An ESIPT solvatochromic fluorescent and colorimetric probe for sensitive and selective detection of copper ions in environmental samples and cell lines.

Copper is one of the most important transition metals in our body for various physiological functions. An imbalance in the homeostasis of copper in our body can lead to various neurological disorders such as Alzheimer's disease, Parkinson's, and Menkes disease. As a result, there is a need for the detection of excess copper present in the environment and the human system. In this work, we have designed a quinoline-based fluorescent/colorimetric probe (QHS) for rapid and selective detection of copper ions via quenching of fluorescence/color change from yellow to peach which is visible to the naked eye. The probe displayed high selectivity towards copper(II), i.e., Cu(II) in the presence of different metal analytes in water samples. The sensing mechanism of the probe was confirmed by NMR, HRMS, IR spectroscopy, and SEM. The detection limit of Cu(II) ions was found to be 0.493 µM which is lower than the tolerable limit of copper (20 µM) in drinking water as per the EPA. The probe was also utilized for the visualization of Cu(II) in cell lines. The probe was also demonstrated for its application in real-time detection of aqueous samples using portable paper strips.

Intracellular DAMPs in Neurodegeneration and Their Role in Clinical Therapeutics.

Neuroinflammation is the major implication of neurodegeneration. This is a complex process which initiates from the cellular injury triggering the innate immune system which gives rise to damage-associated molecular patterns (DAMPs) which are also recognized as endogenous danger indicators. These originate from various compartments of the cell under pathological stimulus. These are very popular candidates having their origin in the intracellular compartments and organelles of the cell and may have their site of action itself in the intracellular or at the extracellular spaces. Under the influence of the pathological stimuli, they interact with the pattern-recognition receptor to initiate their pro-inflammatory cascade followed by the cytokine release. This provides a good opportunity for diagnostic and therapeutic interventions creating better conditions for repair and reversal. Since the major contributors arise from the intracellular compartment, in this review, we have attempted to focus on the DAMP molecules arising from the intracellular compartments and their specific roles in the neurodegenerative events explaining their downstream mediators and signaling. Moreover, we have tried to cover the latest interventions in terms of DAMPs as clinical biomarkers which can assist in detecting the disease and also target it to reduce the innate-immune activation response which can help in reducing the sterile neuroinflammation having an integral role in the neurodegenerative processes.

Age-dependent assessment of selenium nanoparticles: biodistribution and toxicity study in young and adult rats.

Aim: To study the biodistribution and toxicology of selenium nanoparticles (SeNPs) versus their bulk counterpart in young and adult male rats in a 28-day study. Methods: SeNPs were synthesized and conjugated with indocyanine green to assess comparative biodistribution by in vivo imaging and further characterized by transmission electron microscopy, Fourier transform infrared, scanning electron microscopy/energy dispersive x-ray spectroscopy, UV and ζ-analysis. The toxicity of bulk selenium was evaluated relative to its nano form by hematology indices, redox, inflammatory markers and histopathology. Results: Indocyanine green-conjugated nanoparticles showed preferential accumulation in the liver, followed by testis and kidney. The protective effect of SeNPs was more significantly observed in young livers than in adults compared with the bulk counterpart. Conclusion: Age-dependent monitoring and diagnosis of toxicity may need different biomarkers of selenium and may also provide better understanding of SeNPs as therapeutics.

Selenium is an essential element in the body. Its bioactive properties can protect against neurological conditions, diabetes, cancer and other chronic disorders. However, selenium in various biological forms (bulk) can be toxic. Selenium nanoparticles (SeNPs) have unique properties which might prevent this toxicity, providing a potential alternative for selenium supplementation and therapy. However, more studies are needed to see where SeNPs localize in the body, as well as comparing their toxicology with conventional forms of selenium in different age groups. We synthesized and characterized SeNPs of 70­90 nm, then injected them into young and adult rats to see where they distributed in the body. This was compared with rats injected with bulk selenium. SeNPs showed preferential accumulation in the liver, followed by the testes and kidneys. Next, the toxicity profiles of SeNPs and bulk selenium were established by measuring a series of health markers in the liver. It was found that the protection against toxicity provided by SeNPs was more significant in younger rats. Our results demonstrate that the same dose may behave differently in different age groups and that bulk selenium induces different toxicities in young and adult rats compared with SeNPs, highlighting the importance of different indicators of health for the monitoring of selenium-related toxicity when designing selenium-based therapeutics.

