The role of LPCAT in regulating cellular inflammatory responses to toll-like receptor ligands

A novel lipid modifying enzyme, lysophosphatidylcholine acyltransferase (LPCAT), has been shown to regulate pro-inflammatory cytokine responses to lipopolysaccharide (LPS), a key molecule in initiating sepsis. Current research has shown that phospholipid metabolism may influence cell signalling, since that is dependent on the clustering of the receptor molecules into membrane microdomains (lipid rafts). LPCAT therefore, may be a target for novel anti-sepsis therapies. This study aimed to determine if LPCAT is essential in the regulation of pro-inflammatory cytokine responses to Gram-positive cell bacterial components, such as peptidoglycan (PG), lipoteichoic acid (LTA) and synthetic tripalmitoylated lipopeptide Pam3CSK4 in monocytes and lung epithelial cells. Inhibition of LPCAT activity consistently reduced TNF-α, IL-6 and IL-8 protein and mRNA levels in monocytes stimulated with all microbial ligands tested. Furthermore, cytokine production observed in monocytes primed with IFN-γ prior to ligand stimulation was twice that seen in unprimed cells, yet still inhibition of LPCAT significantly decreased the amplified inflammatory response. Lung epithelial cells, BEAS-2B, were more immunologically responsive to Pam3CSK4 where it consistently induced a high secretion of IL-8 and IL-6. However, whilst inhibition of LPCAT demonstrated a reduction in cytokine secretion, it was not a predominant as observed in monocytes indicating that LPCAT may have a lesser role in these cells.The secondary aim was to characterise the expression levels of 5 LPCAT enzymes in diverse cell types to ascertain if there is differential expression of the LPCATs, which may help to explain the altered effects that LPCAT inhibition produces on inflammatory cytokine production between cell types. Data in this report did not demonstrate one particular LPCAT iso-form to be highly expressed by monocytes, however literature has suggested that LPCAT2 might be the enzyme to modulate the inducible phospholipid remodeling pathways in innate immune cells. Further studies on inducible LPCAT expression in human cell lines is required to support these ideas.Previous studies have observed LPCAT to regulate translocation of TLR4 into membrane lipid raft domains and subsequent down-stream inflammatory responses. Although the translocation of TLR2 into membrane lipid rafts has been researched, its regulation by LPCAT has not, thus it was investigated in this study. Complications with experiments, however, did not allow this to be adequately assessed but, literature suggests this is the likely mechanism that LPCAT elicits its immuno-regulatory effect in response to microbial stimuli.The present study provided evidence that LPCAT influences the complex network of cell signalling involved in microbial responses, underlying it’s importance in inflammatory responses and potentially offering a target for novel anti-sepsis therapies

Family-focused treatment for childhood depression: model and case illustrations

Although the evidence base for treatment of depressive disorders in adolescents has strengthened in recent years, less is known about the treatment of depression in middle to late childhood. A family-based treatment may be optimal in addressing the interpersonal problems and symptoms frequently evident among depressed children during this developmental phase, particularly given data indicating that attributes of the family environment predict recovery versus continuing depression among depressed children. Family-Focused Treatment for Childhood Depression (FFT-CD) is designed as a 15-session family treatment with both the youth and parents targeting two putative mechanisms involved in recovery: (a) enhancing family support, specifically decreasing criticism and increasing supportive interactions; and (b) strengthening specific cognitive-behavioral skills within a family context that have been central to CBT for depression, specifically behavioral activation, communication, and problem solving. This article describes in detail the FFT-CD protocol and illustrates its implementation with three depressed children and their families. Common themes/challenges in treatment included family stressors, comorbidity, parental mental health challenges, and inclusion/integration of siblings into sessions. These three children experienced positive changes from pre- to posttreatment on assessor-rated depressive symptoms, parent- and child-rated depressive symptoms, and parent-rated internalizing and externalizing symptoms. These changes were maintained at follow-up evaluations 4 and 9 months following treatment completion.K23 MH101238 - NIMH NIH HHS; R01 MH082856 - NIMH NIH HHS; R01 MH082861 - NIMH NIH HH

Stress rapidly suppresses in vivo LH pulses and increases activation of RFRP-3 neurons in male mice

