An amperometric nanobiosensor using a biocompatible conjugate for early detection of metastatic cancer cells in biological fluid.

Metastasis is the major cause of cancer-associated death in humans, and its early diagnosis will help clinicians to develop suitable therapeutic strategies which may save life of cancer patients. In this direction, we designed an amperometric biosensor using a biocompatible conjugate to diagnose cancer metastasis by detecting epithelial cell adhesion molecule expressing metastatic cancer cells (Ep-MCCs). The sensor probe is fabricated by immobilizing monoclonal capture antibody (CapAnti) on the gold nanoparticles (AuNPs)/conducting polymer composite layer. The detection relies on a sandwich-type approach using a bioconjugate composed of reporter antibody (RepAnti), nanostructured collagen (nCOL), AuNPs, and hydrazine (Hyd) which served as a nonenzymatic electrocatalyst for the reduction of H2O2. The binding of Ep-MCCs with the sensor probe was confirmed using electrochemical impedance spectroscopy, cyclic voltammetry, and chronoamperometry. A dynamic range for the Ep-MCCs detection is determined between 45 and 100,000 Ep-MCCs/mL with the detection limit of 28±3 Ep-MCCs/mL. The proposed immunosensor is successfully applied to detect Ep-MCCs in serum and mixed cell samples and interferences due to nontarget cells and molecules present in the real sample matrix are also examined. The early stage of Ep-MCCs was examined by fluorescence-activated cell sorting assay, which confirms that the developed biosensor has detected Ep-MCCs in its early stage.

Ultrasensitive detection of drug resistant cancer cells in biological matrixes using an amperometric nanobiosensor.

Multidrug resistance (MDR) is a key issue in the failure of cancer chemotherapy and its detection will be helpful to develop suitable therapeutic strategies for cancer patients and overcome the death rates. In this direction, we designed a new amperometric sensor (a medical device prototype) to detect drug resistant cancer cells by sensing "Permeability glycoprotein (P-gp)". The sensor probe is fabricated by immobilizing monoclonal P-gp antibody on the gold nanoparticles (AuNPs) conducting polymer composite. The detection relies on a sandwich-type approach using a bioconjugate, where the aminophenyl boronic acid (APBA) served as a recognition molecule which binds with the cell surface glycans and hydrazine (Hyd) served as an electrocatalyst for the reduction of H2O2 which are attached on multi-wall carbon nanotube (MWCNT) (APBA-MWCNT-Hyd). A linear range for the cancer cell detection is obtained between 50 and 100,000 cells/mL with the detection limit of 23±2 cells/mL. The proposed immunosensor is successfully applied to detect MDR cancer cells (MDRCC) in serum and mixed cell samples. Interferences by drug sensitive (SKBr-3 and HeLa), noncancerous cells (HEK-293 and OSE), and other chemical molecules present in the real sample matrix are examined. The sensitivity of the proposed immunosensor is excellent compared with the conventional reporter antibody based assay.

Nutraceutical and pharmacological implications of marine carbohydrates.

Current day's research has been focusing much on the potential pharmacological or nutraceutical agents of selective health benefits with less toxicity. As a consequence of increased demand of nutritional supplements of great medicinal values, development of therapeutic agents from natural sources, in particular, marine environment are being considered much important. A diverse array of marine natural products containing medicinally useful nutritional substances, i.e., marine nutraceuticals have been focused to the benefit of mankind. Carbohydrates, by being constituted in considerable amount of many marine organisms display several nutraceutical and pharmaceutical behavior to defend from various diseases. Moreover, the carbohydrates from algae as well as from shellfish wastes, like chitosan and its derivatives, showed tremendous applications in biology and biomedicine. In the current chapter, several of marine carbohydrates from various marine flora and fauna have been covered with their applications and prospects in the development of nutraceuticals and pharmaceuticals.

Applications of carbon nanomaterials in bone tissue engineering.

