Bioaccumulation of Titanium in diatom Cyclotella atomus Hust.

Diatomaceous earth or diatomite is a fossil rock deposit of diatoms made up of silica and other minerals. A distinguishing feature of diatoms that placed them in the single class of microalgae Bacillariophyceae, is the frustule, a transparent, hard-shelled cell wall. It's interesting to note that the diatom has specific proteins and enzymes for heavy metal detoxification and can intake and store more heavy metals in its frustule. Consequently, an attempt has been made in this study to determine the bioaccumulation of metals in the frustules of the diatom. Hence, a centric diatom was isolated from the freshwater sample collected from the Adyar River, Chennai, Tamil Nadu. The diameter of the cell was 5-7.5 µm and 20-23 striations with radial arrangement. A single, dark off-center fultoportula and marginal fultoportula on the striae are found in the diatom. Additionally, one rimoportula between two marginal fultoportula distributed on the striae between the costa was also seen. As a result, the isolated diatom was morphologically identified as Cyclotella atomus Hust. Simultaneously, the bioaccumulation study reveals that the Titanium (Ti) was found accumulated in the frustules of the diatom incubated in the Ti-supplemented culture medium based on the scanning electron microscope-energy-dispersive X-ray analysis (SEM-EDAX). Therefore, the biogenic accumulation and fabrication of Titanium frustules in diatom have advantages in enhancing the efficiency of solar cells.

Response of Saponaria officinalis L. hairy roots to the application of TiO2 nanoparticles in terms of production of valuable polyphenolic compounds and SO6 protein.

Saponaria officinalis L. is a perennial plant from the Caryophyllaceae family whose various parts are used in traditional medicine as the treatment agent of skin diseases, blood purifier, diuretic, sudorific, and bile purifier. The cultivation system of hairy roots is a proper alternative for improving the valuable pharmaceutical compounds production compared to other in-vitro methods. The extensive nanotechnology applications in hairy roots cultivation is a sustainable production foundation to produce such active elements. In this study, the effect of various concentrations of titanium dioxide nanoparticles (TiO2 NPs) (0, 10, 20, 30, 50 mg L-1) with two treatments (24 and 48 h) was examined on the growth level, antioxidant capacity, total phenol and flavonoid contents, antioxidant enzyme activities, certain polyphenol compounds and SO6 protein in hairy roots of S. officinalis. According to the results, the maximum (3.09 g) and minimum (0.96 g) fresh weight (FW) of hairy roots were observed in treated culture media with 10 and 20 mg L-1 of TiO2 NPs after 24 and 48 h of exposure times, respectively. The highest rate of total phenol (9.79 mg GLA g-1 FW) and total flavonoid contents (1.06 mg QE g-1 FW) were obtained in the treated hairy roots with 50 and 30 mg L-1 of nano elicitor in 24 and 48 h of treatments, respectively. The maximum level of most polyphenols, such as rosmarinic acid, cinnamic acid, and rutin, was produced in 24 h of treatment. The use of TiO2 NP for 48 h with 50 mg L-1 concentration showed the highest production level of SO6 protein.

Melatonin alleviates titanium nanoparticles induced osteolysis via activation of butyrate/GPR109A signaling pathway.

BACKGROUND: Inflammatory osteolysis after total joint replacement (TJR) may cause implant failure, periprosthetic fractures, and be a severe threat to global public health. Our previous studies demonstrated that melatonin had a therapeutic effect on wear-particles induced osteolysis. Gut microbiota is closely related to bone homeostasis, and has been proven to be affected by melatonin. However, whether melatonin could play its anti-osteolysis effects through reprogramming gut microbiota remains elusive. RESULTS: Here, we demonstrated that melatonin could alleviate Ti-particles induced osteolysis, while this therapeutic effect was blocked by antibiotic cocktail treatment. Interestingly, transplantation of fecal microbiota from mice treated with melatonin reappeared the same beneficial effect. Analysis of the 16S rRNA revealed that melatonin could reverse dysbacteriosis triggered by osteolysis, and elevate the relative abundance of some short chain fatty acid (SCFA) producing bacteria. Moreover, butyrate was enriched by exogenous melatonin administration, while acetate and propionate did not show an evident difference. This was consistent with the results of the metagenomic approach (PICRUSt2) analysis, which revealed a general increase in the synthetic enzymes of butyrate. More importantly, direct supplementation of butyrate could also recapitulate the anti-osteolysis effect of melatonin. Further analysis identified that butyrate alleviated osteolysis via activating its receptor GPR109A, and thus to suppress the activation of NLRP3 inflammasome triggered by Ti-particles. CONCLUSIONS: Taken together, our results suggested that the benefits of melatonin mainly depend on the ability of modulating gut microbiota and regulating butyrate production.

