Acclimation of photosynthetic proteins to rising atmospheric CO2.
In this review we discuss how the photosynthetic apparatus, particularly Rubisco, acclimates to rising atmospheric CO2 concentrations (ca). Elevated ca alters the control exerted by different enzymes of the Calvin cycle on the overall rate of photosynthetic CO2 assimilation, so altering the requirement for different functional proteins. A decreased flux of carbon through the photorespiratory pathway will decrease requirements for these enzymes. From modeling of the response of CO2 uptake (A) to intracellular CO2 concentration (ci) it is shown that the requirement for Rubisco is decreased at elevated ca, whilst that for proteins limiting ribulose 1,5 bisphosphate regeneration may be increased. This balance may be altered by other interactions, in particular plasticity of sinks for photoassimilate and nitrogen supply; hypotheses on these interactions are presented. It is speculated that increased accumulation of carbohydrate in leaves developed at elevated ca may signal the 'down regulation' of Rubisco. The molecular basis of this 'down regulation' is discussed in terms of the repression of photosynthetic gene expression by the elevated carbohydrate concentrations. This molecular model is then used to predict patterns of acclimation of perennials to long term growth in elevated ca.


Topographic reorganization in area 18 of adult cats following circumscribed monocular retinal lesions in adolescence.
Circumscribed laser lesions were made in the nasal retinae of one eye in adolescent cats. Ten to sixteen months later, about 80 % of single neurones recorded in the lesion projection zone (LPZ) of contralateral area 18 (parastriate cortex, area V2) were binocular but when stimulated via the lesioned eye had ectopic discharge fields (displaced to normal retina in the vicinity of the lesion). Although the clear majority of binocular cells recorded from the LPZ responded with higher peak discharge rates to stimuli presented via the non-lesioned eye, the orientation and direction selectivities as well as preferred and upper cut-off velocities for stimuli presented through either eye were very similar. Furthermore, the sizes of the ectopic discharge fields of binocular cells recorded from the LPZ were not significantly different from those of their counterparts plotted via the non-lesioned eye. Thus, monocular retinal lesions performed in adolescent cats induce topographic reorganization in the LPZ of area 18. Although a similar reorganization occurs in area 17 (striate cortex, area V1) of cats in which monocular retinal lesions were made either in adulthood or adolescence, in view of the very different velocity response profiles of ectopic discharge fields in areas 17 and those in area 18, it appears that ectopic discharge fields in area 17 are largely independent of excitatory feedback input from area 18.


The concepts of rash impulsiveness and reward sensitivity in substance use disorders.
According to recent theories of addiction, the commonly used term impulsivity comprises two factors: rash impulsiveness and reward sensitivity. The present study addresses the relevance and generalizability of this two-factor model in a clinical sample of substance use disorder patients. This was examined by examining both internal and external validity. In addition, a comparison was made between self-reported and behavioral measures reflecting reward sensitivity and rash impulsiveness. Results provide evidence for the existence of the two hypothesized impulsivity factors in a clinical sample of substance dependent patients. Meaningful relationships between the model and drug use characteristics have been found, providing further evidence for the validity of the two-factor model. Furthermore, it is suggested that behavioral and self-report measures of impulsivity represent different constructs.


Meta-analysis of microarray data identifies GAS6 expression as an independent predictor of poor survival in ovarian cancer.
Seeking new biomarkers for epithelial ovarian cancer, the fifth most common cause of death from all cancers in women and the leading cause of death from gynaecological malignancies, we performed a meta-analysis of three independent studies and compared the results in regard to clinicopathological parameters. This analysis revealed that GAS6 was highly expressed in ovarian cancer and therefore was selected as our candidate of choice. GAS6 encodes a secreted protein involved in physiological processes including cell proliferation, chemotaxis, and cell survival. We performed immunohistochemistry on various ovarian cancer tissues and found that GAS6 expression was elevated in tumour tissue samples compared to healthy control samples (P < 0.0001). In addition, GAS6 expression was also higher in tumours from patients with residual disease compared to those without. Our data propose GAS6 as an independent predictor of poor survival, suggesting GAS6, both on the mRNA and on the protein level, as a potential biomarker for ovarian cancer. In clinical practice, the staining of a tumour biopsy for GAS6 may be useful to assess cancer prognosis and/or to monitor disease progression.


Seiridium (Sporocadaceae): an important genus of plant pathogenic fungi.
The genus Seiridium includes multiple plant pathogenic fungi well-known as causal organisms of cankers on Cupressaceae. Taxonomically, the status of several species has been a topic of debate, as the phylogeny of the genus remains unresolved and authentic ex-type cultures are mostly absent. In the present study, a large collection of Seiridium cultures and specimens from the CBS and IMI collections was investigated morphologically and phylogenetically to resolve the taxonomy of the genus. These investigations included the type material of the most important Cupressaceae pathogens, Seiridium cardinale, S. cupressi and S. unicorne. We constructed a phylogeny of Seiridium based on four loci, namely the ITS rDNA region, and partial translation elongation factor 1-alpha (TEF), β-tubulin (TUB) and RNA polymerase II core subunit (RPB2). Based on these results we were able to confirm that S. unicorne and S. cupressi represent different species. In addition, five new Seiridium species were described, S. cupressi was lectotypified and epitypes were selected for S. cupressi and S. eucalypti.


Progression of disability in multiple sclerosis: A study of factors influencing median time to reach an EDSS value.
The extended disability severity scale (EDSS) is clinically useful in assessing disability in multiple sclerosis (MS) patients. It is also being used in studies to determine how genes and environment influence disability. However, since it has a complex relationship with functional scores and mobility and is strongly determined by disease duration its use can be limiting. Study associations of variables with progression described by time from disease onset until EDSS. We used a variable based on below/above median time from MS onset to reach a single EDSS value to define slow or fast progression. We compared patient categorization using this variable and MSSS, and in 533 patients (EDSS 1-8) and 242 of these patients with EDSS1-4, studied associations with skin type, gender, ultraviolet radiation and MC1R Asp294His. Classifying patients into quartiles of slow/fast progression showed mean MSSS increased with faster progression (p<0.001). For EDSS 1-8: MSSS, late onset age and childhood sunburning were associated with fast and MC1R CG/GG(294) with slow progression. Combinations of skin type (1/2 or 3/4) with childhood weekend exposure (< or ≥median) or sunburning (yes/no) were not associated with progression. However, in patients with EDSS1-4, relative to other combinations, those with no sunburning history and types 1/2 demonstrated slow progression (odds ratio=0.15, 95% CI=0.04, 0.57). This method, though a pilot, allows study of associations of variables with EDSS. It is based on local patients and could substitute for MSSS. In patients with EDSS1-4 but not 1-8, skin type 1/2 with no history of childhood sunburning was associated with slow progression. This is compatible with the view that disability develops through a first stage dependent on inflammation.


