A multicenter, phase 2 study to evaluate the efficacy and safety of osilodrostat, a new 11ß-hydroxylase inhibitor, in Japanese patients with endogenous Cushing's syndrome other than Cushing's disease.

This phase 2, single-arm, open-label, dose-titration, multicenter study evaluated osilodrostat (11ß-hydroxylase inhibitor) in Japanese patients with endogenous Cushing's syndrome (CS) caused by adrenal tumor/hyperplasia or ectopic adrenocorticotropic hormone syndrome. The primary endpoint was percent change from baseline to week 12 in mean urinary free cortisol (mUFC) at the individual patient level. Of the nine patients enrolled in the study, seven completed the 12-week core treatment period and two discontinued at or prior to week 12 due to adverse events (AEs). Of the seven patients who completed 12 weeks of study treatment, two completed 48 weeks of study treatment. Median osilodrostat exposure was 12 weeks. Median (range) average dose including dose interruption (0 mg/day) was 2.143 (1.16-7.54) mg/day. Median (range, population) percentage change in mUFC was -94.47% (-99.0% to -52.6%, n = 7) at week 12. At week 12, 6/9 patients were complete responders (mUFC ≤ upper limit of normal [ULN]) and 1/9 was a partial responder (mUFC > ULN but decreased by ≥50% from baseline). Most frequent AEs were adrenal insufficiency (n = 7), gamma-glutamyl transferase increase, malaise, and nasopharyngitis (n = 3 each). Serious AEs were seen in four patients. No deaths occurred in this study. In conclusion, osilodrostat treatment led to a reduction in mUFC in all nine patients with endogenous CS other than Cushing's disease (CD), regardless of disease type, with >80% reduction seen in 6/7 patients at week 12. The safety profile was consistent with previous reports in CD patients, and the reported AEs were manageable.

Effects of skin surface cooling before exercise on lactate accumulation in cool environment.

PURPOSE: We assessed whether plasma lactate accumulation increased and the lactate threshold (LT) declined when the skin temperature was lowered by whole body skin surface cooling before exercise in cool, but not temperate, conditions, and whether the lowered LT was associated with sympathetic activation or lowered plasma volume (PV) by cold-induced diuresis. METHODS: Ten healthy subjects performed a graded maximal cycling exercise after pre-conditioning under three different conditions for 60 min. Ambient temperature (using an artificial climatic chamber) and water temperature in a water-perfusion suit controlled at 25 and 34 °C in temperate-neutral (Temp-Neut); 25 and 10 °C in temperate-cool (Temp-Cool); and at 10 and 10 °C in cool-cool (Cool-Cool) conditions, respectively. Esophageal (Tes) and skin temperatures were measured; plasma lactate ([Lac]p) and noradrenaline concentrations ([Norad]p), and relative change in PV (%ΔPV) were determined before and after pre-conditioning and during exercise, and LT was determined. RESULTS: After pre-conditioning, Tes was not different among trials, whereas the mean skin temperature was lower in Cool-Cool and Temp-Cool than in Temp-Neut (P < 0.001). During exercise, [Lac]p and [Norad]p were higher (P = 0.009 and P < 0.001, respectively) and LT was lower (P = 0.013) in Cool-Cool than in the other trials. The %ΔPV was not different among trials. LT was correlated with [Norad]p during exercise (R = 0.50, P = 0.005). CONCLUSIONS: Whole body skin surface cooling before exercise increases lactate accumulation and decreases LT with sympathetic activation when exercise is performed in a cool, but not in a temperate, environment.

Thermal sensation during mild hyperthermia is modulated by acute postural change in humans.

