SUMMARY Multiple angiogenesis inhibitors have been therapeutically validated in preclinical cancer models, and several in clinical trials. Here we report that angiogenesis inhibitors targeting the VEGF pathway demonstrate antitumor effects in mouse models of pancreatic neuroendocrine carcinoma and glioblastoma but concomitantly elicit tumor adaptation and progression to stages of greater malignancy, with heightened invasiveness and in some cases increased lymphatic and distant metastasis. Increased invasiveness is also seen by genetic ablation of the Vegf-A gene in both models, substantiating the results of the pharmacological inhibitors. The realization that potent angiogenesis inhibition can alter the natural history of tumors by increasing invasion and metastasis warrants clinical investigation, as the prospect has important implications for the development of enduring antiangiogenic therapies.

Zinc (Zn) is an essential trace mineral that regulates the expression and activation of biological molecules such as transcription factors, enzymes, adapters, channels, and growth factors, along with their receptors. Zn deficiency or excessive Zn absorption disrupts Zn homeostasis and affects growth, morphogenesis, and immune response, as well as neurosensory and endocrine functions. Zn levels must be adjusted properly to maintain the cellular processes and biological responses necessary for life. Zn transporters regulate Zn levels by controlling Zn influx and efflux between extracellular and intracellular compartments, thus, modulating the Zn concentration and distribution. Although the physiological functions of the Zn transporters remain to be clarified, there is growing evidence that Zn transporters are related to human diseases, and that Zn transporter-mediated Zn ion acts as a signaling factor, called "Zinc signal". Here we describe critical roles of Zn transporters in the body and their contribution at the molecular, biochemical, and genetic levels, and review recently reported disease-related mutations in the Zn transporter genes.

Cellular oxygen consumption is a determinant of intracellular oxygen levels. Because of the high demand of mitochondrial respiration during insulin secretion, pancreatic ␤-cells consume large amounts of oxygen in a short time period. We examined the effect of insulin secretion on cellular oxygen tension in vitro. We confirmed that Western blotting of pimonidazole adduct was more sensitive than immunostaining for detection of cellular hypoxia in vitro and in vivo. The islets of the diabetic mice but not those of normal mice were hypoxic, especially when a high dose of glucose was loaded. In MIN6 cells, a pancreatic ␤-cell line, pimonidazole adduct formation and stabilization of hypoxia-inducible factor-1␣ (HIF-1␣) were detected under mildly hypoxic conditions. Inhibition of respiration rescued the cells from becoming hypoxic. Glucose stimulation decreased cellular oxygen levels in parallel with increased insulin secretion and mitochondrial respiration. The cellular hypoxia by glucose stimulation was also observed in the isolated islets from mice. The MIN6 cells overexpressing HIF-1␣ were resistant to becoming hypoxic after glucose stimulation. Thus, glucose-stimulated ␤-cells can become hypoxic by oxygen consumption, especially when the oxygen supply is impaired.

Within the limitations of this study, from the viewpoint of energy absorption ability, the minimum thickness required for a mouthguard is 4 mm, which is generally too large from the viewpoint of player comfort. This finding indicates the necessity of improving the impact absorption ability of mouthguards by considering new designs and developing new materials.

Most impact force and impact energy absorption tests for mouthguards have used a steel ball in a drop-ball or the pendulum device. However, in reality most sports-related trauma is caused by objects other than the steel ball, e.g. various sized balls, hockey puck, or bat or stick. Also, the elasticity, the velocity and the mass of the object could change the degree and the extent of injuries. In this study, we attempted to measure the impact force from actual sports equipment in order to clarify the exact mechanism of dental-related sports injuries and the protective effects of mouthguards. The present study was conducted using the pendulum impact device and load cell. Impact objects were removable. Seven mobile impact objects were selected for testing: a steel ball, baseball, softball, field hockey ball, ice hockey puck, cricket ball, and wooden baseball bat. The mouthguard material used in this study was a 3-mm-thick Drufosoft (Dreve-Dentamid GmbH, Unna, Germany), and test samples were made of the one-layer type. The peak transmitted forces without mouthguard ranged from the smallest (ice hockey stick, 46.9 kgf) to the biggest (steel ball, 481.6 kgf). The peak transmitted forces were smaller when the mouthguard was attached than without it for all impact materials but the effect was significantly influenced by the object type. The steel ball showed the biggest (62.1%) absorption ability while the wooden bat showed the second biggest (38.3%). The other balls or the puck showed from 0.6 to 6.0% absorbency. These results show that it is important to test the effectiveness of mouthguards on specific types of sports equipment. In future, we may select different materials and mouthguard designs suitable for specific sports.

