Regional differences in store-operated Ca2+ entry in the epithelium of the intact human lens

An elevated level of Ca2+ is an important factor in cataract, yet precisely how Ca2+ enters the lens is unknown. Lens epithelial cells contain a range of G-protein–coupled receptors and receptor tyrosine kinases that induce increases in intracellular Ca2+. Receptor-associated Ca2+ influx is, therefore, likely to be an important route for Ca2+ influx to the lens. The authors investigated stimulated and passive Ca2+ influx in in situ human lens epithelium. Ca2+ changes in equatorial (E) and central anterior (CA) epithelial cells were monitored with the use of a Ca2+ indicator (Fluo4) and confocal microscopy. Gene expression was monitored by RT-PCR and immunoblotting. Adenosine triphosphate (ATP) induced Ca2+ responses that were smaller in CA than E. Ca2+ store depletion, using ATP (100 µM) or thapsigargin (1 µM), revealed greater relative store capacity and Ca2+ influx in E. Ca2+ influx was blocked by La3+ (0.5 µM) in both regions. Unstimulated Ca2+ influx was greater in E than CA. Greater expression of Orai1 and STIM1 was detected in E than in CA. Greater Ca2+ store capacity and Ca2+ influx in E compared with CA reflects underlying differences in proliferation and differentiation between the regions. The relatively small resting Ca2+ influx in CA epithelium suggests that store-operated Ca2+ entry (SOCE) is the main route of Ca2+ influx in these cells. Greater resting influx and SOCE in E cells suggests that these are a major route for Ca2+ influx into the lens. Increased expression of Orai1 and STIM1 in E could account for the differences in Ca2+ entry. Receptor activation will modulate Ca2+ influx, and inappropriate activity may contribute to cortical cataract

Intrusive effects of semantic information on visual selective attention

Every object contains semantic information in extension to its low-level properties. It is well documented that such information biases attention when it is necessary for an ongoing task. However, whether semantic relationships influence attentional selection when they are irrelevant to the ongoing task remains an open question. The ubiquitous nature of semantic information suggests that it could bias attention even when these properties are irrelevant. In the present study, three objects appeared on screen, two of which were semantically related. After a varying time interval, a target or distractor appeared on top of each object. The objects’ semantic relationships never predicted target location. Despite this, a semantic bias on attentional allocation was observed with an initial, transient bias to semantically related objects. Further experiments demonstrated that this effect was contingent on the objects being attended: if an object never contained the target, it no longer exerted a semantic influence. In a final set of experiments, we demonstrate that semantic bias is robust and appears even in the presence of more predictive cues (spatial probability). The results suggest that as long as an object is attended, its semantic properties bias attention, even if it is irrelevant to an ongoing task and there are more predictive factors available

Intrusive effects of task-irrelevant information on visual selective attention: semantics and size

Attentional selection is a mechanism by which incoming sensory information is prioritized for further, detailed and more effective, processing. Given that attended information is privileged by the sensory system, understanding and predicting what information is granted prioritization becomes an important endeavor. It has been argued that salient events as well as information that is related to the current goal of the organism (i.e., task-relevant) receive such priority. Here, we propose that attentional prioritization is not limited to task-relevance, and discuss evidence showing that task-irrelevant, non-salient, high-level properties of unattended objects, namely object meaning and size, influence attentional allocation. Such intrusion of non-salient task-irrelevant high-level information points to the need to re-conceptualize and formally modify current models of attentional guidance

The Effect of GnRH at Time of Insemination on Initiation of LH Pulses and Subsequent Progesterone