Low-Cost Zeta Potentiometry Using Solute Gradients.
The zeta potential is an electric potential in the Debye screening layer of an electrolyte, which represents a key physicochemical surface property in various fields ranging from electrochemistry to pharmaceuticals. Thus, characterizing the zeta potential is essential for many applications, but available measurement techniques are limited. Electrophoretic light scattering is typically used to measure the zeta potential of particles in suspension, whereas zeta potential measurements of a solid wall in solution rely on either streaming potential or electroosmotic mobility measurement techniques, both of which are expensive and sophisticated. Here, a simple, robust method to simultaneously measure the zeta potential of particles in suspension and solid walls is presented. The method uses solute gradients to induce particle and fluid motions via diffusiophoresis and diffusioosmosis, respectively, which are both sensitive to the zeta potential of the particle and the wall. By visualizing the particle dynamics, both zeta potentials can be determined independently. Finally, a compact microscope is used to demonstrate low-cost zeta potentiometry that allows measurement of both particle and wall zeta potentials, which suggests a cost-effective tool for pharmaceuticals as well as for educational purposes.


Ethical Considerations in End-of-life Care in the Face of Clinical Futility.
Management of patients with terminal brain disorders can be medically, socially, and ethically complex. Although a growing number of feasible treatment options may exist, there are times when further treatment can no longer meaningfully improve either quality or length of life. Clinicians and patients should discuss goals of care while patients are capable of making their own decisions. However, because such discussions can be challenging, they are often postponed. These discussions are then conducted with patients' health care proxies after patients lose the capacity to make their own decisions. Disagreements may arise when a patient's surrogate desires continued aggressive interventions that are either biologically futile (incapable of producing the intended physiologic result) or potentially inappropriate (potentially capable of producing the patient's intended effect but in conflict with the medical team's ethical principles). This article explores best practices in addressing these types of conflicts in the critical care unit, but these concepts also broadly apply to other sites of care.


Anti-diabetic effect of citrus pectin in diabetic rats and potential mechanism via PI3K/Akt signaling pathway.
This study was performed to investigate the anti-diabetic effect of citrus pectin in type 2 diabetic rats and its potential mechanism of action. The results showed that fasting blood glucose levels were significantly decreased after 4 weeks of citrus pectin administration. Citrus pectin improved glucose tolerance, hepatic glycogen content and blood lipid levels (TG, TC, LDL-c and HDL-c) in diabetic rats. Citrus pectin also significantly reduced insulin resistance, which played an important role in the resulting anti-diabetic effect. Moreover, after the pectin treatment, phosphorylated Akt expression was upregulated and GSK3β expression was downregulated, indicating that the potential anti-diabetic mechanism of citrus pectin might occur through regulation of the PI3K/Akt signaling pathway. Together, these results suggested that citrus pectin could ameliorate type 2 diabetes and potentially be used as an adjuvant treatment.


Recent trends in economic burden of acute myocardial infarction in South Korea.
In 2010, ischemic heart disease was the leading cause of disability-adjusted life-years (DALYs) worldwide. More specially, the prevalence of acute myocardial infarctions (AMI) is increasing in the aged population as mortality decreases; South Korea is no exception. This study aims to examine the economic burden of AMI in the Korean population between 2007 and 2012. AMI-related costs were assessed from a societal perspective. A prevalence-based cost-of-illness framework was used for this analysis. The subjects included all South Koreans with AMI-related ICD-10 codes (I21, I22, I23, I25.0, and I25.1). Data on direct (medical and non-medical) costs and indirect (productivity loss due to AMI-associated morbidity and mortality) costs were collected from the Korean National Health Insurance Service's claims data. The human capital approach was used to calculate indirect costs. The total estimated cost of AMI in 2012 was $1,177,649,323 USD. The majority (52%) of this amount was made up of medical costs, followed by productivity losses due to mortality and morbidity (42% of annual cost). Although the total cost declined by approximately 18% compared to 2007 ($1,427,643,854 USD), the cost of AMI in the over 60 age group amounted to 47% of the total cost of AMI in 2012. AMI led to a high economic burden in 2012. This study, which identified not only the size, but also the trends of AMI-related costs, will provide information to evaluate effects of governmental health projects and the effective allocation of public research funds.


Oxygen extraction index measured by near infrared spectroscopy--a parameter for monitoring tissue oxygenation?
The objective was to assess the ability of near infrared spectrophotometry (NIRS) to detect changes in tissue oxygenation due to alterations in oxygen delivery. Ten hemodynamically stable preterm neonates with a median gestational age of 27.9 weeks (range 25.1-31.2), a median birth weight of 840g (range 690-1310), and a postnatal age of 29 days (range 2-45) were included in this prospective trial. Tissue oxygenation of the lower leg was measured by NIRS and the oxygen extraction index (OEI) was calculated prior and after a transfusion of 10-20 ml/kg body weight packed red blood cells. The OEI decreased from 0.31 (range 0.13-0.39) to 0.24 (range 0.12-0.36, p<0.005). This decrease correlated positively with the weight matched amount of packed red cell transfusion (r2=0.40, p<0.05) and with the increase in hematocrit (r2=0.58, p<0.005). The OEI obtained by a NIRS may allow to monitor changes in tissue oxygenation.


Benznidazole blocks NF-kappaB activation but not AP-1 through inhibition of IKK.
Previously, we demonstrated that benznidazole (BZL), known for its antiparasitic action on Trypanosoma cruzi, modulates pro-inflammatory cytokines and NO release in macrophages by inhibiting NF-kappaB. We now proceeded to elucidate the molecular mechanisms by which BZL exerts its inhibitory action on NF-kappaB. We demonstrated that the inhibitory effect of BZL is not extended to other macrophage responses, since it did not inhibit other typical hallmarks of macrophage activation such as phagocytosis, MHC-II molecules expression or production of reactive oxygen species (ROS) by NADPH oxidase. BZL was able to interfere specifically with the activation of NF-kappaB pathway without affecting AP-1 activation in RAW 264.7 macrophages, not only in LPS-mediated activation, but also for other stimuli, such as pro-inflammatory cytokines (IL-1beta, TNF-alpha), PMA or H(2)O(2). Also, BZL delayed the activation of p38 MAPK, but not that of ERK1/2 and JNK. Finally, treatment with BZL inhibited IkappaBalpha phosporylation and hence its degradation, whereas it did not block IkappaB kinase (IKK) alpha/beta phosphorylation. Collectively, BZL behaves as a broad range specific inhibitor of NF-kappaB activation, independently of the stimuli tested.


Motivational enhancement therapy to improve treatment utilization and outcome in pregnant substance users.
Pregnant substance users can benefit significantly from substance abuse treatment, but treatment retention can be challenging. Two hundred pregnant substance users entering outpatient substance abuse treatment at one of four treatment programs were randomized to receive either three individual sessions of Motivational Enhancement Therapy for pregnant substance users (MET-PS) or the first three individual sessions normally provided by the program. All participants were encouraged to participate in all other treatment offered by the program. Outcome measures included treatment utilization according to clinic records, qualitative urine toxicology measures, and self-report of substance use. One hundred sixty-two (81%) participants completed the 1-month active phase. Participants attended 62% of scheduled treatment on average and reported decreased substance use during the first month of treatment, with no differences between MET-PS and treatment-as-usual (TAU) participants. There was some evidence that the efficacy of MET-PS varied between sites and that MET-PS might be more beneficial than TAU in decreasing substance use in minority participants. These results suggest that MET-PS is not more effective than TAU for pregnant substance users in general but that there might be particular subgroups or treatment programs for which MET-PS might be more or less effective than TAU.


