DNA vaccines encoding IL-2 linked to HPV-16 E7 antigen generate enhanced E7-specific CTL responses and antitumor activity

DNA vaccination has emerged as a promising strategy for cancer immunotherapy. However, since DNA vaccines have low immunogenicity, various strategies have been developed to enhance the potency of DNA vaccines. In the current study, we aim to determine whether the potency of the DNA vaccine encoding human papillomavirus type 16 (HPV-16) E7 antigen can be enhanced by IL-2. We have generated a DNA vaccine encoding IL-2 linked to HPV-16 E7 antigen. Our results indicate that the DNA vaccine encoding a fusion of IL-2 and E7 proteins generated the highest frequency of E7-specific CD8(+) T cells. We also found that the DNA vaccine encoding a fusion of IL-2 and E7 proteins generated the strongest protective as well as therapeutic anti-tumor effect against E7-expressing tumors. In addition, it was observed that CD8(+) T cells were mainly responsible for the antitumor effect generated by the DNA vaccine encoding a fusion of IL-2 and E7 proteins. Thus, we conclude that the linkage of IL-2 to HPV-16 E7 antigen significantly enhances the DNA vaccine potency against E7-expressing tumors. Our strategy may potentially be used in other antigenic systems to control infectious diseases and/or cancer.     

Long Working Hours and Sleep Disturbances: The Whitehall II Prospective Cohort Study

Study Objective:

To examine whether exposure to long working hours predicts various forms of sleep disturbance; short sleep, difficulty falling asleep, frequent waking, early waking and waking without feeling refreshed.

Design:

Prospective study with 2 measurements of working hours (phase 3, 1991–1994 and phase 5, 1997–1999) and 2 measurements of subjective sleep disturbances (phase 5 and phase 7, 2002–2004).

Setting:

The Whitehall II study of British civil servants.

Participants:

Full time workers free of sleep disturbances at phase 5 and employed at phases 5 and 7 (n = 937–1594) or at phases 3, 5, and 7 (n = 886–1510).

Measurements and Results:

Working more than 55 hours a week, compared with working 35–40 hours a week, was related to incident sleep disturbances; demographics-adjusted odds ratio (95% CI) 1.98 (1.05, 3.76) for shortened sleeping hours, 3.68 (1.58, 8.58) for difficulty falling asleep; and 1.98 (1.04, 3.77) for waking without feeling refreshed. Repeat exposure to long working hours was associated with odds ratio 3.24 (1.45, 7.27) for shortened sleep, 6.66 (2.64, 16.83) for difficulty falling asleep, and 2.23 (1.16, 4.31) for early morning awakenings. Some associations were attenuated after adjustment for other risk factors. To a great extent, similar results were obtained using working hours as a continuous variable. Imputation of missing values supported the findings on shortened sleep and difficulty in falling asleep.

Conclusion:

Working long hours appears to be a risk factor for the development of shortened sleeping hours and difficulty falling asleep.

Citation:

Virtanen M; Ferrie JE; Vahtera J; Elovainio M; Singh-Manoux A; Marmot MG; Kivimäki M. Long working hours and sleep disturbances: the whitehall II prospective cohort study. SLEEP 2009;32(6):737–745.     

Palmar arch modulation in patients with hemiparesis after a stroke

Hand shape modulation has traditionally been studied within the framework of reach-to-grasp tasks by examining the control of arm transport, grip aperture scaling, and finger joint excursions. However, global parameters characterizing arm and hand movement can be enhanced by additional knowledge of biomechanical changes in the hand. We previously examined palmar arch modulation during grasping in healthy subjects by identifying thenar and hypothenar displacement. This method was used to characterize hand shape modulation in 10 stroke survivors with mild hand paresis, as assessed by the Chedoke-McMaster clinical scale, during two types of grasps (spherical, cylindrical). Palmar arch modulation was examined during the three phases of prehensile movement: transport shaping (P1), preshaping (P2), and contact shaping (P3). Compared to the control group, the stroke survivors showed significant differences (spherical: F _2,18 = 12.025, P < 0.001; cylindrical: F _2,18 = 9.054, P < 0.001) in palmar arch modulation particularly during P3 wherein fine adjustments are made to the grip in preparation for object manipulation. While control subjects completed most of hand shape modulation early in the task, stroke survivors took longer to complete each phase. Furthermore, stroke survivors started with a flatter hand which required relatively more arch modulation during the latter part of the task, thereby reflecting a temporal and spatial concurrency between the phases. Stroke survivors with well-recovered hand grasping ability tended to incorporate compensations/adaptations in hand posture during specific grasping phases. Palmar arch analysis provides us with a more complete understanding about how hand biomechanics, specifically palmar concavity articulation, is changed post-stroke. This will allow us to better identify the motor compensations used for grasping and to re-focus rehabilitation interventions to reduce compensations and improve functional motor recovery.     

