Distal Events Research Articles

Percutaneous endoscopic gastrostomy and gastro-oesophageal reflux in neurologically impaired children

To investigate the effects of percutaneous endoscopic gastrostomy (PEG) feeding on gastro-oesophageal reflux (GOR) in a group of these children using combined intraluminal pH and multiple intraluminal impedance (pH/MII). Ten neurologically impaired children underwent 12 h combined pH/MII procedures at least 1 d before and at least 12 d after PEG placement. Prior to PEG placement (pre-PEG) a total of 183 GOR episodes were detected, 156 (85.2%) were non-acidic. After PEG placement (post-PEG) a total of 355 episodes were detected, 182 (51.3%) were non-acidic. The total number of distal acid reflux events statistically significantly increased post-PEG placement (pre-PEG total 27, post-PEG total 173, P = 0.028) and the mean distal pH decreased by 1.1 units. The distal reflux index therefore also significantly increased post-PEG [pre-PEG 0.25 (0-2), post-PEG 2.95 (0-40)]. Average proximal pH was lower post-PEG but the within subject difference was not statistically significant (P = 0.058). Median number of non-acid GOR, average reflux height, total acid clearance time and total bolus clearance time were all lower pre-PEG, but not statistically significant. PEG placement increases GOR episodes in neurologically impaired children.

CD3γ-independent pathways in TCR-mediated signaling in mature T and iNKT lymphocytes

Antigen recognition by T-lymphocytes through the T-cell antigen receptor, TCR–CD3, is a central event in the initiation of an immune response. CD3 proteins may have redundant as well as specific contributions to the intracellular propagation of TCR-mediated signals. However, to date, the relative role that each CD3 chain plays in signaling is controversial. In order to examine the roles of CD3γ chain in TCR signaling, we analyzed proximal and distal signaling events in human CD3γ−/− primary and Herpesvirus saimiri (HVS)-transformed T cells. Following TCR–CD3 engagement, certain early TCR signaling pathways (ZAP-70, ERK, p38 and mTORC2 phosphorylation, and actin polymerization) were comparable with control HVS-transformed T cells. However, other signaling pathways were affected, such TCRζ phosphorylation, indicating that the CD3γ chain contributes to improve TCR signaling efficiency and survival. On the other hand, CD3γ−/− primary invariant NKT cells (iNKT cells) showed a normal expansion in response to alpha-galactosylceramide (α-GalCer) and TCRVβ11bright iNKT cells were preferentially selected in this in vitro culture system, perhaps as a consequence of selective events in the thymus. Our results collectively indicate that a TCR lacking CD3γ can propagate a number of signals through the remaining invariant chains, likely the homologous CD3δ chain, which replaces it at the mutant TCR.

TNF Receptor-Associated Factor 3 Is Required for T Cell-Mediated Immunity and TCR/CD28 Signaling

We recently reported that TNFR-associated factor (TRAF)3, a ubiquitously expressed adaptor protein, promotes mature B cell apoptosis. However, the specific function of TRAF3 in T cells has remained unclear. In this article, we report the generation and characterization of T cell-specific TRAF3(-/-) mice, in which the traf3 gene was deleted from thymocytes and T cells. Ablation of TRAF3 in the T cell lineage did not affect CD4 or CD8 T cell populations in secondary lymphoid organs or the numbers or proportions of CD4(+),CD8(+) or double-positive or double-negative thymocytes, except that the T cell-specific TRAF3(-/-) mice had a 2-fold increase in FoxP3(+) T cells. In striking contrast to mice lacking TRAF3 in B cells, the T cell TRAF3-deficient mice exhibited defective IgG1 responses to a T-dependent Ag, as well as impaired T cell-mediated immunity to infection with Listeria monocytogenes. Surprisingly, we found that TRAF3 was recruited to the TCR/CD28 signaling complex upon costimulation and that TCR/CD28-mediated proximal and distal signaling events were compromised by TRAF3 deficiency. These findings provide insights into the roles played by TRAF3 in T cell activation and T cell-mediated immunity.