Evaluation of the neurotoxic effects of engineered nanomaterials in C57BL/6J mice in 28-day oral exposure studies.

In recent years, concerns have emerged about the potential neurotoxic effects of engineered nanomaterials (NMs). Titanium dioxide and silver are among the most widely used types of metallic NMs. We have investigated the effects of these NMs on behaviour and neuropathology in male and female C57BL/6J mice following 28-day oral exposure with or without a 14-day post-exposure recovery. The mice were fed ad libitum with food pellets dosed with 10 mg/g TiO2, 2 mg/g polyvinylpyrrolidone-coated Ag or control pellets. Behaviour was evaluated by X-maze, open field, string suspension and rotarod tests. Histological alterations were analysed by immunohistochemistry and brain tissue homogenates were investigated for markers of oxidative stress, inflammation and blood-brain barrier disruption. Effects of the NMs on tyrosine and serine/threonine protein kinase activity in mouse brains were investigated by measuring kinase activity on peptide microarrays. Markers of inflammation, oxidative stress and blood-brain barrier integrity were not significantly affected in the male and female mice following exposure to Ag or TiO2. Both types of NMs also revealed no consistent significant treatment-related effects on anxiety and cognition. However, in the Ag NM exposed mice altered motor performance effects were observed by the rotarod test that differed between sexes. At 1-week post-exposure, a diminished performance in this test was observed exclusively in the female animals. Cortex tissues of female mice also showed a pronounced increase in tyrosine kinase activity following 28 days oral exposure to Ag NM. A subsequent Inductively Coupled Plasma - Mass Spectrometry (ICP-MS) based toxicokinetic study in female mice revealed a rapid and persistent accumulation of Ag in various internal organs including liver, kidney, spleen and the brain up to 4 weeks post-exposure. In conclusion, our study demonstrated that subacute exposure to foodborne TiO2 and Ag NMs does not cause substantial neuropathological changes in mice. However, the toxicokinetic and specific toxicodynamic findings indicate that long-term exposures to Ag NM can cause neurotoxicity, possibly in a sex-dependent manner.

TiO2 nanoparticles dose, application method and phosphorous levels influence genotoxicity in Rice (Oryza sativa L.), soil enzymatic activities and plant growth.

The present study focused on investigating the effect of titanium dioxide nanoparticles (TiO2NPs) on rice (Oryza sativa L.) growth and changes in soil health in two contrasting soil textures (silt-loam and clay). Moreover, response of rice to different methods of TiO2NPs application and phosphorous fertilizer levels were also evaluated. For toxicity assessment, pot experiment was carried out. TiO2NPs (0, 500, 750 mg kg-1) were applied and plants were grown till vegetative stage. After harvesting, physiological parameters, stress assay, soil microbial and enzymatic activities were determined. Based on the results of toxicity study, impact of three methods of TiO2NPs application (foliar, irrigation, soil) and four phosphorous fertilizer levels (0, 10, 20, 40 mg kg-1) on rice growth were assessed. During the 1st phase, results showed an adverse effect of TiO2NPs on plant growth and soil microorganisms in both soil textures at 750 mg kg-1. The H2O2 production, lipid peroxidation and leaf membrane injury index were increased by 4.3-, 2.4-, and 1.9-folds in clay soil upon 750 mg kg-1 TiO2NPs application. Likewise, at the same level of TiO2NPs; microbial biomass, dehydrogenase, and respiration were decreased by 0.91-, 0.79-, and 0.78- folds respectively. In 2nd phase, maximum shoot length, biomass, phosphorous uptake and rice grain protein content were observed under application of TiO2NPs (500 mg kg-1) through irrigation method in combination with 40 mg P kg-1. However, 20 and 40 mg P kg-1 performed equally well upon TiO2NPs application and the results were not statistically significant. The results suggest that 750 mg kg-1 of TiO2NPs negatively affect plant growth and soil enzymatic activities. Moreover, combined application of TiO2NPs (500 mg kg-1) through irrigation and 20 mg P kg-1 is recommended to be the optimum for growth of rice plant.

