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Survival of organisms requires the ability to adapt to changes in the environment. Adaptation of oxidative metabolism is essential for meeting increased energy demands in response to stressors, such as exposure to cold temperatures or increased physical activity. Adaptive changes in metabolism are often achieved at the level of gene expression, and nuclear receptors have prevalent roles in mediating such responses. Estrogen-related receptor [alpha] (ERR[alpha]) was the first orphan nuclear receptor to be identified, and yet its physiologic function remains unknown. Here, we show that mice lacking ERR[alpha] are unable to maintain body temperature when exposed to cold. Surprisingly, the inability to adapt to cold is not due to defects in the acute transcriptional induction of genes important for thermogenesis. Rather, we show that ERR[alpha] is needed for the high levels of mitochondrial biogenesis and oxidative capacity characteristic of brown adipose tissue (BAT), and thus for providing the energy necessary for thermogenesis. ERR[alpha] fulfills this role by acting directly at genes important for mitochondrial function, parallel to other factors controlling mitochondrial gene expression, such as NRF1 and NRF2/GABPA. Our findings demonstrate that ERR[alpha] is a key regulator of mitochondrial biogenesis and oxidative metabolism, and essential for adaptive thermogenesis. mitochondrial biogenesis | oxidative metabolism | brown adipose tissue

Estrogen-related receptor [alpha](ERR[alpha]) is one of the first orphan nuclear receptors to be identified, yet its physiological functions are still unclear. We show here that ERR[alpha] is an effector of the transcriptional coactivator PGC-1[alpha][peroxisome proliferator-activated receptor [gamma] (PPAR[gamma]) coactivator 1[alpha]], and that it regulates the expression of genes involved in oxidative phosphorylation and mitochondrial biogenesis. Inhibition of ERR[alpha] compromises the ability of PGC-1[alpha] to induce the expression of genes encoding mitochondrial proteins and to increase mitochondrial DNA content. A constitutively active form of ERR[alpha] is sufficient to elicit both responses. ERR[alpha] binding sites are present in the transcriptional control regions of ERR[alpha]/PGC-1[alpha]-induced genes and contribute to the transcriptional response to PGC-1[alpha]. The ERR[alpha]-regulated genes described here have been reported to be expressed at reduced levels in humans that are insulin-resistant. Thus, changes in ERR[alpha] activity could be linked to pathological changes in metabolic disease, such as diabetes.

Electron transport parallel to dislocation lines is not affected by the scattering at charged dislocation lines, according to research based on a model developed to analyze the low transverse mobility of GaN by scattering of electrons at charged dislocation lines. The repulsive band bending around dislocations and the directional dependence of the dislocation scattering prevent vertical devices being affected by the scattering of electrons at threading dislocation lines.

We experimentally demonstrate the formation of GaN nanotip pyramids by selective and anisotropic etching of N-polar GaN in KOH solution. For samples grown with adjacent Ga- and N-polar regions on the same wafer, the KOH solution was found to selectively etch only the N-polar surface while leaving the Ga-polar surface intact. An aggregation of hexagonal pyramids with well defined {10 &1macr; &1macr;} facets and very sharp tips with diameters less than ∼20 nm were formed. The density of the pyramids can be controlled by varying the KOH concentration, solution temperature or the etch duration. The GaN etching activation energy is estimated to be E[sub a] ≈ 0.587 eV. Dense GaN pyramids with sharp tips have applications in both electronic and photonic devices. [ABSTRACT FROM AUTHOR]

The role of [SCF.sub.Cdc4] as an E3 ubiquitin ligase for regulating peroxisome coactivator-1 (PGC-1) through ubiquitin-mediated proteolysis is discussed. Results suggest that the attenuation of [SCF.sub.Cdc4] dependent proteasomal degradation of PGC-1 plays an important role in the mediation of the PGC-1-dependent transcriptional response to oxidative stress.