Deciphering Magma-Mixing Processes Using Zoned Titanite from the Ross of Mull Granite, Scotland

A textural and geochemical investigation of titanite from the Ross of Mull Granite, Scotland, reveals that zoning in trace elements strongly correlates with field evidence of magma-mixing processes within the pluton. Diorite enclaves contain a range of textures indicative of variable states of interaction with the host granite and suggest that a number of discrete mixing events have occurred in the pluton, both in situ and prior to emplacement at the present level. Single titanites within both the enclaves and the host are compositionally zoned and provide a detailed record of magmatic processes. Zoning includes concentric fine-scale oscillatory types and major discontinuities linked to dissolution events associated with changes in titanite stability. The zones reflect growth during changing melt chemistry and correlate with variations in rare earth element and high field strength element contents. Geothermometry using the Zr content of titanite indicates that titanite crystallization occurs at temperatures of c. 750–700°C. Titanites respond to changes in melt composition, oxygen fugacity and temperature caused by mixing and the introduction of fresh diorite magma into the granite host. Mixing processes recorded by the titanites include complete mixing of crystal-free melts, crystal transfer between partially crystallized magmas, melt segregation and transfer from crystalline enclaves and late-stage diffusive exchange. Titanite chemistry responds to local activity adjacent to sites of mixing but is also influenced by more distal magma-mixing events. In this respect titanite is capable of revealing many otherwise hidden events within the magma chamber.

The Serotonin Transporter Gene Length Polymorphism (5-HTTLPR) and Life Events: No Evidence for an Interaction Effect on Neuroticism and Anxious Depressive Symptoms

The finding of a significant gene by environment interaction effect on depression of the serotonin transporter length polymorphism (5-HTTLPR) and the Number of experienced Life Events (NLE) was not replicated in two large meta-analyses (Munafo et al., 2009; Risch et al., 2009). These meta-analyses have been criticized on the grounds that large studies that get most weight in meta-analyses have the poorest measurement quality of life events and, as a consequence, do not find an effect. Another issue is the time frame across which the NLE are measured. Proximal life events appear to be better predictors of depression than more distal events. We present the results of analyses of the 5-HTTLPR × NLE effect on anxious depression and neuroticism scores in a sample of 1,155 twins and their parents and siblings from 438 families. The interaction effect was tested separately for NLE experienced across the life span and NLE experienced in the past year. There was a significant main effect of NLE on anxious depression and neuroticism, especially when these were experienced in the past year. No interaction with 5-HTTLPR was found for NLE either experienced across the life span or across the past year. Our results support the two recent meta-analyses. Given recent insights from genome wide association studies, it seems more useful to focus on the joint effect of several genes, that are, for example, part of the same biological pathway, in interaction with the environment, than on one candidate gene.

The Effect of Homebound Status on Older Persons

To examine the effect of homebound status of older persons in Israel on mortality, mental health and function, future homebound status, and institutionalization. Cross-sectional and longitudinal analysis using existing data sets of a national survey. A national survey. The analyzed sample was drawn from a representative cohort of 1,191 older persons in Israel (mean age 83.1±5.3) in the first wave of the Cross-Sectional and Longitudinal Aging Study and 621 participants in the second wave of the study. Homebound status, health, function, environment, mental health, distal events, mortality, activities daily living (ADLs), instrumental activities of daily living (IADLs), Orientation-Memory-Concentration Test, and Center for Epidemiologic Studies Depression Scale. Mortality data were recorded from the Israeli National Population Registry. Homebound participants had a significantly higher risk of mortality than their non-homebound counterparts, even after controlling for background variables, health, and function (risk ratio=1.33, 95% confidence interval=1.08-1.63). In cross-sectional analysis, homebound status was related to depressed affect even after controlling for demographics, health, and function. In longitudinal analysis, homebound status predicted future depressed affect and ADL and IADL difficulties when controlling for demographics and health, but only IADL prediction was statistically significant when baseline levels of the outcome variable were entered into the regression. The results highlight the detrimental effects of homebound status, underscoring the importance of preventing this state, of interventions to assist those who are homebound, and of future research to examine the efficacy and coverage of services to this population.

