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PG-dendritic amide-linked amphiphilic micelles were studied to increase the solubility of hydrophobic molecules. The encapsulation study was done by utilising hydrophobic pyrene dye using U.V. tech...

PLGA (Lactic- co-glycolic acid) coated chitosan microspheres loaded with hydroxyapatite and doxycycline hyclate complex were developed in the present study for periodontal delivery. A modified single emulsion method was adopted for the development of microspheres. Formulation was optimized on the basis of particle size, drug loading and encapsulation efficiency with the central composite design using 23 factorial design. Microspheres were optimized and electron microscopy revealed their spherical shape and porous nature. In-vitro study showed initial burst and then sustained release behavior of the formulation for 14 days. Further, in-vitro antibacterial study performed on E. coli (ATCC-25922) and S. aureus (ATCC-29213) revealed concentration dependent activity. Also, in-vitro cyto-toxicity assessment ensures biocompatibility of the formulation with the fibroblast’s cells. Overall, the quality by design assisted PLGA microspheres, demonstrated the desired attributes and were found suitable for periodontal drug delivery.

Phagocytosis of particulate vaccine delivery systems is a critical immune mechanism involved in antigen capture and processing by macrophages and dendritic cells. The internalization and degradation of the particles involve a complex sequence of events. This process coordinates lipids, signaling proteins, and the cytoskeleton. Dynamic changes in the actin cytoskeleton are essential for phagocytosis and antigen presentation. Knowledge regarding the correlation of surface properties, attached ligand density and geometric size of particles with the efficiency of phagocytosis may facilitate their design and application. To investigate this, polylactide biodegradable particles with different diameters (2-4 μm and 200-300 nm) were exposed to murine macrophages and dendritic cells and the effect of size on a series of cellular responses was evaluated. Cellular uptake studies using microscopy and flow cytometry showed size dependent internalization of particles, with nanoparticles accumulating in cells at a faster rate. The particles induced homoaggregation of cells and also showed cytoskeletal remodeling that could be inhibited by cytochalasin-D. Scanning electron microscopy images showed the time dependent formation of phagocytic cups and invaginations that promote particle uptake. The particles were observed to co-localized with the endo-lysosomal compartments after phagocyotosis. In our experiments, particle mediated immunoactivation, antigen processing and cytokine secretion have shown a good correlation with the uptake process. These findings would allow a better understanding of the process of particle uptake and may be instrumental in the rational design of optimal vaccine delivery systems.

The therapeutic potential of plant-based natural products is well known, yet it is actually limited by poor water solubility, low bioavailability and rapid metabolism. These issues can be solved by formulation with silica, gold, iron and silver nanoparticles. This manuscript discusses the synthesis and toxicity of inorganic nanoparticles used for therapeutic applications. The use of silver nanoparticles in the 0.4–30 µg dose range is optimal without any severe short-term toxicity. The bio-distribution of gold nanoparticles are controlled by their size; for instance, 5–15 nm particles show broader organ distribution, whereas 50–100 nm particles were mainly found in the liver and spleen. Curcumin-loaded silver nanoparticles decrease four times the minimum effective concentration against gram-positive and gram-negative bacteria. Mesoporous silica nanoparticles enhance the solubility of resveratrol up to about 95%. Mesoporous silica nanoparticles delivered 2.68 fold higher paclitaxel concentrations in lungs as compared to the free drug.

Cancer is one of the most lethal disease that affects humans. Many anticancer agents have been developed but have downsides such as toxicity, affordability and limits of the administered dose. Alternatively, natural anticancer products are promising due to their non-toxic nature. Indeed, natural anticancer products have doubled the human life span in twentieth century. Moreover, over 60% of approved drugs and drug candidates against cancer are of natural origin. Yet, natural drugs have limitations such as non-specific action, low aqueous solubility and reduced bioavailability. These limitations can be circumvented by delivering drugs using polymeric nanocarriers. For instance, polylactic acid and polylactic-co-glycolic acid have been used as carriers in the medical field for a very long time. About 20 products based on these polymers have been approved by the Food and Drug Administration and European Medicine. Polymeric nanocarriers are versatile and have potential to meet actual challenges in the delivery of natural products for cancer treatment. Here we review natural products in treating cancer with focus on polymeric nanocarriers.

