A Wirelessly Powered 4-Channel Neurostimulator for Reconstructing Walking Trajectory.

A wirelessly powered four-channel neurostimulator was developed for applying selective Functional Electrical Stimulation (FES) to four peripheral nerves to control the ankle and knee joints of a rat. The power of the neurostimulator was wirelessly supplied from a transmitter device, and the four nerves were connected to the receiver device, which controlled the ankle and knee joints in the rat. The receiver device had functions to detect the frequency of the transmitter signal from the transmitter coil. The stimulation site of the nerves was selected according to the frequency of the transmitter signal. The rat toe position was controlled by changing the angles of the ankle and knee joints. The joint angles were controlled by the stimulation current applied to each nerve independently. The stimulation currents were adjusted by the Proportional Integral Differential (PID) and feed-forward control method through a visual feedback control system, and the walking trajectory of a rat's hind leg was reconstructed. This study contributes to controlling the multiple joints of a leg and reconstructing functional motions such as walking using the robotic control technology.

Fabrication of Flexible and Transparent Conductive Nanosheets by the UV-Irradiation of Gold Nanoparticle Monolayers.

Conductive films that are highly transparent and flexible are extremely attractive for emerging optoelectronic applications. Currently, indium-doped tin oxide films are the most widely used transparent conductive films and much research effort is devoted to developing alternative transparent conductive materials to overcome their drawbacks. In this work, a novel and facile approach for fabricating transparent conductive Au nanosheets from Au nanoparticles (AuNPs) is proposed. Irradiating an AuNP monolayer at the air-water interface with UV light results in a nanosheet with ≈3.5 nm thickness and ≈80% transparency in the UV-visible region. Further, the so-fabricated nanosheets are highly flexible and can maintain their electrical conductivity even when they are bent to a radius of curvature of 0.6 mm. Fourier-transform infrared and X-ray photoelectron spectroscopy characterizations reveal that the transformation of the monolayer of AuNPs into the nanosheet is induced by the photodecomposition and/or photodetachment of the dodecanethiol ligands capping the AuNPs. Further, the UV-irradiation of a hybrid monolayer consisting of AuNPs and silica particles affords the patterning of Au nanosheets with periodic hole arrays.

Hyperferritinemia is associated with short survival time in dogs with multicentric lymphoma.

In the present study, we examined the relationship between serum ferritin concentration before treatment and survival time in dogs with multicentric lymphoma. Eighteen dogs with multicentric lymphoma were enrolled in the study. When the dogs were classified into high and low ferritin groups on the basis of their serum ferritin concentration (3,000 ng/ml cut-off value), the median survival time of dogs with high concentrations (≥3,000 ng/ml, n=7) was 40 days, whereas it was 360 days among dogs with low concentrations (<3,000 ng/ml, n=11). This difference was statistically significant (P=0.001). This finding suggests that the initial high level of serum ferritin indicates short survival time in dogs with multicentric lymphoma. Large-scale research is necessary to confirm this finding.

Erythrophagocytosis enhances heme-dependent cytotoxicity of antimalarial drugs in canine histiocytic sarcoma cell line DH82.

Antimalarial drugs, dihydroartemisinin (DHA) and artesunate (ATS), exhibit iron-dependent cytotoxicity in tumor cells. We hypothesized that erythrophagocytic uptake of heme-iron enhances the cytotoxicity of DHA and ATS. Erythrophagocytic (EP) treatment of the canine histiocytic sarcoma cell line DH82 markedly increased the cytotoxicity of DHA and ATS compared to controls. Succinyl acetone, an inhibitor of intracellular heme synthesis, decreased the cytotoxicity of DHA and ATS in normal cells, but this change was not observed in EP cells. These results suggest that exogenous heme derived from erythrocytes can enhance the cytotoxicity of DHA and ATS. Furthermore, our study suggests that heme could be a novel component of tumor treatment in veterinary medicine.

Factors influencing measurement of serum iron concentration in dogs: diurnal variation and hyperferritinemia.

We evaluated diurnal variation and hyperferritinemia as factors that influence the values of serum iron concentration in dogs, using the International Committee for Standardization in Hematology (ICSH) colorimetric method. Serum iron levels were significantly higher in the morning than in the evening in 6 clinically healthy beagle dogs, and the maximum decrease in serum iron concentration was 47.3%. Moreover, the change in serum iron concentrations in 22 clinical canine cases with various serum ferritin levels was evaluated by immunoprecipitation of ferritin. The rate of decline in the serum iron concentrations positively correlated with serum ferritin levels (r=0.48, P=0.024). These results show that it is necessary to consider the sampling time and serum ferritin level for accurate interpretation of serum iron concentrations in dogs.

Hyperferritinemia in dogs with splenic hemangiosarcoma.

