The motion of single- and two-cavitation bubbles generated by laser beams directly beneath a free surface is studied experimentally, using high-speed photography, and theoretically using the highly accurate boundary integral method. Favorable comparisons of bubble shape history and centroid motion are observed while the numerical calculations provide information on the pressure field surrounding the bubbles. A range of responses, including the null impulse state, is obtained for the two bubbles depending on the bubble size ratio and the interbubble and bubble-free surface distances, although in all cases reported in this article, the bubble nearest the free surface yields a high-speed liquid jet directed away from the free surface. It is also found that when the free-surface–bubble interaction is strong, a fast free-surface spike is formed for both the single- and two-bubble cases.

Reactive oxygen species (ROS) act as intracellular signaling molecules in the regulation of receptor activator of nuclear factor-κB ligand (RANKL)-dependent osteoclast differentiation, but they also have cytotoxic effects that include peroxidation of lipids and oxidative damage to proteins and DNA. Cellular protective mechanisms against oxidative stress include transcriptional control of cytoprotective enzymes by the transcription factor, nuclear factor E2-related factor 2 (Nrf2). This study investigated the relationship between Nrf2 and osteoclastogenesis. Stimulation of osteoclast precursors (mouse primary peritoneal macrophages and RAW 264.7 cells) with RANKL resulted in the up-regulation of kelch-like ECH-associated protein 1 (Keap1), a negative regulator of Nrf2. It also decreased the Nrf2/Keap1 ratio, and it down-regulated cytoprotective enzymes (heme oxygenase-1, γ-glutamylcysteine synthetase, and glucose-6-phosphate dehydrogenase). Nrf2 overexpression up-regulated the expression of cytoprotective enzymes, decreased ROS levels, decreased the number of tartrate-resistant acid phosphatase-positive multinucleated cells, reduced marker genes for osteoclast differentiation, and attenuated bone destruction in both in vitro and in vivo models. Overexpression of Keap1 or RNAi knockdown of Nrf2 exerted the opposite actions. In addition, in vivo local Nrf2 overexpression attenuated lipopolysaccharide-mediated RANKL-dependent cranial bone destruction in vivo. This is the first study to show that the Keap1/Nrf2 axis regulates RANKL-dependent osteoclastogenesis through modulation of intracellular ROS signaling via expression of cytoprotective enzymes. This raises the exciting possibility that the Keap1-Nrf2 axis may be a therapeutic target for the treatment of bone destructive disease.

Appropriate ischemia models are required for successful studies of therapeutic angiogenesis. While collateral routes are known to be present within the innate vasculature, there are no reports describing the detailed vascular anatomy of the murine hindlimb. In addition, differences in the descriptions of anatomical names and locations in the literature impede understanding of the circulation and the design of hindlimb ischemia models. To understand better the collateral circulation in the whole hindlimb, clarification of all the feeding arteries of the hindlimb is required. The aim of this study is to reveal the detailed arterial anatomy and collateral routes in murine hindlimb to enable the appropriate design of therapeutic angiogenesis studies and to facilitate understanding of the circulation in ischemia models. Arterial anatomy in the murine hindlimb was investigated by contrast-enhanced X-ray imaging and surgical dissection. The observed anatomy is shown in photographic images and in a schema. Previously unnoticed but relatively large arteries were observed in deep, cranial and lateral parts of the thigh. The data indicates that there are three collateral routes through the medial thigh, quadriceps femoris, and the biceps femoris muscles. Furthermore, anatomical variations were found at the origins of the three feeding arteries. The detailed arterial anatomy of murine hindlimb and collateral routes deduced from the anatomy are described. Limitations on designs of ischemia models in view of anatomical variations are proposed. These observations will contribute to the development of animal studies of therapeutic angiogenesis using murine hindlimb ischemia models.