Molecular symmetry change of perfluoro-n-alkanes in 'Phase I' monitored by infrared spectroscopy.

Phase diagram of polytetrafluoroethylene (PTFE) comprises four regions. Phases II and IV are characterized by twisted perfluoroalkyl (R) chains having different twisting rate of 13/6 and 15/7, respectively, while Phase III is characterized by a planer trans-zigzag molecular skeleton like a normal alkyl chain. These are confirmed by X-ray and electron diffraction and have already been established.

Effect of quick simple exercise on non-specific low back pain in Japanese workers: a randomized controlled trial.

Background: We designed a quick simple exercise program that can be performed in a short period of time in real-world occupational health settings and investigated the effects of three months of program implementation on non-specific low back pain (NSLBP).

Methods: Participants were 136 individuals working in the manufacturing industry. The quick simple exercise program was designed to be doable in three minutes and consisted of two exercises: a hamstring stretch and a lumbar spine rotation with forward, backward, and lateral flexion.

Reliable identification of cardiac conduction abnormalities in drug discovery using automated patch clamp II: Best practices for Nav1.5 peak current in a high throughput screening environment.

Introduction: For reliable identification of cardiac safety risk, compounds should be screened for activity on cardiac ion channels in addition to hERG, including Na1.5 and Ca1.2.

Mechanism of hERG inhibition by gating-modifier toxin, APETx1, deduced by functional characterization.

Background: Human ether-à-go-go-related gene potassium channel 1 (hERG) is a voltage-gated potassium channel, the voltage-sensing domain (VSD) of which is targeted by a gating-modifier toxin, APETx1. APETx1 is a 42-residue peptide toxin of sea anemone Anthopleura elegantissima and inhibits hERG by stabilizing the resting state. A previous study that conducted cysteine-scanning analysis of hERG identified two residues in the S3-S4 region of the VSD that play important roles in hERG inhibition by APETx1.

Reliable identification of cardiac liability in drug discovery using automated patch clamp: Benchmarking best practices and calibration standards for improved proarrhythmic assessment.

Introduction: Screening compounds for activity on the hERG channel using patch clamp is a crucial part of safety testing. Automated patch clamp (APC) is becoming widely accepted as an alternative to manual patch clamp in order to increase throughput whilst maintaining data quality. In order to standardize APC experiments, we have investigated the effects on IC values under different conditions using several devices across multiple sites.

Automated Patch Clamp Meets High-Throughput Screening: 384 Cells Recorded in Parallel on a Planar Patch Clamp Module.

We have developed an automated patch clamp module for high-throughput ion channel screening, recording from 384 cells simultaneously. The module is incorporated into a laboratory pipetting robot and uses a 384-channel pipettor head for application of cells and compounds. The module contains 384 amplifier channels for fully parallel recordings using a digital amplifier.

Electrophysiological analysis of mammalian cells expressing hERG using automated 384-well-patch-clamp.

Background: An in vitro electrophysiological assay system, which can assess compound effects and thus show cardiotoxicity including arrhythmia risks of test drugs, is an essential method in the field of drug development and toxicology.

Methods: In this study, high-throughput electrophysiological recordings of human embryonic kidney (HEK 293) cells and Chinese hamster ovary (CHO) cells stably expressing human ether-a-go-go related gene (hERG) were performed utilizing an automated 384-well-patch-clamp system, which records up to 384 cells simultaneously. hERG channel inhibition, which is closely related to a drug-induced QT prolongation and is increasing the risk of sudden cardiac death, was investigated in the high-throughput screening patch-clamp system.

Effect of terfenadine and pentamidine on the HERG channel and its intracellular trafficking: combined analysis with automated voltage clamp and confocal microscopy.

The effects of terfenadine and pentamidine on the human ether-a-go-go related gene (hERG) channel current and its intracellular trafficking were evaluated. Green fluorescent protein (GFP)-linked hERG channels were expressed in HEK293 cells, and the membrane current was measured by an automated whole cell voltage clamp system. To evaluate drug effects on channel trafficking to the cell membrane, the fraction of channel present on the cell membrane was quantified by current measurement after drug washout and confocal microscopy.

A novel DCTN1 mutation with late-onset parkinsonism and frontotemporal atrophy.

Background: Depression, parkinsonism, and hypoventilation (Perry syndrome) or familial motor neuron disease have been linked to mutations in dynactin P150(Glued) (DCTN1).

Methods: We employed genealogic, clinical, neurologic, and MRI investigations, as well as analysis of genes implicated in parkinsonism. Cellular transfection, immunocytochemistry, and immunoprecipitation analysis of wild-type (WT) and mutant DCTN1 were also performed.

Degeneration and dysfunction of retinal neurons in acute ocular hypertensive rats: involvement of calpains.