Research has indicated that luteinizing hormone (LH) pulses play a vital role in corpus luteum (CL) formation and subsequent progesterone concentrations. Therefore, our objectives were to determine: 1) when LH pulses begin following onset of estrus, 2) the effect an injection of gonadotropin releasing hormone (GnRH) would have on initiation of LH pulses, and 3) the effect LH pulse initiation had on subsequent plasma progesterone concentrations. Cows were synchronized with the Select Synch + Controlled Internal Drug Releasing device (CIDR) protocol (d -7 100 μg GnRH and CIDR; d 0 25 mg prostaglandin (PG) and removal of CIDR; estrus detected with HeatWatch). Following detection in estrus, a jugular catheter was inserted in each cow (n = 10). Based on initiation of estrus, cows were allotted into two treatments: 1) GnRH given 12 h (12.5 ± 1.2 h) after the initiation of estrus (n = 5; 100 μg) and 2) Control (n = 5). Blood samples were collected at 15-min intervals for 6 h at 12 h (bleed 1), 26 h (bleed 2), 40 h (bleed 3), 54 h (bleed 4), and 68 h (bleed 5) after the onset of estrus. The interval from onset of estrus to bleed 1 and ovulation was similar between treatments. The GnRH cows tended to have a greater area under the LH curve for bleed 1 compared to control cows. No differences were detected in bleeds 2, 3, 4, or 5. Average concentration of LH for GnRH cows in bleed 1 tended to be greater than control. No differences were detected in bleeds 2, 3, 4, or 5. No differences were detected in pulse frequency between treatments in bleeds 1, 3, 4, or 5, but in bleed 2, control tended to have more pulses than GnRH (2.5 ± 0.5 vs 1.4 ± 0.4). The GnRH-treated cows tended to have greater subsequent progesterone concentrations; however, GnRH-treated cows that had no LH pulses during bleed 2 had lower progesterone concentrations than cows with pulses (control or GnRH). In summary, injecting cows with GnRH approximately 12 h after the onset of estrus tended to reduce LH pulses 26-32 h following initiation of estrus, and elimination of LH pulses between 26-32 h resulted in decreased concentrations of progesterone during the subsequent cycle

Neural Correlates of Suspiciousness and Interactions with Anxiety During Emotional and Neutral Word Processing

Suspiciousness is usually classified as a symptom of psychosis, but it also occurs in depression and anxiety disorders. Though how suspiciousness overlaps with depression is not obvious, suspiciousness does seem to overlap with anxious apprehension and anxious arousal (e.g., verbal iterative processes and vigilance about environmental threat). However, suspiciousness also has unique characteristics (e.g., concern about harm from others and vigilance about social threat). Given that both anxiety and suspiciousness have been associated with abnormalities in emotion processing, it is unclear whether it is the unique characteristics of suspiciousness or the overlap with anxiety that drive abnormalities in emotion processing. Event-related brain potentials were obtained during an emotion-word Stroop task. Results indicated that suspiciousness interacts with anxious apprehension to modulate initial stimulus perception processes. Suspiciousness is associated with attention to all stimuli regardless of emotion content. In contrast, anxious arousal is associated with a later response to emotion stimuli only. These results suggest that suspiciousness and anxious apprehension share overlapping processes, but suspiciousness alone is associated with a hyperactive early vigilance response. Depression did not interact with suspiciousness to predict response to emotion stimuli. These findings suggest that it may be informative to assess suspiciousness in conjunction with anxiety in order to better understand how these symptoms interact and contribute to dysfunctional emotion processing

Differential effects of tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 on atherosclerosis and monocyte/macrophage invasion