Restraint stress is a psychosocial stressor that suppresses reproductive status, including LH pulsatile secretion, but the neuroendocrine mechanisms underlying this inhibition remains unclear. Reproductive neural populations upstream of gonadotropin-releasing hormone (GnRH) neurons, such as kisspeptin, neurokinin B and RFRP-3 (GnIH) neurons, are possible targets for psychosocial stress to inhibit LH pulses, but this has not been well examined, especially in mice in which prior technical limitations prevented assessment of in vivo LH pulse secretion dynamics. Here, we examined whether one-time acute restraint stress alters in vivo LH pulsatility and reproductive neural populations in male mice, and what the time-course is for such alterations. We found that endogenous LH pulses in castrated male mice are robustly and rapidly suppressed by one-time, acute restraint stress, with suppression observed as quickly as 12–18 min. This rapid LH suppression parallels with increased in vivo corticosterone levels within 15 min of restraint stress. Although Kiss1, Tac2 and Rfrp gene expression in the hypothalamus did not significantly change after 90 or 180 min restraint stress, arcuate Kiss1 neural activation was significantly decreased after 180 min. Interestingly, hypothalamic Rfrp neuronal activation was strongly increased at early times after restraint stress initiation, but was attenuated to levels lower than controls by 180 min of restraint stress. Thus, the male neuroendocrine reproductive axis is quite sensitive to short-term stress exposure, with significantly decreased pulsatile LH secretion and increased hypothalamic Rfrp neuronal activation occurring rapidly, within minutes, and decreased Kiss1 neuronal activation also occurring after longer stress durations

The sub-millimeter properties of broad absorption line quasars

We have carried out the first systematic survey of the sub-millimeter properties of broad absorption line (BAL) quasars. 30 BAL quasars drawn from a homogeneously selected sample from the Sloan Digital Sky Survey at redshifts 2<z<2.6 were observed with the SCUBA array at the JCMT to a typical rms sensitivity of 2.5 mJy. Eight quasars were detected at > 2 sigma significance, four of which are at > 3 sigma significance. The far-infrared luminosities of these quasars are > 10^{13} L_solar. There is no correlation of sub-millimeter flux with either the strength of the broad absorption feature or with absolute magnitude in our sample. We compare the sub-millimeter flux distribution of the BAL quasar sample with that of a sample of quasars which do not show BAL features in their optical spectra and find that the two are indistinguishable. BAL quasars do not have higher sub-millimeter luminosities than non-BAL quasars. These findings are consistent with the hypothesis that all quasars would contain a BAL if viewed along a certain line-of-sight. The data are inconsistent with a model in which the BAL phenomenon indicates a special evolutionary stage which co-incides with a large dust mass in the host galaxy and a high sub-millimeter luminosity. Our work provides constraints on alternative evolutionary explanations of BAL quasars.Comment: 8 pages, 2 figures, ApJ, in pres

Stability of extemporaneously prepared ophthalmic solutions for mydriasis

Purpose Results of an evaluation of the physical and chemical stability of extemporaneously prepared adult and pediatric ophthalmic solutions containing combinations of phenylephrine, tropicamide, and cyclopentolate are reported. Methods A stability study was conducted to help determine the feasibility of innovative formulations to meet an unmet clinical need for combination mydriatic ophthalmic eyedrops. An adult mydriatic ophthalmic solution containing phenylephrine hydrochloride 2.5% and tropicamide 1.0% and a pediatric formulation containing phenylephrine hydrochloride 2.5%, tropicamide 0.5%, and cyclopentolate hydrochloride 0.5% were prepared using proper aseptic techniques. Triplicate samples of each formulation were stored for 60 days at refrigeration temperatures (2–8 °C) and analyzed on day 0 and days 7, 14, 28, and 60. At each time point, the stability samples were assessed by visual inspection, pH measurement, and stability-indicating high-performance liquid chromatography (HPLC) analysis. Results Over the 60-day storage period, there was no significant change in the visual appearance or pH level of any of the adult or pediatric solution samples. The results of HPLC analysis indicated that all samples retained 97–102% of the initial drug concentrations for up to 60 days. Conclusion Both adult and pediatric ophthalmic formulations containing combinations of phenylephrine, tropicamide, and cyclopentolate were stable physically and chemically for up to 60 days when stored at refrigeration temperatures (2–8 °C)

Real-time Assessment of Right and Left Ventricular Volumes and Function in Children Using High Spatiotemporal Resolution Spiral bSSFP with Compressed Sensing