Mouthguards are expected to reduce sports-related orofacial injuries. Numerous studies have been conduced to improve the shock absorption ability of mouthguards using air cells, sorbothane, metal wire, or hard material insertion. Most of these were shown to be effective; however, the result of each study has not been applied to clinical use. The aim of this study was to develop mouthguards that have sufficient prevention ability and ease of clinical application with focus on a hard insertion and space. Ethylene vinyl acetate (EVA) mouthguard blank used was Drufosoft and the acrylic resin was Biolon (Dreve-Dentamid GMBH, Unna, Germany). Three types of mouthguard samples tested were constructed by means of a Dreve Drufomat (Type SO, Dreve-Dentamid) air pressure machine: the first was a conventional laminated type of EVA mouthguard material; the second was a three layer type with acrylic resin inner layer (hard-insertion); the third was the same as the second but with space that does not come into contact with tooth surfaces (hard + space). As a control, without any mouthguard condition (NOMG) was measured. A pendulum type impact testing machine with interchangeable impact object (steel ball and baseball) and dental study model (D17FE-NC.7PS, Nissin, Tokyo, Japan) with the strain gages (KFG-1-120-D171-11N30C2: Kyowa, Tokyo, Japan) applied to teeth and the accelerometer to the dentition (AS-A YG-2768 100G, Kyowa) were used to measure transmitted forces. Statistical analysis (anova, P < 0.01) showed significant differences among four conditions of NOMG and three different mouthguards in both objects and sensor. About acceleration: in a steel ball which was a harder impact object, shock absorption ability of about 40% was shown with conventional EVA and hard-insertion and about 50% with hard + space. In a baseball that was softer compared with steel ball, a decrease rate is smaller, reduction (EVA = approximately 4%, hard-insertion = approximately 12%, hard + space = approximately 25%) was admitted in the similar order. A significant difference was found with all the combinations except for between EVA and hard-insertion with steel ball (Tukey test). About distortion: both buccal and lingual, distortions had become small in order of EVA, hard-insertion, and hard + space, too. The decrease rate is larger than acceleration, EVA = approximately 47%, hard-insertion = 80% or more, and hard +space = approximately 98%, in steel ball. EVA = approximately 30%, hard-insertion = approximately 75%, and hard + space = approximately 98% in baseball. And a significant difference was found with all the combinations (Tukey test). Especially, hard + space has decreased the distortion of teeth up to several percentages. Acceleration of the maxilla and distortions of the tooth became significantly smaller when wearing any type of mouthguard, in both impact objects. But the effect of mouthguard was clearer in the distortion of the tooth and with steel ball. Considering the differences of mouthguards, the hard-insertion and the hard + space...

The success of pancreatic β-cells transplantation to treat type 1 diabetes has been hindered by massive β-cell dysfunction and loss of β-cells that follows the procedure. Hypoxia-mediated cell death has been considered one of the main difficulties that must be overcome for transplantation to be regarded as a reliable therapy. Here we have investigated the mechanisms underlying β-cell death in response to hypoxia (1% O2). Our studies show that mouse insulinoma cell line 6 (Min6) cells undergo apoptosis with caspase-3 activation occurring as early as 2 h following exposure to hypoxia. Hypoxia induces endoplasmic reticulum stress in Min6 cells leading to activation of the three branches of the unfolded protein response pathway. In response to hypoxia the pro-apoptotic transcription factor C/EBP homologous protein (CHOP) is upregulated. The important role of CHOP in the apoptotic process was highlighted by the rescue of Min6 cells from hypoxia-mediated apoptosis observed in CHOP-knockdown cells. Culturing isolated pancreatic mouse islets at normoxia showed intracellular hypoxia with accumulation of hypoxia-inducible factor-1α and upregulation of CHOP, the latter one occurring as early as 4 h after isolation. Finally, we observed that pancreatic islets of type 2 db/db diabetic mice were more hypoxic than their counterpart in normoglycemic animals. This finding indicates that hypoxia-mediated apoptosis may occur in type 2 diabetes.

Mouthguards have been tested for impact energy absorption using drop-ball and/or pendulum devices. While all reports show efficiency of the mouthguard, the impact absorption abilities reported differ considerably. This difference has been attributed to differences of mouthguard material, design, and the impact force used. However, it is also possibly because of the difference in the sensors used in the experiments. The purpose of this study was to test three types of sensors and to assess which type was most appropriate for measurement of the impact absorption ability of mouthguards. A pendulum-type testing equipment and steel ball, wooden bat, baseball, field-hockey ball were used as the impact object. For all sensors or impact objects, the mouthguard decreased the impact forces. However, the absorption ability of the mouthguard varied according to the sensor or impact object. The absorbency values became smaller with the strain gauge, the accelerometer, and the load cell, respectively. With the steel ball as the impact object, 80.3% of impact absorption was measured with the strain gauge and the accelerometer but, only 62.1% with the load cell sensor. With the wooden bat, impact absorption was 76.3% with the strain gauge and 38.8% for the load cell. For the baseball ball, the absorption measurement decreased from 46.3% with the strain gauge to 4.36 with the load cell and for the field-hockey ball, the decrease in measurement values were similar (23.6% with the strain gauge and 2.43% with the load cell). It is clear that the sensor plays an important role in the measurement values reported for absorbency of mouthguard materials and a standard sensor should be used for all experiments.