Albumin-Based Nanocarriers for the Simultaneous Delivery of Antioxidant Gene and Phytochemical to Combat Oxidative Stress.

Human serum albumin (HSA) nanoparticles are promising biocompatible, nontoxic, and non-immunogenic platforms for biomedical applications such as bioimaging and drug and gene delivery. The development of nonviral gene delivery vectors is a great challenge for efficient and safe gene therapy. Sulforaphane (SF) can stimulate the expression of antioxidant genes via activation of a nuclear transcription factor, the erythroid-2 related factor 2 (Nrf-2). Here, we use polyethyleneimine (PEI)-stabilized HSA nanoparticles to stimulate endogenous antioxidant defense mechanisms in lung epithelial cells L-132 through the combinatorial effect of SF drug and antioxidant superoxide dismutase 1 gene (pSOD1 plasmid) delivered by HSA-PEI-SF-pSOD1 nanocomposites (NCs). The developed NCs demonstrated high biocompatibility (L-132 viability, >95%, MTT assay) and high antioxidant activity because of efficient entry of the SOD1 gene and SF-loaded NCs at a very low (3 µg) dose in L-132 cells. A high transfection efficiency of L-132 cells (∼66%, fluorescent microscopy) was obtained with the GFP-tagged transgene SOD1-GFP. We speculate that the antioxidant activity of HSA-PEI-SF-pSOD1 NCs in L-132 cells is due to the initial release of SF followed by subsequent SOD1 gene expression after three to four days of incubation. Hence, the developed HSA-based NCs can be efficient biocompatible nanocarriers for safe and effective drug and gene delivery applications to treat diseases with high oxidative stress due to combinatorial SF and SOD1 gene mechanisms.

Formulation development of tocopherol polyethylene glycol nanoengineered polyamidoamine dendrimer for neuroprotection and treatment of Alzheimer disease.

Amyloid-beta (Aß) aggregates deposition at extra neuronal sites induces neurotoxicity and major hallmarks of Alzheimer's disease (AD). To reduce the Aß fibril toxicity, multi-functional polyamidoamine (PAMAM) dendrimer was conjugated with tocopheryl polyethylene glycol succinate-1000 (TPGS) which acts as a carrier matrix for the delivery of neuroprotective molecule piperine (PIP). This PIP-TPGS-PAMAM dendrimer was fabricated to mitigate the Aß1-42 fibril toxicity on SHSY5Y cells. TPGS-PAMAM was fabricated through carbodiimide coupling reaction, and PIP was encapsulated in dendrimer through solvent injection method to prepare PIP-TPGS-PAMAM. Antioxidant assay of PIP-TPGS-PAMAM showed 90.18% inhibition of 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radicals compared to free PIP, which was 28.27%. The SHSY5Y cells showed 37.25% for negative control group and 82.55% cell viability for PIP-TPGS-PAMAM treated group against Aß1-42 toxicity. PIP-TPGS-PAMAM reduced the ROS activity to 15.21% and 48.5% for free PIP treated in cell group. Similarly, extent of Aß1-42-induced apoptosis also reduced significantly from 38.2% to 12.36% in PIP-TPGS-PAMAM treated group. In addition, PIP-TPGS-PAMAM also disaggregated the Aß1-42 fibril in SHSY5Y cells. Our findings suggested that PIP-TPGS-PAMAM showed mitigation of Aß1-42-induced toxicity in neuronal cells, which can offer excellent prospect of neuroprotection and AD therapy.

Alpha-Lipoic Acid Protects Co-Exposure to Lead and Zinc Oxide Nanoparticles Induced Neuro, Immuno and Male Reproductive Toxicity in Rats.

