Surface-Functionalized Nanoceria: Dual Action in Diabetes Management via Glucose-Responsive Insulin Delivery and Oxidative Stress Mitigation.

Nanoceria (NC) is gaining scientific attention due to its widespread drug delivery efficacy and modulation of oxidative stress. Herein, we developed dextran (Dex) capped insulin (INS)-loaded phenylboronic acid (PBA)-functionalized nanoceria (NC-PBA-INS-Dex) for glucose-responsive insulin delivery and mitigating excessive ROS production to regulate both hyperglycemia and oxidative stress in diabetes mellitus (DM). The prepared nanoparticle showed favorable loading capacity and excellent encapsulation efficiency of insulin. Glucose-responsive insulin release from NC-PBA-INS-Dex was observed initially in the cell-free mode when subjected to varying glucose concentrations (5.5, 11, and 25 mM). Interestingly, under in vitro setting, promising insulin release from NC-PBA-INS-Dex was found in muscle cells (major glucose storage cells) compared to lung cells against exposure to different glucose concentration suggesting a glucose-sensitive intracellular insulin delivery. NC-PBA-INS-Dex treatment further upregulated GLUT4 translocation and glucose uptake/utilization in sodium palmitate-exposed muscle cells, and results were significantly higher compared to NC or INS alone treated cells. Studies in diabetic animals demonstrated the maintenance of normoglycemia for up to 12 h upon gavaging a single dose of NC-PBA-INS-Dex compared to INS alone treatment (subcutaneous/oral). Oral administration of NC-PBA-INS-Dex also increased insulin bioavailability (in both serum and muscle tissue) compared with either subcutaneous or oral insulin administration. NC-PBA-INS-Dex further exhibited ROS scavenging (superoxide radical) potential in cell-free, in vitro, and in vivo systems, and results were comparable to treatment with NC alone. NC-PBA-INS-Dex could effectively regulate the expression of occludin and induce the reversible opening of a tight junction in intestinal epithelial cells, allowing the particle transport through the intestinal mucosa. Treatment with NC-PBA-INS-Dex did not exhibit any toxicity to in vitro and in vivo models. The NC-based drug delivery system will mimic the physiological regulation of insulin secretion in a noninvasive manner, offering improved patient compliance, reduced risk of hyperglycemia, and enhanced overall management of DM.

A comparative evaluation of the effect of eugenol exposure time on the bond strength of an etch-and-rinse and a self-etch adhesive to dentin: An in vitro study.

Aim: The study aimed to comparatively evaluate the effect of eugenol exposure time on the micro-shear bond strength (µ-SBS) of etch-and-rinse and a self-etch adhesive to dentin. Materials and Methods: One hundred and twelve teeth samples were prepared from bisectioning 56 freshly extracted human mandibular molars and were randomly divided into 14 subgroups of 8 samples each (n = 8). Three subgroups containing eugenol and a noneugenol-based restorative material were placed on the dentin surface and left for 24 h, 7 days, and 14 days, respectively, and were compared to a control. Two bonding systems were evaluated: one being etch-and-rinse and the other self-etch adhesive. The µ-SBS were calculated and expressed in MPa. Statistical Analysis: The data were analyzed using mixed model analysis of variance. The level of statistical significance was set at 5%. Results: There was a statistically significant reduction in the µ-SBS values when the self-etch adhesive was used, after the removal of eugenol-containing cement placed for 24 h. However, the reduction in the µ-SBS values after 7 days or 14 days was not significant. Conclusion: Exposure to eugenol containing temporary cement for 24 h significantly reduces the µ-SBS of self-etching adhesives to dentin. However, exposure for 1 week or more has minimal effects.

Antibacterial and Antioxidant Compounds from the Root Extract of Aloe debrana.

