Annexin A5 regulates surface αvß5 integrin for retinal clearance phagocytosis.

Diurnal clearance phagocytosis by the retinal pigment epithelium (RPE) is a conserved efferocytosis process whose binding step is mediated by αvß5 integrin receptors. Two related annexins, A5 (ANXA5) and A6 (ANXA6), share an αvß5 integrin-binding motif. Here, we report that ANXA5, but not ANXA6, regulates the binding capacity for spent photoreceptor outer segment fragments or apoptotic cells by fibroblasts and RPE. Similar to αvß5-deficient RPE, ANXA5-/- RPE in vivo lacks the diurnal burst of phagocytosis that follows photoreceptor shedding in wild-type retina. Increasing ANXA5 in cells lacking αvß5 or increasing αvß5 in cells lacking ANXA5 does not affect particle binding. Association of cytosolic ANXA5 and αvß5 integrin in RPE in culture and in vivo further supports their functional interdependence. Silencing ANXA5 is sufficient to reduce levels of αvß5 receptors at the apical phagocytic surface of RPE cells. The effect of ANXA5 on surface αvß5 and on particle binding requires the C-terminal ANXA5 annexin repeat but not its unique N-terminus. These results identify a novel role for ANXA5 specifically in the recognition and binding step of clearance phagocytosis, which is essential to retinal physiology.This article has an associated First Person interview with the first author of the paper.

Neural retina and MerTK-independent apical polarity of alphavbeta5 integrin receptors in the retinal pigment epithelium.

The apical plasma membrane domain of retinal pigment epithelial (RPE) cells in the eye faces the outer segment portions of rods and cones and the interphotoreceptor matrix in the subretinal space. Two important receptor-mediated interactions between the apical surface of the retinal pigment epithelium (RPE) and adjacent photoreceptors are adhesion ensuring outer segment alignment and diurnal phagocytosis of shed outer segment fragments contributing to outer segment renewal. Both depend on the apical distribution of the integrin family adhesion receptor alphavbeta5 as lack of alphavbeta5 in mice causes weakened retinal adhesion and asynchronous phagocytosis. With age, lack of alphavbeta5 leads to accumulation of harmful lipofuscin in the RPE and to vision loss. Here, we discuss three different possible mechanisms that could generate the exclusive apical distribution of alphavbeta5 integrin receptors in the RPE. (1) alphavbeta5 could be apical in the RPE because RPE attachment to neural retina generally or alphavbeta5 ligands specifically in the subretinal space stabilize apical but not basolateral alphavbeta5 surface receptors. (2) alphavbeta5 could be apical in the RPE because it resides in a complex with other components of the phagocytic machinery that assembles at the apical, phagocytic surface of the RPE. (3) alphavbeta5 could be apical due to mechanisms intrinsic to this receptor protein and specifically to its beta5 integrin subunit.

Proteomic profiling of murine oocyte maturation.

In an effort to better understand oocyte function, we utilized two-dimensional (2D) electrophoresis and mass spectrometry to identify proteins that are differentially expressed during murine oocyte maturation. Proteins from 500 germinal vesicle (GV) and metaphase II-(MII) arrested oocytes were extracted, resolved on 2D electrophoretic gels, and stained with silver. Analysis of the gels indicated that 12 proteins appeared to be differentially expressed between the GV and MII stage. These proteins were then cored from the 2D gels and identified by mass spectrometry as: transforming acidic coiled-coil protein 3 (TACC3), heat shock protein 105 (HSP105), programmed cell death six-interacting protein (PDCD6IP), stress-inducible phosphoprotein (STI1), importin alpha2, adenylsuccinate synthase (ADDS), nudix, spindlin, lipocalin, lysozyme, translationally controlled tumor protein (TCTP), and nucleoplasmin 2 (NPM2). Interestingly, PDCD6IP, importin alpha2, spindlin, and NPM2 appear slightly larger in mass and more acidic on the MII oocyte gel compared to the GV oocyte gel, suggesting that they may be post-translationally modified during oocyte maturation. Given NPM2 is an oocyte-restricted protein, we chose to further investigate its properties during oocyte maturation and preimplantation development. Real-Time RT-PCR showed that NPM2 mRNA levels rapidly decline at fertilization. Indirect immunofluorescence analysis showed that, with the exception of cortical localization in MII-arrested oocytes, NPM2 is localized to the nucleus of both GV stage oocytes and all stages of preimplantation embryos. We then performed one-dimensional (1D) western blot analysis of mouse oocytes and preimplantation embryos and found that, as implicated by the 2D gel comparison, NPM2 undergoes a phosphatase-sensitive electrophoretic mobility shift during the GV to MII transition. The slower migrating NPM2 form is also present in pronuclear embryos but by the two-cell stage, the majority of NPM2 exists as the faster migrating form, which persists to the blastocyst stage.

