Limestone and yellow gypsum can reduce cadmium accumulation in groundnut (Arachis hypogaea): A study from a three-decade old landfill site.

Cadmium (Cd) toxicity has cropped up as an important menace in the soil-plant system. The use of industrial by-products to immobilise Cd in situ in polluted soils is an interesting remediation strategy. In the current investigation, two immobilizing amendments of Cd viz., Limestone (traditionally used) and Yellow gypsum (industrial by-product) have been used through a green-house pot culture experiment. Soil samples were collected from four locations based on four graded levels of DTPA extractable Cd as Site 1 (0.43 mg kg-1), Site 2 (0.92 mg kg-1), Site 3 (1.77 mg kg-1) and Site 4 (4.48 mg kg-1). The experiment was laid out in a thrice replicated Factorial Complete Randomized Design, with one factor as limestone (0, 250, 500 mg kg-1) and the other being yellow gypsum (0, 250, 500 mg kg-1) on the collected soils and groundnut was grown as a test crop. Results revealed that the DTPA-extractable Cd content in soil and Cd concentration in plants decreased significantly with the increasing doses of amendments irrespective of initial soil available Cd and types of amendment used. The effect of amendment was soil specific and in case of Site 1 (low initial Cd) the effect was more prominent. The reduction in DTPA-extractable Cd in combined application of limestone and yellow gypsum @500 mg kg-1 over the absolute control in soil under groundnut for the sites was by far the highest with the values of 83.72%, 77.17%, 48.59% and 40.63% respectively. With the combined application, Target Cancer Risk (TCR) of Cd was also reduced. Hence, combined application of limestone and yellow gypsum can be beneficial in the long run for mitigating Cd pollution.

Cisplatin-Conjugated Polyurethane Capsule for Dual Drug Delivery to a Cancer Cell.

This paper describes the synthesis of a polymer-prodrug conjugate, its aqueous self-assembly, noncovalent encapsulation of a second drug, and stimuli-responsive intracellular dual drug delivery. Condensation polymerization between a functionalized diol and a commercially available diisocyanate in the presence of poly(ethylene glycol) hydroxide (PEG-OH) as the chain stopper produces an ABA-type amphiphilic block copolymer (PU-1) in one pot, with the middle hydrophobic block being a polyurethane containing a pendant tert-butyloxycarbonyl (Boc)-protected amine in every repeating unit. Deprotection of the Boc group, followed by covalent attachment of the Pt(IV) prodrug using the pendant amine groups, produces the polymer-prodrug conjugate PU-Pt-1, which aggregates to nanocapsule-like structures in water with a hydrophilic interior. In the presence of sodium ascorbate, the Pt(IV) prodrug can be detached from the polymer backbone, producing the active Pt(II) drug. Cell culture studies show appreciable cell viability by the parent polymer. However, the polymer-prodrug conjugate nanocapsules exhibit cellular uptake and intracellular release of the active drug under a reducing environment. The capsule-like aggregates of the polymer-prodrug conjugate were used for noncovalent encapsulation of a second drug, doxorubicin (Dox), and Dox-loaded PU-Pt-1 aggregate showed a significantly superior cell killing efficiency compared to either of the individual drugs, highlighting the promising application of such a dual-drug-delivery approach.

Polyglutamate-based nanoconjugates for image-guided surgery and post-operative melanoma metastases prevention.