This study was conducted to isolate and identify the chemical compounds from the roots of Aloe debrana (L.) and evaluate their antioxidant and antibacterial activities. From the acetone (99.5%) extract of the roots of this plant, four anthraquinones, such as chrysophanol (1), asphodeline (2), aloesaponarin I (5), and laccaic acid D-methyl ester (6), and a new catechol derivative, 5-allyl-3-methoxybenzene-1,2-diol (3), were isolated and elucidated by different chromatographic and spectroscopic methods together with linoleic acid (4), respectively. Compounds 2, 3, and 4 were reported here for the first time from this plant and compound 3 from the genus Aloe. The compounds were evaluated for their antioxidant activity using H2O2 and DPPH assays and bactericidal activity against S. aureus and E. coli. Compounds 3 and 6 showed highest antioxidant activities with IC50 values of 19.38 ± 0.64 and 32.81 ± 0.78 µg/mL in DPPH, and 28.52 ± 1.08 and 27.31 ± 1.46 µg/mL in H2O2, respectively. The isolated compounds also demonstrated considerable activity towards S. aureus. Among these compounds, compound 3 exhibited the highest activity (91.20 ± 0.12% and 9.14 ± 0.93 mm at 1.0 mg/mL) against this bacterium. The overall results suggest that the isolated compounds may be considered as potential sources of the bioactive agents to be used in the pharmacological, food, and other industries. Moreover, their high sensitivity against S. aureus may also support the use of A. debrana plant in the traditional medicine to treat wounds. Therefore, the isolated compounds are responsible for medicinal properties of this plant.

Recent Contributions of Mass Spectrometry-Based "Omics" in the Studies of Breast Cancer.

Breast cancer (BC) is one of the most heterogeneous groups of cancer. As every biotype of BC is unique and presents a particular "omic" signature, they are increasingly characterized nowadays with novel mass spectrometry (MS) strategies. BC therapeutic approaches are primarily based on the two features of human epidermal growth factor receptor 2 (HER2) and estrogen receptor (ER) positivity. Various strategic MS implementations are reported in studies of BC also involving data independent acquisitions (DIAs) of MS which report novel differential proteomic, lipidomic, proteogenomic, phosphoproteomic, and metabolomic characterizations associated with the disease and its therapeutics. Recently many "omic" studies have aimed to identify distinct subsidiary biotypes for diagnosis, prognosis, and targets of treatment. Along with these, drug-induced-resistance phenotypes are characterized by "omic" changes. These identifying aspects of the disease may influence treatment outcomes in the near future. Drug quantifications and characterizations are also done regularly and have implications in therapeutic monitoring and in drug efficacy assessments. We report these studies, mentioning their implications toward the understanding of BC. We briefly provide the MS instrumentation principles that are adopted in such studies as an overview with a brief outlook on DIA-MS strategies. In all of these, we have chosen a model cancer for its revelations through MS-based "omics".

Antibacterial and antibiofilm effects of Pseudomonas aeruginosa derived outer membrane vesicles against Streptococcus mutans.

Antimicrobial resistance (AMR) is a serious and most urgent global threat to human health. AMR is one of today's biggest difficulties in the health system and has the potential to harm people at any stage of life, making it a severe public health issue. There must be fewer antimicrobial medicines available to treat diseases given the rise in antibiotic-resistant organisms. If no new drugs are created or discovered, it is predicted that there won't be any effective antibiotics accessible by 2050. In most cases, Streptococcus increased antibiotic resistance by forming biofilms, which account for around 80 % of all microbial infections in humans. This highlights the need to look for new strategies to manage diseases that are resistant to antibiotics. Therefore, development alternative, biocompatible and high efficacy new strategies are essential to overcome drug resistance. Recently, bacterial derived extracellular vesicles have been applied to tackle infection and reduce the emergence of drug resistance. Therefore, the objective of the current study was designed to assess the antibacterial and antibiofilm potential of outer membrane vesicles (OMVs) derived from Pseudomonas aeruginosa againstStreptococcus mutans. According to the findings of this investigation, the pure P. aeruginosa outer membrane vesicles (PAOMVs) display a size of 100 nm. S. mutans treated with PAOMVs showed significant antibacterial and antibiofilm activity. The mechanistic studies revealed that PAOMVs induce cell death through excessive generation of reactive oxygen species and imbalance of redox leads to lipid peroxidation, decreased level of antioxidant markers including glutathione, superoxide dismutase and catalase. Further this study confirmed that PAOMVs significantly impairs metabolic activity through inhibiting lactate dehydrogenase activity (LDH), adenosine triphosphate (ATP) production, leakage of proteins and sugars. Interestingly, combination of sub-lethal concentrations of PAOMVs and antibiotics enhances cell death and biofilm formation of S. mutans. Altogether, this work, may serve as an important basis for further evaluation of PAOMVs as novel therapeutic agents against bacterial infections.