We evaluated the neuro-, immuno-, and male reproductive toxicity of zinc oxide nanoparticles (ZnO NPs) alone and in combination with lead acetate. We also studied the therapeutic role of α-lipoic acid postexposure. Lead (10 mg/kg, body weight), ZnO NPs (100 mg/kg, bwt) alone, and their combination were administered orally in Wistar rats for 28 days, followed by the administration of α-lipoic acid (15 mg/kg, bwt) for the next 15 days. Our results demonstrated protective effects of α-lipoic acid on lead and ZnO NP-induced biochemical alterations in neurological, immunological, and male reproductive organs in rats. The altered levels of blood δ-aminolevulinic acid dehydratase (ALAD), immunoglobulins (IgA, IgG, IgM, and IgE), interleukins (IL-1ß, IL-4, and IL-6), caspase-3, and tumor necrosis factor (TNF-α) were attenuated by lipoic acid treatment. Lead and ZnO NP-induced oxidative stress was decreased by lipoic acid treatment, while a moderate recovery in the normal histoarchitecture of the brain section (cortex and hippocampus) and testes further confirmed the neuro- and male reproductive toxicity of lead and ZnO NPs. We also observed a significant decrease in the blood metal content in the animals treated with lipoic acid compared to the lead-administered group, indicating the moderate chelating property of lipoic acid. It may thus be concluded that lipoic acid might be a promising protective agent against lead and ZnO NP-induced alterations in the neurological, immunological, and reproductive parameters.

Dose-dependent hepatic toxicity and oxidative stress on exposure to nano and bulk selenium in mice.

Selenium is an essential mineral naturally found in soil, water, and some of the food and is required as essential elements in human and animal body. Se supplementation is required especially for those having Se deficiency. Food supplement of selenium has several forms such as selenocysteine, selenite, selenomethionine, and selenate. Recently, Se supplement as selenium nanoparticles (SeNPs) has gained worldwide attention due to its bioactivities and properties. In the present study, we determined the potential hepatotoxicity of nano and bulk selenium using low and high doses in mice. Twenty-five Swiss albino mice (n=5) were randomly divided into 5 groups and treated orally for 28 days: Group 1: sterile saline (0.9%) as a control; Group 2: sodium selenite (1mg/kg); Group 3: sodium selenite (4mg/kg); Group 4: selenium nanoparticles (1mg/kg); and Group 5: selenium nanoparticles (4mg/kg). Administration of nano-selenium (70-90 nm) led to an increase in the activities of serum transaminases (ALT and AST), while no significant effects were noted on biochemical variables indicative of changes in heme synthesis pathway and oxidative stress like blood δ-aminolevulinic acid dehydratase (δ-ALAD), hepatic reactive oxygen species (ROS), catalase activity, superoxide dismutase (SOD), malondialdehyde assay (MDA), reduced glutathione (GSH) and oxidized glutathione (GSSG), glutathione peroxidase (GPx) compared to controls, and a high dose of sodium selenite. Our results suggest that nano-selenium at low dose (1mg/kg) exhibited antioxidant effects in the liver compared to the high dose (4mg/kg) of SeNPs and sodium selenite (1 and 4 mg/kg). The data from the present study might be useful for pharmacologists and toxicologists in providing future directions while designing selenium-based therapeutic strategies.

Bio-engineered palladium nanoparticles: model for risk assessment study of automotive particulate pollution on macrophage cell lines.

The surge in vehicular activity in densely populated areas has led to an increased concentration of airborne palladium nanoparticles (PdNPs) in the environment. Recent toxicity data have indicated that PdNPs exhibit adverse effects in in vitro and in vivo models, however, their effect on the immune system is not fully understood. Therefore, in the present study, we aimed to evaluate possible toxic effects of bio-engineered palladium nanoparticles on the murine macrophage cell line (J774). Here we prepared palladium nanoparticles using aqueous leaf extract of Parthenium hysterophorus and characterized them by UV-Vis spectroscopy, XRD, FT-IR spectroscopy, HR-TEM, EDX, SEM and zeta potential. Toxicity parameters such as cell viability, cell membrane integrity, induction of apoptosis and ROS production were assessed on J774 cell lines. Spherical palladium nanoparticles of mean size ∼4 nm, when subjected to time and dose-dependent cytotoxicity assay, showed cell viability was >95% at lower doses (25, 200 µg mL-1) and <50% at higher doses of palladium nanoparticles (400, 500 µg mL-1) after 24 hours of incubation. We also observed cell membrane injury at higher doses by lactate dehydrogenase assay. The induction of apoptosis observed was moderate. H2DCFDA assay revealed visible cell damage which could be due to modest levels of ROS generation. The detection of Pd in the road-dust samples of New Delhi using inductively coupled plasma-mass spectroscopy (ICP-MS) technique was also investigated.