Rationale: Cutaneous melanoma is the most aggressive and deadliest of all skin malignancies. Complete primary tumor removal augmented by advanced imaging tools and effective post-operative treatment is critical in the prevention of tumor recurrence and future metastases formation. Methods: To meet this challenge, we designed novel polymeric imaging and therapeutic systems, implemented in a two-step theranostic approach. Both are composed of the biocompatible and biodegradable poly(α,L-glutamic acid) (PGA) nanocarrier that facilitates extravasation-dependent tumor targeting delivery. The first system is a novel, fluorescent, Turn-ON diagnostic probe evaluated for the precise excision of the primary tumor during image-guided surgery (IGS). The fluorescence activation of the probe occurs via PGA degradation by tumor-overexpressed cathepsins that leads to the separation of closely-packed, quenched FRET pair. This results in the emission of a strong fluorescence signal enabling the delineation of the tumor boundaries. Second, therapeutic step is aimed to prevent metastases formation with minimal side effects and maximal efficacy. To that end, a targeted treatment containing a BRAF (Dabrafenib - mDBF)/MEK (Selumetinib - SLM) inhibitors combined on one polymeric platform (PGA-SLM-mDBF) was evaluated for its anti-metastatic, preventive activity in combination with immune checkpoint inhibitors (ICPi) αPD1 and αCTLA4. Results: IGS in melanoma-bearing mice led to a high tumor-to-background ratio and reduced tumor recurrence in comparison with mice that underwent surgery under white light (23% versus 33%, respectively). Adjuvant therapy with PGA-SLM-mDBF combined with ICPi, was well-tolerated and resulted in prolonged survival and prevention of peritoneal and brain metastases formation in BRAF-mutated melanoma-bearing mice. Conclusions: The results reveal the great clinical potential of our PGA-based nanosystems as a tool for holistic melanoma treatment management.

Meet me halfway: Are in vitro 3D cancer models on the way to replace in vivo models for nanomedicine development?

The complexity and diversity of the biochemical processes that occur during tumorigenesis and metastasis are frequently over-simplified in the traditional in vitro cell cultures. Two-dimensional cultures limit researchers' experimental observations and frequently give rise to misleading and contradictory results. Therefore, in order to overcome the limitations of in vitro studies and bridge the translational gap to in vivo applications, 3D models of cancer were developed in the last decades. The three dimensions of the tumor, including its cellular and extracellular microenvironment, are recreated by combining co-cultures of cancer and stromal cells in 3D hydrogel-based growth factors-inclusive scaffolds. More complex 3D cultures, containing functional blood vasculature, can integrate in the system external stimuli (e.g. oxygen and nutrient deprivation, cytokines, growth factors) along with drugs, or other therapeutic compounds. In this scenario, cell signaling pathways, metastatic cascade steps, cell differentiation and self-renewal, tumor-microenvironment interactions, and precision and personalized medicine, are among the wide range of biological applications that can be studied. Here, we discuss a broad variety of strategies exploited by scientists to create in vitro 3D cancer models that resemble as much as possible the biology and patho-physiology of in vivo tumors and predict faithfully the treatment outcome.

Evaluation of Furcraea foetida (L.)Haw. for phytoremediation of cadmium contaminated soils.

In the present study, we evaluated Furcraea foetida for the phytoremediation of cadmium (Cd)-contaminated soils. We selected F. foetida because it is a drought-resistant plant, produces high biomass, and needs minimum maintenance. It belongs to the leaf fiber group of plants and therefore has economic importance. Since it is a non-edible crop, there is no danger of food chain contamination. Despite possessing these ideal characteristics, surprisingly, to date, the plant is underutilized for phytoremediation purposes. Therefore, to evaluate the phytoremediation potential of the plant, we exposed it to five levels of cadmium (0, 25, 50, 100, and 200 mg Cd kg-1 soil) and studied its influence on growth, dry matter production, uptake, and translocation efficiency. The plant showed good tolerance to Cd 200 mg kg-1 soil without exhibiting any visible toxicity symptoms. The metal mainly accumulated in the roots (233 µg g-1dw), followed by leaf (51 µg g-1 dw). The bioconcentration factor was > 1, but the translocation factor was < 1. The plant was not classified as a hyperaccumulator of Cd; however, because of its high uptake (897 µ g-1 plant) and translocation efficiency (78%), we concluded that the plant could be utilized for phytoextraction of Cd from soils with low to moderately contaminated soils.

Disulfide chemistry in responsive aggregation of amphiphilic systems.