Chitosan-PEI passivated carbon dots for plasmid DNA and miRNA-153 delivery in cancer cells.

These days carbon dots have been developed for multiple biomedical applications. In the current study, the transfection potential of synthesized carbon dots from single biopolymers such as chitosan, PEI-2kDa, and PEI-25kDa (CS-CDs, PEI2-CDs, and PEI25-CDs) and by combining two biopolymers (CP2-CDs and CP25-CDs) through a bottom-up approach have been investigated. The characterization studies revealed successful synthesis of fluorescent, positively charged carbon dots <20 nm in size. Synthesized carbon dots formed a stable complex with plasmid DNA (EGFP-N1) and miRNA-153 that protected DNA/miRNA from serum-induced degradation. In-vitro cytotoxicity analysis revealed minimal cytotoxicity in cancer cell lines (A549 and MDA-MB-231). In-vitro transfection of EGFP-N1 plasmid DNA with PEI2-CDs, PEI25-CDs and CP25-CDs demonstrated that these CDs could strongly transfect A549 and MDA-MB-231 cells. The highest EGFP-N1 plasmid transfection efficiency was observed with PEI2-CDs at a weight ratio of 32:1. PEI25-CDs polyplex showed maximum transfection at a weight ratio of 8:1 in A549 at a weight ratio of 16:1 in MDA-MB-231 cells. CP25-CDs exhibited the highest transfection at a weight ratio of 16:1 in both cell lines. The in-vitro transfection of target miRNA, i.e., miR-153 in A549 and MDA-MB-231 cells with PEI2-CDs, PEI25-CDs, and CP25-CDs suggested successful transfer of miR-153 into cells which induced significant cell death in both cell lines. Importantly, CS-CDs and CP2-CDs could be tolerated by cells up to 200 µg/mL concentration, while PEI2-CDs, PEI25-CDs, and CP25-CDs showed non-cytotoxic behavior at low concentrations (25 µg/mL). Together, these results suggest that a combination of carbon dots synthesized from chitosan and PEI (CP25-CDs) could be a novel vector for transfection nucleic acids that can be utilized in cancer therapy.

In silico study to unravel molecular networking of melatonin in the regulation of gametogenesis.

Melatonin, a pineal hormone, has potential role on steroidogenesis, growth and maturation of sperm and ovum during gametogenesis. The possible use of this indolamine as an antioxidant in the production of good quality gametes opens up a new area of current research. Nowadays, a large number of reproductive dysfunctions like infertility and failure in fertilization due to gametic malformations are major concern worldwide. So, understanding molecular mechanisms including interacting genes and their action is a prerequisite to the therapeutic approach against these issues. The aim of present bioinformatic study is the detection of molecular network concerning therapeutic potential of melatonin in gametogenesis. It includes target genes identification, gene ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, network analysis, prediction of signalling pathways and molecular docking. We obtained common top 52 targets of melatonin in the process of gametogenesis. They are involved in biological processes related to the development of gonads and primary sexual characteristics and sex differentiation. We took top 10 pathways out of total 190 enriched pathways for further analysis. Subsequently, principal component analysis also revealed that among top ten hub targets (TP53, CASP3, MAPK1, JUN, ESR1, CDK1, CDK2, TNF, GNRH1 and CDKN1A), only TP53, JUN and ESR1were significantly interacted with melatonin on the basis of squared cosine value. So, present in silico investigation provides considerable information on the interactive network between therapeutic targets of melatonin along with the involvement of intracellular signalling cascade regulating biological processes associated with the gametogenesis. This novel approach may be pertinent in improving modern research on reproductive dysfunctions associated abnormalities.

Rhythmicity in testicular melatonin and its correlation with the dynamics of spermatogenic cells in an annual reproductive cycle of Clarias batrachus under natural photo-thermal conditions.