Increased beat-to-beat QT variability in patients with congestive cardiac failure.

BACKGROUND: QT interval on the surface electrocardiogram reflects the time for repolarization of myocardium. Prolongation of rate-corrected QT interval, QTc is strongly associated with sudden cardiac death. Recent studies using novel techniques on beat-to-beat QT interval variability have shown that an increase in QT interval variability is associated with increased sympathetic activity and is a predictor of sudden cardiac death. We studied QT variability in patients with congestive cardiac failure, as it is associated with an increase in cardiac sympathetic activity and also sudden death. METHODS AND RESULTS: We compared beat-to-beat heart rate and QT interval data in 2 3 patients with congestive cardiac failure and 19 age-matched normal controls. The electrocardiographic data were acquired in lead II configuration at a sampling rate of 1000 Hz. Heart rate variability was found to be significantly lower while QT variability measures were significantly higher in patients compared to controls. QTvi (a common log ratio of QT variability normalized for mean QT interval squared divided by heart rate variability normalized for mean heart rate squared) was also significantly higher in patients compared to controls. Clinical improvement in some of these patients is associated with a decrease in QTvi, due mainly to an increase in cardiac vagal function. CONCLUSIONS: Our results suggest a decrease in cardiac vagal and an increase in cardiac sympathetic functions in patients with congestive cardiac failure. QTvi may prove to be a useful surrogate end point to evaluate treatment effect in these patients.

Beat-to-beat heart rate and QT variability in patients with congestive cardiac failure: blunted response to orthostatic challenge.

BACKGROUND: Congestive cardiac failure is associated with increased sympathetic activity and impaired baroreflex function. We sought to test the hypothesis that these patients also have blunted response of beat-to-beat QT interval variability during orthostatic challenge. METHODS: We compared beat-to-beat heart rate and QT interval data in 17 patients with congestive cardiac failure and 17 age-matched normal controls in supine normal breathing, supine controlled breathing, and standing controlled breathing conditions. The ECG data were acquired in lead II configuration at a sampling rate of 1000 Hz. RESULTS: Supine controlled breathing was associated with an increase in spectral HF power (0.15-0.5 Hz) of HR and QT interval time series compared to spontaneous breathing condition only in controls. While there were significant changes in HR, HR LF power, HR LF/HF ratios, and QT variability measures in standing posture in controls, there were no such changes in patients. CONCLUSIONS: This impairment of postural changes of HR variability is most likely due to an impaired baroreceptor function in patients with congestive heart failure. The etiology of this is likely due to an increased cardiac sympathetic and a decreased vagal function. However, the relationship of postural changes in beat-to-beat QT interval variability and baroreflex need further investigation.

Linear and nonlinear measures of blood pressure variability: increased chaos of blood pressure time series in patients with panic disorder.