Impaired insulin-induced site-specific phosphorylation of TBC1 domain family, member 4 (TBC1D4) in skeletal muscle of type 2 diabetes patients is restored by endurance exercise-training

Insulin-mediated glucose disposal rates (R(d)) are reduced in type 2 diabetic patients, a process in which intrinsic signalling defects are thought to be involved. Phosphorylation of TBC1 domain family, member 4 (TBC1D4) is at present the most distal insulin receptor signalling event linked to glucose transport. In this study, we examined insulin action on site-specific phosphorylation of TBC1D4 and the effect of exercise training on insulin action and signalling to TBC1D4 in skeletal muscle from type 2 diabetic patients. During a 3 h euglycaemic-hyperinsulinaemic (80 mU min⁻¹ m⁻²) clamp, we obtained M. vastus lateralis biopsies from 13 obese type 2 diabetic and 13 obese, non-diabetic control individuals before and after 10 weeks of endurance exercise-training. Before training, reductions in insulin-stimulated R (d), together with impaired insulin-stimulated glycogen synthase fractional velocity, Akt Thr³⁰⁸ phosphorylation and phosphorylation of TBC1D4 at Ser³¹⁸, Ser⁵⁸⁸ and Ser⁷⁵¹ were observed in skeletal muscle from diabetic patients. Interestingly, exercise-training normalised insulin-induced TBC1D4 phosphorylation in diabetic patients. This happened independently of increased TBC1D4 protein content, but exercise-training did not normalise Akt phosphorylation in diabetic patients. In both groups, training-induced improvements in insulin-stimulated R(d) (~20%) were associated with increased muscle protein content of Akt, TBC1D4, α2-AMP-activated kinase (AMPK), glycogen synthase, hexokinase II and GLUT4 (20-75%). Impaired insulin-induced site-specific TBC1D4 phosphorylation may contribute to skeletal muscle insulin resistance in type 2 diabetes. The mechanisms by which exercise-training improves insulin sensitivity in type 2 diabetes may involve augmented signalling of TBC1D4 and increased skeletal muscle content of key insulin signalling and effector proteins, e.g., Akt, TBC1D4, AMPK, glycogen synthase, GLUT4 and hexokinase II.

Genetic and Pharmacological Evaluation of Cathepsin S in a Mouse Model of Asthma

Cathepsin S (Cat S) is predominantly expressed in antigen-presenting cells and is up-regulated in several preclinical models of antigen-induced inflammation, suggesting a role in the allergic response. Prophylactic dosing of an irreversible Cat S inhibitor has been shown to attenuate pulmonary eosinophilia in mice, supporting the hypothesis that Cat S inhibition before the initiation of airway inflammation is beneficial in airway disease. In addition, Cat S has been shown to play a role in more distal events in the allergic response. To determine where Cat S inhibition may affect the allergic response, we used complementary genetic and pharmacological approaches to investigate the role of Cat S in the early and downstream allergic events in a murine model of antigen-induced lung inflammation. Cat S knockout mice did not develop ovalbumin-induced pulmonary inflammation, consistent with a role for Cat S in the development of the allergic response. Alternatively, wild-type mice were treated with a reversible, highly selective Cat S inhibitor in prophylactic and therapeutic dosing paradigms and assessed for changes in airway inflammation. Although both treatment paradigms resulted in potent Cat S inhibition, only prophylactic Cat S inhibitor dosing blocked lung inflammation, consistent with our findings in Cat S knockout mice. The findings indicate that although Cat S is up-regulated in allergic models, it does not appear to play a significant role in the downstream effector inflammatory phase in this model; however, our results demonstrate that Cat S inhibition in a prophylactic paradigm would ameliorate airway inflammation.

BIOSTRATINOMY OF THE LATE EDIACARAN PYRITIZED GAOJIASHAN LAGERSTATTE FROM SOUTHERN SHAANXI, SOUTH CHINA: IMPORTANCE OF EVENT DEPOSITS

Abstract The upper Ediacaran Gaojiashan Lagerstatte (551–541 Ma) in southern Shaanxi Province, South China, hosts a variety of soft-bodied or lightly biomineralized tubular fossils (Shaanxilithes, Conotubus, Gaojiashania, Sinotubulites, and Cloudina) and calcareous microfossils (Protolagena). This study focuses on the fossiliferous middle Gaojiashan Member at the Gaojiashan section, where pyritization of soft-bodied or lightly skeletonized tubular fossils is the primary mode of preservation. Integrated paleoecological, sedimentological, and taphonomic analysis shows that event deposition played an important role in the biostratinomy of the Gaojiashan Lagerstatte. Gaojiashan fossils, particularly pyritized fossils, preferentially occur in mm-thick, normally graded calcisiltite-siltstone layers, interpreted as distal event deposits, but are rarely present in calcilutite-mudstone beds. Two taphofacies are recognized in the middle Gaojiashan calcisiltites-siltstones, on the basis of biostratinomic data (e.g.,...