An investigation of vitamin D status in alopecia areata

Alopecia areata (AA) is a type of non-scarring, recurrent patchy loss of hair in hair-bearing areas and is mostly of autoimmune origin. Previous studies have suggested that some autoimmune diseases were found to be associated with vitamin D deficiency. The current study was designed to assess the levels of serum 25-hydroxy vitamin D and C-reactive protein in AA, as compared with controls and to further identify the association between vitamin D levels and disease severity in patients with AA. This cross-sectional study included 45 patients with AA and 45 healthy volunteers. Clinical and anthropometric parameters were recorded, according to a pre-designed proforma. Serum 25-hydroxy vitamin D and high-sensitivity C-reactive protein were estimated using ELISA kits. The severity of AA was determined using Severity of Alopecia Tool (SALT) score. We observed a significant rise in systemic inflammation as seen by elevated high-sensitive C-reactive protein levels and lowered 25-hydroxy vitamin D levels in patients with alopecia areata, compared to controls (p?=?0.001). The levels of 25-hydroxy vitamin D showed a significant negative correlation with disease severity, while hs-CRP levels showed a significant positive correlation with disease severity (ρ?=???0.714, p?=?0.001 and ρ?=?0.818, p?=?0.001). Our results suggest significant systemic inflammation and vitamin D deficiency in alopecia areata, more so with increasing disease severity. This gains particular importance in the treatment of alopecia areata in future, as supplementing vitamin D to AA patients would result in reducing the disease severity and inducing remission.     

Sleep duration and sleep disturbances partly explain the association between depressive symptoms and cardiovascular mortality: the Whitehall II cohort study

Depressive symptoms are associated with an increased risk of death, but most of this association remains unexplained. Our aim was to explore the contribution of sleep duration and disturbances to the association between depressive symptoms, all‐cause and cardiovascular disease mortality. A total of 5813 (4220 men and 1593 women) aged 50–74 years at baseline, participants of the British Whitehall II prospective cohort study, were included. Depressive symptoms, sleep duration and disturbances were assessed in 2003–04. Mortality was ascertained through linkage to the national mortality register until August 2012, with a mean follow‐up of 8.8 years. Depressive symptoms were associated with an increased risk of mortality from all causes [hazard ratio (HR) = 1.51; 95% confidence interval (CI): 1.16–1.97)] and cardiovascular diseases (HR = 1.63; 95% CI: 1.01–2.64) after adjustment for sociodemographic characteristics. Further adjustment for sleep duration and disturbances reduced the association between depressive symptoms and cardiovascular mortality by 21% (HR = 1.53; 95% CI: 0.91–2.57). Sleep seems to have a role, as a mediator or confounder, in explaining the association between depressive symptoms and cardiovascular mortality. These findings need replication in larger studies with longer follow‐up.     

