Non-secretion of mutant proteins of the glaucoma gene myocilin in cultured trabecular meshwork cells and in aqueous humor

Until recently, very little was known about the molecular mechanisms responsible for the development of glaucoma, a leading cause of blindness worldwide. Mutations in the glaucoma gene myocilin (MYOC, GLC1A) are associated with elevated intraocular pressure and the development of autosomal dominant juvenile glaucoma and a subset of adult-onset glaucoma. MYOC is expressed in the trabecular meshwork (TM), a tissue responsible for drainage of aqueous humor from the eye, and the tissue involved in elevated intraocular pressure associated with glaucoma. To better understand the role of MYOC in glaucoma pathogenesis, we examined the expression of normal and mutant myocilin in cultured ocular (TM) and non-ocular cells as well as in the aqueous humor of patients with and without MYOC glaucoma. Normal myocilin was secreted from cultured cells, but very little to no myocilin was secreted from cells expressing five different mutant forms of MYOC. In addition, no mutant myocilin was detected in the aqueous humor of patients harboring a nonsense MYOC mutation (Q368X). Co-transfection of cultured cells with normal and mutant myocilin led to suppression of normal myocilin secretion. These studies suggest that MYOC glaucoma is due either to insufficient levels of secreted myocilin or to compromised TM cell function caused by congestion of the TM secretory pathway.     

Motives for the adoption of protective health behaviours for men and women: an evaluation of the psychosocial-appraisal health model

The present study has been designed to: (1) evaluate the adequacy of a psychosocial-appraisal health model posited to explain the decision-making processes associated with the intention as well as the (non)practice of protective behaviours; and (2) test for sex differences in the hypothesized structural relations (both direct and indirect) between the model's exogenous (psychosocial) and endogenous (cognitive appraisal) factors in representing the way health decisions are made. Results obtained from path analysis (N=1269) provided overall support for the 'fit' of the hypothesized health model. The implications of the findings with regard to differences in male and female health status are discussed.     

Bacterial Clearance and Cytokine Profiles in a Murine Model of Postsurgical Nosocomial Pneumonia

The development of a nosocomial pneumonia is facilitated by alterations in host innate pulmonary antibacterial defenses following surgical trauma, which can result in decreased pulmonary bacterial clearance and increased morbidity and mortality. In a murine model of postoperative nosocomial infection, surgical stress (laparotomy) decreased Escherichia coli clearance from the lungs of animals that underwent surgery. Consistent with previous studies, (i) pulmonary levels of tumor necrosis factor alpha at 6 h and of interleukin-1β (IL-1β), IL-6, and gamma interferon (IFN-γ) at 24 h post-bacterial infection (PBI) were decreased in animals that underwent laparotomy 24 h prior to E. coli infection (LAP/E. coli) compared to animals that received E. coli only; (ii) KC and macrophage inhibitory protein 2 were elevated at 6 h PBI in LAP/E. coli animals compared to E. coli-only animals; however, at 24 h PBI, levels were higher in the E. coli-only group; (iii) at 24 h PBI, monocyte chemoattractant protein 1 was lower in the LAP/E. coli group compared to the E. coli-only group; (iv) IL-10 levels were unaffected at all time points evaluated; and (v) the total number of neutrophils present in the lungs of LAP/E. coli animals at 6 h PBI was decreased in comparison to that in E. coli-only animals, resulting in decreased bacterial clearance and increased mortality in LAP/E. coli animals by 24 h PBI. Similar changes in cytokine profiles, pulmonary bacterial clearance, and mortality were consistent with reported findings in patients following surgical trauma. This model, therefore, provides a clinically relevant system in which the molecular and cellular mechanisms that lead to the development of nosocomial pneumonia can be further explored.     

Expression of fragile X chromosome in human-rodent somatic cell hybrids

The fragile X chromosome, associated with a common form of X-linked mental retardation, is cytologically observed most often as a gap or fragile site near the distal end of the long arm in band Xq28. Expression of this site is variable and dependent upon lowered thymidylate pools. In order to examine the behavior of this fragile site in a foreign genetic background, interspecific somatic cell hybrids were isolated from crosses of hamster cells and lymphoblastoid cells derived from male patients with fragile X-linked mental retardation. Three hybrid cell lines containing the human X chromosome were analyzed. Following induction with 5-fluorodeoxyuridine, all three hybrids expressed the fragile site in approximately 10% of the metaphases examined. Our data indicate that expression of the fragile site in band Xq27 is dependent neither on the integrity of the human genome nor on the expression of human autosomal genes.     