Serum ferritin concentration increases in dogs in association with various diseases. In this study, we measured serum ferritin levels in dogs with splenic masses, using a sandwich ELISA assay. Eleven dogs with hemangiosarcoma (HSA), six with hematoma, 1 with hemangioma and 3 with lymphoma were enrolled. All dogs with HSA had serum ferritin concentrations above the normal limit (1,357 ng/ml, mean + 2× standard deviation of normal). Increased serum ferritin concentrations have also been observed in few cases of hematoma, hemangioma and lymphoma. Therefore, hyperferritinemia is not specific for splenic HSA, but may have clinical usefulness as a sensitive test for the disease. Further evaluation of serum ferritin concentrations in dogs with splenic HSA is needed.

Change in serum ferritin concentration in experimentally induced anemia of chronic inflammation in dogs.

In veterinary medicine, hyperferritinemia is often observed in dogs with various diseases (e.g., histiocytic sarcoma and immune-mediated hemolytic anemia) without evidence of iron overload. The mechanism underlying hyperferritinemia development is not well understood. Anemia caused by inflammation is termed as anemia of chronic disease (ACD), and experimentally induced ACD is known to cause slight hyperferritinemia. However, almost all these studies were based on short-term acute inflammation. Hepcidin, a protein mainly produced by hepatocytes, is thought to be a key regulator in iron release from reticuloendothelial cells (RECs), and its expression is related to ACD. We hypothesized that in the case of long-term ACD, iron deposition in RECs increases through hepcidin, causing a diachronic increase in serum ferritin levels. In the present study, we used a canine model with repeated subcutaneous administration of turpentine oil every 3 days over a period of 42 days (15 injections) and induced long-term inflammatory conditions; furthermore, we evaluated the change in serum ferritin concentration. Hypoproliferative anemia, bone marrow iron deposition and hypoferremia, which are characteristic of ACD, were observed on administering the turpentine injections. Hepatic iron content, hepatic hepcidin mRNA expression and serum ferritin concentration increased during the early period after turpentine injection, but returned to normal levels later. These results show that experimentally induced long-term ACD caused hypoproliferative anemia without sustained increase in hepcidin expression and did not cause systemic iron overload. Thus, chronic inflammation may not contribute greatly to increase in hyperferritinemia.

Development of a sandwich enzyme-linked immunosorbent assay to detect and measure serum levels of canine ferritin.

We established a homologous sandwich enzyme-linked immunosorbent assay (ELISA) to measure serum levels of canine ferritin. Our assay uses a rabbit anti-canine heart ferritin polyclonal antibody, and canine heart ferritin as a standard. Serum ferritin concentration in healthy dogs (n=163) was 789 ± 284 ng/ml (mean ± standard deviation), a value higher than reported previously. Confidence levels relating to repeatability, dilution and recovery for this method were high. Therefore, we believe our developed sandwich ELISA will be effective in evaluating serum levels of canine ferritin.

Molecular cloning and expression analysis of feline α1-microglobulin.

Full-length cDNA that encodes feline α1-microglobulin (Feα1m)-bikunin was obtained from a feline liver and cloned using an oligo-capping method. The Feα1m-bikunin cDNA was found to contain 1284 nucleotides, and Feα1m was found to include an open reading frame encoding a polypeptide of 201 amino acids. The deduced amino acid sequence of Feα1m showed varying amino acid identity when compared with the published sequences of the related α1-m of other species, ranging from 71.1 to 82.1%. Feα1m mRNA expression was confirmed by reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR analysis in the cerebrum, cerebellum, lung, heart, liver, spleen, pancreas, kidney, adrenal gland, and testicle. The highest Feα1m mRNA level was found in the liver.

Molecular cloning, expression in Escherichia coli, and development of monoclonal antibodies to feline cystatin C.

Cystatin C is a member of the cystatin superfamily of low-molecular-weight proteins that inhibit the activity of cysteine protease. The full-length cDNA that encodes feline cystatin C (FeCysC) has been cloned from feline white blood cells by the oligo-capping method. The cDNA consists of 796 nucleotides and includes an open reading frame encoding a polypeptide of 146 amino acids. Next, we developed several mouse anti-FeCysC monoclonal antibodies (mAbs). The recombinant FeCysC (rFeCysC) was expressed in Escherichia coli (E. coli) BL21 and used as an antigen to immunise mice. The reactivity of the mAbs to native FeCysC was examined by western blot analysis against the urinary protein from cats with chronic kidney disease (CKD). The three anti-rFeCysC mAbs (3-9G, 7-7C, and 9-12F) were able to recognise native FeCysC.

Convenient synthesis and evaluation of glycosidase inhibitory activity of alpha- and beta-galactose-type valienamines, and some N-alkyl derivatives.

Valienamine analogues having alpha- and beta-galactose-type structures were synthesized by racemic modification from (1SR,2RS,3SR)-6-methylenecyclohex-4-ene-1,2,3-triol. Four N-alkyl derivatives of the beta-anomer were readily prepared selectively by treatment of key intermediate 2,6-di-O-acetyl-3,4-O-isopropylidene-5a-carba-alpha- and beta-l-arabino-hex-5(5a)-enopyranosyl bromides with alkyl amines. All compounds were assayed for inhibitory activity against six glycosidases, and the N-dodecyl derivative was shown to be a very strong inhibitor of beta-galactosidase (IC(50) 0.01 microM, bovine liver).