Thermal sensation represents the primary stimulus for behavioral and autonomic thermoregulation. We assessed whether the sensation of skin and core temperatures for the driving force of behavioral thermoregulation was modified by postural change from the supine (Sup) to sitting (Sit) during mild hyperthermia. Seventeen healthy young men underwent measurements of noticeable increase and decrease (±0.1 °C/s) of skin temperature (thresholds of warm and cold sensation on the skin, 6.25 cm2 of area) at the forearm and chest and of the whole-body warm sensation in the Sup and Sit during normothermia (NT; esophageal temperature (Tes), ∼36.6 °C) and mild hyperthermia (HT; Tes, ∼37.2 °C; lower legs immersion in 42 °C of water). The threshold for cold sensation on the skin at chest was lower during HT than NT in the Sit (P < 0.05) but not in Sup, and at the forearm was lower during HT than NT in the Sup and further in Sit (both, P < 0.05), with interactive effects of temperature (NT vs. HT) × posture (Sup vs. Sit) (chest, P = 0.08; forearm, P < 0.05). The threshold for warm sensation on the skin at both sites remained unchanged with changes in body posture or temperature. The whole-body warm sensation was higher during HT than NT in both postures and higher in the Sit than Sup during both NT and HT (all, P < 0.05). Thus, thermal sensation during mild hyperthermia is modulated by postural change from supine to sitting to sense lesser cold on the skin and more whole-body warmth.

Lower thermal sensation in normothermic and mildly hyperthermic older adults.

PURPOSE: It is important to know how thermal sensation is affected by normal aging under conditions that elevate core body temperature for the prevention of heat-related illness in older people. We assessed whether thermal sensation under conditions of normothermia (NT) and mild hyperthermia (HT) is lowered in older adults. METHODS: Seventeen younger (23 ±  3 years) and 12 older (71 ±  3 years) healthy men underwent measurements of the cold and warmth detection thresholds ( ± 0.1 °C/s) of their chest and forearm skin, and whole body warmth perception under NT (esophageal temperature, T es, ~36.5 °C) and HT (T es, ~37.3 °C; lower legs immersed in 42 °C water) conditions. RESULTS: Warmth detection threshold at the forearm was increased in older compared with younger participants under both NT (P = 0.006) and HT (P = 0.004) conditions. In contrast, cold detection threshold at the forearm was decreased in older compared with younger participants under NT (P = 0.001) but not HT (P = 0.16). Mild hyperthermia decreased cold detection threshold at forearm in younger participants (P = 0.001) only. There were no effects of age and condition on warmth and cold detection thresholds at chest. Whole body warmth perception increased during HT compared with NT in both groups (both, P < 0.001), and older participants had lower values than the younger group under NT (P = 0.001) and HT (P = 0.051). CONCLUSIONS: Skin warmth detection thresholds at forearm and whole body warmth perception under NT and HT and skin cold detection thresholds at forearm under NT deteriorated with aging.

Thermoregulatory responses are attenuated after fructose but not glucose intake.

PURPOSE: We examined whether plasma hyperosmolality induced by oral monosaccharide intake attenuated thermoregulatory responses and whether the responses were different between fructose and glucose. METHODS: Ten healthy young subjects performed three trials in a sitting position in an artificial climate chamber (ambient temperature, 28°C; relative humidity, 40%). After resting for 10 min, the subjects drank 300 mL of water alone (control), or 300 mL of water supplemented with 75 g fructose or 75 g glucose. Twenty minutes later, they were heated passively by immersing the lower legs in water at 42°C for 60 min. Plasma osmolality (Posm), sodium ([Na+]p) and insulin concentrations ([Ins]p), and percent change in plasma volume (%ΔPV) were measured, and esophageal temperature (Tes) thresholds for cutaneous vasodilation (THCVC) and sweating (THSR) at the forearm were determined. RESULTS: Posm was significantly increased by fructose and glucose intake compared with water alone, although %ΔPV and [Na+]p were not significantly different among the three trials. [Ins]p was significantly higher after glucose intake than after fructose or water alone. THCVC and THSR were significantly higher after fructose intake than after glucose intake, which showed similar values to water intake. CONCLUSIONS: These results suggest that the Tes threshold for thermoregulation is elevated after fructose intake, indicating the attenuation of thermoregulatory responses, whereas it is not attenuated after glucose intake. These results provide a novel insight to better determine the carbohydrate component of oral rehydration fluids for preventing dehydration and/or heat disorders.

Feasibility study of B16 melanoma therapy using oxidized ATP to target purinergic receptor P2X7.