Etelcalcetide (AMG 416) is a novel synthetic peptide calcium-sensing receptor activator in clinical development as an intravenous calcimimetic for the treatment of secondary hyperparathyroidism in patients with chronic kidney disease (CKD) on hemodialysis. Etelcalcetide is composed of seven D-aminoacids with an L-cysteine linked to a D-cysteine by a disulfide bond. A single intravenous dose of [14C]etelcalcetide (10 mg; 26.3 kBq; 710 nCi) was administered to patients with CKD on hemodialysis to elucidate the pharmacokinetics, biotransformation, and excretion of etelcalcetide in this setting. Blood, dialysate, urine, and feces were collected to characterize the pharmacokinetics, biotransformation product profiles, mass balance, and formation of anti-etelcalcetide antibodies. Accelerator mass spectrometry was necessary to measure the microtracer quantities of C-14 excreted in the large volumes of dialysate and other biomatrices. An estimated 67 % of the [14C]etelcalcetide dose was recovered in dialysate, urine, and feces 176 days after dose administration. Etelcalcetide was primarily cleared by hemodialysis, with approximately 60 % of the administered dose eliminated in dialysate. Minor excretion was observed in urine and feces. Biotransformation resulted from disulfide exchange with endogenous thiols, and preserved the etelcalcetide D-amino acid backbone. Drug-related radioactivity circulated primarily as serum albumin peptide conjugate (SAPC). Following removal of plasma etelcalcetide by hemodialysis, re-equilibration occurred between SAPC and L-cysteine present in blood to partially restore the etelcalcetide plasma concentrations between dialysis sessions. No unanticipated safety signals or anti-etelcalcetide or anti-SAPC antibodies were detected. [ABSTRACT FROM AUTHOR]

The article covers a study using a single microtracer intravenous dose of [14C]etelcalcetide that indicated that 60% of the administered dose was eliminated in dialysate. It describes how etelcalceditde can be used to treat hyperparathyroidism in patients diagnosed with chronic kidney disease.

Objective: To evaluate the time-independent effect of the early administration of epinephrine (EPI) on favorable neurological outcome (as CPC [cerebral performance category] 1-2) at 1 month in patients with out-of-hospital cardiac arrest.Materials and Methods: A total of 119 639 witnessed cardiac arrest patients from 2008 to 2012 were eligible for this nationwide, prospective, population-based observational study. Patients were divided into EPI group (n = 20 420) and non-EPI group (n = 99 219). To determine the time-dependent effects of EPI, EPI-administered patients were divided into 4 groups as follows: early EPI (5-18 min), intermediate EPI (19-23 min), late EPI (24-29 min), and very late EPI (30-62 min), respectively. Multiple logistic regression analyses and adjusted odds ratios (AORs) were determined for CPC 1-2 at 1 month (primary outcome) and field return of spontaneous circulation (as secondary outcome) among the groups.Results: The EPI and non-EPI group had identical background, but EPI group shows higher incidence public access defibrillation and emergency medical technician defibrillation delivered than the non-EPI group. The differences were clinically negligible. Higher return of spontaneous circulation rate (18.0%) and lower CPC 1-2 (2.9%) shown in the EPI group than in the non-EPI group (9.4% and 5.2%). In the time dependent analysis, CPC 1 to 2 was greatest in the early EPI group (AOR, 2.49; 95% confidence interval [CI], 1.90-3.27), followed by the intermediate EPI group (AOR, 1.53; 95% CI, 1.14-2.05) then the late EPI group (AOR, 0.71; 95% CI, 0.47-1.08) as reference.Conclusion: Early EPI administration within 19 minutes after emergency medical service call independently improved the neurological outcome compared with late EPI (24-29 minutes) administration in patients with out-of-hospital cardiac arrest. [ABSTRACT FROM AUTHOR]