Chronic exposure to multi-metals on testicular toxicity in rats.

Despite the availability of sufficient data on the effects of individual metal exposure on living organisms, a critical knowledge gap still exists in predicting effects of multi-metals particularly on the pituitary-testicular axis. Thus, the aim of the present study was to check the effects of individual or combined (binary and ternary) exposure to aluminum, copper, and zinc on (i) sperm and testosterone levels (ii) oxidative stress and (iii) structural changes in testis of male Wistar rats. Animals were exposed to aluminum, copper, and zinc either individually (20 mg/kg, orally, once, daily), binary (10 mg/kg each, orally, once daily) or in ternary combination (5 mg/kg, each, orally, once daily) for 24 weeks. The exposure to aluminum, copper individually and in combination led to a significant decrease in sperm counts and an increased oxidative stress compared to the control group. Exposure to zinc caused significant decrease in oxidative stress and an increase in different sperm variables. The exposure to zinc with aluminum or copper had no toxic effects on testis while concomitant exposure to aluminum, copper, and zinc produced more pronounced testicular injury. In summary, while co-exposure to zinc with aluminum or copper produced reproductive toxicity the co-exposure to all the three metals may lead to a significant testicular toxicity and these changes were related to increase in oxidative stress in rats.

Comparative efficacy of Nano and Bulk Monoisoamyl DMSA against arsenic-induced neurotoxicity in rats.

Chelation therapy is considered as a safe and effective strategy to combat metal poisoning. Arsenic is known to cause neurological dysfunctions such as impaired memory, encephalopathy, and peripheral neuropathy as it easily crosses the blood-brain barrier. Oxidative stress is one of the mechanisms suggested for arsenic-induced neurotoxicity. We prepared Solid Lipid nanoparticles loaded with Monoisoamyl 2, 3-dimercaptosuccinic acid (Nano-MiADMSA), and compared their efficacy with bulk MiADMSA for treating arsenic-induced neurological and other biochemical effects. Solid lipid nanoparticles entrapping MiADMSA were synthesized and particle characterization was carried out by transmission electron microscopy (TEM) and dynamic light scattering (DLS). An in vivo study was planned to investigate the therapeutic efficacy of MiADMSA-encapsulated solid lipid nanoparticles (Nano-MiADMSA; 50 mg/kg orally for 5 days) and compared it with bulk MiADMSA against sodium meta-arsenite exposed rats (25 ppm in drinking water, for 12 weeks) in male rats. The results suggested the size of Nano-MiADMSA was between 100-120 nm ranges. We noted enhanced chelating properties of Nano-MiADMSA compared with bulk MiADMSA as evident by the reversal of oxidative stress variables like blood δ-aminolevulinic acid dehydratase (δ-ALAD), Reactive Oxygen Species (ROS), Catalase activity, Superoxide Dismutase (SOD), Thiobarbituric Acid Reactive Substances (TBARS), Reduced Glutathione (GSH) and Oxidized Glutathione (GSSG), Glutathione Peroxidase (GPx), Glutathione-S-transferase (GST) and efficient removal of arsenic from the blood and tissues. Recoveries in neurobehavioral parameters further confirmed nano-MiADMSA to be more effective than bulk MiADMSA. We conclude that treatment with Nano-MiADMSA is a better therapeutic strategy than bulk MiADMSA in reducing the effects of arsenic-induced oxidative stress and associated neurobehavioral changes.

Nanotechnology: A Promising Approach for Delivery of Neuroprotective Drugs.

Central nervous system (CNS) disorders especially neurodegenerative disorders are the major challenge for public health and demand the great attention of researchers to protect people against them. In past few decades, different treatment strategies have been adopted, but their therapeutic efficacy are not enough and have only shown partial mitigation of symptoms. Blood-brain barrier (BBB) and blood-cerebrospinal fluid barrier (BSCFB) guard the CNS from harmful substances and pose as the major challenges in delivering drugs into CNS for treatment of CNS complications such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), stroke, epilepsy, brain tumors, multiple sclerosis (MS), and encephalitis, etc. Nanotechnology has come out as an exciting and promising new platform of treating neurological disorders and has shown great potential to overcome problems related to the conventional treatment approaches. Molecules can be nanoengineered to carry out multiple specific functions such as to cross the BBB, target specific cell or signaling pathway, respond to endogenous stimuli, and act as a vehicle for gene delivery, support nerve regeneration and cell survival. In present review, the role of nanocarrier systems such as liposomes, micelles, solid lipid nanoparticles (SLNPs), dendrimers, and nanoemulsions for delivery of various neurotherapeutic agents has been discussed, besides this, their mechanism of action, and nanoformulation of different neuroprotective agents like curcumin, edaravone, nerve growth factors in CNS disorders like Alzheimer's, Parkinsonism, epilepsy, stroke, and brain tumors has been reviewed.