Arterial blood pressure (BP) variability increases progressively with the development of hypertension and an increase in BP variability is associated with end organ damage and cardiovascular morbidity. On the other hand, a decrease in heart rate (HR) variability is associated with significant cardiovascular mortality. There is a strong association between cardiovascular mortality and anxiety. Several previous studies have shown decreased HR variability in patients with anxiety. In this study, we investigated beat-to-beat variability of systolic and diastolic BP (SBP and DBP) in normal controls and patients with panic disorder during normal breathing and controlled breathing at 12, and 20 breaths per minute using linear as well as nonlinear techniques. Finger BP signal was obtained noninvasively using Finapres. Standing SBPvi and DBP BPvi (log value of BP variance corrected for mean BP divided by HR variance corrected for mean HR) were significantly higher in patients compared to controls. Largest Lyapunov exponent (LLE) of SBP and DBP, a measure of chaos, was significantly higher in patients in supine as well as standing postures. The ratios of LLE (SBP/HR) and LLE (DBP/HR) were also significantly higher (P<.001) in patients compared to controls. These findings further suggest dissociation between HR and BP variability and a possible relative increase in sympathetic function in anxiety. This increase in BP variability may partly explain the increase in cardiovascular mortality in this group of patients.

Approximate entropy of symptoms of mood: an effective technique to quantify regularity of mood.

OBJECTIVES: Several psychiatric conditions are associated with lability of affect. In this study, we investigated regularity of mood using APEN (Approximate Entropy) on daily subjective ratings using a Visual Analog Scale with 11 items pertaining to mood. METHODS: APEN was applied to the data in a double-blind placebo controlled investigation on the effects of fluoxetine (n=19), pemoline (n=18) and placebo (n=20) in normal controls. These subjects rated their subjective feelings daily at the end of each day. We analysed 56 data point time series (each value was obtained on each day) after the three experimental conditions. RESULTS: While the mean or the SD of all the 56 points was not significantly different among the three conditions, APEN was highly and significantly lower for pemoline compared with fluoxetine and placebo. There was no significant correlation between average APEN and mean or SD (standard deviation). The one symptom that explained most of this difference among the groups after drug administration was the feeling of 'happiness'. CONCLUSIONS: This result indicates that there was a more consistent subjective sense of happiness during the 8-week period of pemoline administration compared with the other two drugs. Though this study was not designed to address the efficacy of mood stabilizing drugs, such daily subjective ratings may be useful in future studies that evaluate the stability of mood. APEN has been used in several different fields of research with small data sets and this study extends its possible use to evaluate changes in mood in certain populations such as patients with bipolar disorders.

Major depression with ischemic heart disease: effects of paroxetine and nortriptyline on measures of nonlinearity and chaos of heart rate.

Depression is associated with increased cardiovascular mortality in patients with preexisting cardiac illness. A decrease in cardiac vagal function as suggested by a decrease in heart rate variability (HRV) or heart period variability has been linked to sudden death in patients with cardiac disease as well as in normal controls. Recent studies have shown decreased vagal function in cardiac patients with depression as well as in depressed patients without cardiac illness. In this study, we compared 20 h awake and sleep heart period nonlinear measures using quantification of nonlinearity and chaos in two groups of patients with major depression and ischemic heart disease (mean age 59-60 years) before and after 6 weeks of treatment with paroxetine or nortriptyline. Patients received paroxetine, 20-30 mg/day or nortriptyline targeted to 190-570 nmol/l for 6 weeks. For HRV analysis, 24 patients were included in the paroxetine treatment study and 20 patients in the nortriptyline study who had at least 20000 s of awake data. The ages of these groups were 60.4 +/- 10.5 years for paroxetine and 60.8 +/- 13.4 years for nortriptyline. There was a significant decrease in the largest Lyapunov exponent (LLE) after treatment with nortriptyline but not paroxetine. There were also significant decreases in nonlinearity scores on S(netPR) and S(netGS) after nortriptyline, which may be due to a decrease in cardiac vagal modulation of HRV. S(netGS) and awake LLE were the most significant variables that contributed to the discrimination of postparoxetine and postnortriptyline groups even with the inclusion of time and frequency domain measures. These findings suggest that nortriptyline decreases the measures of chaos probably through its stronger vagolytic effects on cardiac autonomic function compared with paroxetine, which is in agreement with previous clinical and preclinical reports. Nortriptyline was also associated with a significant decrease in nonlinearity scores, which may be due to anticholinergic and/or sympatholytic effects. As depression is associated with a strong risk factor for cardiovascular mortality, one should be careful about using any drug that adversely affects cardiac vagal function.