AIMS: MMPs contribute to atherosclerotic plaque progression and instability, but the relative potency of their endogenous tissue inhibitors of metalloproteinases (TIMPs) as protective factors has not been defined. We therefore investigated the impact of TIMP-1 and TIMP-2 knockout on atherosclerotic plaque burden and composition in apolipoprotein E-knockout (Apoe(−/−)) mice and studied the underlying effects on monocyte/macrophage behaviour. METHODS AND RESULTS: Analysis of brachiocephalic artery plaques revealed comparable atherosclerotic lesion areas between TIMP-1(−/−) Apoe(−/−) or TIMP-2(−/−) Apoe(−/−) double deficient mice and relevant age-matched, strain-matched Apoe(−/−) controls after 8 weeks of high-fat feeding. However, lesions from TIMP-2(−/−) Apoe(−/−) mice had higher levels of markers associated with plaque vulnerability, including increased macrophage: vascular smooth muscle cell ratios, larger necrotic core areas, reduced collagen contents, increased macrophage proliferation, and apoptosis frequencies, compared with TIMP-1(−/−)Apoe(−/−) and controls. In contrast, TIMP-1(−/−) Apoe(−/−) animals only had a significant reduction in vascular smooth muscle cell content compared with Apoe(−/−) controls. In vitro and in vivo findings implicated heightened monocyte/macrophage invasion in the detrimental effects observed on atherosclerotic plaque composition in TIMP-2(−/−) Apoe(−/−) mice. Moreover, TIMP-2 specifically decreased MMP-14-dependent monocyte/macrophage infiltration into sites of experimentally induced inflammation and established atherosclerotic lesions. CONCLUSION: Our data demonstrate that TIMP-2 plays a greater protective role than TIMP-1 during the pathogenesis of atherosclerosis, in part by suppressing MMP-14-dependent monocyte/macrophage accumulation into plaques

Disparate Effects of MMP and TIMP Modulation on Coronary Atherosclerosis and Associated Myocardial Fibrosis

Matrix metalloproteinase (MMP) activity is tightly regulated by the endogenous tissue inhibitors (TIMPs), and dysregulated activity contributes to extracellular matrix remodelling. Accordingly, MMP/TIMP balance is associated with atherosclerotic plaque progression and instability, alongside adverse post-infarction cardiac fibrosis and subsequent heart failure. Here, we demonstrate that prolonged high-fat feeding of apolipoprotein (Apo)e-deficient mice triggered the development of unstable coronary artery atherosclerosis alongside evidence of myocardial infarction and progressive sudden death. Accordingly, the contribution of select MMPs and TIMPs to the progression of both interrelated pathologies was examined in Apoe-deficient mice with concomitant deletion of Mmp7, Mmp9, Mmp12, or Timp1 and relevant wild-type controls after 36-weeks high-fat feeding. Mmp7 deficiency increased incidence of sudden death, while Mmp12 deficiency promoted survival, whereas Mmp9 or Timp1 deficiency had no effect. While all mice harboured coronary disease, atherosclerotic burden was reduced in Mmp7-deficient and Mmp12-deficient mice and increased in Timp1-deficient animals, compared to relevant controls. Significant differences in cardiac fibrosis were only observed in Mmp-7-deficient mice and Timp1-deficient animals, which was associated with reduced capillary number. Adopting therapeutic strategies in Apoe-deficient mice, TIMP-2 adenoviral-overexpression or administration (delayed or throughout) of a non-selective MMP inhibitor (RS-130830) had no effect on coronary atherosclerotic burden or cardiac fibrosis. Taken together, our findings emphasise the divergent roles of MMPs on coronary plaque progression and associated post-MI cardiac fibrosis, highlighting the need for selective therapeutic approaches to target unstable atherosclerosis alongside adverse cardiac remodelling while negating detrimental adverse effects on either pathology, with targeting of MMP-12 seeming a suitable target

Highly active and selective endopeptidases with programmed substrate specificities

A family of engineered endopeptidases has been created that is capable of cleaving a diverse array of peptide sequences with high selectivity and catalytic efficiency (kcat/KM > 104 M?1 s?1). By screening libraries with a selection-counterselection substrate method, protease variants were programmed to recognize amino acids having altered charge, size and hydrophobicity properties adjacent to the scissile bond of the substrate, including Glu?Arg, a specificity that to our knowledge has not been observed among natural proteases. Members of this artificial protease family resulted from a relatively small number of amino acid substitutions that (at least in one case) proved to be epistatic