Kir Channel Molecular Physiology, Pharmacology, and Therapeutic Implications.

For the past two decades several scholarly reviews have appeared on the inwardly rectifying potassium (Kir) channels. We would like to highlight two efforts in particular, which have provided comprehensive reviews of the literature up to 2010 (Hibino et al., Physiol Rev 90(1):291-366, 2010; Stanfield et al., Rev Physiol Biochem Pharmacol 145:47-179, 2002). In the past decade, great insights into the 3-D atomic resolution structures of Kir channels have begun to provide the molecular basis for their functional properties. More recently, computational studies are beginning to close the time domain gap between in silico dynamic and patch-clamp functional studies. The pharmacology of these channels has also been expanding and the dynamic structural studies provide hope that we are heading toward successful structure-based drug design for this family of K+ channels. In the present review we focus on placing the physiology and pharmacology of this K+ channel family in the context of atomic resolution structures and in providing a glimpse of the promising future of therapeutic opportunities.

Catalpol inhibits LPS plus IFN-γ-induced inflammatory response in astrocytes primary cultures.

A large body of evidence suggests that the inflammatory reaction plays an important role in the pathogenesis of neurodegenerative diseases. Our previous studies described the neuroprotective effects of catalpol in lipopolysaccharide (LPS)-induced inflammatory models, in which catalpol was shown to prevent mesencephalic neuron death and ameliorate cognitive ability animals. To further investigate the protective effect and underlying mechanism of catalpol, astrocytes were pretreated with low (0.1mM) and high dose (0.5mM) catalpol for 1h prior to LPS plus interferon-γ stimulation. Biochemical analyses showed that NO and ROS production and iNOS activity were significantly reduced by catalpol. Data at transcriptional level also demonstrated that catalpol potently attenuated gene expressions involved in inflammation, such as iNOS, COX-2 and TLR4. In addition, our exploration further revealed that the suppressive action of catalpol on inflammation was mediated via inhibiting nuclear factor-κB (NF-κB) activation. Collectively, these results suggest that catalpol can exert inhibitory effects on the inflammatory reaction in astrocytes and that inactivation of NF-κB could be the major determinant for its anti-inflammatory mechanism. Therefore, catalpol may potentially be a highly effective therapeutic agent in treating neurodegenerative diseases associated with inflammation.

Alterations in gene expression in response to compression of nucleus pulposus cells.

BACKGROUND CONTEXT: It is clear that mechanical forces are involved in initiating disc degeneration but also have the potential to exert beneficial effects. However, the signaling pathways initiated by mechanical stress and thresholds for these responses have not been elucidated. We have developed a metabolically active compression system with the advantages of having the ability to test cells in vitro as well as within their native matrix and control exposure to environmental factors. We hypothesized that nucleus pulposus cells would respond to compressive stress with different thresholds for alterations in catabolic and anabolic gene expression. PURPOSE: The purpose of the study was to establish the utility of a novel compression chamber and examine the effects of various magnitudes and durations of compression on nucleus pulposus inflammatory, catabolic, and anabolic gene expression. STUDY DESIGN: In vitro controlled examination of intervertebral disc cell responses to compression. METHODS: A chamber capable of imparting 0 to 20 MPa of hydrostatic compression onto nucleus pulposus cells was fabricated. Healthy rabbit nucleus pulposus cells were cultured in alginate beads and exposed to static compression at 0.7, 2, and 4 MPa for 4 or 24 hours. Gene expression analysis (real-time polymerase chain reaction) was performed to compare markers of inflammation (inducible nitric oxide synthase, cyclooxygenase-2), matrix catabolism (matrix metalloproteinase-3), and anticatabolic/anabolic metabolism (tissue inhibitor of metalloproteinase-1, aggrecan) in control and compressed cells. RESULTS: Compression resulted in magnitude- and duration-dependent changes in gene expression. Increasing magnitudes showed more anticatabolic gene expression changes, whereas increasing duration resulted in increases in procatabolic gene expression. CONCLUSION: These data demonstrate favorable effects of compression in relation to genes involved in matrix homeostasis and procatabolic gene expression in response to sustained loading levels, consistent with traumatic effects. These data provide an improved understanding of how compression affects cell signaling, which has the potential to be exploited to initiate repair and prevent matrix breakdown.