Purpose: Retinal ischemic diseases primarily lead to damage of the inner retinal neurons. Electrophysiological studies also suggest impairment of the inner retinal neurons. Our recent studies with acute ocular hypertensive rats confirmed damage predominantly in the inner retinal layer along with the ganglion cell layer, changes that are ameliorated by the calpain inhibitor SNJ-1945.

Effects of the antitussive drug cloperastine on ventricular repolarization in halothane-anesthetized guinea pigs.

Cloperastine is an antitussive drug, which can be received as an over-the-counter cold medicine. The chemical structure of cloperastine is quite similar to that of the antihistamine drug diphenhydramine, which is reported to inhibit hERG K⁺ channels and clinically induce long QT syndrome after overdose. To analyze its proarrhythmic potential, we compared effects of cloperastine and diphenhydramine on the hERG K⁺ channels expressed in HEK293 cells.

A novel calpain inhibitor, ((1S)-1-((((1S)-1-Benzyl-3-cyclopropylamino-2,3-di-oxopropyl)amino)carbonyl)-3-methylbutyl)carbamic acid 5-methoxy-3-oxapentyl ester (SNJ-1945), reduces murine retinal cell death in vitro and in vivo.

We examined whether ((1S)-1-((((1S)-1-benzyl-3-cyclopropylamino-2,3-di-oxopropyl)amino)carbonyl)-3-methylbutyl)carbamic acid 5-methoxy-3-oxapentyl ester (SNJ-1945), a new orally available calpain inhibitor, might reduce retinal cell death in vivo and/or in vitro. Retinal cell damage was induced in vivo in mice by intravitreal injection of N-methyl-d-aspartate (NMDA), and SNJ-1945 was intraperitoneally or orally administered twice. NMDA-induced calpain activity (measured as the cleaved products of alpha-spectrin) and its substrate, p35 (a neuron-specific activator for cyclin-dependent kinase 5), in the retina were examined by immunoblotting.

Comparison of the L-glutamate level in mouse hippocampal slices under tetraethylammonium chloride stimulation as measured with a glass capillary sensor and a patch sensor.

The concentration level of L-glutamate released from the region CA1 of mouse hippocampal slices under tetraethylammonium chloride (TEA) stimulation was measured by two independent methods, i.e., a glass capillary-based enzyme sensor and a patch sensor, and compared with each other for different slice preparations.

Implantation of a glass capillary-based enzyme electrode in mouse hippocampal slices for monitoring of L-glutamate release.

A glass capillary-based enzyme electrode (tip size approximately 10 microm) was implanted in the target neuronal region, i.e., dentate gyrus (DG) or cornu ammonis 1 (CA1), of acute brain slices at a depth of approximately 10 microm from the slice surface in order to allow the monitoring of chemical stimulant-induced changes in L-glutamate levels.

Aqueous humor dynamics associated with the phorbol ester-induced decrease in intraocular pressure in the rabbit.

Purpose: To determine the effects of injection of the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) into the anterior chamber of the eye on intraocular pressure (IOP) and aqueous humor dynamics.

Methods: IOP was measured for 24 h after intracameral injection of PMA (3 to 50 pmol) in unanesthetized rabbits. Aqueous humor dynamics (aqueous flow, total outflow facility, and uveoscleral outflow) were determined approximately 6 h after injection of 50 pmol of PMA in animals pretreated with indomethacin.

Contribution of calpains to photoreceptor cell death in N-methyl-N-nitrosourea-treated rats.

The purpose of the present study was to determine if proteolysis by the calcium-dependent enzyme calpains (EC 3.4.22.

Presence of calpain-induced proteolysis in retinal degeneration and dysfunction in a rat model of acute ocular hypertension.

The purpose of this study was to determine if calpain-induced proteolysis was associated with retinal degeneration or dysfunction in the rat acute ocular hypertensive model. Acute glaucoma was produced by elevation of IOP to 120 mm Hg for 1 hr. Retinal degeneration was evaluated by H&E staining and apoptosis was determined by TUNEL staining in histologic sections of retina.

Involvement of cyclooxygenase-2 in rat models of conjunctivitis.

Purpose: Using two animal models to determine which isoform of cyclooxygenase (COX), constitutive COX-1 or inducible COX-2, is involved in the progression of anterior ocular inflammation.

Methods: Lambda-carrageenan (500 mg/eye) or bacterial lipopolysaccharide (LPS; 3 mg/eye) was injected into rat conjunctiva to induce conjunctivitis. Vascular permeability in inflamed conjunctiva was measured by uptake of systemic Evans blue.

Novel pathway for utilization of cyclopropanecarboxylate by Rhodococcus rhodochrous.

A new strain isolated from soil utilizes cyclopropanecarboxylate as the sole source of carbon and energy and was identified as Rhodococcus rhodochrous (H. Nishihara, Y. Ochi, H.