Autosomal-dominant nystagmus,foveal hypoplasia and presenile cataract associated with a novel PAX6 mutation

Autosomal-dominant idiopathic infantile nystagmus has been linked to 6p12 (OMIM 164100), 7p11.2 (OMIM 608345) and 13q31-q33 (OMIM 193003). PAX6 (11p13, OMIM 607108) mutations can also cause autosomal-dominant nystagmus, typically in association with aniridia or iris hypoplasia. We studied a large multigenerational white British family with autosomal-dominant nystagmus, normal irides and presenile cataracts. An SNP-based genome-wide analysis revealed a linkage to a 13.4-MB region on chromosome 11p13 with a maximum lod score of 2.93. A mutation analysis of the entire coding region and splice junctions of the PAX6 gene revealed a novel heterozygous missense mutation (c.227C>G) that segregated with the phenotype and is predicted to result in the amino-acid substitution of proline by arginine at codon 76 p.(P76R). The amino-acid variation p.(P76R) within the paired box domain is likely to destabilise the protein due to steric hindrance as a result of the introduction of a polar and larger amino acid. Eye movement recordings showed a significant intrafamilial variability of horizontal, vertical and torsional nystagmus. High-resolution in vivo imaging of the retina using optical coherence tomography (OCT) revealed features of foveal hypoplasia, including rudimentary foveal pit, incursion of inner retinal layers, short photoreceptor outer segments and optic nerve hypoplasia. Thus, this study presents a family that segregates a PAX6 mutation with nystagmus and foveal hypoplasia in the absence of iris abnormalities. Moreover, it is the first study showing detailed characteristics using eye movement recordings of autosomal-dominant nystagmus in a multigenerational family with a novel PAX6 mutation.     

Effect of Retirement on Sleep Disturbances: the GAZEL Prospective Cohort Study

Objectives:

Changes in health following retirement are poorly understood. We used serial measurements to assess the effect of retirement on sleep disturbances.

Design:

Prospective cohort study.

Setting:

The French national gas and electricity company.

Participants:

Fourteen thousand seven hundred fourteen retired employees (79% men).

Measurements and Results:

Annual survey measurements of sleep disturbances ranging from 7 years before to 7 years after retirement (a mean of 12 measurements). Before retirement 22.2% to 24.6% of participants reported having disturbed sleep. According to repeated-measures logistic-regression analysis with generalized estimating equations estimation, the odds ratio (OR) for having a sleep disturbance in the postretirement period was 0.74 (95% confidence interval 0.71-0.77), compared with having a sleep disturbance in the preretirement period. The postretirement improvement in sleep was more pronounced in men (OR 0.66 [0.63-0.69]) than in women (OR 0.89 [0.84-0.95]) and in higher-grade workers than lower-grade workers. Postretirement sleep improvement was explained by the combination of preretirement risk factors suggesting removal of work-related exposures as a mechanism. The only exception to the general improvement in sleep after retirement was related to retirement on health grounds. In this group of participants, there was an increase in sleep disturbances following retirement.

Conclusions:

Repeated measurements provide strong evidence for a substantial and sustained decrease in sleep disturbances following retirement. The possibility that the health and well-being of individuals are significantly worse when in employment than following retirement presents a great challenge to improve the quality of work life in Western societies in which the cost of the aging population can only be met through an increase in average retirement age.

Citation:

Vahtera J; Westerlund H; Hall M; Sjösten N; Kivimäki M; Salo P; Ferrie JE; Jokela M; Pentti J; Singh-Manoux A; Goldberg M; Zins M. Effect of retirement on sleep disturbances: the GAZEL prospective cohort study. SLEEP 2009;32(11):1459-1466.     

Microfluidic system for formation of PC-3 prostate cancer co-culture spheroids

The niche microenvironment in which cancer cells reside plays a prominent role in the growth of cancer. It is therefore imperative to mimic the in vivo tumor niche in vitro to better understand cancer and enhance development of therapeutics. Here, we engineer a 3D metastatic prostate cancer model that includes the types of surrounding cells in the bone microenvironment that the metastatic prostate cancer cells reside in. Specifically, we used a two-layer microfluidic system to culture 3D multi-cell type spheroids of fluorescently labeled metastatic prostate cancer cells (PC-3 cell line), osteoblasts and endothelial cells. This method ensures uniform incorporation of all co-culture cell types into each spheroid and keeps the spheroids stationary for easy tracking of individual spheroids and the PC-3's residing inside them over the course of at least a week. This culture system greatly decreased the proliferation rate of PC-3 cells without reducing viability and may more faithfully recapitulate the in vivo growth behavior of malignant cancer cells within the bone metastatic prostate cancer microenvironment.