc, C3bBbP, and C5b-9. In conclusion, this work provides new insights into the diverse aspects and (non-)immunoglobulin nature of factors causing CP convertase overactivity in C3G/IC-MPGN.

Beneficial Effect of Quercetin on Erythrocyte Properties in Type 2 Diabetic Rats

Diabetes mellitus is characterized by tissue oxidative damage and impaired microcirculation, as well as worsened erythrocyte properties. Measurements of erythrocyte deformability together with determination of nitric oxide (NO) production and osmotic resistance were used for the characterization of erythrocyte functionality in lean (control) and obese Zucker diabetic fatty (ZDF) rats of two age categories. Obese ZDF rats correspond to prediabetic (younger) and diabetic (older) animals. As antioxidants were suggested to protect erythrocytes, we also investigated the potential effect of quercetin (20 mg/kg/day for 6 weeks). Erythrocyte deformability was determined by the filtration method and NO production using DAF-2DA fluorescence. For erythrocyte osmotic resistance, we used hemolytic assay. Erythrocyte deformability and NO production deteriorated during aging—both were lower in older ZDF rats than in younger ones. Three-way ANOVA indicates improved erythrocyte deformability after quercetin treatment in older obese ZDF rats only, as it was not modified or deteriorated in both (lean and obese) younger and older lean animals. NO production by erythrocytes increased post treatment in all experimental groups. Our study indicates the potential benefit of quercetin treatment on erythrocyte properties in condition of diabetes mellitus. In addition, our results suggest potential age-dependency of quercetin effects in diabetes that deserve additional research.

4-Chlorothymol Exerts Antiplasmodial Activity Impeding Redox Defense System in Plasmodium falciparum

Malaria remains one of the major health concerns due to the resistance of Plasmodium species toward the existing drugs warranting an urgent need for new antimalarials. Thymol derivatives were known to exhibit enhanced antimicrobial activities; however, no reports were found against Plasmodium spp. In the present study, the antiplasmodial activity of thymol derivatives was evaluated against chloroquine-sensitive (NF-54) and -resistant (K1) strains of Plasmodium falciparum. Among the thymol derivatives tested, 4-chlorothymol showed potential activity against sensitive and resistant strains of P. falciparum. 4-Chlorothymol was found to increase the reactive oxygen species and reactive nitrogen species level. Furthermore, 4-chlorothymol could perturb the redox balance by modulating the enzyme activity of GST and GR. 4-Chlorothymol also showed synergy with chloroquine against chloroquine-resistant P. falciparum. 4-Chlorothymol was found to significantly suppress the parasitemia and increase the mean survival time in in vivo assays. Interestingly, in in vivo assay, 4-chlorothymol in combination with chloroquine showed higher chemosuppression as well as enhanced mean survival time at a much lower concentration as compared to individual doses of chloroquine and 4-chlorothymol. These observations clearly indicate the potential use of 4-chlorothymol as an antimalarial agent, which may also be effective in combination with the existing antiplasmodial drugs against chloroquine-resistant P. falciparum infection. In vitro cytotoxicity/hemolytic assay evidently suggests that 4-chlorothymol is safe for further exploration of its therapeutic properties.

Evaluation of Hemolysis Activity of Zerumbone on RBCs and Brine Shrimp Toxicity

Zerumbone is a well-known compound having anti-cancer, anti-ulcer, anti-inflammatory and anti-hyperglycemic effects. During its use for the disease treatment, the membrane of erythrocyte can be affected by consumption of this bioactive compound. The current study was the first report of investigation of the hemolytic activities on human erythrocytes and cytotoxic profile of zerumbone. The toxicity of zerumbone on human erythrocytes was determined by in vitro hemolytic assay. Brine shrimp lethality assay was used to evaluate the cytotoxic effect of zerumbone at concentrations 10, 100 and 1000 μg/mL. The human erythrocyte test showed no significant toxicity at low concentrations, whereas hemolytic effect was amplified up to 17.5 % at the highest concentration. The half lethal concentration (LC50) value of zerumbone against brine shrimp was less than 1000 µg /mL (LC50=190 µg/ml) showing the significant toxic nature of this compound. These results provide a baseline in terms of the toxicity of therapeutic formulations from this compound to membrane erythrocytes with a great attention to the highest concentrations, which paves promise for drug development.

Evaluation of Cytotoxic Effect, Anti-Cholinesterase, Hemolytic and Antibacterial activities of the Species Scabiosa stellata L.