Plant isoflavones can affect accumulation and impact of silver and titania nanoparticles on ovarian cells.

Objectives. The application of nanoparticles is experiencing a rapid growth, but it faces a problem of their toxicity, especially adverse effects on female reproduction. Food and medicinal plants and their isoflavones can be protectors against environmental stressors, but their ability to abate the adverse effects of nanoparticles has not been studied yet. In the present study, we examined the effect of silver (AgNPs) and titanium dioxide (titania, TiO2NPs) nanoparticles alone or in combination with plant phytoestrogens/antioxidants (resveratrol, diosgenin, and quercetin) on accumulation of nanoparticles, and progesterone release by cultured porcine ovarian granulosa cells.Methods. Porcine granulosa cells were incubated in the presence of AgNPs or TiO2NPs (0.1, 1, 10 or 100 µg/ml) alone or in combination with resveratrol, diosgenin or quercetin (10 µg/ml) for 48 h. The accumulation of tested nanoparticles by granulosa cells was assessed under light microscope. Progesterone concentration in culture media was measured by ELISA kit.Results. Cells accumulated both AgNPs and TiO2NPs in a dose-dependent manner. AgNPs, but not TiO2NPs, at highest dose (100 µg/ml) resulted in a destruction of cell monolayer. Both Ag-NPs and TiO2NPs reduced progesterone release. Resveratrol, diosgenin, and quercetin promoted accumulation of both AgNPs and TiO2NPs in ovarian cells and inhibited the progesterone output. Furthermore, resveratrol and diosgenin, but not quercetin, prevented the suppressive action of both AgNPs, and TiO2NPs on progesterone release.Conclusions. These observations (1) demonstrate accumulation of AgNPs and TiO2NPs in ovarian cells, (2) confirm the toxic impact of AgNPs, and TiO2NPs on these cells, (3) confirm the inhibitory effects of plant polyphenols/phytoestrogens on ovarian steroidogenesis, (4) show the ability of these isoflavones to increase the accumulation of AgNPs and TiO2NPs, and (5) show their ability to reduce the suppressive effect of AgNPs and TiO2NPs on ovarian progesterone release. The suppressive effect of AgNPs and TiO2NPs on ovarian functions should be taken into account by their exposition. However, these adverse effects could be mitigated by some plant isoflavones.

Evaluation of soft-tissue response around laser microgrooved titanium percutaneous devices.

Percutaneous devices are prone to epidermal downgrowth and sinus tract formation, which can serve as a nidus for bacterial colonization and increase the risk of peri-prosthetic infection. A laser microgrooved topography has been shown to limit gingival epidermal downgrowth around dental implants. However, the efficacy of this laser microgrooved topography to limit epidermal downgrowth around nongingival percutaneous devices is yet to be investigated. In this study, devices with a porous-coated subdermal component and a percutaneous post were designed and manufactured. The proximal 2 mm section of the percutaneous post were left smooth, or were textured with either a porous coating, or with the laser microgrooved topography. The smooth and porous topographies served as controls. The devices were tested in a hairless guinea pig back model, where 18 animals were randomly assigned into three groups, with each group receiving one implant type (n = 6/group). Four weeks postimplantation, the devices with surrounding soft-tissues were harvested and processed for histological analyses. Results indicated that the laser microgrooved topography failed to prevent epidermal downgrowth (23 ± 4%) around percutaneous posts in this model. Furthermore, no significant differences (p = 0.70) in epidermal downgrowth were present between the three topographies, with all the groups exhibiting similar measures of downgrowth. Overall, these findings suggest that the laser microgrooved topography may not halt downgrowth around percutaneous devices for dermal applications.