Melatonin, the pineal hormone, is synthesized and secreted rhythmically in accordance with various environmental cues especially photo-thermal conditions. The reproductive physiology of seasonal breeders is synchronized with the surroundings by melatonin as a neuroendocrine mediator to acts as an important factor in fish reproduction. However, the data on the participation of melatonin in male reproduction and the putative interaction with the process of spermatogenesis in fish is scarce till date. So, major objectives of the current study are to determine for the first time, the relationship, if any, between seasonal levels of melatonin and testicular development and maturation of the germ cells, and also the involvements of specific meteorological parameters in spermatogenesis under natural photo-thermal conditions. We measured the concentration of circulatory and testicular melatonin; value of gonadosomatic index (GSI), relative percentages of different developing spermatogenic cells, area and perimeter (size and shape) of seminiferous lobules along with the level/duration of rainfall, water temperature and day length in six reproductive phases throughout an annual cycle in adult male catfish (Clarias batrachus). Intra-testicular and serum melatonin concentration showed a similar seasonal pattern with a peak during "functional maturity" phase and trough during "slow spermatogenesis" phase. Correlation as well as regression analyses also supported this positive relationship. Interestingly, intra-testicular melatonin also showed a significant positive correlation with GSI and relative percentage as well as lobular size of mature stages (spermatid and spermatozoa) of germ cells in an annual cycle. Furthermore, meteorological factors exhibited as critical cues to regulate the dynamics (in %) of spermatogenic cells and the level of testicular melatonin throughout the annual gonadal cycle. Our results corroborated by principal component (PC) analysis and showed very clearly that active "functional maturity" state is characterized by GSI, testicular melatonin, relative abundance and lobular size of mature spermatogenic stages as key internal oscillators; and studied environmental variables as the external clues for the regulation of spawning process. Collectively, the present data revealed that there is a relationship between melatonin levels and testicular growth and development of germ cells in Clarias batrachus under natural photo-thermal conditions.

pH-responsive and targeted delivery of chrysin via folic acid-functionalized mesoporous silica nanocarrier for breast cancer therapy.

Cancer is a disease of global importance. In order to mitigate conventional chemotherapy-related side effects, phytochemicals with inherent anticancer efficacy have been opted. However, the use of nanotechnology is essential to enhance the bioavailability and therapeutic efficacy of these phytochemicals. Herein, we have formulated folic acid conjugated polyacrylic acid capped mesoporous silica nanoparticles (∼47.6 nm in diameter) for pH-dependent targeted delivery of chrysin to breast cancer (MCF-7) cells. Chrysin loaded mesoporous silica nanoparticles (Chr- mSiO2@PAA/FA) have been noted to induce apoptosis in MCF-7 cells through oxidative insult and mitochondrial dysfunction with subsequent G1 arrest. Further, in tumor bearing mice, intravenous incorporation of Chr-mSiO2@PAA/FA has been noticed to enhance the anti-neoplastic effects of chrysin via tumor site-specific accumulation. Enhanced cytotoxicity of chrysin contributed towards in vivo tumor regression, restoration of normalized tissue architecture and maintenance of healthy body weight. Besides, no serious systemic toxicity was manifested in response to Chr-mSiO2@PAA/FA administration in vivo. Thus, the study evokes about the anticancer potentiality of chrysin and its increased therapeutic activity via incorporation into folic acid conjugated mesoporous silica nanoparticles, which may hold greater impact in field of future biomedical research.

The eco-friendly treatment of rubber industry effluent by using adsorbent derived from Moringa oleifera bark and Pseudomonas sp, cultured from effluent.

Rubber processing generates a large volume of wastewater containing rubber latex residues and chemicals. Remediation of the wastewater needs a cost-effective and environment-friendly treatment method. For this study, Moringa oleifera stem bark and Pseudomonas sp. bacteria were used for adsorption and microbial treatment of the effluent. The adsorbent surface was mostly amorphous with crystallinity index 37.9% and the BET surface area was 6.622 m2/g. FTIR analysis indicated involvement of O-H stretching, ketone α, ß-unsaturated, C-H stretching, carboxylic acid and derivatives O-C stretching functional groups in the adsorption process. The assessment of the above two agents was based on their reduction capabilities of the toxic parameters, such as total suspended and dissolved solids, total solids, biological and chemical oxygen demand, sulphate, ammonium, dissolved oxygen, phosphate, pH, electrical conductivity, turbidity, and oxidation reduction potential from the wastewater. A comparative study of the present work revealed that both the agents were effective in reduction of most of the above parameters below the safe discharge limits. However, the adsorption using Moringa oleifera stem bark was better compared to the biodegradation by Pseudomonas sp. bacteria. The main challenges that typically accompany biodegradation include microbe handling and a lower removal percentage than adsorption.