MMP12, lung function, and COPD in high-risk populations.
Genetic variants influencing lung function in children and adults may ultimately lead to the development of chronic obstructive pulmonary disease (COPD), particularly in high-risk groups. We tested for an association between single-nucleotide polymorphisms (SNPs) in the gene encoding matrix metalloproteinase 12 (MMP12) and a measure of lung function (prebronchodilator forced expiratory volume in 1 second [FEV(1)]) in more than 8300 subjects in seven cohorts that included children and adults. Within the Normative Aging Study (NAS), a cohort of initially healthy adult men, we tested for an association between SNPs that were associated with FEV(1) and the time to the onset of COPD. We then examined the relationship between MMP12 SNPs and COPD in two cohorts of adults with COPD or at risk for COPD. The minor allele (G) of a functional variant in the promoter region of MMP12 (rs2276109 [-82A-->G]) was positively associated with FEV(1) in a combined analysis of children with asthma and adult former and current smokers in all cohorts (P=2x10(-6)). This allele was also associated with a reduced risk of the onset of COPD in the NAS cohort (hazard ratio, 0.65; 95% confidence interval [CI], 0.46 to 0.92; P=0.02) and with a reduced risk of COPD in a cohort of smokers (odds ratio, 0.63; 95% CI, 0.45 to 0.88; P=0.005) and among participants in a family-based study of early-onset COPD (P=0.006). The minor allele of a SNP in MMP12 (rs2276109) is associated with a positive effect on lung function in children with asthma and in adults who smoke. This allele is also associated with a reduced risk of COPD in adult smokers.


Analyzing movement trajectories using a Markov bi-clustering method.
In this study we treat scribbling motion as a compositional system in which a limited set of elementary strokes are capable of concatenating amongst themselves in an endless number of combinations, thus producing an unlimited repertoire of complex constructs. We broke the continuous scribblings into small units and then calculated the Markovian transition matrix between the trajectory clusters. The Markov states are grouped in a way that minimizes the loss of mutual information between adjacent strokes. The grouping algorithm is based on a novel markov-state bi-clustering algorithm derived from the Information-Bottleneck principle. This approach hierarchically decomposes scribblings into increasingly finer elements. We illustrate the usefulness of this approach by applying it to human scribbling.


How long should the elderly take antidepressants? A double-blind placebo-controlled study of continuation/prophylaxis therapy with dothiepin. Old Age Depression Interest Group.
Of 219 elderly patients with a major depressive disorder (meeting RDC), 69 recovered sufficiently and consented to enter a two-year double-blind placebo-controlled trial of dothiepin. Survival analysis revealed that dothiepin reduced the relative risk of relapse by two and a half times. Past but not current serious physical illness was also associated with a favourable outcome, whereas a prolonged index depressive illness trebled the relative risk of relapse. In the light of previous research on prognosis it is suggested that elderly persons who recover from a major depressive illness should continue with antidepressant medication for at least two years, if not indefinitely.


Supplementation based on protein or energy ingredients to beef cattle consuming low-quality cool-season forages: I. Forage disappearance parameters in rumen-fistulated steers and physiological responses in pregnant heifers.
Two experiments evaluated the influence of supplement composition on ruminal forage disappearance, performance, and physiological responses of Angus × Hereford cattle consuming a low-quality cool-season forage (8.7% CP and 57% TDN). In Exp. 1, 6 rumen-fistulated steers housed in individual pens were assigned to an incomplete 3 × 2 Latin square design containing 2 periods of 11 d each and the following treatments: 1) supplementation with soybean meal (PROT), 2) supplementation with a mixture of cracked corn, soybean meal, and urea (68:22:10 ratio, DM basis; ENER), or 3) no supplementation (CON). Steers were offered meadow foxtail (Alopecurus pratensis L.) hay for ad libitum consumption. Treatments were provided daily at 0.50 and 0.54% of shrunk BW/steer for PROT and ENER, respectively, to ensure that PROT and ENER intakes were isocaloric and isonitrogenous. No treatment effects were detected on rumen disappearance parameters of forage DM (P ≥ 0.33) and NDF (P ≥ 0.66). In Exp. 2, 35 pregnant heifers were ranked by initial BW on d -7 of the study, allocated into 12 feedlot pens (4 pens/treatment), and assigned to the same treatments and forage intake regimen as in Exp. 1 for 19 d. Treatments were fed once daily at 1.77 and 1.92 kg of DM/heifer for PROT and ENER, respectively, to achieve the same treatment intake as percent of initial BW used in Exp. 1 (0.50 and 0.54% for PROT and ENER, respectively). No treatment effects (P = 0.17) were detected on forage DMI. Total DMI was greater (P < 0.01) for PROT and ENER compared with CON and similar between PROT and ENER (P = 0.36). Accordingly, ADG was greater (P = 0.01) for PROT compared with CON, tended to be greater for ENER compared with CON (P = 0.08), and was similar between ENER and PROT (P = 0.28). Heifers receiving PROT and ENER had greater mean concentrations of plasma glucose (P = 0.03), insulin (P ≤ 0.09), IGF-I (P ≤ 0.04), and progesterone (P = 0.01) compared to CON, whereas ENER and PROT had similar concentrations of these variables (P ≥ 0.15). A treatment × hour interaction was detected (P < 0.01) for plasma urea N (PUN), given that PUN concentrations increased after supplementation for ENER and PROT (time effect, P < 0.01) but did not change for CON (time effect, P = 0.62). In conclusion, beef cattle consuming low-quality cool-season forages had similar ruminal forage disappearance and intake, performance, and physiological status if offered supplements based on soybean meal or corn at 0.5% of BW.


A systematic review of variables used to assess clinically acceptable alignment of unilateral transtibial amputees in the literature.
Prosthetic alignment is a subjective concept which lacks reliability. The outcome responsiveness to prosthetic alignment quality could help to improve subjective and instrument assisted prosthetic alignment. This study was aimed to review variables used to assess clinically acceptable alignment in the literature. The search was done in some databases including: Google Scholar, PubMed, EBSCO, EMBASE, ISI Web of Knowledge and Scopus. The first selection criterion was based on abstracts and titles to address the research questions of interest. The American Academy of Orthotics and Prosthetics checklists were used for paper risk of bias assessment. A total of 25 studies were included in this study. Twenty-four studies revealed the critics of standing position or walking to locate clinically acceptable alignment, only one study measured outcomes in both situations. A total of 253 adults with transtibial amputations and mean age of 48.71 years participated in included studies. The confidence level of included studies was low to moderate, and before-after trial was the most common study design (n = 19). The joint angle, load line location with respect to joints and center of pressure-related parameters were reported as sensitive outcomes to prosthetic alignment quality in standing posture. The amount of forces at various parts of gait cycle and time of events were sensitive to prosthetic alignment quality during walking. Standing balance and posture and temporal parameters of walking could help to locate clinically acceptable alignment.