In the biomedical field, remarkable advancements have been made in artificial biomaterials for treating bone loss or defects. A variety of synthetic polymers, natural polymers and bioceramics are being used to develop artificial bones. Many natural and synthetic biomaterials, which are being investigated for their physiochemical role in vivo, are currently in the clinical trial stage. Carbon-based prostheses are promising materials that mimic the natural function of bone, e.g., mechanical strength. Recently, carbon-based bone materials, such as carbon nanotubes and graphene, have been widely investigated as potential solutions to several biomedical problems. This review summarizes the biophysicochemical and biomedical properties of carbon nanomaterials composed of polymer and ceramic structures and discusses their functionality in bone tissue engineering.

Synthesis and evaluation of the cytotoxic activities of some isatin derivatives.

A series of isatin derivatives, 1-butyl-5/7-chloro/fluoro-3-((4-methoxybenzyl)imino)indolin-2-ones (3a-d), 6-butyl-chloro/fluoro-6H-indolo[2,3-b]quinoxalines (4a-h), and 5/7-chloro/fluoro-3-((4-methoxybenzyl)imino)indolin-2-ones (5a-h) were synthesized and characterized by using Fourier transform (FT)-IR, (1)H- and (13)C-NMR spectroscopy, mass spectrometric and elemental analysis. The substances were further subjected to in vitro cytotoxicity evaluation against HeLa, SK-BR-3, and MCF-7 cells. The results showed that quinoxalines 4d, 4e, and 4g; and indolin-2-one 5f display significant in vitro cytotoxic activities against HeLa cells and further the compound 4d has resulted in highest cytotoxicity in the entire series studied. In addition, 5f was shown to display substantial activity against all the three cell lines used in the current study.

Spatial variation of potentially toxic elements in different grain size fractions of marine sediments from Gulf of Mannar, India.

Marine sediments of the Gulf of Mannar (GoM), India are contaminated by potential toxic elements (PTEs) due to anthropogenic activities posing a risk to the existing fragile coral ecosystem and human health. The current study aimed to assess the distribution of PTEs (arsenic--As; cobalt--Co; copper--Cu, molybdenum--Mo; lead--Pb; and zinc--Zn) in marine sediments of different grain size fractions, viz., medium sand (710 µm), fine sand (250 µm), and clay (<63 µm) among the different coastal regions of Pamban, Palk Bay, and Rameswaram coasts of GoM, using grain size as one of the key factor controlling their concentrations. The concentrations of PTEs were measured in the different size fractions of sediment using inductively coupled plasma mass spectrophotometer. The order of accumulation of all PTEs in the three fractions was ranked as Zn > Cu > Pb > As > Co > Mo and in the three locations as Rameswaram > Palk Bay > Pamban. The concentration of PTEs in Palk Bay and Rameswaram coast was significantly different (P < 0.05), when compared to Pamban coast. Measured geoaccumulation index (I(geo)) and contamination factor (CF) indicated significant enrichment of Co and Pb from Rameswaram coast when compared to other two coasts. Although the concentration of Co was low but the measured I(geo) and CF values indicated significant enrichment of this PTE in Rameswaram coast. The increased input of PTEs in the coastal regions of GoM signifies the need to monitor the coast regularly using suitable monitoring tools such as sediments to prevent further damage to the marine ecosystem.

Chitosan-amylopectin/hydroxyapatite and chitosan-chondroitin sulphate/hydroxyapatite composite scaffolds for bone tissue engineering.

Over the past few decades, artificial graft materials for bone tissue engineering are gaining much importance. In this study, tri-component scaffolds of chitosan/natural hydroxyapatite with chondroitin sulfate (chitosan-CS/HAp) and amylopectin (chitosan-AP/HAp) have been developed for the first time via freeze-drying method and were characterized physicochemically for bone grafting substitutes. Chemical interactions and dispersion of HAp, CS and AP in the chitosan matrix have been evaluated by various analytical techniques. The porosity and water uptake/retention ability of these composite scaffolds decreased whereas thermal stability increased when compared to the chitosan scaffold. The pore size of the chitosan/HAp, chitosan-CS/HAp and chitosan-AP/HAp scaffolds varied from 60 to 180 µm, 60 to 400 µm and 80 to 500 µm, respectively. Cell proliferation, alkaline phosphatase activity and type-1 collagen production was evaluated in vitro using MG-63 cell line, which was observed to be higher in the composite scaffolds. Excellent interconnected porosity, controlled biodegradation and enhanced cell proliferation of the novel chitosan-CS/HAp and chitosan-AP/HAp scaffolds suggests that these scaffolds are promising biomaterials for bone tissue engineering.