Fabrication of phenyl boronic acid modified pH-responsive zinc oxide nanoparticles as targeted delivery of chrysin on human A549 cells.

Recently, different natural bioactive compounds have been used as anticancer agents for their various therapeutic benefits and non-toxic nature to other organs. However, they have various restrictions in preclinical and clinical studies due to their non-targeting nature and insufficient bioavailability. As a result, a zinc oxide nanoparticle (ZnO) based drug delivery medium was constructed which has good bio-compatibility and bio-degradability. It also displays cancer cell-specific drug delivery in a targeted and controlled way. In the present study, phenylboronic acid (PBA) tagged ZnO nanoparticles (ZnO-PBA) was fabricated and in the next step, chrysin (a natural bio-active molecule) was loaded to it to form the nanoconjugate (ZnO-PBA-Chry). Different characterization techniques were used to confirm the successful fabrication of ZnO-PBA-Chry. PBA-tagging to the nanoparticle helps in targeted delivery of chrysin in lung cancer cells (A549) as PBA binds with sialic acid receptors which are over-expressed on the surface of A549 cells. As ZnO dissociates in acidic pH, it shows stimuli-responsive release of chrysin in tumor microenvironment. Application of ZnO-PBA-Chry nanohybrid in lung cancer cell line A549 caused oxidative stress mediated intrinsic cell death and cell cycle arrest. ZnO-PBA-Chry downregulated MMP-2 and VE-Cadherin, thereby inhibiting metastasis and the invasive property of A549 cells.

An eco-friendly removal of Cd(II) utilizing banana pseudo-fibre and Moringa bark as indigenous green adsorbent and modelling of adsorption by artificial neural network.

Heavy metal-contaminated water can be effectively treated using adsorbents made from abundantly available biomass. The present investigation was carried out to adsorb Cd(II) from synthetic solution by banana pseudo-stem (BP) and Moringa oleifera stem bark (MB). Adsorption efficiencies of both adsorbents were studied in the batch reactor by conducting experiments to determine the consequences of changes of pH, adsorbent dosages, initial Cd(II) concentrations, incubation time, and temperature. The process parameters were tuned to attain the highest possible removal percentage. The characterization of the adsorbents was performed by utilizing Fourier-transform infrared spectroscopy (FTIR), field emission scanning electron microscopy (FESEM), and energy-dispersive X-ray (EDX) for the fresh and metal-loaded adsorbents. Atomic absorption spectroscopy (AAS) was employed to calculate the amount of Cd(II) in an aqueous solution. The experimental data were entirely consistent with the pseudo-second-order model for BP and MB. The findings of the study illustrated the better adsorption efficiency of BP-derived adsorbent (≈ 99%) at optimum conditions over the MB (≈ 97%), and the corresponding adsorption capacities were 11.98 and 7.04 mg/g, respectively. The 4 (four) well-known isotherm models were attempted both in linear and non-linear forms. BP (R2 =0.995) and MB (R2 =0.994) were found to be best described by the Freundlich isotherm, which was selected based on the highest R2 value. In thermodynamic studies, ΔH and ΔS were calculated for both the adsorbents. Cd(II) adsorption on BP and MB was endothermic, as evidenced by the positive ΔH. Finally, the prediction of the removal percentage was made by the artificial neural network (ANN) modelling. The present work developed regionally derived waste materials which are helpful for small-scale industrial units for their waste management in an economical and sustainable way.

Carnosic acid attenuates doxorubicin-induced cardiotoxicity by decreasing oxidative stress and its concomitant pathological consequences.