Pupillary response to chromatic flicker.
There is significant evidence for higher-level cortical control of pupillary responses to visual stimuli, suggesting that factors other than luminance changes may induce a pupillary response. In the present study, the pupillary responses to equiluminant flickering stimuli in a range of 3-13 Hz were examined. Flicker stimuli included color-black (luminance-modulated) and color-color (hue-modulated) flicker. Equiluminance was determined both by objective luminance measures as well as by subjective, perceptual equiluminance for each subject. For both objectively and subjectively equiluminant flicker, significant, sustained pupillary constrictions were recorded. The magnitude of these responses was sensitive to both color and frequency parameters; red-blue color-paired flicker consistently produced the strongest constrictions. These responses occurred even when the flicker was of a lower luminance, both physically and perceptually, than a preceding nonflickering color, indicating that chromatic rather than luminance-sensitive mechanisms are involved in this response. Interestingly, the color- and frequency-sensitivity of constriction parallels those of flickers which maximally stimulate photosensitive epileptic patients, raising the possibility that chromatic response may be a factor in photosensitivity.


Evaluating Innovative In-Ear Pulse Oximetry for Unobtrusive Cardiovascular and Pulmonary Monitoring During Sleep.
Homecare is healthcare based on the principle "outpatient before inpatient," with the aim of moving at least some care-delivery to the home. But reliable determination of vital signs at home requires new, smart sensors, which can be used by the patients themselves. We present a novel pulse oximetry sensor worn in the ear channel. It was previously shown that measurement of heart rate, arterial oxygen saturation and related respiratory information can be performed with reliable accuracy under laboratory conditions. The present study explores the clinical feasibility of the sensor system for cardiovascular monitoring during sleep, with the aim to diagnose sleep apnea. For this, human trials were performed in a sleep laboratory including patients with a clinical suspicion of sleep apnea. Besides a general analysis of the sensor's signal quality during sleep, the evaluation focuses on heart rate dynamics and time-variant oxygen saturation. In addition, several methods to derive respiration rate from photoplethysmographic signals are examined and discussed. Results from the in-ear sensor are compared with standard polysomnography monitoring and demonstrate that this novel system allows long-term nocturnal measurement of heart rate, oxygen saturation and respiratory rate with sufficient accuracy.


Factors affecting attendance to cervical cancer screening among women in the Paracentral Region of El Salvador: a nested study within the CAPE HPV screening program.
Cervical cancer is the third most commonly occurring cancer among women and the fourth leading cause of cancer-related deaths in women worldwide, with more than 85 % of these cases occurring in developing countries. These global disparities reflect the differences in cervical cancer screening rates between high-income and medium- and low-income countries. At 19 %, El Salvador has the lowest reported screening coverage of all Latin American countries. The purpose of this study is to identify factors affecting public sector HPV DNA-based cervical cancer screening participation in El Salvador. This study was nested within a public sector screening program where health promoters used door-to-door outreach to recruit women aged 30-49 years to attend educational sessions about HPV screening. A subgroup of these participants was chosen randomly and questioned about demographic factors, healthcare utilization, previous cervical cancer screening, and HPV knowledge. Women then scheduled screening appointments at their public health clinics. Screening participants were adherent if they attended their scheduled appointment or rescheduled and were screened within 6 months. The association between non-adherence and demographic variables, medical history, history of cancer, sexual history, birth control methods, and screening barriers was assessed using Chi-square tests of significance and logistic regression. All women (n = 409) enrolled in the study scheduled HPV screening appointments, and 88 % attended. Non-adherence was associated with a higher number of lifetime partners and being under-screened-defined as not having participated in cervical cancer screening within the previous 3 years (p = 0.03 and p = 0.04, respectively); 22.8 % of participants in this study were under-screened. Adherence to cervical cancer screening after educational sessions was higher than expected, in part due to interactions with the community-based health promoters as well as the educational session itself. More effective recruitment methods targeted toward under-screened women are required.


Improved resin-to-dentin bond strength and durability via non-thermal atmospheric pressure plasma drying of etched dentin.
This study aimed to evaluate the improvement in strength and durability of the bond between dentin and composite resins following plasma drying of the etched dentin surface using non-thermal atmospheric pressure plasma. Plasma drying was applied to the etched dentin before applying adhesive. Conventional wet-bonding and helium (He) gas-dried bonding schemes were used as control groups. The bond strength of the composite resin to dentin was measured as the microtensile bond strength at 24 h after bonding and after 10,000 cycles of thermocycling. Hybrid layer formation was observed using micro-Raman spectroscopy and scanning electron microscopy. Although the bond-strength values were not statistically different either at 24 h after bonding or after thermocycling, the bond strength of the plasma-dried bonding group was significantly higher than the conventional wet-bonding group and He gas-dried bonding group. Micro-Raman spectral analysis revealed effective penetration of the adhesive and an improved polymerization rate of the adhesive after plasma drying. Plasma drying increased the penetration of hydrophobic resin into the collagen mesh structure, which improved mechanical bonding and long-term durability between dentin and composite resin.


Scandcleft randomised trials of primary surgery for unilateral cleft lip and palate: 4. Speech outcomes in 5-year-olds - velopharyngeal competency and hypernasality.
Adequate velopharyngeal function and speech are main goals in the treatment of cleft palate. The objective was to investigate if there were differences in velopharyngeal competency (VPC) and hypernasality at age 5 years in children with unilateral cleft lip and palate (UCLP) operated on with different surgical methods for primary palatal repair. A secondary aim was to estimate burden of care in terms of received additional secondary surgeries and speech therapy. Three parallel group, randomised clinical trials were undertaken as an international multicentre study by 10 cleft teams in five countries: Denmark, Finland, Sweden, Norway, and the UK. Three different surgical protocols for primary palatal repair were tested against a common procedure in the total cohort of 448 children born with a non-syndromic UCLP. Speech audio and video recordings of 391 children (136 girls, 255 boys) were available and perceptually analysed. The main outcome measures were VPC and hypernasality from blinded assessments. There were no statistically significant differences between the prevalences in the arms in any of the trials. VPC: Trial 1, A: 58%, B: 61%; Trial 2, A: 57%, C: 54%; Trial 3, A: 35%, D: 51%. No hypernasality: Trial 1, A: 54%, B: 44%; Trial 2, A: 47%, C: 51%; Trial 3, A: 34%, D: 49%. No differences were found regarding VPC and hypernasality at age 5 years after different methods for primary palatal repair. The burden of care in terms of secondary pharyngeal surgeries, number of fistulae, and speech therapy visits differed. ISRCTN29932826.