Evaluation of titanium dioxide and chromic oxide as digestibility markers in ponies fed alfalfa hay in relation to marker dosing frequency.

In equines, Cr2O3 is widely accepted as an indigestible marker, but there are health concerns regarding the carcinogenic properties of Cr2O3. Recently, TiO2 has been suggested to be an alternative digestibility marker in equines. However, a comparison between Cr2O3 and TiO2 has not been made in equines. Six Welsh pony geldings (initial BW: 254±3 kg; 7 years of age) fed chopped alfalfa hay were used to evaluate the use of TiO2 (Ti) and Cr2O3 (Cr) as markers for calculating apparent digestibility and to investigate the effect of frequency of marker administration on the measurement of digestibility values. Diets contained 4.65 kg dry matter (DM) chopped alfalfa hay supplemented with minerals, vitamins, TiO2 (3.3 g Ti/day) and Cr2O3 (3.2 g Cr/day). Ponies were dosed with either 3.3 g Ti and 3.2 g Cr once daily (DF1) or with 1.65 g Ti and 1.60 g Cr twice daily (DF2). After adaptation to the diets and procedures for 14 days, voluntary voided faeces were collected quantitatively over 7 days and analysed for moisture, ash, Ti and Cr. Apparent total tract DM digestibility (DMD) and organic matter digestibility (OMD) were calculated using the total faecal collection (TFC) and marker method (Ti and Cr). The overall mean cumulative faecal recovery of Cr and Ti (as % of intake) were 102.0% and 96.6%, respectively. Mean daily faecal recoveries of Cr as well as of Ti were not different (P=0.323; P=0.808, respectively) between treatments. Overall daily faecal recovery of Cr differed (P=0.019) from 100% when the marker was dosed once daily, whereas overall daily faecal recovery was similar to 100% for both administration frequencies when Ti was used as a marker. For both markers, the coefficient of variation of the mean faecal marker recovery between horses was lower when the markers were administrated twice per day. Across treatments, cumulative DMD and OMD estimated with Ti were similar (P=0.345; P=0.418, respectively) compared with those values determined by TFC method. When Cr was used, the calculated cumulative DMD tended (P=0.097) to be greater compared with those estimated with TFC, and cumulative OMD values were overestimated (P=0.013). Orally supplemented Ti recovery in the faeces of ponies fed chopped alfalfa hay with Ti administered once or twice daily was close to 100%, making it the preferred marker for digestibility trials in equines.

A comparison of inverted and upright laser-activated titanium nitride micropyramids for intracellular delivery.

The delivery of biomolecules into cells relies on porating the plasma membrane to allow exterior molecules to enter the cell via diffusion. Various established delivery methods, including electroporation and viral techniques, come with drawbacks such as low viability or immunotoxicity, respectively. An optics-based delivery method that uses laser pulses to excite plasmonic titanium nitride (TiN) micropyramids presents an opportunity to overcome these shortcomings. This laser excitation generates localized nano-scale heating effects and bubbles, which produce transient pores in the cell membrane for payload entry. TiN is a promising plasmonic material due to its high hardness and thermal stability. In this study, two designs of TiN micropyramid arrays are constructed and tested. These designs include inverted and upright pyramid structures, each coated with a 50-nm layer of TiN. Simulation software shows that the inverted and upright designs reach temperatures of 875 °C and 307 °C, respectively, upon laser irradiation. Collectively, experimental results show that these reusable designs achieve maximum cell poration efficiency greater than 80% and viability greater than 90% when delivering calcein dye to target cells. Overall, we demonstrate that TiN microstructures are strong candidates for future use in biomedical devices for intracellular delivery and regenerative medicine.