Polysaccharide antigens do not promote antibody class switching and memory antibody response, thus require conjugation with a T cell dependent carrier protein to generate protective immune response. The intensity of immune responses varies with the carrier proteins for the same carbohydrate antigen and most of the carrier proteins do not generate strong immune responses. Vi polysaccharide and r-flagellin of Salmonella typhi were conjugated and formulated in PLA particles as nanoglycoconjugate which not only generated strong immune response but also promoted antibody class switching and elicited memory antibody response from single point immunization. Nanoglycoconjugate immunization also modulate anti-inflammatory property of Vi polysaccharide with an enhance secretion of pro-inflammatory cytokine TNF-α and IL-6. This was with concomitant decrease of IFN-γ production, antibody class switching from IgG3 to IgG2 with memory antibody generation against Vi polysaccharide. Antibody elicited by nanoglycoconjugate showed better opsonization and clearance of Salmonella typhi in THP-1 macrophages as compared to Vi-flagellin glycoconjugate and Vi TT (Typhbar®). Delivery of glycoconjugate through nanoparticles provides a platform technology for improving the immunogenicity of polysaccharide based vaccines.

To manage population of dogs (Canis familiaris), the efficacy of recombinant proteins-based contraceptive vaccines to inhibit fertility has been evaluated in female beagle dogs.Female beagle dogs (n = 4) were immunized with physical mixture of Escherichia coli-expressed recombinant porcine ZP3 with promiscuous T cell epitope of tetanus toxoid (TT-KK-pZP3) and porcine ZP4 with promiscuous T cell epitope of bovine RNase (bRNase-KK-pZP4), or with a fusion protein encompassing dog ZP3 fragment and two copies of GnRH with appropriate promiscuous T cell epitopes (dZP3-GnRHActive immunization of female dogs following a three injections schedule at 4-week intervals with a physical mixture of TT-KK-pZP3 + bRNase-KK-pZP4 as well as dZP3-GnRHThese studies revealed the potential of recombinant dZP3-GnRH

Expression of heterologous proteins in E. coli often leads to the formation of protein aggregates known as inclusion bodies (IBs). Inclusion body aggregates pose a major hurdle in the recovery of bioactive proteins from E. coli. Usage of strong denaturing buffers for solubilization of bacterial IBs results in poor recovery of bioactive protein. Structure-function understanding of IBs in the last two decades have led to the development of several mild solubilization buffers, which improve the recovery of bioactive from IBs. Recently, combinatorial mild solubilization methods have paved the way for solubilization of wide range of inclusion bodies with appreciable refolding yield. Here, we describe a simple protocol for solubilization and refolding of an inclusion body protein with appreciable recovery.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel coronavirus that presented in Wuhan, China in December of 2019. SARS-CoV-2-mediated coronavirus disease 2019 (COVID-19) is strongly associated with the rampant of infection with catastrophic morbidity. This pandemic has jiggled the society with its widespread social, psychologic, and economic distress. Society immunity or herd immunity is the most crucial and natural prophylactic vaccination interference against any infectious disease around the globe. It postulates protection at both the individual and population levels against pathogen-borne infections. The accomplishment of herd immunity is reliant upon the transmission efficiency of pathogens and the emerging natural immunity within a society. This is an exclusively pertinent mechanism to control any pandemic. The microbiome plays a crucial role in the coevolution of the body's immune cells, which are cardinal for bolstering a proficient vaccination process, ensuring herd immunity. The dynamics of interaction among microbiota, nutrients, and individual immunity determines the avidity of vaccines against several pathogens. Synchronization of microbial symbiosis preserves pathogen transmissibility and the engrossment of vaccination among different clusters based on the age and gender. Imbalance of nutrients refutes microbiome harmony, develops environmental enteropathy, and exacerbates immunity, which perturbs the efficacy of the vaccination process. Furthermore, discrepancies in the protective response of many vaccines in developing countries than developed countries are due to differences in healthy microbiota among the individuals in a particular cohort. We suggested that the rigorous pan-India oral poliovirus vaccination program for the past 30 years has been finally capable of inducing herd immunity against poliovirus infection among societies, which may also inspire the commencement of natural heterologous immunity against SARS-CoV-2 infection. However, this anamnestic recall is somewhat counterintuitive, as antibody generation against original antigens of SARS-CoV-2 will be restrained due to original antigenic sin.

Protein based vaccines are the most safe and affordable strategy to combat pneumococcal disease circumventing the limitations of conventional polysaccharide-based vaccines like serotype dependence, high cost and inability to be administered to immunocompromised. SP0845 is a highly conserved vaccine candidate shown to provide protection against heterologous strains of Streptococcus pneumoniae, the primal cause of pneumonia. However, the associated poor immunogenicity warrants the need for adjuvants and multiple doses to mount desired responses. The present study relates to improve the immunogenicity of pneumococcal protein SP0845 by use of poly lactic acid biodegradable polymer microparticles. The immunization studies showed that microparticles elicited higher antibody response compared to alum adjuvanted protein and this immunopotentiation was achieved without the use of any additional adjuvant. They were also capable of eliciting secondary antibody response upon boosting after four months. Further, the particles upon storage at 25 and 37 °C for one month were still capable of mounting an immune response equivalent to those stored in cold chain. Thus, using microparticles entrapping SP0845 for immunization not only improve the immunogenicity but also offer better temperature stability. This can greatly reduce the cost and increase access of protein-based vaccine to resource limited settings.

