Autophagy is an intracellular degradation pathway conserved in eukaryotes. Among core autophagy-related (Atg) proteins, mammalian Atg9A is the sole multi-spanning transmembrane protein, and both of its N-and C-terminal domains are exposed to the cytoplasm. It is known that Atg9A travels through the trans-Golgi network (TGN) and the endosomal system under nutrient-rich conditions, and transiently localizes to the autophagosome upon autophagy induction. However, the significance of Atg9A trafficking for autophagosome formation remains elusive. Here, we identified sorting motifs in the N-terminal cytosolic stretch of Atg9A that interact with the adaptor protein AP-2. Atg9A with mutations in the sorting motifs could not execute autophagy and was abnormally accumulated at the recycling endosomes. The combination of defects in autophagy and Atg9A accumulation in the recycling endosomes was also found upon the knockdown of TRAPPC8, a specific subunit of the TRAPPIII complex. These results show directly that the trafficking of Atg9A through the recycling endosomes is an essential step for autophagosome formation.

Several types of myeloid cell are resident in the CNS. In the steady state, microglia are present in the CNS parenchyma, whereas macrophages reside in boundary regions of the CNS, such as perivascular spaces, the meninges and choroid plexus. In addition, monocytes infiltrate into the CNS parenchyma from circulation upon blood-brain barrier breakdown after CNS injury and inflammation. Although several markers, such as CD11b and ionized calcium-binding adapter molecule 1 (Iba1), are frequently used as microglial markers, they are also expressed by other types of myeloid cell and microglia-specific markers were not defined until recently. Previous transcriptome analyses of isolated microglia identified a transmembrane lectin, sialic acid-binding immunoglobulin-like lectin H (Siglec-H), as a molecular signature for microglia; however, this was not confirmed by histological studies in the nervous system and the reliability of Siglec-H as a microglial marker remained unclear. Here, we demonstrate that Siglec-H is an authentic marker for microglia in mice by immunohistochemistry using a Siglec-H-specific antibody. Siglec-H was expressed by parenchymal microglia from developmental stages to adulthood, and the expression was maintained in activated microglia under injury or inflammatory condition. However, Siglec-H expression was absent from CNS-associated macrophages and CNS-infiltrating monocytes, except for a minor subset of cells. We also show that the Siglech gene locus is a feasible site for specific targeting of microglia in the nervous system. In conclusion, Siglec-H is a reliable marker for microglia that will allow histological identification of microglia and microglia-specific gene manipulation in the nervous system.

A CoFe binary alloy film with 24 kG of saturation magnetic flux density, B s , was prepared by electrodeposition. The B s value of the CoFe alloy film prepared by electrodeposition from the conventional bath is usually equal to 21 kG. With the addition of trimethylamineborane, a well-known reducing agent, the B s value was found to increase to 23 kG. Furthermore, with the use of a separated compartment dual cell system, a B s value as high as 24 kG was reached. For obtaining the highest B s , it was found to be essential to avoid the oxidation of ferrous ion to ferric ion in the plating bath. The coercivity, H c , values were 15, 14, and 15 Oe for the films with 20, 23, and 24 kG of B s , respectively. The key for obtaining high B s materials is to avoid the oxidation of ferrous ions in the bath. In addition, it was found that the H c value of 15 Oe for CoFe film with the highest B s value of 24 kG could be lowered to 8 Oe by annealing in an applied magnetic field of 500 Oe.To meet the strong demand for high performance write-heads to be used for high density magnetic recording, 1 soft magnetic materials with high saturation magnetic density, B s , are being developed by many researchers. 2-6 We previously developed a CoNiFe alloy with B s ϭ 20-21 kG using electrodeposition 7,8 and it has been in practical use. The CoFe binary alloy is known as a material with B s of about 24 kG, which is close to the limiting value achievable with ferromagnetic alloys. 9 Several soft magnetic thin films with the highest B s value of 24 kG have recently been prepared by using a dry sputtering process, e.g., NiFe/CoFe-N/NiFe trilayer film, 10-12 CoFe-O granular film, 13 and CoFe-Al-O granular film. 14,15 However, thin films containing only Co and Fe do not exhibit soft magnetic properties. 16 On the other hand, the technique of electrodeposition is useful as an industrial process, and write-head cores are now being fabricated by electrodeposition. Thus, electrodeposited soft magnetic thin films of CoFe binary alloy with the highest B s value of 24 kG are now in strong demand.We have succeeded in preparing soft magnetic materials of CoFe binary alloy film with B s ϭ 24 kG by electrodeposition. 17 The key requirement for obtaining the high B s CoFe material in this binary alloy system is the prevention of oxidation of ferrous ion, Fe 2ϩ , to ferric ion, Fe 3ϩ , in the plating bath. In this paper, we discuss this key issue in fabricating the soft magnetic material with B s ϭ 24 kG. ExperimentalBinary CoFe alloy films were electrodeposited using a rotating disk electrode ͑RDE͒ system, in which the anode was made for Pt and the electrodeposited substrate was removable to determine properties of the electrodeposited film. The thickness of the electrodeposited alloy films was about 0.7 m. The substrate was a circular glass plate with a diam of 10 mm, coated with sputtered NiCr ͑5 nm͒ and NiFe ͑100 nm͒ films, and a Cu foil with a diam of 10 mm and a thickness of 8 m. The plating bath composition and operating conditions are shown in Table...

BACKGROUND:Several companies offer direct-toconsumer (DTC) genetic testing to evaluate ancestry and wellness. Massive-scale testing of thousands of single-nucleotide polymorphisms (SNPs) is not error free, and such errors could translate into misclassification of risk and produce a false sense of security or unnecessary anxiety in an individual. We evaluated 3 DTC services and a genomics service that are based on DNA microarray or solution genotyping with hydrolysis probes (TaqMan analysis) and compared the test results obtained for the same individual.

In the present work we numerically simulated the electrogenerated chemiluminescence (ECL) from a Ru­(bpy)3 2+-doped silica nanoparticle (Ru-DSNP) in buffer containing tripropylamine (TPrA). An experimental study reported from Zanarini et al. showed that ECL intensity for the Ru-DSNP/TPrA system exhibits two emission waves, while the potential of the working electrode is swept in the positive direction. The first ECL wave with a peak at ∼0.9 V (vs Ag|AgCl) is triggered by TPrA oxidation and is governed by the deprotonation equilibrium of TPrA cation radical (TPrA•+ = TPrA• + H+). We present a model for the description of the first ECL wave, which also takes into consideration the influence on the deprotonation equilibrium of the electrode surface functionality. This model indicated that the detachment of a Ru-DSNP (initially bound to the electrode surface via alkylthiols linkers) from the electrode surface and the subsequent electrode surface oxidation facilitate the radical deprotonation on the electrode surface causing the ECL quenching. The second ECL wave having its peak at ∼1.2 V is triggered by direct Ru­(bpy)3 2+ oxidation. We modeled the second ECL wave as related to the electron hopping mechanism between Ru­(bpy)3 2+ labels inside the Ru-DSNP. The results of the numerical simulations indicate that electrode surface functionality modification, which occurs during potential sweep, and the electron hopping mechanism between Ru­(bpy)3 2+ labels play important roles in defining the Ru-DSNP/TPrA ECL signal.