The Thin But Fat Phenotype is Uncommon at Birth in Indian Babies.

BACKGROUND: Indian babies are hypothesized to be born thin but fat. This has not been confirmed with precise measurements at birth. If it is true, it could track into later life and confer risk of noncommunicable diseases (NCDs). OBJECTIVES: Primarily, to accurately measure percentage of body fat (%BF) and body cell mass (BCM) in Indian babies with normal birth weight, compare them across different gestational ages and sex, and test the hypothesis of the thin but fat phenotype in Indian babies. Secondarily, to examine the relation between body weight and body fat in Indian babies. METHODS: Term newborns (n = 156) weighing ≥2500 g, from middle socioeconomic status mothers were recruited in Bengaluru, India, and their anthropometry, %BF (air displacement plethysmography), and BCM (whole-body potassium counter) were measured. Maternal demography and anthropometry were recorded. The mean %BF and its dispersion were compared with earlier studies. The relation between newborn %BF and body weight was explored by regression analysis. RESULTS: Mean birth weight was 3.0 ± 0.3 kg, with mean %BF 9.8 ± 3.5%, which was comparable to pooled estimates of %BF from published studies (9.8%; 95% CI: 9.7, 10.0; P > 0.05). Appropriate-for-gestational age (AGA) babies had higher %BF (1.8%) compared to small-for-gestational age (SGA) babies (P < 0.01). Mean %BCM of all babies at birth was 35.4 ± 10.5%; AGA babies had higher %BCM compared to SGA babies (7.0%, P < 0.05). Girls in comparison to boys had significantly higher %BF and lower %BCM. Body weight was positively associated with %BF. CONCLUSION: Indian babies with normal birth weight did not demonstrate the thin but fat phenotype. Body weight and fat had positive correlation, such that SGA babies did not show a preservation of their %BF. These findings will have relevance in planning optimal interventions during early childhood to prevent NCDs risk in adult life.

Lactobionic Acid Conjugated Quercetin Loaded Organically Modified Silica Nanoparticles Mitigates Cyclophosphamide Induced Hepatocytotoxicity.

OBJECTIVE: The study was designed to investigate the therapeutic potential of lactobionic acid (LA) conjugated quercetin (Q) loaded organically modified silica nanoparticles (LA-Q-ORMOSIL) with bulk quercetin to mitigate cyclophosphamide (CP) induced liver injury. METHODOLOGY: Q-ORMOSIL nanoparticles were synthesized and characterized using UV-Vis spectroscopy, TEM, Zeta sizer, FTIR and EDX. Further, encapsulation efficiency and in vitro release kinetic study was done. Q-ORMOSIL nanoparticles surface were modified with lactobionic acid, a ligand for the asialoglycoprotein receptor on the hepatocyte surface. The hepatoprotective effects of Q-ORMOSIL and LA-Q-ORMOSIL nanoparticles were evaluated in vivo. Cyclophosphamide (20 mg/kg/day, i.p) was co-administered for seven days with bulk quercetin (50mg/kg/day) and quercetin nanoparticles (50µg/kg/day). After seven days, the number of biomarkers for liver function test and oxidative stress were determined in liver homogenate. Histopathological changes were also analyzed in control and treated liver tissues. RESULTS: Physiochemical characterization of LA-Q-ORMOSIL nanoparticles depicts that the particles formed were of approx. 80 nm, spherical, monodispersed in nature and showed sustain drug release in in vitro study. Our results further suggested that Q-ORMOSIL and LA-Q-ORMOSIL nanoparticles significantly decreased tissue TBARS, ROS levels and ALT, AST, and ALP activities compared to CP induced group. On the other hand, tissue antioxidant levels (GSH, GST, and catalase) showed a significant increase in LA-Q-ORMOSIL treated group compared to the CP treated group confirming its high therapeutic efficacy during liver injury. CONCLUSION: Targeted nanoquercetin demonstrated a significant hepatoprotective effect compared to bulk quercetin against CP-induced hepatotoxicity and it considerably reduced bulk quercetin dose level to many folds. Bulk quercetin has low bioavailability and thus, from obtained data we suggest that LA-Q-ORMOSIL nanoparticles provide high therapeutic value in protecting experimental animals against CP-induced liver injury. We also propose multifunctional dye-doped LA-modified ORMOSIL nanoparticles for future studies in facilitating nanoparticles uptake to hepatocytes for liver diagnosis and treatment.