Identification of susceptibility loci for scoliosis in FIS families with triple curves.

The triple curve pattern (three lateral curvatures of equal severity) has been recognized as a distinct and unique clinical subtype of scoliosis. As part of a large study of familial idiopathic scoliosis (FIS), a subset of five families with a triple curve pattern (at least one member of each family having a triple curve) was evaluated to determine if this curve pattern was linked to any of the markers previously genotyped as part of the STRP-based previous linkage screen. Model independent linkage analysis (SIBPAL, v4.5) of the initial genomic screen identified candidate regions on chromosomes 6 and 10 when FIS was analyzed both as qualitative and quantitative traits in single- and multipoint linkage analyses. Additional fine mapping analyses of this subgroup with SNPs corroborated the findings in these regions (P < 0.001). These regions have been previously linked to FIS, however, this is the first time these regions have been implicated in a clinically well-defined subgroup and may suggest a unique genetic etiology for the formation of a triple curve.

Light-scattering and ultrastructure of healed penetrating corneal wounds.

PURPOSE: To investigate quantitatively for the first time the relationship between light-scattering and ultrastructure of semitransparent scars resulting from penetrating wounds in rabbit cornea. METHODS: Penetrating wounds, 2 mm in diameter, were made in the central cornea and allowed to heal for 3.6 to 4.5 years at which time the rabbits were killed. The scar and cornea thickness outside the scar were measured using ultrasonic pachymetry. Corneas were excised immediately and their transmissivity was measured from 400 to 700 nm. The tissue was then prepared for transmission electron microscopy. Transmission electron micrographs (TEMs) were analyzed to determine fibril positions and radii. Scattering was calculated using the direct summation of fields (DSF) METHOD: RESULTS: Scar thickness averaged 0.26 +/- 0.04 mm, and the scars were flat. Thickness outside the scars averaged 0.40 +/- 0.04 mm. Three scars were moderately transparent, five were less transparent, and one was much less transparent. The wavelength dependence of the measured total scattering cross- section was indicative of the presence of voids (lakes) in the collagen fibril distribution, and lakes were evident in the TEMs. The images showed enlarged fibrils and some showed bimodal distributions of fibril diameters. Calculated scattering was characteristic of that expected from regions containing lakes-a finding consistent with the scattering measurements. CONCLUSIONS: Despite the long healing time, these scars remained highly scattering. A combination of lakes, disordered fibril distributions, and a significant population of enlarged fibrils can explain the scattering. A possible cellular contribution cannot be ruled out.

Sensory versus motor information in the control of predictive saccade timing.

Humans readily make predictive saccades to periodic alternating targets. This predictive behavior depends on internal monitoring of timing error of past saccades in order to determine the time of initiation of future saccades; our earlier studies have confirmed this by finding correlations between latencies of consecutive predictive saccades. It is natural to consider that timing error is determined by visual detection of the difference between the time the target appears and the time the eyes arrive at the target; this in turn implies that saccades must actually be produced in order for their timing errors to be determined and predictive saccade timing to be established. We tested this hypothesis by having subjects view alternating visual targets while fixating a central target in order to eliminate saccade production. After six alternating target presentations, subjects began tracking the alternating targets. Tracking performance was assessed with an error measure that compared saccade latency and inter-saccade interval with desired values (zero and inter-stimulus interval, respectively). Errors in this Prior Viewing paradigm were compared to those from a conventional De Novo paradigm in which saccades began as soon as the alternating targets were presented. Saccades under Prior Viewing reached a low-error steady predictive state more rapidly than under De Novo tracking. The initial saccade under Prior Viewing had a higher latency than the others, suggesting that this saccade was reactive even though the paradigm is predictable; other reasons for this higher latency include time to disengage from the fixation target and time required to pre-program the initial set of saccades. The results show that visual detection of timing error from an actual motor act (saccades) is not necessary to establish predictive saccadic pacing: sensory-only information from viewing the moving targets can help to establish this predictive state.