In this study, an efficient electrochemical biosensor for Vitamin-D3 detection using gadolinium oxide nanorods (Gd2O3NRs) has been reported. Gd2O3NRs were hydrothermally synthesized and functionalized with aspartic acid (Asp-Gd2O3NRs). Asp functionalization did not change the phase, shape and structure of Gd2O3NRs. The high-resolution transmission electron microscopy study revealed the diameter of Asp-Gd2O3NRs as 14.26 ± 0.13 nm with enhanced dispersivity. The Gd2O3NRs and Asp-Gd2O3NRs showed zeta potential of +29 and +24 mV, respectively. Asp-Gd2O3NRs exhibited enhanced hydrophilicity and electrochemical property than the bare Gd2O3NRs. A thin film of Asp-Gd2O3NRs was deposited on a glass substrate coated with indium-tin-oxide (ITO) by electrophoretic deposition. The immobilization of Vitamin-D3 monoclonal antibody (Ab-VD) was done on the surface of Asp-Gd2O3NRs/ITO electrode for determination of Vitamin-D3. The study of BSA/Ab-VD/Asp-Gd2O3NRs/ITO immunoelectrode response with different Vitamin-D3 concentration was investigated using differential pulse voltammetry technique. The results of response study exhibited an improved sensitivity value of 0.38 μA ng−1 mL cm−2 with a linear range of 10–100 ng mL−1 for Vitamin-D3 detection while the detection limit of 0.10 ng mL−1 was obtained. This immunosensor showed a satisfactory response to commercially available Vitamin-D3 oral solution. Besides this, in vitro study of Gd2O3NRs and Asp-Gd2O3NRs was performed on RAW 264.7 and MCF-7 cells. Keywords: Gadolinium oxide, Nanorods, Aspartic acid, Vitamin-D3, Electrochemical immunosensor

Infectious diseases are globally associated with high mortality in spite of the availability of therapeutic agents against most pathogenic microorganisms. This is due to the emergence of new infectious diseases and novel pathogen strategies to evade host defenses. There is thus a need to develop potent therapeutics and techniques for effective delivery of vaccines and drugs. Particles based on poly(lactic acid) and poly(lactic-co-glycolic acid) polymer-based particles are suitable delivery systems due to their biodegradable and biocompatible nature. They can be tailored to display various properties such as sustained release, dose sparing, bioactivity maintenance and targeted delivery. This review focuses on polymeric particle-based delivery systems to develop novel vaccines or drugs. Cellular interactions of particulate systems and the mechanism of action in animal models are also discussed.

Formation of protein aggregates as inclusion bodies (IBs) still poses a major hurdle in the recovery of bioactive proteins from E. coli. Despite the development of many mild solubilization buffers in last two decades, high-throughput recovery of functional protein from wide range of IBs is still a challenge at an academic and industrial scale. Herein, a novel formulation for improved recovery of bioactive protein from variety of bacterial IBs is developed. This novel formulation is comprised of 20% trifluoroethanol, 20% n-propanol and 2 M urea at pH 12.5 which disrupts the major dominant forces involved in protein aggregation. An extensive comparative study of novel formulation conducted on different IBs demonstrates its high solubilization and refolding efficiency. The overall yield of bioactive protein from human growth hormone expressed as bacterial IBs is reported to be around 50%. This is attributed to the capability of novel formulation to disrupt the tertiary structure of the protein while protecting the secondary structure of the protein, thereby reducing the formation of soluble aggregates during refolding. Thus, the formulation can eliminate the need of screening and optimizing various solubilization formulation and will improve the efficiency of recovering bioactive protein from variety of IB aggregates.