This work aimed to reveal the protective mechanism of CA against Dox (doxorubicin)-induced cardiotoxicity. In isolated murine cardiomyocytes, CA showed a concentration-dependent cytoprotective effect against Dox. Dox treatment significantly (p < 0.01) increased the formation of reactive oxygen species (ROS), increased NO levels, activated NADPH oxidase, and inactivated the cellular redox defense mechanism in cardiac cells, resulting in augmented oxidative stress in cardiomyocytes and rat hearts. Dox-induced oxidative stress significantly (p < 0.01) upregulated several pathogenic signal transductions, which induced apoptosis, inflammation, and fibrosis in cardiomyocytes and murine hearts. In contrast, CA significantly (p < 0.05-0.01) reciprocated Dox-induced cardiac apoptosis, inflammation, and fibrosis by suppressing oxidative stress and interfering with pathological signaling events in both isolated murine cardiomyocytes and rat hearts. CA treatment significantly (p < 0.05-0.01) countered Dox-mediated pathological changes in blood parameters in rats. Histological examinations backed up the pharmacological findings. In silico chemometric investigations predicted potential interactions between CA and studied signal proteins, as well as the drug-like features of CA. Thus, it would be concluded that CA has the potential to be regarded as an effective agent to alleviate Dox-mediated cardiotoxicity in the future.

Synthesis of carbon dots from taurine as bioimaging agent and nanohybrid with ceria for antioxidant and antibacterial applications.

Here we have synthesized water soluble and biocompatible carbon dots (CDs) from taurine via thermal decomposition method. The CDs showed nearly spherical shape with diameter less than 10 nm. The CDs exhibited excitation dependent fluorescence emission and could be used for mammalian cell imaging. The CDs showed excellent DPPH and hydrogen peroxide radical scavenging activity in cell free system. Besides, the CDs also displayed significant intracellular radical scavenging activity in human normal kidney epithelial (NKE) cells. Furthermore, nanohybrids consisting of both CDs and nanoceria (CeO2) were prepared and tested for their biomedical applications. The nanohybrids showed significant antioxidant activities in both cell free and intracellular conditions. The CDs and nanohybrids possessed very little toxicity upto the concentration of 100 µg/mL when treated for 24 hours in human NKE cells. The CDs as well as nanohybrids further displayed significant bacterial growth inhibition against both gram-positive and gram-negative bacteria under dark as well as light illumination condition via the bacterial membrane damage. However, under the light illumination, the bacterial growth inhibition of CDs and nanohybrids was further enhanced due to the generation of reactive oxygen radicals and subsequent DNA degradation. A higher dose-dependent intracellular antioxidant and antibacterial activities of the nanohybrid is attributed to the synergistic effect of nanoceria and CDs. All these results clearly reflected that our synthesized CDs and their nanohybrids can be used for several biomedical applications.

Enhancement of anti-neoplastic effects of cuminaldehyde against breast cancer via mesoporous silica nanoparticle based targeted drug delivery system.

AIMS: Synthesis of novel drug delivery system for targeted delivery of cuminaldehyde to breast cancer cells and the subsequent analyses of anti-neoplastic potential of the drug. MAIN METHODS: 3-carboxy-phenyl boronic acid (PBA) conjugated and polyacrylic acid (PAA) gated mesoporous silica nanoparticles (MSNs) were synthesized for the targeted delivery of cuminaldehyde (CUM) to breast cancer cells. Enhancement of anti-neoplastic effects of cuminaldehyde (4-isopropylbenzaldehyde) by the nanoconjugates was assessed. KEY FINDINGS: The anti-cancer effects of non-targeted and targeted drug-nanoconjugates were examined in vitro and in vivo. The targeted drug-nanoconjugates caused cell cycle arrest and induced the intrinsic pathway of apoptosis in MCF-7 cells through mitochondrial damage. In vivo intravenous injection of the targeted drug-nanoconjugates led to effective reduction in growth of 4 T1 induced mammary pad tumor in female BALB/c mice via augmented accumulation of cuminaldehyde. The drug-nanoconjugates did not exhibit any systemic toxicity. SIGNIFICANCE: Therefore, MSN-PBA-CUM-PAA represents a potent therapeutic model for breast cancer treatment.