Variation of axial and oblique astigmatism with accommodation across the visual field.
In this study we investigated the impact of accommodation on axial and oblique astigmatism along 12 meridians of the central 30° of visual field and explored the compensation of corneal first-surface astigmatism by the remainder of the eye's optical system. Our experimental evidence revealed no systematic effect of accommodation on either axial or oblique astigmatism for two adult populations (myopic and emmetropic eyes). Although a few subjects exhibited systematic changes in axial astigmatism during accommodation, the dioptric value of these changes was much smaller than the amount of accommodation. For most subjects, axial and oblique astigmatism of the whole eye are both less than for the cornea alone, which indicates a compensatory role for internal optics at all accommodative states in both central and peripheral vision. A new method for determining the eye's optical axis based on visual field maps of oblique astigmatism revealed that, on average, the optical axis is 4.8° temporal and 0.39° superior to the foveal line-of-sight in object space, which agrees with previous results obtained by different methodologies and implies that foveal astigmatism includes a small amount of oblique astigmatism (0.06 D on average). Customized optical models of each eye revealed that oblique astigmatism of the corneal first surface is negligible along the pupillary axis for emmetropic and myopic eyes. Individual variation in the eye's optical axis is due in part to misalignment of the corneal and internal components that is consistent with tilting of the crystalline lens relative to the pupillary axis.


Direct visualization of alpha-synuclein oligomers reveals previously undetected pathology in Parkinson's disease brain.
Oligomeric forms of alpha-synuclein are emerging as key mediators of pathogenesis in Parkinson's disease. Our understanding of the exact contribution of alpha-synuclein oligomers to disease is limited by the lack of a technique for their specific detection. We describe a novel method, the alpha-synuclein proximity ligation assay, which specifically recognizes alpha-synuclein oligomers. In a blinded study with post-mortem brain tissue from patients with Parkinson's disease (n = 8, age range 73-92 years, four males and four females) and age- and sex-matched controls (n = 8), we show that the alpha-synuclein proximity ligation assay reveals previously unrecognized pathology in the form of extensive diffuse deposition of alpha-synuclein oligomers. These oligomers are often localized, in the absence of Lewy bodies, to neuroanatomical regions mildly affected in Parkinson's disease. Diffuse alpha-synuclein proximity ligation assay signal is significantly more abundant in patients compared to controls in regions including the cingulate cortex (1.6-fold increase) and the reticular formation of the medulla (6.5-fold increase). In addition, the alpha-synuclein proximity ligation assay labels very early perikaryal aggregates in morphologically intact neurons that may precede the development of classical Parkinson's disease lesions, such as pale bodies or Lewy bodies. Furthermore, the alpha-synuclein proximity ligation assay preferentially detects early-stage, loosely compacted lesions such as pale bodies in patient tissue, whereas Lewy bodies, considered heavily compacted late lesions are only very exceptionally stained. The alpha-synuclein proximity ligation assay preferentially labels alpha-synuclein oligomers produced in vitro compared to monomers and fibrils, while stained oligomers in human brain display a distinct intermediate proteinase K resistance, suggesting the detection of a conformer that is different from both physiological, presynaptic alpha-synuclein (proteinase K-sensitive) and highly aggregated alpha-synuclein within Lewy bodies (proteinase K-resistant). These disease-associated conformers represent previously undetected Parkinson's disease pathology uncovered by the alpha-synuclein proximity ligation assay.


Distribution of Archaea in Japanese patients with periodontitis and humoral immune response to the components.
There is controversy regarding the existence of archaeal pathogens. Periodontitis is one of the human diseases in which Archaea have been suggested to have roles as pathogens. This study was performed to investigate the distribution of Archaea in Japanese patients with periodontitis and to examine the serum IgG responses to archaeal components. Subgingival plaque samples were collected from 111 periodontal pockets of 49 patients (17 with aggressive periodontitis and 32 with chronic periodontitis), and 30 subgingival plaque samples were collected from 17 healthy subjects. By PCR targeting the 16S rRNA gene, Archaea were detected in 15 plaque samples (13.5% of total samples) from 11 patients (29.4% of patients with aggressive periodontitis and 18.8% of patients with chronic periodontitis). Archaea were detected mostly (14/15) in severe diseased sites (pocket depth > or =6 mm), while no amplicons were observed in any samples from healthy controls. Sequence analysis of the PCR products revealed that the majority of Archaea in periodontal pockets were a Methanobrevibacter oralis-like phylotype. Western immunoblotting detected IgG antibodies against M. oralis in eight of the 11 sera from patients. These results suggest the potential of Archaea (M. oralis) as an antigenic pathogen of periodontitis.


Sapphire fibers: optical attenuation and splicing techniques.
The optical attenuation in sapphire fibers was examined. Attenuation was found to depend heavily on injection conditions. A number of techniques for making sapphire-silica fiber splices were attempted, with an effort toward optimizing injection conditions in the sapphire fiber. The most successful of these techniques, interior capillary-tube splicing, produced robust splices with an attenuation of less than 1 dB.


Determination of molecular orientation at the surface of an aerosol particle by morphology-dependent photoselection.
The orientation of fluorescent molecules on the surface of an aerosol particle has been determined by using a new technique, morphology-dependent photoselection. The excitation spectrum of the angularly averaged fluorescence from a single levitated particle is measured by using an integrating sphere levitator. By utilizing the relative intensities of resonant peaks in this spectrum and knowledge of the polarization properties of different resonances, we show that the orientation of molecules relative to the surface normal may be obtained.


A reexamination of active and passive tumor targeting by using rod-shaped gold nanocrystals and covalently conjugated peptide ligands.
The targeted delivery of nanoparticles to solid tumors is one of the most important and challenging problems in cancer nanomedicine, but the detailed delivery mechanisms and design principles are still not well understood. Here we report quantitative tumor uptake studies for a class of elongated gold nanocrystals (called nanorods) that are covalently conjugated to tumor-targeting peptides. A major advantage in using gold as a "tracer" is that the accumulated gold in tumors and other organs can be quantitatively determined by elemental mass spectrometry (gold is not a natural element found in animals). Thus, colloidal gold nanorods are stabilized with a layer of polyethylene glycols (PEGs) and are conjugated to three different ligands: (i) a single-chain variable fragment (ScFv) peptide that recognizes the epidermal growth factor receptor (EGFR); (ii) an amino terminal fragment (ATF) peptide that recognizes the urokinase plasminogen activator receptor (uPAR); and (iii) a cyclic RGD peptide that recognizes the a(v)β(3) integrin receptor. Quantitative pharmacokinetic and biodistribution data show that these targeting ligands only marginally improve the total gold accumulation in xenograft tumor models in comparison with nontargeted controls, but their use could greatly alter the intracellular and extracellular nanoparticle distributions. When the gold nanorods are administered via intravenous injection, we also find that active molecular targeting of the tumor microenvironments (e.g., fibroblasts, macrophages, and vasculatures) does not significantly influence the tumor nanoparticle uptake. These results suggest that for photothermal cancer therapy, the preferred route of gold nanorod administration is intratumoral injection instead of intravenous injection.