Adverse life events, area socio-economic disadvantage, and adolescent psychopathology: The role of closeness to grandparents in moderating the effect of contextual stress

The study, using data from 801 11–16-year-olds clustered in 68 schools across England and Wales, tested whether closeness to grandparents moderates the association between contextual stress and adolescent psychopathology and prosocial behavior, measured with the strengths and difficulties questionnaire (SDQ). Contextual stress was measured at both school area level (assessed with the index of multiple deprivation) and child level (assessed, as life stress, with the number of proximal and distal adverse life events experienced). At baseline, area stress (multiple deprivation) was unrelated to psychopathology (SDQ), and although both proximal (during the last 12 months) and distal (before the last 12 months) life stress was associated with broad and specific child psychopathology, the association with proximal life stress was stronger. Closeness to the most significant grandparent moderated both the effect of proximal life stress on hyperactivity and broad psychopathology, and the effect of the interaction between distal and proximal life stress on broad and externalizing psychopathology. These findings suggest that the role of grandparents deserves further attention in future investigations of the development of resilience in youth.

Wetland Vegetation in Manzala Lagoon, Nile Delta Coast, Egypt: Rapid Responses of Pollen to Altered Nile Hydrology and Land Use

The pollen record in a sediment core from Manzala lagoon on the Nile delta coastal margin of Egypt, deposited from ca. AD 1860 to 1990, indicates rapid coastal wetland vegetation responses to two primary periods of human activity. These are associated with artificially altered Nile hydrologic regimes in proximal areas and distal sectors located to ∼1200 km south of Manzala. Freshwater wetland plants that were dominant, such as Typha and Phragmites, decreased rapidly, whereas in the early 1900s, brackish water wetland species (e.g., Amaranthaceae) increased. This change occurred after closure of the Aswan Low Dam in 1902. The second major modification in the pollen record occurred in the early 1970s, after Aswan High Dam closure from 1965 to 1970, when Typha pollen abundance increased rapidly. Massive population growth occurred along the Nile during the 130 years represented by the core section. During this time, the total volume of lagoon water decreased because of conversion of wetland areas to agricultural land, and input of organic-rich sediment, sewage (municipal, agricultural, industrial), and fertilizer in Manzala lagoon increased markedly. Although the wetland plant community has continued to respond to increasingly intensified and varied human-induced pressures in proximal sectors, the two most marked changes in Manzala pollen best correlate with distal events (i.e., closure of the two dams at Aswan). The study also shows that the two major vegetation changes in Manzala lagoon each occurred less than 10 years after closure upriver of the Low and High dams that markedly altered the Nile regime from Upper Egypt to the coast.

Correlation versus causation in multisensory perception

Events are often perceived in multiple modalities. The co-occurring proximal visual and auditory stimuli events are mostly also causally linked to the distal event, which makes it difficult to evaluate whether learned correlation or perceived causation guides binding in multisensory perception. Piano tones are an interesting exception: They are associated with the act of the pianist striking keys, an event that is visible to the perceiver, but directly results from hammers hitting strings, an event that typically is not visible to the perceiver. We examined the influence of seeing the hammer or the keystroke on auditory temporal order judgments (TOJs). Participants judged the temporal order of a dog bark and a piano tone, while seeing the piano stroke shifted temporally relative to its audio signal. Visual lead increased "piano-first" responses in auditory TOJ, but more so if the associated keystroke was visible than if the sound-producing hammer was visible, even though both were equally visually salient. This provides evidence for a learning account of audiovisual perception.