Objective: In this study, cytotoxic effect, anticholinesterase, hemolytic and antibacterial activities of crude extracts (petroleum ether, ethyl acetate and n-butanol) obtained from the plant Scabiosa stellata L. were evaluated. Methods: The cytotoxicity of extracts was tested by Brine shrimp lethality method; the acetylcholinesterase inhibitory activity was performed using Ellman's colorimetric method and the hemolytic activity was assessed by spectrophotometric method towards human erythrocytes. Furthermore, the antibacterial activity was estimated by agar disk diffusion assay against ten bacterial strains. Results: The phytochemical screening of the extracts revealed the presence of several types of secondary metabolites. A significant cytotoxic effect was observed for the n-butanolic extract with 57.2 ± 0.2 % of mortality at 80 μg/mL, the ethyl acetate extract had a moderate anticholinesterase activity at 200 μg/mL. The hemolytic assay exhibited that n-butanolic and ethyl acetate extracts induce hemolysis in dose-dependent manner with values of EC50 at 37.3 ± 0.5 and 106.6 ± 0.3 μg/mL, respectively. All the crude extracts showed antibacterial activity against most tested strains, with zones of inhibition ranging from 9 to 20 mm. Conclusion: The results indicate that the extracts obtained from S. stellata can be an important source of therapeutic agents against pathological damage due to free radicals inducing neurodegenerative and infectious diseases, while n-butanolic extract could be used as a good source of alternative natural antiproliferative compounds.

C4 nephritic factor in patients with immune-complex-mediated membranoproliferative glomerulonephritis and C3-glomerulopathy

Background Acquired or genetic abnormalities of the complement alternative pathway are the primary cause of C3glomerulopathy(C3G) but may occur in immune-complex-mediated membranoproliferative glomerulonephritis (IC-MPGN) as well. Less is known about the presence and role of C4nephritic factor(C4NeF) which may stabilize the classical pathway C3-convertase. Our aim was to examine the presence of C4NeF and its connection with clinical features and with other pathogenic factors. Results One hunfe IC-MPGN/C3G patients were enrolled in the study. C4NeF activity was determined by hemolytic assay utilizing sensitized sheep erythrocytes. Seventeen patients were positive for C4NeF with lower prevalence of renal impairment and lower C4d level, and higher C3 nephritic factor (C3NeF) prevalence at time of diagnosis compared to C4NeF negative patients. Patients positive for both C3NeF and C4NeF had the lowest C3 levels and highest terminal pathway activation. End-stage renal disease did not develop in any of the C4NeF positive patients during follow-up period. Positivity to other complement autoantibodies (anti-C1q, anti-C3) was also linked to the presence of nephritic factors. Unsupervised, data-driven cluster analysis identified a group of patients with high prevalence of multiple complement autoantibodies, including C4NeF. Conclusions In conclusion, C4NeF may be a possible cause of complement dysregulation in approximately 10–15% of IC-MPGN/C3G patients.

Cytotoxic Activity of Different Extracts from Brown Marine Macroalgae from the El Jadida Region (Morocco)

The therapeutic use of extraordinary virtues of algae in healthcare is very ancient and has evolved throughout the history of humanity. However, low or high doses and/or prolonged administration of algae-derived biologically active substances may trigger adverse or even harmful effects such as hemolytic properties. In this study, hemolytic activities of the brown algae Cystoseira myriophylloides (F/Sargassaceae) from El Jadida coast (Moroccan Atlantic coast) on human erythrocyte were investigated. The toxicity of chloroform-methanolic, methanolic, chloroformic, isoprpanolic and butanolic extracts and fractions derived from Cystoseira myriophylloides on human erythrocytes was measured by in vitro hemolytic assay. The results obtained show variable cytotoxicity of C. myriophyloides against the red blood cells. Indeed, the crude extract has a fairly high hemolytic activity that is very marked when the erythrocytes are incubated in the presence of the chloroformic extract. However, the cytotoxic activity is greatly increased in the presence of the butanolic fraction derived by partitioning of the isopropanolic extract. Moreover, the hemolysis activity is dose-dependent and depending on the evolution of incubation time. Thus, these results show that the majority of extracts and fractions of brown macroalgae Cystoseira myriophylloides manifest hemolytic activity.

Morphological properties of almond oil constituted nanofibrous scaffold for bone tissue engineering

One of the greatest challenges in the bone remodeling is to fabricate the structure resembling the extracellular matrix. This research aims to fabricate a novel bone scaffold comprising polyurethane (PU) added with almond nanofibers via electrospinning technique. The PU/almond oil nanocomposites showed smaller fiber diameter (629 ± 148.92 nm) compared to the pristine PU (890 ± 116.91 nm). The interaction of PU with almond oil was confirmed in the infrared spectrum by the strong formation of a hydrogen bond. The wettability analysis showed that the prepared PU/almond oil nanocomposites were hydrophobic in nature (107° ± 1) compared with the pure PU (100° ± 0.5774). Thermal analysis revealed enhancement of the thermal stability with the addition of the almond oil. The addition of almond oil into the PU matrix increased the surface roughness and blood compatibility properties. Further, the fabricated PU/almond oil nanocomposites showed less toxicity to red blood cells (RBCs), as indicated in the hemolytic assay. Hence, the novel fabricated scaffold possesses better physicochemical properties and is nontoxic to the RBCs, which may be utilized for bone tissue regeneration.