Occurrence and bioconcentration of micropollutants in Silver Perch (Bidyanus bidyanus) in a reclaimed water reservoir.
This study examined the occurrence of 49 micropollutants in reclaimed water and Silver Perch (Bidyanus bidyanus) living in a reclaimed water reservoir. The numbers of micropollutants detected in reclaimed water, Silver Perch liver, and Silver Perch flesh were 20, 23, and 19, respectively. Concentrations of all micropollutants in reclaimed water, except benzotriazole, were well below the Australian Guideline for Recycled Water (AGRW) values for potable purposes. The concentration of benzotriazole in reclaimed water was 675 ± 130 ng/L while the AGRW value for this compound was 7 ng/L. Not all micropollutants detected in the water phase were identified in the Silver Perch flesh and liver tissues. Likewise, not all micropollutants detected in the Silver Perch flesh and liver were identified in the reclaimed water. In general, micropollutant concentrations in the liver were higher than in the flesh. Perfluorooctane sulfonate (PFOS) was detected at a trace level in reclaimed water well below the AGRW guideline value for potable purposes, but showed a high and medium bioconcentration factor in Silver Perch liver and flesh, respectively. In addition, the risk quotient for PFOS was medium and high when considering its concentration in Silver Perch liver and flesh, respectively. Results reported here highlight the need to evaluate multiple parameters for a comprehensive risk assessment. The results also single out PFOS as a notable contaminant of concern for further investigation.


[Seasonal influence on the microbiological colonization on surfaces of milk liners and flushing adapters in a newly installed milking parlor].
A field trial was conducted to evaluate long term and seasonal effects on the development of mictobiological colonization in a newly installed milking parlor. The milking equipment was a 2 x 34 side-by-side milking parlor in a commercial dairy herd in Germany. The 1st batch of samples was taken before beginning of milking and in weekly intervals for a period of two months in winter 2002. The 2nd batch of samples was collected in summer 2003. After sampling the diagnostic material was kept in Amies medium at 4 degrees C until analysis. Samples were analyzed from the mouthpiece, the shank of the liner and the flushing adapter. The samples were collected immediately after disinfection of the milking equipment. They were analyzed qualitatively and quantitatively according to the official German guidelines (Amtliche Sammlung von Untersuchungsverfahren, paragraph 35, LMBG). Mesophil aerobe total plate count, S. aureus, E. coli, Coliforms, Streptococci, Yeast and Lactobacilli were considered. Results indicate a relationship of microbiological colonization of the surface of the milking technique with season and location of sampling. The microbiological colonization was higher in summer than in winter. There were more mesophil aerobe pathogens on the flushing adapters than on mouthpiece and shank of the liner.


Crying following stroke. A report on 30 cases.
Following stroke, many patients experience a heightened tendency to cry. The aim of the present study was to provide a detailed description of this behavior and its accompaniments. Thirty stroke patients who had cried in the previous month were studied using a detailed structured clinical interview. There was a wide range of frequency of crying. The components of crying combine in a more complex and varied way than commonly held stereotyped views suggest. No distinct subtypes were identified. Further research is required to determine the relationship between emotionalism and the frequency and severity of symptoms of depression.


On soliton propagation in biomembranes and nerves.
The lipids of biological membranes and intact biomembranes display chain melting transitions close to temperatures of physiological interest. During this transition the heat capacity, volume and area compressibilities, and relaxation times all reach maxima. Compressibilities are thus nonlinear functions of temperature and pressure in the vicinity of the melting transition, and we show that this feature leads to the possibility of soliton propagation in such membranes. In particular, if the membrane state is above the melting transition solitons will involve changes in lipid state. We discuss solitons in the context of several striking properties of nerve membranes under the influence of the action potential, including mechanical dislocations and temperature changes.


Feto-Maternal Trafficking of Exosomes in Murine Pregnancy Models.
Timing and initiation of labor are well-orchestrated by signals communicated between the fetal and maternal compartments; however, how these signals are communicated is not completely understood. Fetal exosomes, intercellular signaling vesicles, may play a key role in the process. The objective of this study was to evaluate exosome trafficking in vivo from fetal to maternal compartments. Pregnant CD-1 mice were intra-amniotically injected on gestational day 16 and 17 with exosomes isolated from primary human amnion epithelial cells fluorescently labeled with the lipophilic dye 1,1-dioctadecyl-3,3,3,3-tetramethylindotricarbocyanine iodide (DiR). All our analyses were performed on samples collected on Day 18. After 24 h, mice were imaged using Bruker MS FX PRO In vivo Imager and tissues were collected. In vivo imaging of mouse showed fluorescence in the uterus, on the exosome-injected side whereas the uterine tissues from the uninjected side and saline and dye alone injected animals remained negative. Histological analysis of placenta showed exosome migration from the fetal to the maternal side of the placenta. Fluorescence released from exosomes was seen in maternal blood samples as well as in maternal uterus and kidneys. This study demonstrates that exosomal cargo can be carried through systemic route from the fetal to the maternal side of the uterine tissues during pregnancy, supporting the idea that fetal signals can be delivered via exosomes.


Data Resource Profile: the Survey of Health, Ageing and Retirement in Europe (SHARE).
SHARE is a unique panel database of micro data on health, socio-economic status and social and family networks covering most of the European Union and Israel. To date, SHARE has collected three panel waves (2004, 2006, 2010) of current living circumstances and retrospective life histories (2008, SHARELIFE); 6 additional waves are planned until 2024. The more than 150 000 interviews give a broad picture of life after the age of 50 years, measuring physical and mental health, economic and non-economic activities, income and wealth, transfers of time and money within and outside the family as well as life satisfaction and well-being. The data are available to the scientific community free of charge at www.share-project.org after registration. SHARE is harmonized with the US Health and Retirement Study (HRS) and the English Longitudinal Study of Ageing (ELSA) and has become a role model for several ageing surveys worldwide. SHARE's scientific power is based on its panel design that grasps the dynamic character of the ageing process, its multidisciplinary approach that delivers the full picture of individual and societal ageing, and its cross-nationally ex-ante harmonized design that permits international comparisons of health, economic and social outcomes in Europe and the USA.


Evolutionary perspectives on African North American genetic diversity: Origins and prospects for future investigations.
African-descended peoples of the Americas represent an amalgamation of West, Central, and Southeast African regional and ethnic groups with modest gene flow from specific non-African populations. Despite 16+ generations of residence in the Americas, there is a deficit of evolutionary knowledge about these populations. Focusing on Legacy African American, the African North American descendants of survivors of the transatlantic trade in enslaved Africans, we report on emic evolutionary perspectives of their self-identity gleaned from our interviews of 600 individuals collected over 2 years. Gullah-Geechee peoples of Carolina Coastal regions are a model case study due to their historical antiquity, substantial African retentions, relative geospatial isolation, and proposed progenitor status to other Legacy African American microethnic groups. We identify salient research questions for future studies that will begin to bridge the evolutionary gaps in our knowledge of these diverse peoples and the historical evidence for specific evolutionary processes.


