The Role of Savoring in Young Adult Cannabis Use and Associated Consequences: A Replication Study.

Young adults exhibit high rates of cannabis use and are at heightened risk of experiencing negative cannabis-associated consequences. The purpose of the present study was to replicate and extend to prior work on savoring, the ability to experience positive experiences/emotions, and cannabis use frequency on cannabis-associated consequences. Young adults (18-25 years old,  = 122, 36.

Optical control of PIEZO1 channels.

PIEZO proteins are unusually large, mechanically-activated trimeric ion channels. The central pore features structural similarities with the pore of other trimeric ion channels, including purinergic P2X receptors, for which optical control of channel gating has been previously achieved with photoswitchable azobenzenes. Extension of these chemical optogenetics methods to mechanically-activated ion channels would provide tools for specific manipulation of pore activity alternative to non-specific mechanical stimulations.

A retrospective examination of changes in drinking motives during the early COVID-19 pandemic.

Objective: The COVID-19 pandemic has led to substantial changes in college student alcohol use. Changes in drinking motives may explain some of these changes in drinking patterns. The purpose of the present study is to explore how drinking motives and alcohol use have changed amongst college students considering the timeframes before and after the onset of COVID-19 pandemic (i.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

P2X7 Receptors and TMEM16 Channels Are Functionally Coupled with Implications for Macropore Formation and Current Facilitation.

P2X7 receptors (P2X7) are cationic channels involved in many diseases. Following their activation by extracellular ATP, distinct signaling pathways are triggered, which lead to various physiological responses such as the secretion of pro-inflammatory cytokines or the modulation of cell death. P2X7 also exhibit unique behaviors, such as "macropore" formation, which corresponds to enhanced large molecule cell membrane permeability and current facilitation, which is caused by prolonged activation.

The Middle Ear Muscle Reflex in Rat: Developing a Biomarker of Auditory Nerve Degeneration.

Objectives: The long-term goal of this research is to determine whether the middle ear muscle reflex can be used to predict the number of healthy auditory nerve fibers in hearing-impaired ears. In this study, we develop a high-impedance source and an animal model of the middle ear muscle reflex and explore the influence of signal frequency and level on parameters of the reflex to determine an optimal signal to examine auditory nerve fiber survival.

Design: A high-impedance source was developed using a hearing aid receiver attached to a 0.

On the permeation of large organic cations through the pore of ATP-gated P2X receptors.

Pore dilation is thought to be a hallmark of purinergic P2X receptors. The most commonly held view of this unusual process posits that under prolonged ATP exposure the ion pore expands in a striking manner from an initial small-cation conductive state to a dilated state, which allows the passage of larger synthetic cations, such as -methyl-d-glucamine (NMDG). However, this mechanism is controversial, and the identity of the natural large permeating cations remains elusive.

Photo-switchable tweezers illuminate pore-opening motions of an ATP-gated P2X ion channel.

P2X receptors function by opening a transmembrane pore in response to extracellular ATP. Recent crystal structures solved in apo and ATP-bound states revealed molecular motions of the extracellular domain following agonist binding. However, the mechanism of pore opening still remains controversial.

Optical control of an ion channel gate.

The powerful optogenetic pharmacology method allows the optical control of neuronal activity by photoswitchable ligands tethered to channels and receptors. However, this approach is technically demanding, as it requires the design of pharmacologically active ligands. The development of versatile technologies therefore represents a challenging issue.

Intermediate closed channel state(s) precede(s) activation in the ATP-gated P2X2 receptor.

The molecular mechanism underlying channel opening in response to agonist binding remains a challenging issue in neuroscience. In this regard, many efforts have been recently undertaken in ATP-gated P2X receptors. Among those efforts, we have provided evidence in the P2X2 receptor that tightening of ATP sites upon agonist binding induces opening of the ion channel.

Tightening of the ATP-binding sites induces the opening of P2X receptor channels.

The opening of ligand-gated ion channels in response to agonist binding is a fundamental process in biology. In ATP-gated P2X receptors, little is known about the molecular events that couple ATP binding to channel opening. In this paper, we identify structural changes of the ATP site accompanying the P2X2 receptor activation by engineering extracellular zinc bridges at putative mobile regions as revealed by normal mode analysis.

Agonist trapped in ATP-binding sites of the P2X2 receptor.

ATP-gated P2X receptors are trimeric ion channels, as recently confirmed by X-ray crystallography. However, the structure was solved without ATP and even though extracellular intersubunit cavities surrounded by conserved amino acid residues previously shown to be important for ATP function were proposed to house ATP, the localization of the ATP sites remains elusive. Here we localize the ATP-binding sites by creating, through a proximity-dependent "tethering" reaction, covalent bonds between a synthesized ATP-derived thiol-reactive P2X2 agonist (NCS-ATP) and single cysteine mutants engineered in the putative binding cavities of the P2X2 receptor.

Retrochalcone derivatives are positive allosteric modulators at synaptic and extrasynaptic GABA(A) receptors in vitro.

Background And Purpose: Flavonoids, important plant pigments, have been shown to allosterically modulate brain GABA(A) receptors (GABA(A)Rs). We previously reported that trans-6,4'-dimethoxyretrochalcone (Rc-OMe), a hydrolytic derivative of the corresponding flavylium salt, displayed nanomolar affinity for the benzodiazepine binding site of GABA(A)Rs. Here, we evaluate the functional modulations of Rc-OMe, along with two other synthetic derivatives trans-6-bromo-4'-methoxyretrochalcone (Rc-Br) and 4,3'-dimethoxychalcone (Ch-OMe) on GABA(A)Rs.

A putative extracellular salt bridge at the subunit interface contributes to the ion channel function of the ATP-gated P2X2 receptor.