The dynamic nature of the disulfide bond has enhanced the potential for disulfide based amphiphiles in the emerging biomedical field. Disulfide containing amphiphiles have extensively been used for constructing wide ranging soft nanostructures as potential candidates for delivery of drugs, proteins and genes owing to their degradable nature in the presence of intracellular glutathione (present in a many fold excess compared to the extracellular milieu). This degradable nature of amphiphiles is not only useful to deliver therapeutics but it also eliminates the toxicity issues associated with the carrier after delivery of such therapeutics. Therefore, these bioreducible and biocompatible nano-aggregates inspired researchers to use them as vehicles for therapeutic delivery and as a result the literature of disulfide containing amphiphiles has been intensified. This review article highlights the structural diversity in disulfide containing amphiphilic small molecule and polymeric systems, structural effects on their aqueous aggregation, redox-responsive disassembly and biological applications. Furthermore, the use of disulfide chemistry towards the design of cell penetrating polymers has also been discussed. Finally a brief perspective on some future opportunities of these systems is provided.

Dendronized fluorosurfactant for highly stable water-in-fluorinated oil emulsions with minimal inter-droplet transfer of small molecules.

Fluorosurfactant-stabilized microfluidic droplets are widely used as pico- to nanoliter volume reactors in chemistry and biology. However, current surfactants cannot completely prevent inter-droplet transfer of small organic molecules encapsulated or produced inside the droplets. In addition, the microdroplets typically coalesce at temperatures higher than 80 °C. Therefore, the use of droplet-based platforms for ultrahigh-throughput combination drug screening and polymerase chain reaction (PCR)-based rare mutation detection has been limited. Here, we provide insights into designing surfactants that form robust microdroplets with improved stability and resistance to inter-droplet transfer. We used a panel of dendritic oligo-glycerol-based surfactants to demonstrate that a high degree of inter- and intramolecular hydrogen bonding, as well as the dendritic architecture, contribute to high droplet stability in PCR thermal cycling and minimize inter-droplet transfer of the water-soluble fluorescent dye sodium fluorescein salt and the drug doxycycline.

Synthesis and Self-assembly of a Helical Polymer Grafted from a Foldable Polyurethane Scaffold.

Herein a polyurethane graft poly-l-glutamate amphiphilic copolymer was synthesized from a polyurethane (PU)-based macro-initiator (containing pendant primary amine groups) through the ring opening polymerization of N-carboxy anhydride of γ-benzyl-l-glutamate (BLG-NCA). On average, twenty two l-glutamic acids were grafted from each amino group which was pendant on the polyurethane chain with 10 repeating units. The grafted polymer (PU-PP-1) exhibits self-assembly to produce a hydrogel in a wide pH window ranging from pH 5.0 to 8.0 with a critical gelation concentration (CGC) of 5.0 wt % (w/v) at pH 7.4. Furthermore, circular dichroism study revealed the transition of the α-helix to a random coil upon increasing the pH. Due to the protonation of side chains at pH 4.0, PU-PP-1 adopted an α-helical conformation whereas at pH >8.0 the side-chain carboxylic acid groups of the PLGAs were ionized, leading to the formation of an extended random coil conformation as a result of charge repulsion. Conformational switching was also supported by FTIR spectroscopy.

Biomimetic sulfated polyethylene glycol hydrogel inhibits proteoglycan loss and tumor necrosis factor-α-induced expression pattern in an osteoarthritis in vitro model.

This study aimed to evaluate the potential of an anti-inflammatory polyethylene glycol (PEG) hydrogel for osteoarthritis (OA) management in an OA in vitro model. Freshly isolated porcine chondrocytes were maintained in high-density cultures to form cartilage-like three-dimensional micromasses. Recombinant porcine tumor necrosis factor-alpha (TNF-α) was used to induce OA-like changes. Normal and OA-like micromasses were treated with dendritic polyglycerol sulfate-based PEG hydrogel. Live/dead staining showed that all micromasses remained vital and presented similar morphological characteristics. Safranin-O staining demonstrated a typical depletion of glycosaminoglycans in TNF-α-treated micromasses but not in the presence of the hydrogel. There was no distinct difference in immunohistochemical detection of type II collagen. Microarray data showed that rheumatoid arthritis and TNF signaling pathways were down regulated in hydrogel-treated OA-like micromasses compared to nontreated OA-like micromasses. The hydrogel alone did not affect genes related to OA such as ANPEP, COMP, CXCL12, PTGS2, and TNFSF10, but it prevented their regulation caused by TNF-α. This study provides valuable insights toward a fully synthetic hydrogel for the intra-articular treatment of OA. The findings proved the potential of this hydrogel to prevent the development of TNF-α-induced OA with regard to proteoglycan loss and TNF-α-induced expression pattern without additional signs of differentiation and inflammation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2018. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 490-500, 2019.