Impact of chronic low dose exposure of monocrotophos in rat brain: Oxidative/ nitrosative stress, neuronal changes and cholinesterase activity.

Monocrotophos (MCP) is an organophosphate mainly used as insecticides in agriculture, and veterinary practice to control pests. Exposure to MCP is known to induce significant systemic toxicity in animals and humans. Short term exposure to a high dose of MCP has been reported to cause systemic toxicity, however limited information is available regarding low dose long term exposure in rats. We studied the effects of low dose long term exposure to MCP on oxidative/nitrosative stress, cholinesterase activity and neuronal loss in rat. Male rats were exposed to MCP (0.1 µg or 1 µg/ml) via drinking water for 8 weeks. The pro-oxidant markers such as reactive oxygen species (ROS), lipid peroxidation (MDA), nitrite level and antioxidant markers such as reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and inhibition of cholinesterase activities were measured to evaluate the effects of MCP on brain along with plasma cholinesterase activity. Neuronal loss was analyzed in cortical region using H&E stained slices. The results suggested that exposure to MC even at the low dose, increased reactive oxygen species, thiobarbituric acid reactive substance levels and decreased glutathione, superoxide dismutase, catalase and cholinesterase activities in brain. No significant effect however, was observed on nitrite levels. Histological analysis revealed that low dose MCP exposure lead to structural changes in the cortical neurons in rats. It can be concluded from the study that low dose long term exposure (lower than No Observed Effect Level) of MCP may lead to the generation of oxidative stress by elevation of pro-oxidants markers and depletion of antioxidant enzymes markers along with inhibition of cholinesterase activity. These changes might thus be considered as the possible mechanism of cortical neuronal loss in these animals.

Estimation of protein requirements in Indian pregnant women using a whole-body potassium counter.

BACKGROUND: The 2007 World Health Organization/Food and Agriculture Organization/United Nations University (WHO/FAO/UNU) recommendation for the Estimated Average Requirement (EAR) of additional protein during pregnancy for a gestational weight gain (GWG) of 12 kg (recalculated from a GWG of 13.8 kg) is 6.7 and 21.7 g/d in the second and the third trimester, respectively. This EAR is based on measurements of potassium accretion in high-income country (HIC) pregnant women. It is not known if low- to middle-income country, but well-nourished, pregnant women have comparable requirements. OBJECTIVE: We aimed to estimate total body potassium (TBK) accretion during pregnancy in Indian pregnant women, using a whole-body potassium counter (WBKC), to measure their additional protein EAR. METHODS: Well-nourished pregnant women (20-40 y, n = 38, middle socioeconomic stratum) were recruited in the first trimester of pregnancy. Anthropometric, dietary, and physical activity measurements, and measurements of TBK using a WBKC, were performed at each trimester and at birth. RESULTS: The mid-trimester weight gain was 2.7 kg and 8.0 kg in the second and the third trimester, respectively, for an average 37-wk GWG of 10.7 kg and a mean birth weight of 3.0 kg. Protein accretion was 2.7 and 5.7 g/d, for an EAR of 8.2 and 18.9 g/d in the second and the third trimester, respectively. The additional protein EAR, calculated for a GWG of 12 kg, was 9.1 and 21.2 g/d in the second and the third trimester, respectively. CONCLUSION: The additional protein requirements of well-nourished Indian pregnant women for a GWG of 12 kg in the second and third trimesters were similar to the recalculated 2007 WHO/FAO/UNU requirements for 12 kg.

The development of a whole-body potassium counter for the measurement of body cell mass in adult humans.