Troxerutin (TXR) is a phytochemical reported to possess anti-inflammatory and hepatoprotective effects. In this study, we aimed to exploit anti-arthritic properties of TXR using an adjuvant induced arthritic (AIA) rat model. AIA induced rats showed highest arthritis score at disease onset and by oral administration of TXR (50, 100, 200 mg/kg body weight), reduced to basal level in a dose dependent manner. Isobaric tag for relative and absolute quantitative (iTRAQ) proteomics tool was employed to identify deregulated joint homogenate proteins in AIA and TXR treated rats to decipher probable mechanism of the TXR action in arthritis. iTRAQ analysis identified a set of 434 joint homogenate proteins with 65 deregulated proteins (log2 case/control ≥ 1.5) in AIA. Expressions of a set of important proteins (AAT, T-kininogen, vimentin, desmin, and nucleophosmin) that could classify AIA from healthy were validated using Western blot analysis. Western blot data corroborated proteomics findings. In silico protein-protein interaction study of joint homogenate proteome revealed that complement component 9 (C9), the major building blocks of the membrane attack complex (MAC) responsible for sterile inflammation, gets perturbed in AIA. Our dosimetry study suggests that a TXR dose of 200 mg/kg body weight for 15 days is sufficient to bring the arthritis score to basal levels in AIA rats. We have shown the importance of TXR as an anti-arthritis agent in AIA model and after additional investigation its arthritis ameliorating properties could be exploited for clinical usability.

Vitamin B

Problem The miRNAs show placenta-specific expression patterns, which alter during pregnancy-related complications. In present study, the role of miR-27b-5p during forskolin-mediated BeWo cells fusion has been investigated. Method of study The fusion of BeWo cells in response to forskolin treatment (25 µM) was studied by desmoplakin I+II staining. Expression profile of miR-27b-5p by qRT-PCR and its targets HSD3β1 and WNT2B by qRT-PCR and in Western blot were studied. The effect of overexpression of miR-27b-5p and silencing of HSD3β1 & WNT2B by siRNA on forskolin-mediated BeWo cells fusion and secretion of hCG and progesterone by ELISA was investigated. Results Time-dependent down-regulation in the expression of miR-27b-5p in forskolin-treated BeWo cells has been confirmed by qRT-PCR. Overexpression of miR-27b-5p significantly inhibits forskolin-mediated BeWo cells fusion as well as hCG & progesterone secretion. HSD3β1 and WNT2B were identified as targets of miR-27b-5p and are up-regulated in forskolin-treated BeWo cells. Overexpression of miR-27b-5p in BeWo cells downregulates their expression. Further, luciferase reporter assay revealed that miR-27b-5p directly target expression of both HSD3β1 and WNT2B. Silencing of both HSD3β1 and WNT2B leads to a significant reduction in forskolin-mediated BeWo cells fusion with concomitant decrease in the secretion of progesterone or/and hCG. Decrease in forskolin-mediated cells fusion observed in miR-27b-5p mimic transfected BeWo cells could be rescued by the overexpression of both HSD3β1 and WNT2B. Conclusion These observations suggest that reduced miR-27b-5p in forskolin-treated BeWo cells leads to increased secretion of progesterone and hCG due to loss of repressional control on HSD3β1 and WNT2B.

Annexin V and propidium iodide staining is widely used for determining the cellular death through apoptosis. In the presence of Ca

Annexin V and propidium iodide staining is widely used for determining the cellular death through apoptosis. In the presence of Ca2+ ions, annexin V has a strong binding affinity for phosphatidylserine, a membrane phospholipid that during apoptosis is translocated from the inner side of the cell membrane to its outer side. On the other hand, propidium iodide has ability for DNA binding and it can only enter into necrotic or late apoptotic cells. This chapter describes a commonly used method for detection of apoptosis in a non-small cell lung cancer cell line using annexin V and propidium iodide dye. We describe the detection of different stages of apoptosis in the A549 lung cancer cell line treated with dihydroartemisinin (DHA). This apoptosis detection method can be used to determine the efficacy of different kinds of drugs on cultured cancer cell lines.

Troxerutin (TXR), is a phytochemical reported to possess anti-inflammatory and hepatoprotective effects. In this study, we aimed to exploit anti-arthritic properties of TXR using an adjuvant induced arthritic (AIA) rat model. AIA induced rats showed highest arthritis score at disease onset and by oral administration of TXR (50, 100, 200 mg/kg body weight), reduced to basal level in a dose dependent manner. Isobaric tag for relative and absolute quantitative (iTRAQ) proteomics tool was employed to identify deregulated joint homogenate proteins in AIA and TXR treated rats to decipher probable mechanism of the TXR action in arthritis. iTRAQ analysis identified a set of 434 joint homogenate proteins with 65 deregulated proteins (log2 case/control ≥1.5) in AIA. Expressions of a set of important proteins (AAT, T-kininogen, vimentin, desmin, and nucleophosmin) that could classify AIA from healthy were validated using Western blot analysis. Western blot data corroborated proteomics findings. In silico protein-protein interaction study of joint homogenate proteome revealed that complement component 9, the major building blocks of the membrane attack complex (MAC) responsible for sterile inflammation, gets perturbed in AIA. Our dosimetry study suggests that a TXR dose of 200 mg/kg body weight for 15 days is sufficient to bring the arthritis score to basal levels in AIA rats. We have shown the importance of TXR as an anti-arthritis agent in AIA model and after additional investigation its arthritis ameliorating properties could be exploited for clinical usability.