Enhanced Permeability and Retention-like Extravasation of Nanoparticles from the Vasculature into Tuberculosis Granulomas in Zebrafish and Mouse Models.

The enhanced permeability and retention (EPR) effect is the only described mechanism enabling nanoparticles (NPs) flowing in blood to reach tumors by a passive targeting mechanism. Here, using the transparent zebrafish model infected with Mycobacterium marinum we show that an EPR-like process also occurs allowing different types of NPs to extravasate from the vasculature to reach granulomas that assemble during tuberculosis (TB) infection. PEGylated liposomes and other NP types cross endothelial barriers near infection sites within minutes after injection and accumulate close to granulomas. Although ∼100 and 190 nm NPs concentrated most in granulomas, even ∼700 nm liposomes reached these infection sites in significant numbers. We show by confocal microscopy that NPs can concentrate in small aggregates in foci on the luminal side of the endothelium adjacent to the granulomas. These spots are connected to larger foci of NPs on the ablumenal side of these blood vessels. EM analysis suggests that NPs cross the endothelium via the paracellular route. PEGylated NPs also accumulated efficiently in granulomas in a mouse model of TB infection with Mycobacterium tuberculosis, arguing that the zebrafish embryo model can be used to predict NP behavior in mammalian hosts. In earlier studies we and others showed that uptake of NPs by macrophages that are attracted to infection foci is one pathway for NPs to reach TB granulomas. This study reveals that when NPs are designed to avoid macrophage uptake, they can also efficiently target granulomas via an alternative mechanism that resembles EPR.

Dendritic polyglycerol-poly(ethylene glycol)-based polymer networks for biosensing application.

This work describes the formation of a new dendritic polyglycerol-poly(ethylene glycol)-based 3D polymer network as a matrix for immobilization of the redox enzyme periplasmatic aldehyde oxidoreductase to create an electrochemical biosensor. The novel network is built directly on the gold surface, where it simultaneously stabilizes the enzyme for up to 4 days. The prepared biosensors can be used for amperometric detection of benzaldehyde in the range of 0.8-400 µM.

Mitochondrial DNA copy number and risk of oral cancer: a report from Northeast India.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the sixth most common cancer globally. Tobacco consumption and HPV infection, both are the major risk factor for the development of oral cancer and causes mitochondrial dysfunction. Genetic polymorphisms in xenobiotic-metabolizing enzymes modify the effect of environmental exposures, thereby playing a significant role in gene-environment interactions and hence contributing to the individual susceptibility to cancer. Here, we have investigated the association of tobacco - betel quid chewing, HPV infection, GSTM1-GSTT1 null genotypes, and tumour stages with mitochondrial DNA (mtDNA) content variation in oral cancer patients. METHODOLOGY/PRINCIPAL FINDINGS: The study comprised of 124 cases of OSCC and 140 control subjects to PCR based detection was done for high-risk HPV using a consensus primer and multiplex PCR was done for detection of GSTM1-GSTT1 polymorphism. A comparative ΔCt method was used for determination of mtDNA content. The risk of OSCC increased with the ceased mtDNA copy number (Ptrend  = 0.003). The association between mtDNA copy number and OSCC risk was evident among tobacco - betel quid chewers rather than tobacco - betel quid non chewers; the interaction between mtDNA copy number and tobacco - betel quid was significant (P = 0.0005). Significant difference was observed between GSTM1 - GSTT1 null genotypes (P = 0.04, P = 0.001 respectively) and HPV infection (P<0.001) with mtDNA content variation in cases and controls. Positive correlation was found with decrease in mtDNA content with the increase in tumour stages (P<0.001). We are reporting for the first time the association of HPV infection and GSTM1-GSTT1 null genotypes with mtDNA content in OSCC. CONCLUSION: Our results indicate that the mtDNA content in tumour tissues changes with tumour stage and tobacco-betel quid chewing habits while low levels of mtDNA content suggests invasive thereby serving as a biomarker in detection of OSCC.