BACKGROUND AND OBJECTIVES: Total body potassium (TBK), has a natural radioactive isotope, which can be measured to derive body cell mass (BCM), making it useful in clinical conditions, early growth and pregnancy. The objective was to build a whole-body potassium counter (WBKC), to accurately measure TBK in the body. METHODS AND STUDY DESIGN: A WBKC was designed and constructed using a shadow shield. A cellular four compartment (4C) model of fat free mass (FFM), using estimates of TBK along with total body water (TBW), was compared with a molecular 4C model of the body in twenty healthy adults (10 men and 10 women). The molecular 4C model used measurements of TBW, bone mineral content (BMC), and body volume from deuterium dilution (DD), dual energy x-ray absorptiometry (DXA) and air displacement plethysmography (ADP) respectively. RESULTS: The accuracy and precision of the WBKC were 2.8% and 1.9% with TBK phantoms. The mean estimate of FFM by the molecular 4C model was 40.4±6.8 kg, while it was 41.2±7.3 kg using the cellular 4C model. CONCLUSIONS: A WBKC constructed from base principles, was relatively low cost, efficient, safe and noninvasive, but requires some design considerations. Its measurement of FFM compared well with the molecular 4C model. Once constructed, it offers a relatively costless, accurate and repeatable method to measure body composition in conditions with uncertain hydration status, at all life stages.

Enhanced antineoplastic/therapeutic efficacy using 5-fluorouracil-loaded calcium phosphate nanoparticles.

In the past few decades, the successful theranostic application of nanomaterials in drug delivery systems has significantly improved the antineoplastic potency of conventional anticancer therapy. Several mechanistic advantages of nanomaterials, such as enhanced permeability, retention, and low toxicity, as well as surface engineering with targeting moieties, can be used as a tool in enhancing the therapeutic efficacy of current approaches. Inorganic calcium phosphate nanoparticles have the potential to increase the therapeutic potential of antiproliferative drugs due to their excellent loading efficiency, biodegradable nature and controlled-release behaviour. Herein, we report a novel system of 5-fluorouracil (5-FU)-loaded calcium phosphate nanoparticles (CaP@5-FU NPs) synthesized via a reverse micelle method. The formation of monodispersed, spherical, crystalline nanoparticles with an approximate diameter of 160-180 nm was confirmed by different methods. The physicochemical characterization of the synthesized CaP@5-FU NPs was done with transmission electron microscopy (TEM), dynamic light scattering (DLS), field emission scanning electron microscopy (FE-SEM), Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD). The antineoplastic potential of the CaP@5-FU NPs against colorectal and lung cancer cells was reported. The CaP@5-FU NPs were found to inhibit half the population (IC50) of lung adenocarcinoma (A549) cells at 32 µg/mL and colorectal (HCT-15) cancer cells at 48.5 µg/mL treatment. The apoptotic induction of CaP@5-FU NPs was confirmed with acridine orange/ethidium bromide (AO/EB) staining and by examining the morphological changes with Hoechst and rhodamine B staining in a time-dependent manner. The apparent membrane bleb formation was observed in FE-SEM micrographs. The up-regulated proapoptotic and down-regulated antiapoptotic gene expressions were further confirmed with semiquantitative reverse transcriptase polymerase chain reaction (PCR). The increased intracellular reactive oxygen species (ROS) were quantified via flow cytometry upon CaP@5-FU NP treatment. Likewise, the cell cycle analysis was performed to confirm the enhanced apoptotic induction. Our study concludes that the calcium phosphate nanocarriers system, i.e. CaP@5-FU NPs, has higher antineoplastic potential as compared to 5-FU alone and can be used as an improved alternative to the antimitotic drug, which causes severe side effects when administrated alone.

Niclosamide loaded biodegradable chitosan nanocargoes: an in vitro study for potential application in cancer therapy.