A Quantitative Model of Transcriptional Regulation Reveals the Influence of Binding Location on Expression

Understanding the mechanistic basis of transcriptional regulation has been a central focus of molecular biology since its inception. New high-throughput chromatin immunoprecipitation experiments have revealed that most regulatory proteins bind thousands of sites in mammalian genomes. However, the functional significance of these binding sites remains unclear. We present a quantitative model of transcriptional regulation that suggests the contribution of each binding site to tissue-specific gene expression depends strongly on its position relative to the transcription start site. For three cell types, we show that, by considering binding position, it is possible to predict relative expression levels between cell types with an accuracy approaching the level of agreement between different experimental platforms. Our model suggests that, for the transcription factors profiled in these cell types, a regulatory site's influence on expression falls off almost linearly with distance from the transcription start site in a 10 kilobase range. Binding to both evolutionarily conserved and non-conserved sequences contributes significantly to transcriptional regulation. Our approach also reveals the quantitative, tissue-specific role of individual proteins in activating or repressing transcription. These results suggest that regulator binding position plays a previously unappreciated role in influencing expression and blurs the classical distinction between proximal promoter and distal binding events.

Area and family effects on the psychopathology of the Millennium Cohort Study children and their older siblings

To model and compare contextual (area and family) effects on the psychopathology of children nested in families nested in areas. Data from the first two sweeps of the UK's Millennium Cohort Study were used. The final study sample was 9,630 children clustered in 6,052 families clustered in 1,681 Lower-layer Super Output Areas. The mean age of the children at Sweep 2 was 4.96 (SD = 2.76) years. Contextual risk was measured at area level with the Index of Multiple Deprivation (Sweep 1), and at family level with the number of proximal (Sweep 2) and distal (Sweep 1) adverse life events experienced. Psychopathology was measured at Sweep 2 with the Strengths and Difficulties Questionnaire. At baseline, both proximal and distal family risk and area risk predicted broad psychopathology, although the most parsimonious was the proximal family risk model, and both the family-level and the area-level variability were significant. The area risk/broad psychopathology association remained significant even when family risk was controlled, but not when family socioeconomic status was controlled. The full model added parenting and paternal and maternal psychopathology. When parental qualifications were excluded from the family-level contextual controls the effect of area risk remained significant on both externalizing and internalizing psychopathology. The effect of area on child psychopathology operated via the socioeconomic characteristics of the child's family, not just the adverse characteristics of the neighbors. Multiple family risk predicted child psychopathology directly and independently, and not because it was associated with family socioeconomic status. Family socioeconomic status explained the association between area risk and broad psychopathology.

Predicting Successful Aging in a Population-Based Sample of Georgia Centenarians

Used a population-based sample (Georgia Centenarian Study, GCS), to determine proportions of centenarians reaching 100 years as (1) survivors (43%) of chronic diseases first experienced between 0–80 years of age, (2) delayers (36%) with chronic diseases first experienced between 80–98 years of age, or (3) escapers (17%) with chronic diseases only at 98 years of age or older. Diseases fall into two morbidity profiles of 11 chronic diseases; one including cardiovascular disease, cancer, anemia, and osteoporosis, and another including dementia. Centenarians at risk for cancer in their lifetime tended to be escapers (73%), while those at risk for cardiovascular disease tended to be survivors (24%), delayers (39%), or escapers (32%). Approximately half (43%) of the centenarians did not experience dementia. Psychiatric disorders were positively associated with dementia, but prevalence of depression, anxiety, and psychoses did not differ significantly between centenarians and an octogenarian control group. However, centenarians were higher on the Geriatric Depression Scale (GDS) than octogenarians. Consistent with our model of developmental adaptation in aging, distal life events contribute to predicting survivorship outcome in which health status as survivor, delayer, or escaper appears as adaptation variables late in life.

In Vivo Evaluation of the Phenox CRC Mechanical Thrombectomy Device in a Swine Model of Acute Vessel Occlusion

