Engraftment of allogeneic iPS cell-derived cartilage organoid in a primate model of articular cartilage defect.

Induced pluripotent stem cells (iPSCs) are a promising resource for allogeneic cartilage transplantation to treat articular cartilage defects that do not heal spontaneously and often progress to debilitating conditions, such as osteoarthritis. However, to the best of our knowledge, allogeneic cartilage transplantation into primate models has never been assessed. Here, we show that allogeneic iPSC-derived cartilage organoids survive and integrate as well as are remodeled as articular cartilage in a primate model of chondral defects in the knee joints.

Quality assessment tests for tumorigenicity of human iPS cell-derived cartilage.

Articular cartilage damage does not heal spontaneously and causes joint dysfunction. The implantation of induced pluripotent stem cell (iPSC)-derived cartilage (iPS-Cart) is one candidate treatment to regenerate the damaged cartilage. However, concerns of tumorigenicity are associated with iPS-Cart, because the iPSC reprogramming process and long culture time for cartilage induction could increase the chance of malignancy.

Changes in acetyl-CoA mediate Sik3-induced maturation of chondrocytes in endochondral bone formation.

The maturation of chondrocytes is strictly regulated for proper endochondral bone formation. Although recent studies have revealed that intracellular metabolic processes regulate the proliferation and differentiation of cells, little is known about how changes in metabolite levels regulate chondrocyte maturation. To identify the metabolites which regulate chondrocyte maturation, we performed a metabolome analysis on chondrocytes of Sik3 knockout mice, in which chondrocyte maturation is delayed.

Focal calcium monitoring with targeted nanosensors at the cytosolic side of endoplasmic reticulum.

Ca distribution is spatially and temporally non-uniform inside cells due to cellular compartmentalization. However, Ca sensing with small organic dyes, such as fura-2 and fluo-4, has been practically applied at a single cell level where the averaged signal from freely diffusing dye molecules is acquired. In this study, we aimed to target azide-functionalized fura-2 (N-fura-2) to a specific site of subcellular compartments to realize focal Ca sensing.

A ratiometric fluorescent molecular probe for visualization of mitochondrial temperature in living cells.

Mitochondrial thermodynamics is the key to understand cellular activities related to homeostasis and energy balance. Here, we report the first ratiometric fluorescent molecular probe (Mito-RTP) that is selectively localized in the mitochondria and visualize the temperature. We confirmed that Mito-RTP could work as a ratiometric thermometer in a cuvette and living cells.

A nanoparticle-based ratiometric and self-calibrated fluorescent thermometer for single living cells.

The homeostasis of body temperature and energy balance is one of the major principles in biology. Nanoscale thermometry of aqueous solutions is a challenging but crucial technique to understand the molecular basis of this essential process. Here, we developed a ratiometric nanothermometer (RNT) for intracellular temperature measurement in real time.

Intracellular click reaction with a fluorescent chemical Ca2+ indicator to prolong its cytosolic retention.

The powerful strategy of "intracellular click reaction" was used to retain a chemical Ca(2+) indicator in the cytosol. Specifically, a novel clickable Ca(2+) indicator "N3-fura-2 AM" was coupled with dibenzylcyclooctyl-modified biomacromolecules via copper-free click reaction in living cells and Ca(2+) oscillation was observed for an extended period of time.

Walking nanothermometers: spatiotemporal temperature measurement of transported acidic organelles in single living cells.

We fabricated fluorescent nanoparticles which monitor temperature changes without sensitivity to pH (4-10) and ionic strength (0-500 mM). The nanothermometers spontaneously enter living HeLa cells via endocytosis, enclosed in acidic organelles, i.e.

The roche vault performed by elite gymnasts: somersaulting technique, deterministic model, and judges' scores.

The purpose of the study was to determine the mechanical variables that are related to successful post-flight somersaulting performance of the Roche vault. The 23 Roche vaults performed during the 2000 Olympic Games were filmed by a 16-mm camera operating at 100 Hz. The 2-D direct linear transformation technique was used for spatial calibration.

Somersaulting techniques used in high-scoring and low-scoring Roche vaults performed by male Olympic gymnasts.

The aim of this study was to compare the somersaulting techniques used in the 16 highest-scoring and 16 lowest-scoring Roche vaults. Our hypothesis was that the gymnasts performing the highest-scoring Roche vaults would demonstrate a better technique than those performing the lowest-scoring Roche vaults while on the horse (pushing off the horse more effectively), somersaulting (executing most of the required somersaults higher in flight), and landing (showing a greater control). A 16-mm motion picture camera, operating at 100 Hz, recorded the vaults during the official competition.

Phase-dependent chronotropic response of the heart during running in humans.

Heartbeat modulation by muscle contraction during rhythmic exercise involving a small muscle mass is phase-dependent, reflecting the timing of the muscle contraction within the cardiac cycle, but it remains unclear whether such modulation occurs during whole body exercise. To determine whether phase-dependent chronotropic changes in the heart would occur during running, we investigated the relationship between R-R interval (RRI) and the timing of vastus lateralis muscle contractions within the cardiac cycle. Seven healthy subjects were examined during high intensity running where the target heart rate was 160 beats .

PCR amplification from single DNA molecules on magnetic beads in emulsion: application for high-throughput screening of transcription factor targets.

We have developed a novel method of genetic library construction on magnetic microbeads based on solid-phase single-molecule PCR in a fine and robust water-phase compartment formed in water-in-oil (w/o) emulsions. In this method, critically diluted DNA fragments were distributed over the emulsion as templates, where beads crosslinked with multiple primers and other PCR components were encapsulated to form multiple reaction compartments. The delivered DNA was then amplified and covalently immobilized on the beads in parallel, within individual compartments, to construct a genetic library on beads (GLOBE), which was readily applicable to a genomewide global scanning of genetic elements recognized by a defined DNA-binding protein.

Techniques used in high-scoring and low-scoring 'Roche' vaults performed by elite male gymnasts.

The 16 highest-scored Roche vaults (G1) performed during the 2000 Olympic Games were compared with those receiving the 16 lowest-scores (G2). A 16-mm motion picture camera operating at 100 Hz recorded the vaults during the competition. The results of t tests (p < .

Comparison of cardio-locomotor synchronization during running and cycling.

By comparing the characteristics of cardiac-locomotor synchronization (CLS) in running and cycling individuals, we tested whether the characteristics of CLS occurring during rhythmic exercise adhere to the central origin hypothesis, which postulates a direct interaction between cardiovascular centers in the brain and the pattern generator in the spinal cord. Ten healthy subjects performed both exercises at the same intensity (150 beats.min(-1)) and cadence (150 steps.