Modulation of physiological responses with TiO2 nano-particle in Azolla pinnata R.Br. under 2,4-D toxicity.

The present work is emphasised with the herbicidal tolerance of Azolla pinnata R.Br. and its modulation with TiO2 nano-particle. Both carbohydrate and nitrogen metabolism were effected with 2,4-D as herbicide and in few cases TiO2-NP had recovered few detrimental effects. From the nutrient status in Azolla it recorded the recovery of nitrogen as well as potassium by TiO2-NP but not in case of phosphorus. However, a conversion of nitrate to ammonium was more induced by TiO2-NP under herbicidal toxicity. Similar results were obtained for inter-conversion of amino acid-nitrate pool, but no changes with glutamine synthase activity with TiO2-NP. Initially, the effects of 2,4-D was monitored with changes of chlorophyll content but had not been recovered with nanoparticle. Photosynthetic reserves expressed as both total and reducing sugar were insensitive to TiO2-NP interference but activity of soluble and wall bound invertase was in reverse trend as compared to control. The 2,4-D mediated changes of redox and its oxidative stress was ameliorated in plants with over expressed ADH activity. As a whole the Azolla bio system with TiO2 supplementation may be useful in sustenance against 2,4-D toxicity through recovery of nitrogen metabolism. Thus, Azolla-TiO2-NP bio system would be realised to monitor the herbicidal toxicity in soil and its possible bioremediation.

Effect of exercise on apparent total tract digestibility of nutrients and faecal recovery of ADL and TiO2 in ponies.

Exercise and physical training are known to affect gastrointestinal function and digestibility in horses and can lead to inaccurate estimates of nutrient and energy digestibility when markers are used. The effect of exercise on apparent nutrient digestibility and faecal recoveries of ADL and TiO2 was studied in six Welsh pony geldings subjected to either a low- (LI) or high-intensity (HI) exercise regime according to a cross-over design. Ponies performing LI exercise were walked once per day for 45 min in a horse walker (5 km/h) for 47 consecutive days. Ponies submitted to HI exercise were gradually trained for the same 47 days according a standardized protocol. Throughout the experiment, the ponies received a fixed level of feed and the daily rations consisted of 4.7 kg DM of grass hay and 0.95 kg DM of concentrate. The diet was supplemented with minerals, vitamins and TiO2 (3.0 g Ti/day). Total tract digestibility of DM, organic matter (OM), CP, crude fat, NDF, ADF, starch, sugar and energy was determined with the total faeces collection (TFC) method. In addition, DM and OM digestibility was estimated using internal ADL and the externally supplemented Ti as markers. Urine was collected on the final 2 days of each experimental period. Exercise did not affect apparent digestibility of CP, crude fat, starch and sugar. Digestibility of DM (DMD), OM (OMD), ADF and NDF tended to be lower and DE was decreased when ponies received the HI exercise regime. For all treatments combined, mean faecal recoveries of ADL and Ti were 87.8±1.7% and 99.3±1.7%, respectively. Ti was not detected in the urine, indicating that intestinal integrity was maintained with exercise. Dry matter digestibility estimated with the TFC, ADL and Ti for ponies subjected to LI exercise were 66.3%, 60.3% and 64.8%, respectively, while DMD for HI ponies were 64.2%, 60.3% and 65.2%, respectively. In conclusion, physical exercise has an influence on the GE digestibility of the feed in ponies provided with equivalent levels of feed intake. In addition, the two markers used for estimating apparent DMD and OMD indicate that externally supplemented Ti is a suitable marker to determine digestibility of nutrients in horses performing exercise unlike dietary ADL.

Green synthesis of titanium dioxide (TiO2) nanoparticles by Trigonella foenum-graecum extract and its antimicrobial properties.