The P2X7 receptor is not only involved in cell proliferation, but also acts as an adenosine 5'-triphosphate (ATP)-gated non-selective channel, and its expression is increased in human melanoma. An irreversible antagonist of P2X7, such as oxidized ATP (oxATP), might block P2X7 receptor-mediated ATP release and proliferative signaling. Therefore, we carried out basic studies to test this idea and to examine the feasibility of using oxATP to treat B16 melanoma. We first found that low-pH conditions (mimicking the hypoxia and acidosis commonly seen in solid tumors) induced P2X7 receptor-mediated ATP release from B16 melanoma cells. Then, we compared the proliferation rates of B16 melanoma wild-type cells and B16 P2X7 receptor-knockdown clone (P2X7-KDC) cells in the presence of P2X7 agonists. The proliferation rate, as well as the ATP release, of agonist-treated P2X7-KDC cells was lower than that of agonist-treated wild-type cells. Next, the effect of P2X7 antagonist oxATP on B16 melanoma cell growth was examined in vitro and in vivo. oxATP significantly decreased B16 melanoma cell proliferation in vitro, and also significantly inhibited tumor growth in B16 melanoma-bearing mice. These data indicate that extracellularly released ATP may serve as an intercellular signaling molecule. We propose that the P2X7 receptor is a promising target for treatment of solid tumors.

Lower skin temperature decreases maximal cycling performance.

BACKGROUND: It is known that external cooling of body regions involved in exercise, prior to exercise, decreases anaerobic performance. However, there have been no studies reporting the effects of whole body skin surface cooling before exercise on maximal anaerobic capacity. In order to clarify the effects, we compared power output during the Wingate anaerobic test between preconditioning by exposure to temperature 10 degrees C and 25 degrees C. METHODS: Eight healthy males carried out the Wingate test for 30 seconds, after pre-conditioning for 60 minutes using a perfusion suit with water at a temperature of 10 degrees C or 25 degrees C. We evaluated the peak power (PP) and peak power slope (PS) of the power output. RESULTS: Mean skin temperature (T(sk)) at 60 minutes of pre-conditioning in the 10 degrees C trial was significantly lower than in the 25 degrees C trial (p < 0.05). PP and also PS were significantly lower in the 10 degrees C trial than in the 25 degrees C trial. Changes (Δ) in PP between the 10 degrees C trial and the 25 degrees C trial were strongly correlated with ΔT(sk) and Δ in thigh and leg skin temperature (ΔT(thigh) and ΔT(leg), respectively), whereas ΔPS was strongly correlated with ΔT(sk), but not with ΔT(thigh) and ΔT(leg). CONCLUSIONS: Whole body skin surface cooling prior to exercise restricts anaerobic capacity, especially in the initial phase of exercise.

gamma-Irradiation induces P2X(7) receptor-dependent ATP release from B16 melanoma cells.

BACKGROUND: Ionizing irradiation causes not only growth arrest and cell death, but also release of growth factors or signal transmitters, which promote cancer malignancy. Extracellular ATP controls cancer growth through activation of purinoceptors. However, there is no report of radiation-induced ATP release from cancer cells. Here, we examined gamma-irradiation-induced ATP release and its mechanism in B16 melanoma. METHODS: Extracellular ATP was measured by luciferin-luciferase assay. To investigate mechanism of radiation-induced ATP release, we pharmacologically inhibited the ATP release and established stable P2X(7) receptor-knockdown B16 melanoma cells using two short hairpin RNAs targeting P2X(7) receptor. RESULTS: Cells were exposed to 0.5-8 Gy of gamma-rays. Extracellular ATP was increased, peaking at 5 min after 0.5 Gy irradiation. A selective P2X(7) receptor channel antagonist, but not anion transporter inhibitors, blocked the release of ATP. Further, radiation-induced ATP release was significantly decreased in P2X(7) receptor-knockdown cells. Our results indicate that gamma-irradiation evokes ATP release from melanoma cells, and P2X(7) receptor channel plays a significant role in mediating the ATP release. GENERAL SIGNIFICANCE: We suggest that extracellular ATP could be a novel intercellular signaling molecule released from cancer cells when cells are exposed to ionizing radiation.