SP0845, a pneumococcal surface protein and a potential candidate vaccine for Streptococcus pneumoniae infection, was used to evaluate the role of histidine affinity tag on its biophysical properties and immunogenicity. The protein was expressed in E. coli with and without histidine affinity tag and purified to homogeneity. Size exclusion chromatographic studies revealed that tag free SP0845 was mainly monomeric in solution whereas, histidine tagged SP0845 stayed predominantly in an oligomeric form. Histidine-tagged SP0845 have higher β sheet content than the tag free protein. Removal of histidine tag increased the α-helical content of SP0845 from 35% to 46%. Histidine tagged SP0845 elicited higher serum antibody titer in comparison to the tag free SP0845 in mice. Effect of alum in improving the immunogenicity of tagged SP0845 was low in comparison to that observed with tag free protein. Immunogenicity of tag free SP0845 was enhanced by delivering it using polylactide polymeric particles. The presence of histidine tag thus influences the secondary structure and immunogenicity of protein and need careful consideration before use.

High level expression of recombinant proteins in bacteria often results in their aggregation into inclusion bodies. Formation of inclusion bodies poses a major bottleneck in high-throughput recovery of recombinant protein. These aggregates have amyloid-like nature and can retain biological activity. Here, effect of expression temperature on the quality of Escherichia coli Asparaginase II (a tetrameric protein) inclusion bodies was evaluated. Asparaginase was expressed as inclusion bodies at different temperatures. Purified inclusion bodies were checked for biological activities and analyzed for structural properties in order to establish a structure-activity relationship. Presence of activity in inclusion bodies showed the existence of properly folded asparaginase tetramers. Expression temperature affected the properties of asparaginase inclusion bodies. Inclusion bodies expressed at higher temperatures were characterized by higher biological activity and less amyloid content as evident by Thioflavin T binding and Fourier Transform Infrared (FTIR) spectroscopy. Complex kinetics of proteinase K digestion of asparaginase inclusion bodies expressed at higher temperatures indicated higher extent of conformational heterogeneity in these aggregates.

Purpose: Amikacin, a water-soluble aminoglycoside antibiotic used to treat gram-negative bacillary infections, is a Biopharmaceutics Classification System class III drug having poor permeability and short half-life. It is given parenterally, which limits its use in patients warranting “at-home care.” An oral drug delivery of amikacin is, therefore, imminent. Aim: This work focused on establishing poly d, l-lactide-co-glycolide (PLGA)-based nanoparticles of amikacin with consolidated pharmaceutical attributes capable of circumventing gastrointestinal tract membrane barriers and promoting oral administration of the drug. The partied attributes are suggestive of enhanced uptake of the drug via Peyer’s patches overlaying small intestine and support successful oral delivery. Materials and Methods: To have a robust delivery system, a statistical Box–Behnken experimental design was used and formulation parameters such as homogenization time, probe sonication time, and drug/polymer ratio of amikacin-loaded PLGA nanoparticles (A-NPs) for obtaining monodispersed nanoparticles of adequate size and high drug loading were optimized. Results: The model suggested to use the optimum homogenization time, probe sonication time, and drug/polymer ratio as 30 s, 120 s, and 1:10, respectively. Under these formulation conditions, the particle size was found to be 260.3 nm and the drug loading was 3.645%. Conclusion: Biodegradable PLGA nanoparticulate systems with high payload, optimum size, and low polydispersity index will ensure successful uptake and ultimately leading to better bioavailability. Hence, under the aforementioned optimized conditions, the A-NPs prepared had particle size of 260.3 nm, which is appreciable for its permeability across small intestine, and drug loading of 3.645%.

Amikacin (A), a water soluble aminoglycoside antibiotic is commercially available for intravenous administration only. Present investigation is aimed at the development of poly-lactic-co-glycolic acid (PLGA) nanoparticles (A-NPs). 1 for oral permeability enhancement of amikacin. The pharmaceutical attributes of the A-NPs revealed particle size, 260.3 ± 2.05 nm, zeta potential, −12.9 ± 1.12 mV and drug content, 40.10 ± 1.87 μg/mg with spherical shape and smooth surface. In vitro antibacterial studies showed that the A-NPs were active against P. aeruginosa, K. pneumoniae and E. coli. The permeation study across rat ileum showed 2.6-fold improvement in Papp for A-NPs than A-S 2 This increase in permeability is due to the uptake of nanoparticles by Peyer’s patches of intestinal epithelium and endocytic uptake via enterocytes. Flow cytometric analysis demonstrated 2.2-fold higher uptake of Rh B-NPs 3 than Rh B-S 4 and elucidated the dominance of enterocytes mediated endocytosis of nanoparticles. Furthermore, stability data collected as per ICH guidelines for three months under accelerated conditions had shown that the A-NPs were stable. The purported drug delivery system hence, seems a promising tool to replace successfully the current intravenous therapy and is used to support relevant patient compliance thereby, adding value to the “patient care at home”.