Medicinal foods from marine animals: current status and prospects.

The lifestyle of human being is changing day by day toward the simplified and more convenient way of living. Human wellbeing is majorly dependent on the daily food habits that are in accordance with the habits of individual community and the surrounding environments. Although the food habits are simplified and fashioned according to the current lifestyle, many of the Asians are still showing much importance to the naturally derived and traditional foods. One such medicinally important natural source is the foods from marine organisms, which are an important growing notion for the development of marine nutraceuticals and functional foods. In this context, we have already brought the recent trends and applications of marine algal (macro and micro) foods in my previous volume. The current preliminary chapter of this book volume on marine animals and microbes describes about the prospects of various marine animals and their derived substances/materials as medicinal foods. In addition, this chapter encourages the new researchers as well as various health communities to implement the marine animal-based medicinal foods and their applications.

Biophysicochemical evaluation of chitosan-hydroxyapatite-marine sponge collagen composite for bone tissue engineering.

Tricomponent scaffold systems prepared by natural materials especially of marine origin are gaining much attention nowadays for the application in bone tissue engineering. A novel scaffold (Chi-HAp-MSCol) containing chitosan (Chi), hydroxyapatite (HAp) derived from Thunnus obesus bone and marine sponge (Ircinia fusca) collagen (MSCol) was prepared using freeze-drying and lyophilization method. This biomimetic scaffold, along with the Chi and Chi-HAp scaffolds were characterized biophysicochemically for their comparative significance in bone grafting applications. The structural composition of the chitosan, Chi-Hap, and Chi-HAp-MSCol scaffolds were characterized by Fourier Transform Infrared spectroscopy. The porosity, water uptake, and retention abilities of the composite scaffolds decreased, whereas Thermogravimetric and Differential Thermal Analyses results revealed the increase in thermal stability in the scaffold because of the highly stable HAp and MSCol. Homogeneous dispersion of HAp and MSCol in chitosan matrix with interconnected porosity of 60-180 µm (Chi-HAp) and 50-170 µm (Chi-HAp-MSCol) was observed by Scanning Electron Microscopy, X-ray diffraction, and optical microscopy. Cell proliferation in composite scaffolds was relatively higher than pure chitosan when observed by MTT assay and Hoechst staining in vitro using MG-63 cell line. These observations suggest that the novel Chi-HAp-MSCol composite scaffolds are promising biomaterials for matrix-based bone repair and bone augmentation.

Impact of marine micro- and macroalgal consumption on photoprotection.

The enormousness of species diversity of oceans leads to the isolation and development of health- and beauty-enhancing components from various marine organisms. The significance of these marine-derived compounds or substances has been scientifically well implied for various biological and biomedical parameters. One such important parameter is photoprotectivity, which is the major concern nowadays because of the depletion in ozone layer and the possible high risk of UV irradiation to humans. The marine macro- as well as microalgae and their food products, knowingly and unknowingly, have been used since hundreds of years. These foods possess tremendous implications in defending the highly hazardous UV radiation, thereby facilitating photoprotection to humans. In addition, based on the recent studies, many of the UV-protecting algal species is directed for the use as medicinally valuable foods and food ingredients. This chapter describes certain micro- and macroalgal species along with their photoprotective importance.

Synthesis of polyfunctionalized piperidone oxime ethers and their cytotoxicity on HeLa cells.

A series of twenty 2,6-diarylpiperidin-4-one O-methyloximes were synthesized with fluoro/chloro/bromo/methyl/methoxy/ethoxy/isopropyl substituents on various positions of the phenyl at C-2 and C-6 in association with/without methyl substituent on the secondary amino group and methyl/ethyl/isopropyl substituents on the active methylene centers. Regardless of their substitution all compounds predominantly exist in the chair conformation except 3m, which adopts a twist-boat conformation. All the synthesized compounds were evaluated for their in vitro antiproliferative activity against human cervical carcinoma (HeLa) cell line. The cytotoxicity of the test compounds was determined by measuring the number of live cells after 24 h of treatment by MTT assay method. This preliminary SAR suggests some lead molecules 3c-f, 3j-k, 4d-g, and 4i with a scope of further structural optimization of the piperidone pharmacophore toward the development of anticancer drug synthesis.