The T-cell-dependent antibody response (TDAR) is a functional assay used in immunopharmacology and immunotoxicology to assess ability to mount an antibody response to immunization. Keyhole limpet hemocyanin (KLH) is extensively used as the immunogen of choice in non-clinical and clinical settings. Native KLH is comprised of high molecular weight (HMW; 4-8 MDa) assemblies of KLH subunit dimers (> 600-800 kDa). It is not known how the different forms (HMW vs subunit) and manufacturing processes (commercial sources) may impact the nature of anti-KLH immune responses (e.g. magnitude and inter-animal variability). Anti-KLH IgM and IgG responses were studied in Sprague-Dawley rats immunized with different forms and commercial sources of KLH: 100 µg of HMW KLH from two different sources or subunit KLH from three different sources. Biophysical and biochemical analyses were conducted to characterize the KLH formulations. Anti-KLH IgM and IgG responses were measured using a proprietary indirect quantitative electrochemiluminescence immunoassay. The HMW KLH preparations showed a greater number of sub-visible particles (2-150 µm size range) than the subunit KLH preparations. All HMW KLH and all subunit KLH were equivalent on SEC (hydrodynamic volume), PAGE (size and charge), and SDS-PAGE (molecular radius). Robust primary and secondary anti-KLH responses were detected for both sources of HMW KLH. The subunit KLH immunizations resulted in lower IgG and IgM responses compared to the HMW KLH, with the exception of Stellar Biotechnologies subunit KLH that produced both robust primary and secondary responses, which approached the HMW KLH responses. Inter-animal variability for IgM and IgG responses was lower with HMW KLH than with subunit KLH. In conclusion, different forms and commercial sources of KLH were associated with different magnitudes and inter-animal variability in IgM and IgG responses, a critical finding to take into consideration when designing TDAR studies for robust immunotoxicology or immunopharmacology testing. [ABSTRACT FROM AUTHOR]

Understanding species differences in the placental transfer of monoclonal antibodies is important to inform species selection for nonclinical safety assessment, interpret embryo-fetal changes observed in these studies, and extrapolate their human relevance. Data presented here for a fully human immunoglobulin G2 monoclonal antibody (IgG2X) revealed that, during organogenesis, in both the cynomolgus monkey (gestation day 35 [gd35]) and the rat (gd10) the extent of IgG2X placental transfer (approximately 0.5% maternal plasma concentration, MPC) was similar to the limited published human data for endogenous IgG. At this early gestational stage, IgG2X placental transfer was approximately 6-fold higher in the rabbit (gd10). By the end of organogenesis, rat embryonic plasma concentrations (gd16) exceeded those in the cynomolgus monkey (gd50) by approximately 3-fold. These data suggest that relative to the cynomolgus monkey, the rabbit (and to a lesser extent the rat) may overestimate potential harmful effects to the human embryo during this critical period of development. Beyond organogenesis, fetal IgG2X plasma concentrations increased approximately 10-fold early in the second trimester (gd50-70) in the cynomolgus monkey and remained relatively unchanged thereafter (at approximately 5% MPC). Late gestational assessment was precluded in rabbits due to immunogenicity, but in rats, fetal IgG2X plasma concentrations increased more than 6-fold from gd16 to gd21 (reaching approximately 15% MPC). In rats, maternal exposure consistent with that achieved by ICH S6(R1) high-dose selection criteria resulted in embryonic plasma concentrations, reaching pharmacologically relevant levels during organogenesis. Furthermore, dose proportional exposure in both mothers and embryos indicated that this was unlikely to occur at the lower therapeutic dose levels used in humans [ABSTRACT FROM AUTHOR]

This continuing education course was designed to provide an overview of the immunologic mechanisms involved in immunogenicity and hypersensitivity reactions following administration of biologics in nonclinical toxicity studies, the methods used to determine whether such reactions are occurring, and the associated clinical and anatomic pathology findings. Hypersensitivity reactions have classically been divided into type I, II, III, and IV reactions; type I and III reactions are those most often observed following administration of biologics. A variety of methods can be used to detect these reactions. Antemortem methods include hematology; detection of antidrug antibodies, circulating immune complexes and complement fragments, and immunoglobulin E in serum; tests for serum complement activity; and evaluation of complement receptor 1 on erythrocytes. Postmortem methods include routine light microscopy and electron microscopy, which can demonstrate typical findings associated with hypersensitivity reactions, and immunohistochemistry, which can detect the presence of immune complexes in tissues, including the detection of the test article. A final determination of whether findings are related to a hypersensitivity reaction in individual animals or across the entire study should rely on the overall weight of evidence, as findings indicative of these reactions are not necessarily consistent across all affected animals. [ABSTRACT FROM AUTHOR]