Mechanical thrombectomy in ischemic stroke is of increasing interest as it is a promising strategy for fast and efficient recanalization. Several thrombectomy devices have been introduced to the armentarium of mechanical thrombectomy. Currently, new devices are under development and are continuously added to the neurointerventional tool box. Each device advocated so far has a different design and mechanical properties in terms of thrombus-device interaction. Therefore, a systematic evaluation under standardized conditions in vivo of these new devices is needed. The purpose of this study was to evaluate the efficiency, thrombus-device interaction, and potential complications of the novel Phenox CRC for distal mechanical thrombectomy in vivo. The device was evaluated in an established animal model in the swine. Recanalization rate, thromboembolic events, vasospasm, and complications were assessed. Radiopaque thrombi (2 cm length) were used for the visualization of thrombus-device interaction during retrieval. The Phenox CRC (4 mm diameter) was assessed in 15 vessel occlusions. For every occlusion a maximum of 3 retrieval attempts were performed. Complete recanalization (TICI 3/TIMI 3) was achieved in 86.7% of vessel occlusions. In 66.7% (10/15), the first retrieval attempt was successful, and in 20% (3/15), the second attempt led to complete recanalization of the parent artery. In 2 cases (13.3%) thrombus retrieval was not successful (TICI 0/TIMI 0). In 1 case (6.7%) a minor embolic event occurred in a small side branch. No distal thromboembolic event was observed during the study. Thrombus-device interaction illustrated the entrapment of the thrombus by the microfilaments and the proximal cage of the device. No significant thrombus compression was observed. No vessel perforation, dissection, or fracture of the device occurred. In this small animal study, the Phenox CRC was a safe and effective device for mechanical thrombectomy. The unique design with a combination of microfilaments and proximal cage reduces thrombus compression with a consequently high recanalization and low complication rate.

Effect of pantoprazole in patients with chronic laryngitis and pharyngitis related to gastroesophageal reflux disease: clinical, proximal, and distal pH monitoring results

Few studies had evaluated the results of proton pump inhibitors on distal and proximal pH recording using a dual-channel probe. The aim of this study was to determine the clinical and pH-metric effect of treatment with pantoprazole 80 mg for 8 weeks in patients with ear, nose, and throat (ENT) manifestations of gastroesophageal reflux disease associated with pathological proximal acid exposure. We conducted a prospective open study. Patients included had to have chronic pharyngitis or laryngitis, and a pathological gastroesophagopharyngeal reflux. All patients received treatment with pantoprazole 80 mg daily for 8 weeks. One week after the end treatment, patients had a second ENT examination and a 24-hour pH monitoring using dual-channel probe. We included 33 patients (11 men, 22 women). A pathological distal acid reflux was found in 30 patients (91%). After treatment, the improvement of ENT symptoms was found in 51.5% of patients. Normalization of 24-hour proximal esophageal pH monitoring was observed in 22 patients (66%). After treatment, the overall distal acid exposure, the number of distal reflux events, and the number of reflux during more than 5 minutes were significantly decreased (respectively: 19.4% vs 7.2% [P < 0.0001], 62.7 vs 28.4 [P < 0.0001], and 10.4 vs 3.9 [P < 0.0001] ). Similarly, in proximal level, the same parameters were significantly decreased after treatment (respectively: 6.8% vs 1.6% [P < 0.0001], 32.6 vs 8.1 [P < 0.0001], and 3.4 vs 0.6 [P= 0.005] ). Treatment with pantoprazole reduced the frequency and severity of gastroesophagopharyngeal acid reflux in patients with chronic pharyngitis and laryngitis.

Gene Related to Anergy in Lymphocytes (GRAIL) Expression in CD4+ T Cells Impairs Actin Cytoskeletal Organization during T Cell/Antigen-presenting Cell Interactions

GRAIL (gene related to anergy in lymphocytes), is an E3 ubiquitin ligase with increased expression in anergic CD4+ T cells. The expression of GRAIL has been shown to be both necessary and sufficient for the induction of T cell (T) anergy. To date, several subsets of anergic T cells have demonstrated altered interactions with antigen-presenting cells (APC) and perturbed TCR-mediated signaling. The role of GRAIL in mediating these aspects of T cell anergy remains unclear. We used flow cytometry and confocal microscopy to examine T/APC interactions in GRAIL-expressing T cells. Increased GRAIL expression resulted in reduced T/APC conjugation efficiency as assessed by flow cytometry. Examination of single T/APC conjugates by confocal microscopy revealed altered polarization of polymerized actin and LFA-1 to the T/APC interface. When GRAIL expression was knocked down, actin polarization to the T/APC interface was restored, demonstrating that GRAIL is necessary for alteration of actin cytoskeletal rearrangement under anergizing conditions. Interestingly, proximal TCR signaling including calcium flux and phosphorylation of Vav were not disrupted by expression of GRAIL in CD4+ T cells. In contrast, interrogation of distal signaling events demonstrated significantly decreased JNK phosphorylation in GRAIL-expressing T cells. In sum, GRAIL expression in CD4+ T cells mediates alterations in the actin cytoskeleton during T/APC interactions. Moreover, in this model, our data dissociates proximal T cell signaling events from functional unresponsiveness. These data demonstrate a novel role for GRAIL in modulating T/APC interactions and provide further insight into the cell biology of anergic T cells.