In recent years scientific community has drawn a great deal of attention towards understanding the enigma of cluster of differentiation-44 (CD44) in order to deliver therapeutic agents more selectively towards tumor tissues. Moreover, its over-expression in variety of solid tumors has attracted drug delivery researchers to target this receptor with nanomedicines. Conventional nanomedicines based on biodegradable polymers such as poly(lactide-co-glycolide) (PLGA) are often associated with insufficient cellular uptake by cancer cells, due to lack of active targeting moiety on their surface. Therefore, to address this limitation, CD44 targeted PLGA nanomedicines has gained considerable interest for enhancing the efficacy of chemotherapeutic agents. In this review, we have elaborately discussed the recent progress in the design and synthesis of CD44 targeted PLGA nanomedicines used to improve tumor-targeted drug delivery. We have also discussed strategies based on co-targeting of CD44 with other targeting moieties such as folic acid, human epidermal growth factor 2 (HER2), monoclonal antibodies using PLGA based nanomedicines.

Betulinic acid (BA), a naturally occurring plant-derived pentacyclic triterpenoid, has gained attention in recent years owing to its broad-spectrum biological and medicinal properties. Despite the pharmacological activity of BA, it has been associated with some drawbacks, such as poor aqueous solubility and short half-life in vivo, which limit therapeutic application. To solve these problems, much work in recent years has focused on enhancing BA's aqueous solubility, half-life, and efficacy by using nanoscale drug delivery systems. Several different kinds of nanoscale delivery systems-including polymeric nanoparticles, magnetic nanoparticles, liposomes, polymeric conjugates, nanoemulsions, cyclodextrin complexes, and carbon nanotubes-have been developed for the delivery of BA. Here, we focus on the recent developments of novel nanoformulations used to deliver BA in order to improve its efficacy.

Background:Curcumin and nisin have been widely reported for their antibacterial and anticancer potency. However, their therapeutic applications are hampered by several factors, which necessitate their development into nanosize ranges for improved delivery and activities. Their incorporation into a single nanosynthesized form may suggest desirable efficacy on parasites. The aim of the study was to assess the ovicidal activity of the curcumin-nisin polylactic acid (PLA) entrapped nanoparticle on theFasciolaeggs and its reproductive toxicity.Methods:The nanoparticle was formulated by double emulsion method. The eggs of the adultFasciolaspp. were exposed to different concentrations (0.3125–5 mg/mL) of the nanoparticle to monitor hatchability. Mice were exposed to 0.5 mL of the formulated drug at varying concentrations (10–20 mg/kg) and then sacrificed for sperm morphology assay.Results:The mean particle size, polydispersity index, and drug entrapment efficiency of the formulated drug were 288.4±24.3 nm, 0.232, and 51.7%, respectively. The highest nanoparticulate concentration (5 mg/mL) showed the least percentage egg hatching (41.7%) compared with the other treatment groups and positive control (albendazole) (45.1%). The aberrations observed in sperm cells were not concentration-dependent and no significant differences were observed in the mean aberrations between the nanoparticulate drug-exposed groups and the negative control (p>0.05).Conclusions:The results confirmed the ovicidal activity of the curcumin-nisin nanoparticulate drug against theFasciolaspecies. The formulation also showed no toxicity to sperm cells. More robust studies on anti-fascioliasis activity of the drug on adultFasciolaspp. andin vivoandin vitrotoxicity studies are recommended.

In this study, gadolinium oxide nanorods (Gd2O3 NRs) were hydrothermally synthesized and ex-situ functionalized with aspartic acid (Asp-Gd2O3 NRs). The cytotoxocity studies on two cells namely RAW 264.7 and MCF-7 was assessed in terms of cell viability, morphological changes, and cell cycle analysis. Both types of NRs were well characterized and Asp-Gd2O3 NRs exhibited enhanced hydrophilicity and dispersity of NRs. Cell viability assay revealed enhanced biocompatibility of Asp-Gd2O3 NRs with almost 75% viability even at a higher concentration of 250 µg mL-1. The morphological changes upon internalization of both NRs were done through fluorescent microscopy that revealed no significant change in the morphology of the cell or its nucleus. Further, the cell cycle studies again confirmed the biocompatible nature of these NRs. These results suggest that Asp-Gd2O3 NRs are well suited for therapeutic applications, such as thermal cancer therapy, due to their tunable shape, size, low toxicity, and the possibility of surface modification.