Newly activated CD8+ T cells reprogram their metabolism to meet the extraordinary biosynthetic demands of clonal expansion; however, the signals that mediate metabolic reprogramming remain poorly defined. Here we demonstrate an essential role for sterol regulatory element-binding proteins (SREBPs) in the acquisition of effector-cell metabolism. Without SREBP signaling, CD8+ T cells were unable to blast, which resulted in attenuated clonal expansion during viral infection. Mechanistic studies indicated that SREBPs were essential for meeting the heightened lipid requirements of membrane synthesis during blastogenesis. SREBPs were dispensable for homeostatic proliferation, which indicated a context-specific requirement for SREBPs in effector responses. Our studies provide insights into the molecular signals that underlie the metabolic reprogramming of CD8+ T cells during the transition from quiescence to activation. [ABSTRACT FROM AUTHOR]

Increased colonic Cl- secretion was supposed to be a causative factor of diarrhea in inflammatory bowel diseases. Surprisingly, hyporesponsiveness to Cl- secretagogues was later described in inflamed colon. Our aim was to evaluate changes in secretory responses to cholinergic agonist carbachol in distal and proximal colon during colitis development, regarding secretory activity of enteric nervous system (ENS) and prostaglandins. Increased responsiveness to carbachol was observed in both distal and proximal colon after 3 days of 2% dextran sodium sulfate (DSS) administration. It was measured in the presence of mucosal Ba2+ to emphasize Cl- secretion. The described increase was abolished by combined inhibitory effect of tetrodotoxin (TTX) and indomethacin. Indomethacin also significantly reduced TTX-sensitive current. On the 7th day of colitis development responsiveness to carbachol decreased in distal colon (compared to untreated mice), but did not change in proximal colon. TTX-sensitive current did not change during colitis development, but indomethacin-sensitive current was significantly increased the 7th day. Decreased and deformed current responses to serosal Ba2+ were observed during colitis induction, but only in proximal colon. We conclude that besides inhibitory effect of DSS on distal colon responsiveness, there is an early stimulatory effect that manifests in both distal and proximal colon. [ABSTRACT FROM AUTHOR]

The intestinal transport of nutrients exhibits distinct diurnal rhythmicity, and the enterocytes harbor a circadian clock. However, temporal regulation of the genes involved in colonic ion transport, i.e., ion transporters and channels operating in absorption and secretion, remains poorly understood. To address this issue, we assessed the 24-h profiles of expression of genes encoding the sodium pump (subunits Atp1a1 and Atp1b1), channels (α-, β-, and γ-subunits of Enac and Cftr), transporters (Dra, Ae1, Nkcc1, Kcc1, and Nhe3), and the Na+/H+ exchanger (NHE) regulatory factor (Nherf1) in rat colonic mucosa. Furthermore, we investigated temporal changes in the spatial localization of the clock genes Per1, Per2, and Bmal1 and the genes encoding ion transporters and channels along the crypt axis. In rats fed ad libitum, the expression of Atp1a1, γEnac, Dra, Ae1, Nhe3, and Nherf1 showed circadian variation with maximal expression at circadian time 12, i.e., at the beginning of the subjective night. The peak γEnac expression coincided with the rise in plasma aldosterone. Restricted feeding phase advanced the expression of Dra, Ae1, Nherf, and γEnac and decreased expression of Atp1a1. The genes Atp1b1, Cftr, αEnac, βEnac, Nkcc1, and Kcc1 did not show any diurnal variations in mRNA levels. A low-salt diet upregulated the expression of βEnac and γEnac during the subjective night but did not affect expression of αEnac. Similarly, colonic electrogenic Na+ transport was much higher during the subjective night than the subjective day. These findings indicate that the transporters and channels operating in NaCl absorption undergo diurnal regulation and suggest a role of an intestinal clock in the coordination of colonic NaCl absorption. [ABSTRACT FROM AUTHOR]

Exercise improves the ability of skeletal muscle to metabolise fats and sugars. For these improvements to occur the muscle detects a signal caused by exercise, resulting in changes in genes and proteins that control metabolism. We show that endurance exercise increases the amount of a protein called striated muscle activator of Rho signalling (STARS) as well as several other proteins influenced by STARS. We also show that the amount of STARS can be increased by signals directed from proteins called peroxisome proliferator-activated receptor gamma co-activator 1-α (PGC-1α) and oestrogen-related receptor-α (ERRα). We also observed that when we reduce the amount of STARS in muscle cells, we block the ability of PGC-1α/ERRα to increase a gene called carnitine palmitoyltransferase-1β (CPT-1β), which is important for fat metabolism. Our study has shown that the STARS pathway is regulated by endurance exercise. STARS may also play a role in fat metabolism in muscle. [ABSTRACT FROM AUTHOR]