In recent years, biosynthesis of nanoparticles has received considerable attention due to the growing need to develop clean and nontoxic chemicals, low-cost approaches, eco - friendly solvents and renewable materials. In the current study, the biosynthesis of TiO2 nanoparticles (TiO2NPs) was attained by a chemical and biosynthesized method by using the aqueous leaf extract of Trigonella foenum-graecum (TF-TiO2NP). TiO2 NPs were characterized by FTIR, UV, XRD, HR-TEM and HR-SEM methods. The X-ray diffraction displayed the existence of TF-TiO2NPs which is confirmed by the incidence of peaks at 25.28 corresponds to 101 anatase form. HR-SEM perceptions revealed that synthesized TiO2NPs were spherical in shape and the size of individual nanoparticles as well as a few aggregates was found to be 20-90 nm. The antimicrobial activities of biosynthesized nanoparticles (TF-TiO2NPs) were examined using Kirby-Bauer method. The TF-TiO2 nanoparticles showed significant antimicrobial activity against all the tested microorganisms.

Improved Biocompatibility of Black Phosphorus Nanosheets by Chemical Modification.

Black phosphorus nanosheets (BPs) show great potential for various applications including biomedicine, thus their potential side effects and corresponding improvement strategy deserve investigation. Here, in vitro and in vivo biological effects of BPs with and without titanium sulfonate ligand (TiL4 ) modification are investigated. Compared to bare BPs, BPs with TiL4 modification (TiL4 @BPs) can efficiently escape from macrophages uptake, and reduce cytotoxicity and proinflammation. The corresponding mechanisms are also discussed. These findings may not only guide the applications of BPs, but also propose an efficient strategy to further improve the biocompatibility of BPs.

Growth and Metabolic Responses of Rice (Oryza sativa L.) Cultivated in Phosphorus-Deficient Soil Amended with TiO2 Nanoparticles.

Plants have the natural ability to withstand stress conditions through metabolic adjustments. The present study aimed at investigating the effects of titanium dioxide nanoparticles (TiO2 NPs) application (0, 25, 50, 150, 250, 500, and 750 mg kg-1) in phosphorus-deficient soil in terms of growth responses, P contents, and metabolic alterations in rice. TiO2 NPs application increased shoot length up to 14.5%. Phosphorus contents in rice roots, shoots, and grains were increased by 2.6-, 2.4-, and 1.3-fold, respectively, at 750 mg kg-1 of TiO2 NPs. Gas chromatography-mass spectrometry (GC-MS)-based metabolomics revealed increased levels of amino acids, palmitic acid, and glycerol content in grains resulting from plants grown in 750 mg kg-1 TiO2 NPs-treated soil. Furthermore, no translocation of TiO2 NPs from the treated soil to rice grains was detected by inductively coupled plasma-optical emission spectrometry (ICP-OES), which suggests no risk of TiO2 NPs intake via grain consumption. The observed data indicates the strong relationship among NPs application, P contents, and metabolic alterations.

Trojan-Like Internalization of Anatase Titanium Dioxide Nanoparticles by Human Osteoblast Cells.

Dentistry and orthopedics are undergoing a revolution in order to provide more reliable, comfortable and long-lasting implants to patients. Titanium (Ti) and titanium alloys have been used in dental implants and total hip arthroplasty due to their excellent biocompatibility. However, Ti-based implants in human body suffer surface degradation (corrosion and wear) resulting in the release of metallic ions and solid wear debris (mainly titanium dioxide) leading to peri-implant inflammatory reactions. Unfortunately, our current understanding of the biological interactions with titanium dioxide nanoparticles is still very limited. Taking this into consideration, this study focuses on the internalization of titanium dioxide nanoparticles on primary bone cells, exploring the events occurring at the nano-bio interface. For the first time, we report the selective binding of calcium (Ca), phosphorous (P) and proteins from cell culture medium to anatase nanoparticles that are extremely important for nanoparticle internalization and bone cells survival. In the intricate biological environment, anatase nanoparticles form bio-complexes (mixture of proteins and ions) which act as a kind of 'Trojan-horse' internalization by cells. Furthermore, anatase nanoparticles-induced modifications on cell behavior (viability and internalization) could be understand in detail. The results presented in this report can inspire new strategies for the use of titanium dioxide nanoparticles in several regeneration therapies.