Enzyme immobilization has been achieved on various functional nanostructures to increase their shelf-life, enhancement of productivity, stability, reusability and develop a biocatalyst or biosensor. Current review is ascribing the recent advances in the art of immobilization with great efficiency by adopting logistic and sensible nano-engineering approaches i.e nanoparticles, nanoliposomes, nanotubes, fibers and membranes, beads, sol–gel, metal–organic frameworks, DNA nanostructures, nanowires, nanostars, nanocystals, nanoflowers and nanocages. The activity of various immobilized enzyme i.e. lipase, glucose oxidase, laccase, alcohol dehydrogenase and many more on these newer nanostructures are discussed here demonstrating interaction of biomacromolecules with these nanostructures. Taking Glucose oxidase as an example, it’s immobilization on metallic nanoparticles, nano-liposomes, gold nanostars, carbon nanotubes, hydrogel and hybrid nanoparticles to develop a nano-biocatalyst/biosensor has also been discussed. This review will provide a great insight in the selection of nanostructure with different morphologies and composition so that it can be extended for the multi-enzyme immobilization with multi-functionality, stability and reusability for industrial and clinical translation.

Objective: The aim of this study was to formulate polymer-based artesunate nanoparticles for malaria treatment. Methods: Artesunate was loaded with poly(D,L-lactic-co-glycolic acid) (PLGA) by solvent evaporation from an oil-in-water single emulsion. Nanoparticles were characterized by X-ray diffraction and differential scanning calorimetry analyses. In vivo antimalarial studies at 4 mg/kg were performed on Swiss male albino mice infected with Plasmodium berghei. Hematological and hepatic toxicity assays were performed. In vitro cytotoxicity of free and encapsulated artesunate (Art-PLGA) to cell line RAW 264.7 was determined at concentrations of 7.8–1000 µg/ml. Results: The particle size of the formulated drug was (329.3±21.7) nm and the entrapment efficiency was (38.4±10.1)%. Art-PLGA nanoparticles showed higher parasite suppression (62.6%) compared to free artesunate (58.2%, P

In an attempt to find bio-active heterocyclic analogues, a series of novel 1-(5-(3-(aryl)-1-phenyl-1H-pyrazol-4-yl)-3-(pyridin-3-yl)-4,5-dihydropyrazol-1-yl)ethanones 5a–i and 4-(3-(aryl)-1-phenyl-1H-pyrazol-4-yl)-6-(pyridine-3-yl)pyrimidin-2-amines 6a–i were designed, synthesized, and evaluated for their in vitro cytotoxicity against a panel of human cancer cell lines namely, HeLa (human cervix), NCI-H460 (human lung), PC-3 (human prostate), and NIH-3T3 (mouse embryo fibroblasts) cell lines. Most of these compounds exhibited moderate to good cytotoxicity against the tested cancer cell lines and weak toxicity against normal cell line. Analogs 5f, 5g, 5i, 6b–g showed significant cytotoxicity as compared to the standard drug etoposide. The compound 6g exhibited superior activity with IC50 value of 5.47 ± 0.44 µM against Hela cancer cell line.

Present study explores native L-asparaginase encapsulated long-acting cross-linker-free PLGA-nanoformulation in an Ehrlich ascites tumor model. L-asparaginase-PLGA nanoparticles for tumor were prepared using a double emulsion solvent evaporation technique, optimized and validated by Box-Behnken Design. L-ASN-PNs showed a particle size of 195 nm ± 0.2 nm and a PDI of 0.2. Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) techniques revealed its smooth morphology and elicited an in-vitro release of 80% of the drug, following the Higuchi drug release model. In-vivo studies of L-ASN-PNs on an Ehrlich ascites tumor (EAT) model were completed and compared with the standard medication of 5-fluorouracil (5-FU) treatment. L-ASN-PN treated mice showed a 51.15% decrease in tumor volume and 100% survival rate with no reduction in body weight, no haemotoxicity and no hepatotoxicity, as evident from the hematological parameters, and liver enzyme parameters that were well within the prescribed limits. Chemotherapy has severe side effects and restricted therapeutic success. Henceforth, the purported L-Asparaginase PLGA nanoparticles are a suitable entity for better tumor regression, intra-tumor accumulation and no hematological side-effects.