Behavioral effects induced by mitochondria-targeted antioxidant SkQ1 in Wistar and senescence-accelerated OXYS rats.
Mitochondrial dysfunction is involved in aging and in neurodegenerative diseases and, therefore, pharmacological agents that alleviate mitochondrial dysfunction are expected to have neuroprotective effects. Promising in this respect is mitochondrial-targeted antioxidant plastoquinonyl-decyl-triphenylphosphonium (SkQ1). We investigated the effects of SkQ1 (250 nmol SkQ1/kg x day with food) on behavior in the elevated plus-maze (EPM) and open field (OF) and on spatial memory in a Morris water maze (MWM) in middle-aged (12 mo) Wistar and senescence-accelerated OXYS rats. Given that changes in the behavior of OXYS rats may be associated with visual impairment, the condition of the retina and the lens was evaluated by ophthalmoscopy. 14-month-old as well as 3-month-old OXYS rats had considerably reduced activities in OF, increased anxiety in EPM, and manifested impaired learning abilities in the MWM in comparison with Wistar rats. SkQ1-treated rats of both strains displayed significantly higher locomotor and exploratory activity in the OF and less anxiety in the EPM compared to age-matched controls. SkQ1 significantly improved visual ability of the rats reducing the severity of the developed signs of retinopathy and cataract but had no impact on OXYS rat's spatial memory in the MWM. SkQ1-treated Wistar rats exhibited slower learning in the MWM task comparison to the control group. Thus, SkQ1 had beneficial effects on locomotor and exploratory functions of the rat brain. Nevertheless, SkQ1 did not alter learning performance in the MWM in OXYS rats and slightly reduced it in the Wistar strain, which may be associated with differences in redox homeostasis.


Translating Genetic Research into Preventive Intervention: The Baseline Target Moderated Mediator Design.
In this paper we present and discuss a novel research approach, the baseline target moderated mediation (BTMM) design, that holds substantial promise for advancing our understanding of how genetic research can inform prevention research. We first discuss how genetically informed research on developmental psychopathology can be used to identify potential intervention targets. We then describe the BTMM design, which employs moderated mediation within a longitudinal study to test whether baseline levels of intervention targets moderate the impact of the intervention on change in that target, and whether change in those targets mediates causal impact of preventive or treatment interventions on distal health outcomes. We next discuss how genetically informed BTMM designs can be applied to both microtrials and full-scale prevention trials. We use simulated data to illustrate a BTMM, and end with a discussion of some of the advantages and limitations of this approach.


Hispanic and Asian performance on selection procedures: A narrative and meta-analytic review of 12 common predictors.
Hispanics are both the largest and fastest growing minority group in the U.S. workforce. Asians also make up a substantial and increasing portion of the workforce. Unfortunately, empirical research on how these groups perform selection procedures appears to be lacking. To address this critical gap, we identified and reviewed research from a variety of literatures relevant to Hispanic and/or Asian performance on 12 commonly used staffing procedures. We also contacted authors of studies that included members of these subgroups and requested the relevant data. On the basis of our review, we provide updated estimates of Hispanic-White and Asian-White differences for the predictors, which often differ from existing estimates of these differences. Further, we provide the first known meta-analytic estimates for Hispanics and Asians on many predictors, such as vocational interests and physical ability. We discuss the implications of the findings for staffing research and practice. We also identify critical next steps for future research regarding these 2 important, yet largely neglected, groups. (PsycINFO Database Record


Acute kidney injury in patients with human immunodeficiency virus infection.
Acute kidney injury (AKI) is an important cause of hospitalization and morbidity in human immunodeficiency virus (HIV)-positive patients. However, the data on AKI in such patients is limited. The aim of the present study was to analyze the incidence, causes and outcome of AKI in HIV-positive patients from our antiretroviral therapy centre. All HIV-positive patients were evaluated for evidence of clinical AKI. AKI was noted in 138/3540 (3.9%) patients. Of 138 AKI patients, 96 (69.6%) had acquired immuno deficiency syndrome and 42 (30.4%) were HIV seropositive. Majority of AKI patients belonged to AKI network (AKIN) Stage II (42%) or III (48.5%) at presentation. Prerenal, intrinsic and postrenal AKI were noted in 53.6%, 44.2% and 2.2% of cases, respectively. Hypovolemia (44.2%) and sepsis (14.5%) contributed to AKI in vast majority of cases. AKI was multifactorial (volume depletion, sepsis and drugs) in 39% of patients. Acute tubular necrosis (ATN) was the most common intrinsic lesion. Acute interstitial nephritis and diffuse endocapillary proliferative glomerulonephritis were noted in five and two cases, respectively. In-hospital mortality was 24.64%. Lower CD4 count, decreased serum albumin level and Stage 4 WHO disease were associated with higher mortality. At 3 months or more follow-up complete recovery of renal function, chronic kidney disease Stage 3-5 and progression to end stage renal disease were noted in 58.69%, 14.5% and 2.2% of cases, respectively. Thus, prerenal factors and ischemic ATN were the most common cause of AKI in HIV-infected patients. Recovery of renal function was seen in 59% of cases, but AKI had high in-hospital mortality.


Self-assembly of partially fluorinated hexabenzocoronene derivatives in the solid state.
We report on the synthesis and structural characterization of novel, partially fluorinated hexabenzocoronene (HBC) derivatives. The fluorination of polycyclic aromatic hydrocarbons (PAHs) is a well-established method to enhance the stability of organic semiconductors (OSCs) and render them n-type. For HBC it has been observed that fluorination leads to a modification of the molecular packing motif from a herringbone arrangement to a parallel-packed motif. Here, we study whether this transformation of the molecular packing is also found for the partially fluorinated HBCs 2,5-difluoro-hexa-peri-hexabenzocoronene (F2HBC) and 2,5,8,11-tetrafluoro-peri-hexabenzocoronene (F4HBC). Combining powder diffraction and NEXAFS dichroism measurements, we reveal that indeed all partially fluorinated compounds adopt a parallel molecular packing, hence maximizing their intermolecular contact area. We identify fluorine-hydrogen bonds as the mediating driving force to specifically stabilize this molecular arrangement and direct self-assembly. Furthermore, we show that the relative orientation of the HBCs on the underlying surface can be precisely controlled by varying the substrate materials. Finally, the energetic states of the compounds are analyzed using photoelectron spectroscopy, optical spectroscopy and density functional theory to identify the effects of fluorination on these fundamental electronic characteristics.


Epidermal growth factor receptor distribution in burn wounds. Implications for growth factor-mediated repair.
Epidermal growth factor (EGF) along with several related peptide growth factors has been shown both in vivo and in vitro to accelerate events associated with epidermal wound repair. EGF and transforming growth factor alpha act by binding to a common EGF receptor tyrosine kinase thereby initiating a series of events which ultimately regulate cell proliferation. This study examined the immunohistochemical localization of EGF receptor (EGF-R) in burn wound margins, adjacent proliferating epithelium, and closely associated sweat ducts, sebaceous glands, and hair follicles. Tissue specimens removed during surgical debridement were obtained from full and partial thickness burn wounds in 32 patients with total body surface area burns ranging from 2 to 88%. In the early postburn period (days 2-4), prominent staining for EGF-R was found in undifferentiated, marginal keratinocytes, adjacent proliferating, hypertrophic epithelium, and both marginal and nonmarginal hair follicles, sweat ducts, and sebaceous glands. During the late postburn period (days 5-16), EGF-R was depleted along leading epithelial margins; however, immunoreactive EGF-R remained intensely positive in the hypertrophic epithelium and all skin appendages. Increased detection of immunoreactive EGF-R and the presence of [125I]EGF binding in the hypertrophic epithelium correlated positively with proliferating cell nuclear antigen distributions. Thus, the presence of EGF-R in the appropriate keratinocyte populations suggests a functional role for this receptor during wound repair. Dynamic modulation in EGF receptor distribution during the temporal sequence of repair provides further evidence that an EGF/transforming growth factor alpha/EGF-R-mediated pathway is activated during human wound repair.


