The Age-Sensitive Efficacy of Calorie Restriction on Mitochondrial Biogenesis and mtDNA Damage in Rat Liver

Calorie restriction (CR) is the most efficacious treatment to delay the onset of age-related changes such as mitochondrial dysfunction. However, the sensitivity of mitochondrial markers to CR and the age-related boundaries of CR efficacy are not fully elucidated. We used liver samples from ad libitum-fed (AL) rats divided in: 18-month-old (AL-18), 28-month-old (AL-28), and 32-month-old (AL-32) groups, and from CR-treated (CR) 28-month-old (CR-28) and 32-month-old (CR-32) counterparts to assay the effect of CR on several mitochondrial markers. The age-related decreases in citrate synthase activity, in TFAM, MFN2, and DRP1 protein amounts and in the mtDNA content in the AL-28 group were prevented in CR-28 counterparts. Accordingly, CR reduced oxidative mtDNA damage assessed through the incidence of oxidized purines at specific mtDNA regions in CR-28 animals. These findings support the anti-aging effect of CR up to 28 months. Conversely, the protein amounts of LonP1, Cyt c, OGG1, and APE1 and the 4.8 Kb mtDNA deletion content were not affected in CR-28 rats. The absence of significant differences between the AL-32 values and the CR-32 counterparts suggests an age-related boundary of CR efficacy at this age. However, this only partially curtails the CR benefits in counteracting the generalized aging decline and the related mitochondrial involvement.     

Glutathione depletion in rested and exercised mice: biochemical consequence and adaptation

The effect of chronic in vivo glutathione (GSH) depletion by L-buthionine-[S,R]-sulfoximine (BSO) on intracellular and interorgan GSH regulation was investigated in mice both at rest and after an acute bout of exhaustive swim exercise. BSO treatment for 12 days decreased concentrations of GSH in the liver, kidney, quadriceps muscle, and plasma to 28, 15, 7, and 35%, respectively, compared to GSH-adequate mice. In most tissues, with the exception of the kidney, this decrease was associated with a concomitant decrease of glutathione disulfide (GSSG) such that the GSH/GSSG ratio was maintained. GSH depletion caused adaptive changes in several enzymes related to GSH regulation, such as liver glutathione peroxidase (-25%), kidney gamma-glutamyltranspeptidase (+20%), glutathione disulfide reductase (+131%) and glutathione sulfur-transferase (+53%). There was an apparent down-regulation of muscle gamma-glutamyltranspeptidase (-56%) in the GSH-depleted mice, which contributed to a conservation of plasma GSH. Exhaustive exercise in the GSH-adequate state severely depleted GSH content in the liver (-55%) and kidney (-35%), whereas plasma and muscle GSH levels remained constant. However, exercise in the GSH-depleted state exacerbated GSH deficit in the liver (-57%), kidney (-33%), plasma (-65%), and muscle (-25%) in the absence of adequate reserves of liver GSH. Hepatic lipid peroxidation increased by 220 and 290%, respectively, after exhaustive exercise in the GSH-adequate and -depleted mice. We conclude that GSH homeostasis is essential for the prooxidant-antioxidant balance during prolonged physical exercise.     

Glutathone and glutathione ethyl ester supplementation of mice alter glutathione homeostasis during exercise