The recent crystal structure of the ATP-gated P2X4 receptor revealed a static view of its architecture, but the molecular mechanisms underlying the P2X channels activation are still unknown. By using a P2X2 model based on the x-ray structure, we sought salt bridges formed between charged residues located in a region that directly connects putative ATP-binding sites to the ion channel. To reveal their significance for ion channel activation, we made systematic charge exchanges and measured the effects on ATP sensitivity.

Comparative models of P2X2 receptor support inter-subunit ATP-binding sites.

ATP-gated P2X receptors (P2XRs) are ligand-gated ion channels (LGICs) presumably trimeric. To date, no experimental high-resolution structures are available. Recent X-ray structure of the acid-sensing ion channel 1 (ASIC1) revealed an unexpected trimeric ion channel.

Reactive derivatives for affinity labeling in the ifenprodil site of NMDA receptors.

To prepare thiol-reactive ifenprodil derivatives designed as potential probes for cysteine-substituted NR2B containing NMDA receptors, electrophilic centers were introduced in different areas of the ifenprodil structure. Intermediates and final compounds were evaluated by binding studies and by electrophysiology to determine the structural requirements for their selectivity. The reactive compounds were further tested for their stability and for their reactivity in model reactions; some were found suitable as structural probes to investigate the binding site and the docking mode of ifenprodil in the NR2B subunit.

XRCC1 is phosphorylated by DNA-dependent protein kinase in response to DNA damage.

The two BRCT domains (BRCT1 and BRCT2) of XRCC1 mediate a network of protein-protein interactions with several key factors of the DNA single-strand breaks (SSBs) and base damage repair pathways. BRCT1 is required for the immediate poly(ADP-ribose)-dependent recruitment of XRCC1 to DNA breaks and is essential for survival after DNA damage. To better understand the biological role of XRCC1 in the processing of DNA ends, a search for the BRCT1 domain-associated proteins was performed by mass spectrometry of GST-BRCT1 pulled-down proteins from HeLa cell extracts.

Nursing care of the patient with preterm premature rupture of membranes.

Preterm premature rupture of the membranes (PPROM) is diagnosed when rupture of the amniotic membranes occurs prior to the completion of the 36th week of gestation. PPROM accounts for 25% of all cases of premature rupture of the membranes and is responsible for 30%-40% of all preterm deliveries. In mothers diagnosed with PPROM without evidence of infection, active labor, or fetal compromise, the current standard of care is expectant management.

Validated assay for the quantification of anastrozole in human plasma by capillary gas chromatography-63Ni electron capture detection.

An assay was developed for the quantification of anastrozole [2,2'-[5-(1H-1,2,4-triazol-1-ymethyl)-1,3-phenylene]bis(2-++ +methylpropiononitrile)] in human plasma using liquid-liquid extraction. Anastrozole and an internal standard were chromatographed and detected by gas chromatography with electron capture detection, using a combination temperature-pressure program. The range of the assay is 3 to 100 ng/ml.

Determination of ZD9583, a thromboxane receptor antagonist, in human plasma and urine by liquid chromatography atmospheric pressure chemical ionization tandem mass spectrometry.

A liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry (LC-APCI-MS-MS) method is described for the determination of a thromboxane receptor antagonist (4Z)-6-((2S,4S,5R)-2-(1-(2-cyano-4-methylphenoxy)-1-methylethyl)-4 -(3-pyridyl)-3-dioxan-5-yl)hex-4-enoic acid (ZD9583, I) in human plasma and urine. Proteins in plasma and urine samples are precipitated using acidified acetonitrile. The resulting supernatant is chromatographed on a C8 reversed-phase chromatography column.

Sulfonylurea binding to adipocyte membranes and potentiation of insulin-stimulated hexose transport.

We have previously shown that the sulfonylureas increase insulin-stimulated glucose transport in adipocytes mainly by enhancing the insulin-induced recruitment of glucose transporter from its intracellular storage pool to the plasma membrane (Jacobs, D. B., and Jung, C.

Insulin and phorbol esters affect the maximum velocity rather than the half-saturation constant of 3-O-methylglucose transport in rat adipocytes.

The kinetics of the equilibrium exchange flux of 3-O-methylglucose (MeGlc) were examined in isolated rat adipocytes using a recently described technique (Whitesell, R. R., and Abumrad, N.

Renal cAMP and 1,25(OH)2D3 synthesis in estrogen-treated chick embryos and hens.

Onset of sexual maturity in female chickens or administration of estrogen to mature males or to juveniles of either sex results in increased parathyroid hormone (PTH)-dependent adenylate cyclase activity and increased 25-hydroxyvitamin D3-1-hydroxylase activity in kidney. The relationship between estrogen-mediated alterations of these two enzyme systems was investigated in embryonic and mature, egg-laying chickens treated in vivo with 17 beta-estradiol (E2). Basal and PTH- and forskolin-stimulated adenylate cyclase activity in kidney plasma membrane preparations was not affected by E2 treatment of 19-day-old chick embryos or of 41-wk-old egg-laying females.

Effects of forskolin on bone. Stimulation of cyclic AMP accumulation and calcium efflux from chick embryo tibiae in organ culture.

Forskolin (FSK), an activator of adenylate cyclase in many tissues, was investigated to determine its potential for studying cyclic AMP-mediated, physiological events in bone. Tibiae from 14-day chick embryos were cultured in modified Eagle's medium for up to 8 hr, and the cyclic AMP and Ca2+ efflux responses to parathyroid hormone (PTH) and FSK were observed. FSK caused a concentration-related increase in tissue cyclic AMP, with 10(-6) M FSK producing a cyclic AMP response of similar magnitude to that elicited by 10(-6) M PTH (1-34).