Estrogen-related receptor α (ERRα) is one of the first orphan nuclear receptors to be identified, yet its physiological functions are still unclear. We show here that ERRα is an effector of the transcriptional coactivator PGC-1α [peroxisome proliferator-activated receptor γ(PPARγ) coactivator 1α], and that it regulates the expression of genes involved in oxidative phosphorylation and mitochondrial biogenesis. Inhibition of ERRα compromises the ability of PGC-1α to induce the expression of genes encoding mitochondrial proteins and to increase mitochondrial DNA content. A constitutively active form of ERRα is sufficient to elicit both responses. ERRα binding sites are present in the transcriptional control regions of ERRα/PGC-1α-induced genes and contribute to the transcriptional response to PGC-1α. The ERRα-regulated genes described here have been reported to be expressed at reduced levels in humans that are insulin-resistant. Thus, changes in ERRα activity could be linked to pathological changes in metabolic disease, such as diabetes. [ABSTRACT FROM AUTHOR]

Murine nuclear factor of activated T cells (NFAT)2.α/β differ by 42 and 28 unique amino-terminal amino acids and are differentially expressed. Both isoforms share conserved domains that regulate DNA-binding and subcellular localization. A genetic "one-hybrid" assay was used to define two distinct transactivation (TA) domains: in addition to a conserved TAD present in both isoforms, a second, novel TAD exists within the β-specific amino terminus. Pharmacologic inhibitors Gö6976 and rottlerin demonstrate that both conventional and novel protein kinase C (PKC) family members regulate endogenous mast cell NFAT activity, and NFAT2 TA. Overexpression of dominant active PKCθ (which has been implicated in immune receptor signaling) induces NFAT2.β/β TA. Mutations within the smallest PKCθ-responsive transactivation domain demonstrate that the PKCθ effect is at least partially indirect. Significantly, the β-specific domain confers greater ability to TA in response to treatment with phorbol 12-myristate 13-acetate/ionomycin or lipopolysaccharide, and unique sensitivity to Fc∈RI signaling. Accordingly, overexpression of NFAT2.β results in significantly greater NFAT- and interleukin-4 reporter activity than NFAT2.α. These results suggest that whereas NFAT2 isoforms may share redundant DNA-binding preferences, there are specialized functional consequences of their isoform-specific domains. [ABSTRACT FROM AUTHOR]

We report absorption loss coefficients at 1550 nm of free-standing GaN grown by hydride vapor phase epitaxy. We measured propagation loss values at numerous locations of the facets and observed an average of 2.41 dB/cm and 1.85 dB/cm for transverse electric (TE) and transverse magnetic (TM) polarizations, respectively, in the low loss region of the sample. The minimum loss values in the sample were found to be 0.82 dB/cm (TE) and 0.61 dB/cm (TM). The ability to achieve such low propagation loss coefficients in GaN opens up the possibility of using this material in a variety of integrated optics and nonlinear optics applications. [ABSTRACT FROM AUTHOR]

We report the experimental demonstration of second-harmonic generation in periodically poled GaN by first-order quasiphase matching. The periodically poled structure was grown by plasma-assisted molecular-beam epitaxy. We observed about 9 μW of second-harmonic power from a fundamental input laser wavelength of 1658.6 nm with a normalized conversion efficiency of 12.76% W[sup -1] cm[sup -2]. The ability to perform nonlinear wavelength conversion by periodic poling and the fact that GaN has a very wide window of transparency, pave the way for GaN to be used for nonlinear optical devices in telecommunications as well as a nonlinear light source for biochemical detection in the far infrared and deep ultraviolet. © 2003 American Institute of Physics. [ABSTRACT FROM AUTHOR]

Intersubband optical absorption at λ ∼ 2.1 µm wavelength in doped 17.5 Å wide GaN quantum wells (QWs) with 51 Å wide intermediate AlN barriers is reported. A ∼600 nm thick GaN template and 15 QWs have been grown by molecular beam epitaxy on R-plane sapphire substrate and display pure A-plane orientation. QWs with A-plane orientation are essentially free from spontaneous polarisation, which is expected to greatly simplify the design of GaN/Al(Ga)N devices employing intersubband transitions. [ABSTRACT FROM AUTHOR]