The present study examined the effect of glutathione (GSH) and glutathione ethyl ester (GSH-E) supplementation on GSH homeostasis and exercise-induced oxidative stress. Male Swiss-Webster mice were randomly divided into 4 groups: starved for 24 h and injected with GSH or GSH-E (6 mmol/kg body wt, i.p.) 1 h before exercise, starved for 24 h and injected with saline (S); and having free access to food and injected with saline (C). Half of each group of mice was killed either after an acute bout of exhaustive swimming (E) or after rest (R). Plasma GSH concentration was 100-160% (P < 0.05) higher in GSH mice vs. C or S mice at rest, whereas GSH-E injection had no effect. Plasma GSH was not affected by exercise in C or S mice, but was 44 and 34% lower (P < 0.05) in E vs. R mice with GSH or GSH-E injection, respectively. S, GSH- and GSH-E-treated mice had significantly lower liver GSH concentration and the GSH:glutathione disulfide (GSSG) ratio than C mice. Hepatic and renal GSH and the GSH:GSSG ratio were significantly lower in E vs. R mice in all groups. GSH-E-treated mice had a significantly smaller exercise-induced decrease in GSH vs. C, S, and GSH-treated mice and no difference in the GSH:GSSG ratio in the kidney. Activities of gamma-glutamylcysteine synthetase and gamma-glutamyltranspeptidase in the liver and kidney were not affected by either GSH treatment or exercise. GSH concentration and the GSH:GSSG ratio in quadriceps muscle were not different among C, S and GSH-treated mice, but significantly lower in GSH-E-treated mice (P < 0.05). Hepatic malondialdehyde (MDA) content was greater in exercised mice in all but GSH-E-treated groups. GSH and GSH-E increased MDA levels in the kidney of E vs. R mice, but attenuated exercise-induced lipid peroxidation in muscle. Swim endurance time was approximately 2 h longer in GSH (351 +/- 22 min) and GSH-E (348 +/- 27) than S mice (237 +/- 17). We conclude that 1) acute GSH and GSH-E supplementation at the given doses does not increase tissue GSH content or redox status; 2) both GSH and GSH-E improve endurance performance and prevent muscle lipid peroxidation during prolonged exercise; and 3) while both compounds may impose a metabolic and oxidative stress to the kidney, this side effect is smaller with GSH-E supplementation.     

A low-voltage folded-switching mixer in 0.18-/spl mu/m CMOS

Scaling of CMOS technologies has a great impact on analog design. The most severe consequence is the reduction of the voltage supply. In this paper, a low voltage, low power, AC-coupled folded-switching mixer with current-reuse is presented. The main advantages of the introduced mixer topology are: high voltage gain, moderate noise figure, moderate linearity, and operation at low supply voltages. Insight into the mixer operation is given by analyzing voltage gain, noise figure (NF), linearity (IIP3), and DC stability. The mixer is designed and implemented in 0.18-/spl mu/m CMOS technology with metal-insulator-metal (MIM) capacitors as an option. The active chip area is 160 /spl mu/m/spl times/200 /spl mu/m. At 2.4 GHz a single side band (SSB) noise figure of 13.9 dB, a voltage gain of 11.9 dB and an IIP3 of -3 dBm are measured at a supply voltage of 1 V and with a power consumption of only 3.2 mW. At a supply voltage of 1.8 V, an SSB noise figure of 12.9 dB, a voltage gain of 16 dB and an IIP3 of 1 dBm are measured at a power consumption of 8.1 mW.     

Electrostatic Spray Deposition of Biomimetic Nanocrystalline Apatite Coatings onto Titanium

Titanium (Ti) and its alloys are widely used to manufacture orthopedic and dental implants due to their excellent mechanical properties and corrosion resistance. Although these materials are bioinert, improvement of biological properties (e.g., bone implant contact) can be obtained by the application of a coating made of nanostructured apatite. The aim of this study was to investigate the applicability of the electrostatic spray deposition (ESD) technique for the deposition of nanostructured apatite coatings onto commercially pure (cp) Ti substrates at room temperature. To that end, poorly crystalline, nano‐sized, carbonate‐apatite plate‐like particles with dimensions similar to the nanocrystals present in bone were synthesized using wet‐chemical precipitation techniques and their physicochemical properties were subsequently characterized thoroughly. The apatite suspensions were optimized for the ESD process in terms of dispersion, aggregation, and stability. Furthermore, relevant ESD processing parameters, including nozzle‐to‐substrate distance, relative humidity in the deposition chamber and deposition time were varied in order to study their effects on coating morphology. Porous films made of agglomerates of nano‐sized apatite particles of ≈50 nm were generated, demonstrating the feasibility of the ESD technique for the deposition of thin apatite coatings with a nano‐sized surface morphology onto titanium substrates. The ability of these nanocrystals to bind therapeutic agents for bone diseases and the capability of ESD to produce coating at physiological conditions makes this work a first step toward the set‐up of coatings for bone implants based on surface‐activated apatite with improved functionality.     