Biochemical and biophysical characterization of collagens of marine sponge, Ircinia fusca (Porifera: Demospongiae: Irciniidae).

Collagens were isolated and partially characterized from the marine demosponge, Ircinia fusca from Gulf of Mannar (GoM), India, with an aim to develop potentially applicable collagens from unused and under-used resources. The yield of insoluble, salt soluble and acid soluble forms of collagens was 31.71 ± 1.59, 20.69 ± 1.03, and 17.38 ± 0.87 mg/g dry weight, respectively. Trichrome staining, Scanning & Transmission Electron microscopic (SEM & TEM) studies confirmed the presence of collagen in the isolated, terminally globular irciniid filaments. The partially purified (gel filtration chromatography), non-fibrillar collagens appeared as basement type collagenous sheets under light microscopy whereas the purified fibrillar collagens appeared as fibrils with a repeated band periodicity of 67 nm under Atomic Force Microscope (AFM). The non-fibrillar and fibrillar collagens were seen to have affinity for anti-collagen type IV and type I antibodies raised against human collagens, respectively. The macromolecules, i.e., total protein, carbohydrate and lipid contents within the tissues were also quantified. The present information on the three characteristic irciniid collagens (filamentous, fibrillar and non-fibrillar) could assist the future attempts to unravel the therapeutically important, safer collagens from marine sponges for their use in pharmaceutical and cosmeceutical industries.

Anti-photoaging and photoprotective compounds derived from marine organisms.

Marine organisms form a prominent component of the oceanic population, which significantly contribute in the production of cosmeceutical and pharmaceutical molecules with biologically efficient moieties. In addition to the molecules of various biological activities like anti-bacterial, anti-cancerous, anti-inflammatory and anti-oxidative etc., these organisms also produce potential photoprotective or anti-photoaging agents, which are attracting present day researchers. Continuous exposure to UV irradiation (both UV-A and UV-B) leads to the skin cancer and other photoaging complications, which are typically mediated by the reactive oxygen species (ROS), generated in the oxidative pathways. Many of the anti-oxidative and anti-photoaging compounds have been identified previously, which work efficiently against photodamage of the skin. Recently, marine originated photoprotective or anti-photoaging behavior was observed in the methanol extracts of Corallina pilulifera (CPM). These extracts were found to exert potent antioxidant activity and protective effect on UV-A-induced oxidative stress in human dermal fibroblast (HDF) cells by protecting DNA and also by inhibiting matrix metalloproteinases (MMPs), a key component in photoaging of the skin due to exposure to UV-A. The present review depicts various other photoprotective compounds from algae and other marine sources for further elaborative research and their probable use in cosmeceutical and pharmaceutical industries.

The use of marine sponge, Haliclona tenuiramosa as bioindicator to monitor heavy metal pollution in the coasts of Gulf of Mannar, India.

The results of the present research study indicate that the heavy metal accumulation in the marine sponges provide evidence as an excellent bioindicators for monitoring heavy metal pollution between near and offshore environments of Mandapam coast of "Gulf of Mannar (GoM), India". The heavy metal concentrations in sea water and accumulation in the tissues of Haliclona tenuiramosa were analyzed by ICP-MS (inductively coupled plasma-mass spectrometry). The concentrations of metals in the coastal waters of nearshore (< 0.5 km from shore) were always higher than those in the offshore waters (2-5 km away from shore). Likewise, sponges living in the nearshore accumulated greater concentrations of heavy metals (As, Cd, Co, Cu, Fe, Mn and Ni) ranging from 2 to 17 times higher concentration than the sponges located away from the shore. A positive correlation between concentration levels in water and bioaccumulation in tissues was observed. The bioaccumulation of heavy metals in sponge tissue were in order of Fe > Mn > Ni > Cu > As > Co > Cd in both the near and offshore stations. The present results justified that a more comprehensive monitoring of presence of heavy metals in H. tenuiramosa of surrounding GoM, is necessary to help a better mitigation of the problem.