Autophagy in Human Embryonic Stem Cells

Autophagy (macroautophagy) is a degradative process that involves the sequestration of cytosolic material including organelles into double membrane vesicles termed autophagosomes for delivery to the lysosome. Autophagy is essential for preimplantation development of mouse embryos and cavitation of embryoid bodies. The precise roles of autophagy during early human embryonic development, remain however largely uncharacterized. Since human embryonic stem cells constitute a unique model system to study early human embryogenesis we investigated the occurrence of autophagy in human embryonic stem cells. We have, using lentiviral transduction, established multiple human embryonic stem cell lines that stably express GFP-LC3, a fluorescent marker for the autophagosome. Each cell line displays both a normal karyotype and pluripotency as indicated by the presence of cell types representative of the three germlayers in derived teratomas. GFP expression and labelling of autophagosomes is retained after differentiation. Baseline levels of autophagy detected in cultured undifferentiated hESC were increased or decreased in the presence of rapamycin and wortmannin, respectively. Interestingly, autophagy was upregulated in hESCs induced to undergo differentiation by treatment with type I TGF-beta receptor inhibitor SB431542 or removal of MEF secreted maintenance factors. In conclusion we have established hESCs capable of reporting macroautophagy and identify a novel link between autophagy and early differentiation events in hESC

Manufacture of Micromirror Arrays Using a CMOS-MEMS Technique

In this study we used the commercial 0.35 μm CMOS (complementary metal oxide semiconductor) process and simple maskless post-processing to fabricate an array of micromirrors exhibiting high natural frequency. The micromirrors were manufactured from aluminum; the sacrificial layer was silicon dioxide. Because we fabricated the micromirror arrays using the standard CMOS process, they have the potential to be integrated with circuitry on a chip. For post-processing we used an etchant to remove the sacrificial layer and thereby suspend the micromirrors. The micromirror array contained a circular membrane and four fixed beams set symmetrically around and below the circular mirror; these four fan-shaped electrodes controlled the tilting of the micromirror. A MEMS (microelectromechanical system) motion analysis system and a confocal 3D-surface topography were used to characterize the properties and configuration of the micromirror array. Each micromirror could be rotated in four independent directions. Experimentally, we found that the micromirror had a tilting angle of about 2.55° when applying a driving voltage of 40 V. The natural frequency of the micromirrors was 59.1 kHz

CD4 T-Cell-Independent Antibody Response Reduces Enterovirus 71 Lethality in Mice by Decreasing Tissue Viral Loads

Enterovirus 71 (EV71) has induced fatal encephalitis in hundreds of thousands of infants and young children in the Asia-Pacific region since the past decade. Lymphocyte and antibody responses have been suspected to aggravate EV71-induced neurological symptoms, so anti-inflammatory agents have been used to treat patients with neurological symptoms. In the present study, we found that mice deficient in CD4+ T cells were resistant to EV71 infection as wild-type mice, whereas mice deficient in B cells were highly susceptible to viral infection. Compensation of CD4 T-cell function by other immune cells was not likely, because wild-type mice depleted of CD4+ T cells were also resistant to viral infection. Infected CD4 T-cell-deficient mice produced virus-specific neutralizing antibodies, IgM and IgG. Moreover, adoptive transfer of the virus-specific antibody produced by infected CD4 T-cell-deficient mice protected B-cell-deficient mice from infection by reducing tissue viral loads. Collectively, our results show that the CD4 T-cell-independent antibody response promotes the survival of EV71-infected mice and suggest great potential for the use of vaccines and neutralizing antibodies to reduce fatal symptoms in patients

Reduced expression of alpha-1,2-mannosidase I extends lifespan in Drosophila melanogaster and Caenorhabditis elegans