Adverse life events, area socioeconomic disadvantage, and psychopathology and resilience in young children: the importance of risk factors’ accumulation and protective factors’ specificity

Few studies on resilience in young children model risk appropriately and test theory-led hypotheses about its moderation. This study addressed both issues. Our hypothesis was that for preschool children's emotional/behavioral adjustment in the face of contextual risk protective factors should be located in the cognitive domain. Data were from the first two sweeps of the UK's Millennium Cohort Study. The final study sample was 4,748 three-year-old children clustered in 1,549 Lower layer Super Output Areas in nine strata. Contextual risk was measured at both area (with the Index of Multiple Deprivation) and family (with proximal and distal adverse life events experienced) level. Moderator variables were parenting, verbal and non-verbal ability, developmental milestones, and temperament. Multivariate multilevel models-that allowed for correlated residuals at both individual and area level-and univariate multilevel models estimated risk effects on specific and broad psychopathology. At baseline, proximal family risk, distal family risk and area risk were all associated with broad psychopathology, although the most parsimonious was the proximal family risk model. The area risk/broad psychopathology association remained significant even after family risk was controlled but not after family level socioeconomic disadvantage was controlled. The cumulative family risk was more parsimonious than the specific family risks model. Non-verbal ability moderated the effect of proximal family risk on conduct and emotional problems, and developmental milestones moderated the effect of proximal family risk on conduct problems. The findings highlight the importance of modeling contextual risk appropriately and of locating in the cognitive domain factors that buffer its effect on young children's adjustment.

TNFα activation of PKCδ, mediated by NFκB and ER stress, cross-talks with the insulin signaling cascade

TNFα plays key roles in the regulation of inflammation, cell death, and proliferation and its signaling cascade cross-talks with the insulin signaling cascade. PKCδ, a novel PKC isoform, is known to participate in proximal TNFα signaling events. However, it has remained unclear whether PKCδ plays a role in distal TNFα signaling events. Here we demonstrate that PKCδ is activated by TNFα in a delayed fashion that is temporally associated with JNK activation. To investigate the signaling pathways activating PKCδ and JNK, we used pharmacological and genetic inhibitors of NFκB. We found that inhibition of NFκB attenuated PKCδ and JNK activations. Further analysis revealed that ER stress contributes to TNFα-stimulated PKCδ and JNK activations. To investigate the role of PKCδ in TNFα action, we used 29-mer shRNAs to silence PKCδ expression. A reduction of ~90% in PKCδ protein levels reduced TNFα-stimulated stress kinase activation, including JNK. Further, PKCδ was necessary for thapsigargin-stimulated JNK activation. Because thapsigargin is a potent inducer of ER stress, we determined whether PKCδ was necessary for induction of the UPR. Indeed, a reduction in PKCδ protein levels reduced thapsigargin-stimulated CHOP induction, a hallmark of the UPR, but not BiP/GRP78 induction, suggesting that PKCδ does not globally regulate the UPR. Next, the role of PKCδ in TNFα mediated cross-talk with the insulin signaling pathway was investigated in cells expressing human IRS-1 and a 29-mer shRNA to silence PKCδ expression. We found that a reduction in PKCδ protein levels reversed the TNFα-mediated reduction in insulin-stimulated IRS-1 Tyr phosphorylation, Akt activation, and glycogen synthesis. In addition, TNFα-stimulated IRS protein Ser/Thr phosphorylation and degradation were blocked. Our results indicate that: 1) NFκB and ER stress contribute in part to PKCδ activation; 2) PKCδ plays a key role in the propagation of the TNFα signal; and 3) PKCδ contributes to TNFα-induced inhibition of insulin signaling events.