HNF/HTPB propellants: Influence of HNF particle size on ballistic properties

The burning rate characteristics of solid composite propellants can be modified via different methods. One of these is the application of different oxidizer particle sizes in the propellant. The effect of the use of fine and coarse particles of the oxidizer hydrazinium nitroformate (HNF) in hydroxyl-terminated polybutadiene (HTPB)-based propellants is reported in this paper. The fine HNF particles were obtained by means of a grinding process. The fine and coarse HNF grades were characterized regarding their hazardous and thermal stability properties, as well as the influence of a cured and uncured HTPB binder on these characteristics. The ballistic properties of three (two monomodal and one bimodal) HNF/HTPB-based propellants show that the use of fine HNF (5-10 μm mean size) leads to a lowering of the pressure exponent of the propellant, implying that the propellant burning rate can be tuned by varying the mean size of the HNF grades present in the propellant.     

Electrosprayed Enzyme Coatings as Bioinspired Alternatives to Bioceramic Coatings for Orthopedic and Oral Implants

The biological performance of orthopedic and oral implants can be significantly improved by functionalizing the non‐physiological metallic implant surface through the application of biologically active coatings. In this paper, a cost‐effective alternative to traditional biomedical coatings for bone substitution through exploitation of the specific advantages of the electrospray deposition technique for the immobilization of the enzyme alkaline phosphatase (ALP) onto the implant surface is presented. Since ALP increases the local inorganic phosphate concentration required for physiological mineralization of hard tissues, ALP coatings will enable enzyme‐mediated mineralization onto titanium surfaces. To evaluate the bone‐bioactive capacity of the ALP‐coated titanium surface, soaking experiments are performed. Although the purely inorganic so‐called simulated body fluid is the standard in vitro procedure for predictive studies on potential bone bonding in vivo, an alternative testing solution is proposed that also contains organic phosphates (cell culture medium supplemented with the organic β‐b;‐glycerophosphate (β‐b;‐GP) and serum proteins), thereby resembling the in vivo conditions more closely. Under these physiological conditions, the electrosprayed ALP coatings accelerated mineralization onto the titanium surface as compared to noncoated implant material by means of enzymatic pathways. Therefore, this novel approach toward implant fixation holds significant promise.     

The behavior of osteoblast-like cells on various substrates with functional blocking of integrin-β1 and integrin-β3

This study was designed to examine the influence of integrin subunit-β1 and subunit-β3 on the behavior of primary osteoblast-like cells, cultured on calcium phosphate (CaP)-coated and non coated titanium (Ti). Osteoblast-like cells were incubated with specific monoclonal antibodies against integrin-β1 and integrin-β3 to block the integrin function. Subsequently, cells were seeded on Ti discs, either non coated or provided with a 2 μm carbonated hydroxyapatite coating using Electrostatic Spray Deposition. Results showed that on CaP coatings, cellular attachment was decreased after a pre-treatment with either anti-integrin-β1 or anti-integrin-β3 antibodies. On Ti, cell adhesion was only slightly affected after a pre-treatment with anti-integrin-β3 antibodies. Scanning electron microscopy showed that on both types of substrate, cellular morphology was not changed after a pre-treatment with either antibody. With quantitative PCR, it was shown for both substrates that mRNA expression of integrin-β1 was increased after a pre-treatment with either anti-integrin-β1 or anti-integrin-β3 antibodies. Furthermore, after a pre-treatment with either antibody, mRNA expression of integrin-β3 and ALP was decreased, on both types of substrate. In conclusion, osteoblast-like cells have the ability to compensate to great extent for the blocking strategy as applied here. Still, integrin-β1 and β3 seem to play different roles in attachment, proliferation, and differentiation of osteoblast-like cells, and responses on CaP-coated substrates differ to non coated Ti. Furthermore, the influence on ALP expression suggests involvement of both integrin subunits in signal transduction for cellular differentiation.