Early Detection of Autism (ASD) by a Non-invasive Quick Measurement of Markedly Reduced Acetylcholine & DHEA and Increased ß-Amyloid (1-42), Asbestos (Chrysotile), Titanium Dioxide, Al, Hg & often Coexisting Virus Infections (CMV, HPV 16 and 18), Bacterial Infections etc. in the Brain and Corresponding Safe Individualized Effective Treatment.

A brief historical background on Autism & some of the important symptoms associated with Autism are summarized. Using strong Electro Magnetic Field Resonance Phenomenon between 2 identical molecules with identical weight (which received U.S. Patent) non-invasively & rapidly we can detect various molecules including neurotransmitters, bacteria, virus, fungus, metals & abnormal molecules. Simple non- invasive measurement of various molecules through pupils & head of diagnosed or suspected Autism patients indicated that in Autism patients following changes were often found: 1) Acetylcholine is markedly reduced; 2) Alzheimer's disease markers (i.e. ß-Amyloid (1-42), Tau Protein, Apolipoprotein (Apo E4)) are markedly increased; 3) Chrysotile Asbestos is increased; 4) Titanium Dioxide (TiO2) is moderately increased; 5) Al is moderately increased; 6) Hg is moderately increased; 7) Dopamine, Serotonin & GABA are significantly reduced (up to about 1/10 of normal); 8) Often viral infections (such as CMV, HHV-6, HPV-16, HPV-18, etc.), and Bacterial infections (such as Chlamydia trachomatis, Mycobacterium TB, Borrelia Burgdorferi, etc.) coexist. Research by others on Autism spectrum disorder (ASD) shows that it is a group of complex neurodevelopmental disorders, with about 70% of ASD patients also suffering from gastro-intestinal problems. While Alzheimer disease (AD) is characterized by formation of 1) Amyloid plaques, 2) Neurofibrillary tangles inside of neurons, and 3) Loss of connections between neurons. More than 90% of AD develops in people over the age of 65. These 3 characteristics often progressively worsen over time. Although Autism Spectrum Disorder and Alzheimer's disease are completely different diseases they have some similar biochemical changes. Eight examples of such measurement & analysis are shown for comparison. Most of Autism patients improved significantly by removing the source or preventing intake of Asbestos, TiO2, Al & Hg or enhancing urinary output of above abnormal substances & coexisting infections, if treatment is given early. When HPV-16 & HPV-18 coexist, at triangular central area of the top of head, in addition to inability to talk, severe neuromuscular problems of lower extremity were found to also exist. However, if treatment is given 3-4 years after onset of Autism symptoms, even when successful biochemical reduction of above abnormal substances occurs, clinical improvement is less significant, since permanent damage in brain tissue seems to already exist. Therefore, early diagnosis & early treatment is very important for both Autism & Alzheimer's disease. In addition the optimal doses of Vitamin D3 and Taurine may play an important role in the future treatment of Autism, Alzheimer's Disease and memory disturbances by significantly increasing Acetylcholine and DHEA levels, enhancing the excretion of toxic substances in the urine, as well as having an anticancer effect.

Digestibility marker and ileal amino acid digestibility in phytase-supplemented soybean or canola meals for growing pigs.