Peroxisome proliferator-activated receptor γ (PPARγ) coactivator-1α (PGC-1α) is a highly regulated transcriptional coactivator that coordinates energy metabolism in mammals. Misregulation of PGC-1α has been implicated in the pathogenesis of several human diseases, including diabetes, obesity, and neurological disorders. We identified SCFCdc4 as an E3 ubiquitin ligase that regulates PGC-1α through ubiquitin-mediated proteolysis. PGC-1α contains two Cdc4 phosphodegrons that bind Cdc4 when phosphorylated by Glycogen Synthase Kinase 3² (GSK3²) and p38 MAPK, leading to SCFCdc4-dependent ubiquitylation and proteasomal degradation of PGC-1α. Furthermore, SCFCdc4 negatively regulates PGC-1α-dependent transcription. We demonstrate that RNAi-mediated reduction of Cdc4 in primary neurons results in an increase of endogenous PGC-1α protein, while ectopic expression of Cdc4 leads to a reduction of endogenous PGC-1α protein. Finally, under conditions of oxidative stress in neurons, Cdc4 levels are decreased, leading to an increase in PGC-1α protein and PGC-1α-dependent transcription. These results suggest that attenuation of SCFCdc4-dependent proteasomal degradation of PGC-1α has a role in mediating the PGC-1α-dependent transcriptional response to oxidative stress. [ABSTRACT FROM AUTHOR]

Intersubband absorption in coupled GaN/AlGaN double quantum wells (DQWs) has been measured. The samples were grown by molecular-beam epitaxy on a sapphire substrate and with large (0.65 or 0.9) AlN-mole fraction in the barriers. Peak absorption wavelengths as short as 1.35 and 1.52 μm were measured for a symmetric DQW of 12 Å wide wells coupled by a 10 Å wide barrier, which also showed evidence of excited-state anticrossing. As expected, asymmetric DQWs displayed no such anticrossing, and the ground-state anticrossing energies were found to be much smaller, as a result of the comparatively large effective electron mass, than the energy broadening of individual transitions. Degenerate doping of the DQWs was used to establish a common reference energy at the Fermi level, which allows overcoming uncertainties related to intrinsic internal electric fields. The asymmetric DQWs displayed peak absorption wavelengths between 1.5 and 2.9 μm. © 2001 American Institute of Physics. [ABSTRACT FROM AUTHOR]

Intersubband optical absorption around 1.55 μm has been measured in GaN/AlGaN multiple quantum wells (MQWs) grown by molecular-beam epitaxy. First, peak absorption wavelengths as short as 1.41 μm are reported for ultranarrow, >=11 Å wide, well-doped MQWs with high, 85%, AlN mole-fraction barriers. Second, in order to enable modulation doping as well as the use of lower AlN mole-fraction barriers, we designed and fabricated QWs embedded in barriers consisting of a short period superlattice of narrow GaN QWs and only 65% AlN mole-fraction barriers. The resulting electron Bragg confinement allows peak absorption wavelengths as short as 1.52 μm. Furthermore, the structures can now be modulation doped through doping of the narrow superlattice wells and subsequent charge transfer into the active well. We observe a reduction of the absorption linewidth, from ∼200 to ∼130 meV, for these structures. © 2000 American Institute of Physics. [ABSTRACT FROM AUTHOR]

Intersubband optical absorption in narrow, 15-30 Å wide, GaN/AlGaN quantum wells has been measured. The samples were grown by molecular-beam epitaxy on sapphire substrate and the barrier AlN mole fraction was varied from 0.45 to 0.8. Peak absorption wavelengths ranged from 4.2 μm for 30 Å wide wells to 1.77 μm for a 15 Å wide well. Modeling shows that a large contribution to the considerable spectral width of the absorption of ∼150 meV likely results from monolayer fluctuations. In addition to the composition dependent band offset, the intrinsic electric fields in the wells and barriers are the determining factors for the shortest possible wavelength. © 2000 American Institute of Physics. [ABSTRACT FROM AUTHOR]