Natural products are complex molecules that have been widely used in traditional medicine for therapeutics and diagnostics applications. Despite their long history of use, some challenges are associated with many natural product derived pharmaceuticals, like inadequate stability, poor absorption, distribution, metabolism and excretion. Medicinal chemists have been successful in addressing many of these challenges through structural modifications of the parent compound, but even so, analysis suggests that up to 20% of natural product leads are taken through unchanged as the final drug product. Even modified compounds are a challenge to administer, requiring the use of novel formulations and delivery strategies to enable the launch of an effective natural product derived drug into the market. To outwit these concerns, formulation of these natural product derived bioactive compounds using nanotechnology has been used as a potential tool in diagnostic and therapeutic applications. Compounds of organic or inorganic origin that are prepared from different metals, metal oxides, chitosan, sodium alginate, poly lactic acid, poly lactic co-glycolic acid, synthetic as well as natural origin polymers are amongst commonly used materials for development of natural product nanoformulations.

This book covers nanotechnology based approaches for improving the therapeutic efficacy of natural products. It critically explores lipid nanoarchitectonics, inorganic particles and nanoemulsion based tools for delivering them. With its chapters from eminent experts working in this discipline, it is ideal for researchers and professionals working in the area.

Cancer is a major cause of deaths throughout the world. According to World Health Organization, cancer lead to the loss of about 9.6 million lives in the year 2018 with low and middle income countries accounting for 70% of these deaths. With its ability to grow in an uncontrolled manner and spreading out to different sites in the body, cancer is seen as that dreaded disease with no cure or treatment. Several promising cancer chemotherapeutic advancements have emerged but these came along with unavoidable limitations like toxicity, limits on the administered dosage as well as affordability for the people who most need them. Natural products with anticancer potential have also emerged as an immensely attractive option to treat cancer. The use of natural products has been credited to doubling of human life span in twentieth century. Furthermore, of all the anti-cancer agents approved and pre-new drug application candidates till date, more than 60% are of natural origin or a derivative thereof. However, an estimated 4 in 10 of the new chemical entity proposed against cancer turns out to be lipophilic. Problems of low aqueous solubility, low bioavailability and non-specific action still pose as major stumbling blocks in their advancement. Recently, polymeric nanocarriers have been used to deliver cancer chemotherapeutics to circumvent the associated limitations and improve their efficacy. Polymers like polylactic acid/poly lactic-co-glycolic acid are have a long history of use in medical field. These are safe and biodegradable with about 20 products approved by the Food and Drug Administration and European Medicine Agency for use in humans. The polymeric nanocarriers with their versatility offer solution to many of the hurdles of natural product delivery for cancer chemotherapy.

Recombinant proteins expressed as bacterial inclusion bodies (IBs) are now receiving tremendous attention for many diverse applications in the areas of industrial and medical biotechnology. Understanding the structure–function relationship of protein in IBs has recently created new possibilities in developing innovative isolation, solubilization, refolding, and purification processes for high-throughput recovery of bioactive protein from bacterial IBs. This opinion article describes the advantages, disadvantages, and major challenges presently associated with each of the processing steps. Finally, we conclude with the possible solutions for each operational step and the future direction of the basic and translational research to achieve maximum benefit from IB aggregates.

PROBLEM Requirement of multiple injections of contraceptive vaccines to achieve infertility is one of the important impediments for their application. In the present study, attempts have been made to reduce the number of injections of contraceptive vaccine. METHOD OF STUDY Fusion protein encompassing C-terminus fragment of sperm protein Sp17 (aa residues 76-126) and two copies of gonadotropin-releasing hormone along with T-cell epitopes and dilysine linkers (abbreviated as Sp17C -GnRH2 ) was expressed in Escherichia coli. Its immunogenicity and contraceptive efficacy have been evaluated in female FVB/J mice using different adjuvants and delivery platforms. RESULTS Immunization of female mice with recombinant Sp17C -GnRH2 (25 μg/injection/mouse) emulsified with squalene-arlacel A following two injections schedule led to failure of 88.8% immunized animals to conceive, which was not significantly different from mice immunized with same protein along with alum following three injections schedule. To make single-dose vaccine, poly d,l-lactic acid-based microparticles (PLA-MPs) entrapping Sp17C -GnRH2 were prepared. Immunization of female mice with a combination of soluble Sp17C -GnRH2 (12.5 μg/injection/mouse) along with Sp17C -GnRH2 entrapped in PLA-MPs (12.5 μg/injection/mouse) in alum showed higher antibody titres and contraceptive efficacy as compared to mice immunized with Sp17C -GnRH2 entrapped in PLA-MPs alone in alum. Immunization with recombinant Sp17C -GnRH2 led to long-term infertility as second mating (150 days after immunization) of various groups of immunized mice showed similar infertility as observed during first mating. CONCLUSION Single-dose immunization with PLA-MPs entrapping Sp17C -GnRH2 along with soluble recombinant protein in alum generated long-lasting infertility in female mice.