Background: Real-time (RT) assessment of ventricular volumes and function enables data acquisition during free-breathing. However, in children the requirement for high spatiotemporal resolution requires accelerated imaging techniques. In this study, we implemented a novel RT bSSFP spiral sequence reconstructed using Compressed Sensing (CS) and validated it against the breath-hold (BH) reference standard for assessment of ventricular volumes in children with heart disease. Methods: Data was acquired in 60 children. Qualitative image scoring and evaluation of ventricular volumes was performed by 3 clinical cardiac MR specialists. 30 cases were reassessed for intra-observer variability, and the other 30 cases for inter-observer variability. Results: Spiral RT images were of good quality, however qualitative scores reflected more residual artefact than standard BH images and slightly lower edge definition. Quantification of Left Ventricular (LV) and Right Ventricular (RV) metrics showed excellent correlation between the techniques with narrow limits of agreement. However, we observed small but statistically significant overestimation of LV end-diastolic volume, underestimation of LV end-systolic volume, as well as a small overestimation of RV stroke volume and ejection fraction using the RT imaging technique. No difference in inter-observer or intra-observer variability were observed between the BH and RT sequences. Conclusions: Real-time bSSFP imaging using spiral trajectories combined with a compressed sensing reconstruction is feasible. The main benefit is that it can be acquired during free breathing. However, another important secondary benefit is that a whole ventricular stack can be acquired in ~20 seconds, as opposed to ~6 minutes for standard BH imaging. Thus, this technique holds the potential to significantly shorten MR scan times in children

Marriage length, spousal support, and marital satisfaction in dual-income men and women

The introduction of women into the workforce has led to a steady increase in the number of dual-income couples in the United States. Since dual-income married couples now comprise the majority of couples, they are especially important to study. Approximately half of marriages end in divorce, so an increased knowledge of what makes marriages work is essential. Previous studies indicate that as spousal support increases so does marital satisfaction. This study seeks to confirm the above mentioned relationship for dual-income married couples by studying 143 dual-income couples from varying demographic backgrounds. Also, we will investigate whether couples married for a longer amount of time report greater spousal support and marital satisfaction and whether any gender differences occur

It Takes a Library to Support Distance Learners

Wallace Library\u27s philosophy is to create and provide resources and services that will support all users. Consequently, distance learners and distance faculty have a plethora of online resources available to them, some of which is pushed out, saving time and effort for the library user. The evolvement of Wallace\u27s online resources is continuous, focused on the student or professor\u27s research need from a geographic distance

Attenuated pupillary light responses and downregulation of opsin expression parallel decline in circadian disruption in two different mouse models of Huntington's disease.

Circadian deficits in Huntington’s disease (HD) are recapitulated in both fragment (R6/2) and full-length (Q175) mouse models of HD. Circadian rhythms are regulated by the suprachiasmatic nuclei (SCN) in the hypothalamus, which are primarily entrained by light detected by the retina. The SCN receives input from intrinsically photosensitive retinal ganglion cells (ipRGCs) that express the photopigment melanopsin, but also receive input from rods and cones. In turn, ipRGCs mediate a range of non-image forming responses to light including circadian entrainment and the pupillary light response (PLR). Retinal degeneration/dysfunction has been described previously in R6 /2 mice. We investigated, therefore, whether or not circadian disruption in HD mice is due to abnormalities in retinal photoreception. We measured expression of melanopsin, rhodopsin and cone opsin, as well as other retinal markers (tyrosine hydroxylase, calbindin, PKCα and Brna3 ), in R6/2 and Q175 mice at different stages of disease. We also measured the PLR as a ‘readout’ for ipRGC function and a marker of light reception by the retina. We found that the PLR was attenuated in both lines of HD mice. This was accompanied by a progressive downregulation of cone opsin and melanopsin expression. We suggest that a disease-related change in photoreception by the retina contributes to the progressive dysregulation of circadian rhythmicity and entrainment seen in HD mice. Colour vision is abnormal in HD patients. Therefore, if retinal deficits similar to those seen in HD mice are confirmed in patients, specifically designed light therapy may be an effective strategy to improve circadian dysfunction.This work was supported by a grant from CHDI (Inc.) to AJM. SH is funded by a BBSRC grant (BB/M009998/1). SNP and CAP are funded by a Wellcome Trust strategic grant (098461/Z/12/Z)