Age-related changes in smooth pursuit initiation

Quantitative analysis of eye movements is a useful tool for examining the behavioural effects of ageing. Although the effect of ageing on saccadic eye movement has been examined in some detail, the effect of ageing on a second class of eye movement, smooth pursuit (SP), has received less attention. We examined the initiation of SP in a group of fifteen healthy older people (mean age 72 years) and compared their performance with that of ten young controls (mean age 21 years). Although their performance was qualitatively similar, pursuit latency was increased in the older group. Investigation of the gap effect on pursuit revealed that, while the gap effect was present in the older group, it seemed to be directionally asymmetrical. When the longer absolute latencies were taken into account, although the gap effect in the two groups was identical for leftward tasks, for rightward tasks it was reduced in the older group, although this did not reach statistical significance. The difference between the old and young groups was driven by some of the older subjects. At the longest gap duration employed (400 ms), while there was a clear gap effect for leftward tasks in these subjects, there was no reduction in latency, or increases in latency, for rightward tasks. This asymmetry was not related to chronological age within the older group. These results suggest an age-related alteration in SP initiation that is more complex than general slowing of information processing in ageing. They may be indicative of additional ageing effects specific to the oculomotor or closely related systems.     

Effects of lumbar extensor fatigue and fatigue rate on postural sway

Falls from heights resulting from a loss of balance are a major concern in the occupational setting. Previous studies have documented a deleterious effect of lower extremity fatigue on balance. The purpose of this study was to investigate the effect of lumbar extensor fatigue on balance during quiet standing. Additionally, the effects of fatigue rate on balance and balance recovery rate were assessed. Eight center-of-pressure-based measures of postural sway were collected from 13 participants, both before and after a protocol that fatigued the lumbar extensors to 60% of their unfatigued maximum voluntary exertion force. In addition, postural sway was measured for 30 min after the fatiguing protocol, at 5-min intervals, to quantify balance recovery rate during recovery from fatigue. Two different fatigue rates were achieved by fatiguing participants over either 10 min or 90 min. Results show an increase up to 58% in time-domain postural sway measures with lumbar extensor fatigue, but no change in frequency-domain measures. Fatigue rate did not affect the magnitude of these postural sway increases, nor did it affect the rate of balance recovery following fatigue. Statistical power for the latter result, however, was low. These results show that lumbar extensor fatigue increases postural sway and may contribute to fall-from-height accidents.     

Human TLR-7-, -8-, and -9-mediated induction of IFN-alpha/beta and -lambda Is IRAK-4 dependent and redundant for protective immunity to viruses

Five TLRs are thought to play an important role in antiviral immunity, sensing viral products and inducing IFN-alpha/beta and -lambda. Surprisingly, patients with a defect of IRAK-4, a critical kinase downstream from TLRs, are resistant to common viruses. We show here that IFN-alpha/beta and -lambda induction via TLR-7, TLR-8, and TLR-9 was abolished in IRAK-4-deficient blood cells. In contrast, IFN-alpha/beta and -lambda were induced normally by TLR-3 and TLR-4 agonists. Moreover, IFN-beta and -lambda were normally induced by TLR-3 agonists and viruses in IRAK-4-deficient fibroblasts. We further show that IFN-alpha/beta and -lambda production in response to 9 of 11 viruses tested was normal or weakly affected in IRAK-4-deficient blood cells. Thus, IRAK-4-deficient patients may control viral infections by TLR-3- and TLR-4-dependent and/or TLR-independent production of IFNs. The TLR-7-, TLR-8-, and TLR-9-dependent induction of IFN-alpha/beta and -lambda is strictly IRAK-4 dependent and paradoxically redundant for protective immunity to most viruses in humans.     

The effect of HIPing on the fatigue and tensile strength of a case, porous-coated Co-Cr-Mo alloy

The process required to sinter porous Co-Cr-Mo alloys results in the formation of substrate porosity through carbide dissolution. Since hot isostatic pressing (HIPing) has been shown to eliminate casting porosity in the Co-Cr-Mo alloy, it is possible that it may be equally effective on pores that are generated from the sintering operation. The effect that HIPing a porous-coated Co-Cr-Mo material has on the fatigue and tensile properties was investigated. Fatigue testing was performed on sintered materials as well as sintered and HIPed materials, both with and without a porous coating. Further, the effect of varying coating thickness on the resulting fatigue strength of sintered and HIPed materials was studied. Light microscopy was performed in order to define the microstructural changes brought about by the various thermal cycles. Scanning electron microscopy was utilized to define the crack initiation process. The fatigue strength of uncoated "as sintered" materials was found to be reduced by 34% relative to the "as cast" condition. The same material that was HIPed revealed a fatigue strength slightly lower than the "as cast" condition. It was found that porous coatings created preferential sites for fatigue crack initiation. However, the presence of the coating did not further reduce the fatigue strength of "as sintered" materials because of the already low strength created by the sintering operation. Materials that were sintered exhibited a lowering in both tensile strength and elongation to failure relative to the "as cast" condition. The HIPing of sintered materials improved both fatigue and tensile properties relative to the "as sintered" condition.