Exposure to sub-lethal levels of stress, or hormesis, was a means to induce longevity. By screening for mutations that enhance resistance to multiple stresses, we identified multiple alleles of alpha-1,2-mannosidase I (mas1) which, in addition to promoting stress resistance, also extended longevity. Longevity enhancement is also observed when mas1 expression is reduced via RNA interference in both Drosophila melanogaster and Caenorhabditis elegans. The screen also identified Edem1 (Edm1), a gene downstream of mas1, as a modulator of lifespan. As double mutants for both mas1 and Edm1 showed no additional longevity enhancement, it appeared that both mutations function within a common pathway to extend lifespan. Molecular analysis of these mutants revealed that the expression of BiP, a putative biomarker of dietary restriction (DR), is down-regulated in response to reductions in mas1 expression. These findings suggested that mutations in mas1 may extend longevity by modulating DR

Clonal dissemination of invasive and colonizing clonal complex 1 of serotype VI group B Streptococcus in central Taiwan

Background/PurposeThe aim of this study was to investigate clinical presentation, serotype distribution and genetic correlation of group B streptococcus (GBS) diseases. Since serotype VI prevalence far exceeded that reported in prior studies, genetic relationship of isolates was further analyzed.MethodsGBS isolates obtaining from patients with invasive diseases and pregnant women with colonization between June 2007 and December 2010 were analyzed. All isolates were tested for serotypes by multiplex PCR assay and pulsed-field gel electrophoresis (PFGE). Serotype VI isolates were further analyzed by multilocus sequence typing (MLST).ResultsA total of 134 GBS isolates were recovered from blood of 126 patients with invasive disease (94.0%) and anogenital swabs of 8 pregnant women (6.0%). Most common serotype was Ib (21.6%), followed by V (20.1%), VI (18.7%), III (15.7%), II (11.9 %), Ia (11.2%), and IX (0.7%). Serotype VI was also the leading type in infants with early onset disease (EOD; 3/8, 37.5%) and colonizing pregnant women (3/8, 37.5%). PFGE distinguished 33 pulsotypes, reflecting genetic diversity among GBS isolates. Among 25 serotype VI isolates tested, 14 were ST-1, seven were ST-679, three were ST-678, one was ST-681, and distributed into four PFGE pulsotypes. ST-678, ST-679, and ST-681 were novel sequence types; ST-678 and ST-679 are single-locus variants of ST-1 that belongs to clonal complex (CC) 1.ConclusionCC1 dissemination of serotype VI GBS thus emerges as an important invasive pathogen in infants and nonpregnant adults in central Taiwan. Serotype prevalence of GBS must be continuously monitored geographically to guide prevention strategy of GBS vaccines

VoiceBank-2023: A Multi-Speaker Mandarin Speech Corpus for Constructing Personalized TTS Systems for the Speech Impaired

Services of personalized TTS systems for the Mandarin-speaking speech impaired are rarely mentioned. Taiwan started the VoiceBanking project in 2020, aiming to build a complete set of services to deliver personalized Mandarin TTS systems to amyotrophic lateral sclerosis patients. This paper reports the corpus design, corpus recording, data purging and correction for the corpus, and evaluations of the developed personalized TTS systems, for the VoiceBanking project. The developed corpus is named after the VoiceBank-2023 speech corpus because of its release year. The corpus contains 29.78 hours of utterances with prompts of short paragraphs and common phrases spoken by 111 native Mandarin speakers. The corpus is labeled with information about gender, degree of speech impairment, types of users, transcription, SNRs, and speaking rates. The VoiceBank-2023 is available by request for non-commercial use and welcomes all parties to join the VoiceBanking project to improve the services for the speech impaired.Comment: submitted to 26th International Conference of the ORIENTAL-COCOSD

Levitation by a dipole electric field

The phenomenon of floating can be fascinating in any field, with its presence seen in art, films, and scientific research. This phenomenon is a captivating and pertinent subject with practical applications, such as Penning traps for antimatter confinement and Ion traps as essential architectures for quantum computing models. In our project, we reproduced the 1893 water bridge experiment using glycerol and first observed that lump-like macroscopic dipole moments can undergo near-periodic oscillations that exhibit floating effects and do not need classical bridge form. By combining experimental analysis, neural networks, investigation of Kelvin force generated by the Finite element method, and exploration of discharging, we gain insights into the mechanisms of motion. Our discovery has overturned the previous impression of a bridge floating in the water, leading to a deeper understanding of the new trap mechanism under strong electric fields with a single pair of electrodes.Comment: 5 pages, 5 figure