Two experiments using soybean meal (SBM) or canola meal (CM) were conducted to investigate whether the choice of digestibility marker influenced the apparent ileal digestibility (AID) or standardized ileal digestibility (SID) of N and AA in diets supplemented with phytase. In each experiment, 18 barrows fitted with T-cannulas at the ileocecal junction were assigned to 3 diets consisting of a N-free diet to determine endogenous losses of N and AA, a semipurified diet (SBM in Exp. 1 or CM in Exp. 2), and the semipurified diet supplemented with phytase at 1,000 phytase units/kg. Three digestibility markers including acid-insoluble ash (AIA), chromic oxide (Cr2O3), and titanium dioxide (TiO2) were added to each diet at 3 g/kg. Each diet was fed for 7 d, consisting of a 5-d adjustment and a 2-d collection of ileal digesta. In both studies, basal ileal endogenous losses determined with Cr2O3 as a digestibility marker were lower (P<0.01) than with those determined with AIA or TiO2 digestibility markers. Using SBM as the protein source in Exp. 1, there was no interaction between phytase and digestibility marker on AID or SID of AA. The AID of N and AA in SBM using AIA as a digestibility marker tended to be lower (P<0.1) compared with Cr2O3 or TiO2 digestibility markers. Phytase supplementation increased (P<0.001) the AID of Ca and P. The use of AIA or Cr2O3 digestibility marker tended to be associated with lower (P<0.1) SID values compared with TiO2. Phytase did not affect the SID of N or any AA in SBM except for Met, for which there was an increase (P<0.05) with phytase supplementation. Using CM as the protein source in Exp. 2, there were significant interactions between digestibility marker and phytase. Phytase supplementation had effects (P<0.01) on AID or SID when Cr2O3 or TiO2 was used as the digestibility marker. With Cr2O3 or TiO2 as the digestibility marker in the CM diets, phytase supplementation increased (P<0.05) the SID of N and all AA (except Trp). There was no SID of N or AA response to phytase supplementation of CM when AIA was used as a digestibility marker. In contrast, there were no clear improvements in AA digestibility from phytase supplementation for SBM. Phytase effects on AID or SID of AA were dependent on the digestibility marker used in diets when CM was used as the protein source but not when SBM was used as the protein source. Therefore, AA digestibility response to phytase supplementation may depend on the protein being evaluated as well as the choice of digestibility marker.

Large uptake of titania and iron oxide nanoparticles in the nucleus of lung epithelial cells as measured by Raman imaging and multivariate classification.

It is a challenging task to characterize the biodistribution of nanoparticles in cells and tissue on a subcellular level. Conventional methods to study the interaction of nanoparticles with living cells rely on labeling techniques that either selectively stain the particles or selectively tag them with tracer molecules. In this work, Raman imaging, a label-free technique that requires no extensive sample preparation, was combined with multivariate classification to quantify the spatial distribution of oxide nanoparticles inside living lung epithelial cells (A549). Cells were exposed to TiO2 (titania) and/or α-FeO(OH) (goethite) nanoparticles at various incubation times (4 or 48 h). Using multivariate classification of hyperspectral Raman data with partial least-squares discriminant analysis, we show that a surprisingly large fraction of spectra, classified as belonging to the cell nucleus, show Raman bands associated with nanoparticles. Up to 40% of spectra from the cell nucleus show Raman bands associated with nanoparticles. Complementary transmission electron microscopy data for thin cell sections qualitatively support the conclusions.

Uptake and translocation of Ti from nanoparticles in crops and wetland plants.

Bioavailability of engineered metal nanoparticles affects uptake in plants, impacts on ecosystems, and phytoremediation. We studied uptake and translocation of Ti in plants when the main source of this metal was TiO2 nanoparticles. Two crops (Phaseolus vulgaris (bean) and Triticum aestivum (wheat)), a wetland species (Rumex crispus, curly dock), and the floating aquatic plant (Elodea canadensis, Canadian waterweed), were grown in nutrient solutions with TiO2 nanoparticles (0, 6, 18 mmol Ti L(-1) for P. vulgaris, T. aestivum, and R. crispus; and 0 and 12 mmol Ti L(-1) for E. canadensis). Also examined in E. canadensis was the influence of TiO2 nanoparticles upon the uptake of Fe, Mn, and Mg, and the influence of P on Ti uptake. For the rooted plants, exposure to TiO2 nanoparticles did not affect biomass production, but significantly increased root Ti sorption and uptake. R. crispus showed translocation of Ti into the shoots. E. canadensis also showed significant uptake of Ti, P in the nutrient solution significantly decreased Ti uptake, and the uptake patterns of Mn and Mg were altered. Ti from nano-Ti was bioavailable to plants, thus showing the potential for cycling in ecosystems and for phytoremediation, particularly where water is the main carrier.