Chitosan nanoparticles can advance the pharmacological and therapeutic properties of chemotherapeutic agents by controlling release rates and targeted delivery process, which eliminates the limitations of conventional anti-cancer therapies and they are also safe as well as cost-effective. The aim of present study is to explore the anti-tumour effect of niclosamide in lung and breast cancer cell lines using biocompatible and biodegradable carrier where nanoparticles loaded with hydrophobic drug (niclosamide) were synthesized, characterized and applied as a stable anti-cancer agent. Niclosamide loaded chitosan nanoparticles (Nic-Chi Np's) of size approximately 100-120 nm in diameter containing hydrophobic anti-cancer drug, i.e. niclosamide, were prepared. Physico-chemical characterization confirms that the prepared nanoparticles are spherical, monodispersed and stable in aqueous systems. The therapeutic efficacy of Nic-Chi Np's was evaluated against breast cancer cell line (MCF-7) and human lung cancer cell line (A549). MTT assay reveals the cell viability of the prepared Nic-Chi Np's against A549 and MCF-7 cells and obtained an IC50 value of 8.75 µM and 7.5 µM, respectively. Acridine orange/ethidium bromide dual staining results verified the loss of the majority of the cells by apoptosis. Flow cytometer analysis quantified the generation of intracellular reactive oxygen species (ROS) and signified that exposure to a higher concentration (2 × IC50) of Nic-Chi Np's resulted in elevated ROS generation. Notably, Nic-Chi Np treatment showed more apoptosis and cell death in MCF-7 as compared to A549. Further, the remarkable induction of apoptosis by Nic-Chi Np's was confirmed by semi-quantitative reverse transcription polymerase chain reaction, scanning electron microscopy and cell-cycle analysis. Thus, Nic-Chi Np's may have a great potential even at low concentration for anti-cancer therapy and may replace or substitute more toxic anti-mitotic drugs in the near future.

Antileishmanial activity of nanocurcumin.

BACKGROUND: Leishmaniasis is an endemic disease having a wide spectrum ranging from visceral, cutaneous and mucocutaneous forms caused by unicellular, obligate intracellular parasites of the monocyte-macrophage system. The aim of the present study was to develop an effective, nontoxic and biodegradable polymeric drug-delivery system encapsulating curcumin in its hydrophobic core for the treatment of visceral leishmaniasis. RESULTS: We have reported a co-polymeric micelle of N-isopropyl acrylamide, vinyl pyrrolidone and acrylic acid in 85:10:5 M ratios through free radical polymerization. The characterization of curcumin-loaded nanoparticles (40-50 nm) was done by transmission electron microscopy, dynamic light scattering and spectroscopic methods such as NMR which ensures polymerization and formation of nanoparticles has been achieved. Nanocurcumin was evaluated as an antileishmanial agent through spleenomegaly and delayed-type hypersensitivity experiments. CONCLUSION: Nanocurcumin has shown significantly greater in vivo therapeutic efficacy than pentamidine and free curcumin in an animal model of visceral leishmaniasis. The use of nanocurcumin compared with conventional drugs and free curcumin may prove more feasible and provide a better approach towards treatment of leishmaniasis.

Concentration-dependent toxicity of iron oxide nanoparticles mediated by increased oxidative stress.

Iron oxide nanoparticles with unique magnetic properties have a high potential for use in several biomedical, bioengineering and in vivo applications, including tissue repair, magnetic resonance imaging, immunoassay, drug delivery, detoxification of biologic fluids, cell sorting, and hyperthermia. Although various surface modifications are being done for making these nonbiodegradable nanoparticles more biocompatible, their toxic potential is still a major concern. The current in vitro study of the interaction of superparamagnetic iron oxide nanoparticles of mean diameter 30 nm coated with Tween 80 and murine macrophage (J774) cells was undertaken to evaluate the dose- and time-dependent toxic potential, as well as investigate the role of oxidative stress in the toxicity. A 15-30 nm size range of spherical nanoparticles were characterized by transmission electron microscopy and zeta sizer. MTT assay showed >95% viability of cells in lower concentrations (25-200 µg/mL) and up to three hours of exposure, whereas at higher concentrations (300-500 µg/mL) and prolonged (six hours) exposure viability reduced to 55%-65%. Necrosis-apoptosis assay by propidium iodide and Hoechst-33342 staining revealed loss of the majority of the cells by apoptosis. H2DCFDDA assay to quantify generation of intracellular reactive oxygen species (ROS) indicated that exposure to a higher concentration of nanoparticles resulted in enhanced ROS generation, leading to cell injury and death. The cell membrane injury induced by nanoparticles studied using the lactate dehydrogenase assay, showed both concentration- and time-dependent damage. Thus, this study concluded that use of a low optimum concentration of superparamagnetic iron oxide nanoparticles is important for avoidance of oxidative stress-induced cell injury and death.