Ocular Histopathological Findings in Semi-Domesticated Eurasian Tundra Reindeer (Rangifer tarandus tarandus) with Infectious Keratoconjunctivitis after Experimental Inoculation with Cervid Herpesvirus 2.
Infectious keratoconjunctivitis (IKC) is a common transmissible ocular disease in semi-domesticated Eurasian tundra reindeer (Rangifer tarandus tarandus). In large outbreaks, IKC may affect tens of animals in a herd, with the most severe cases often requiring euthanasia due to the destruction of the affected eyes and permanent blindness. An experimental inoculation with cervid herpesvirus 2 (CvHV2), alone or in combination with Moraxella bovoculi, demonstrated that CvHV2 has the ability to cause clinical signs of IKC in previously unexposed reindeer. Tissues collected from upper and lower eyelids, lacrimal gland and cornea, were processed for light and transmission electron microscopy. Histopathological analysis of the eyes inoculated with CvHV2 showed widespread and severe pathological findings. Mucosal tissues from these eyes showed fibrinous and purulent exudates, hyperemia, hemorrhages, necrosis, vascular thrombosis, vascular necrosis, infiltration of mononuclear cells and neutrophils, and lymphoid follicle reaction, which matches the described histopathology of IKC in reindeer. Characteristic alpha-herpesvirus particles matching the size and morphology of CvHV2 were identified by transmission electron microscopy in the conjunctival tissue. The quantification of viral particles by qPCR revealed high copy numbers of viral DNA in all CvHV2 inoculated eyes, but also in the non-inoculated eyes of the same animals. The histopathology of eye tissues obtained from the CvHV2 inoculated reindeer and the lack of inflammation from bacterial infection, together with the detection of CvHV2 DNA in swabs from the inoculated and non-inoculated eyes of the same animals, verified that CvHV2 was the primary cause of the observed histopathological changes.


Development of novel murine mammary imaging windows to examine wound healing effects on leukocyte trafficking in mammary tumors with intravital imaging.
We developed mammary imaging windows (MIWs) to evaluate leukocyte infiltration and cancer cell dissemination in mouse mammary tumors imaged by confocal microscopy. Previous techniques relied on surgical resection of a skin flap to image the tumor microenvironment restricting imaging time to a few hours. Utilization of mammary imaging windows offers extension of intravital imaging of the tumor microenvironment. We have characterized strengths and identified some previously undescribed potential weaknesses of MIW techniques. Through iterative enhancements of a transdermal portal we defined conditions for improved quality and extended confocal imaging time for imaging key cell-cell interactions in the tumor microenvironment.


Is depression associated with dysfunction of the central reward system?
The neural substrates of MDD (major depressive disorder) are complex and not yet fully understood. In the present review, I provide a short overview of the findings supporting the hypothesis of a dysfunctional dopamine system in the pathophysiology of depression. Because the mesocorticolimbic dopamine system is involved in reward processing, it has been hypothesized that a reduced function of this system could underlie the anhedonia and amotivation associated with depression. This hypothesis is supported by several observations providing indirect evidence for reduced central dopaminergic transmission in depression. However, some of the differences observed between controls and depressed patients in dopamine function seem to be specific to a subsample of patients, and influenced by the methods chosen. Studies that investigated the neural bases of some MDD behavioural symptoms showed that anhedonia, loss of motivation and the diminished ability to concentrate or make decisions could be associated with a blunted reaction to positive reinforcers and rewards on one side, and with a bias towards negative feedback on the other side. Only a few studies have investigated the neural basis of anhedonia and the responses to rewards in MDD subjects, mostly evidencing a blunted response to reward signals that was associated with reduced brain activation in regions associated with the brain reward system. In conclusion, there is evidence for a dysfunction of the dopamine system in depression and for blunted response to reward signals. However, the exact nature of this dysfunction is not yet clear and needs to be investigated in further studies.


Peroxisome proliferator-induced alterations in the expression and modification of rat hepatocyte plasma membrane proteins.
Rats were fed the peroxisome proliferator ciprofibrate (0.025%), and the effects on the expression, modification, and localization of seven domain-specific integral proteins of the rat hepatocyte plasma membrane were assessed using a combination of immunoblotting, -precipitation, and -fluorescence. Ciprofibrate caused the down-regulation of five of the plasma membrane proteins (the epidermal growth factor receptor, the asialoglycoprotein receptor, HA 321, HA 4, and dipeptidylpeptidase IV) and induced the expression of a more basic, lower-Mr isoform of the basolateral plasma membrane protein CE 9. Pulse labeling, chemical deglycosylation, and 125I-wheat germ lectin blotting suggested that the ciprofibrate-induced isoform of CE 9 differed in the posttranslational modification of its oligosaccharides and contained more sialic acid. These changes in hepatocyte surface differentiation were first observed between Days 1 and 5 on the ciprofibrate-containing diet, coincident with other aspects of the pleiotropic response of the hepatocyte to peroxisome proliferators, e.g., the induction of the Mr 78,000 peroxisome proliferation-associated protein. The effects were reversed within 2-3 weeks upon removal of ciprofibrate. The three other peroxisome proliferators tested, di(2-ethylhexyl)phthalate, clofibrate, and Wy-14,643, were found to exert most of these same effects on the expression and modification of the hepatocyte plasma membrane proteins, but the compounds differed in relative potency. The ciprofibrate-induced decreases in the concentrations of the epidermal growth factor receptor, the asialoglycoprotein receptor, HA 321, and HA 4 were similar to the selective down-regulation of these proteins observed transiently during the period of hepatocyte proliferation following two-thirds hepatectomy. Other compounds frequently used in studies of liver enzyme induction and carcinogenesis, the antioxidants ethoxyquin and butylated hydroxyanisole and the liver tumor promoter phenobarbital, were not as effective as ciprofibrate or two-thirds hepatectomy at causing the down-regulation of these proteins. The induction of the lower-Mr isoform of the basolateral plasma membrane protein CE 9 was not observed following two-thirds hepatectomy or upon the feeding of the antioxidants or phenobarbital but was specific to the feeding of the peroxisome proliferators.


Site-specific multipoint fluorescence measurement system with end-capped optical fibers.
We present the development and implementation of a spatially and spectrally resolved multipoint fluorescence correlation spectroscopy (FCS) system utilizing multiple end-capped optical fibers and an inexpensive laser source. Specially prepared end-capped optical fibers placed in an image plane were used to both collect fluorescence signals from the sample and to deliver signals to the detectors. The placement of independently selected optical fibers on the image plane was done by monitoring the end-capped fiber tips at the focus using a CCD, and fluorescence from specific positions of a sample were collected by an end-capped fiber, which could accurately represent light intensities or spectral data without incurring any disturbance. A fast multipoint spectroscopy system with a time resolution of ∼1.5 ms was then implemented using a prism and an electron multiplying charge coupled device with a pixel binning for the region of interest. The accuracy of our proposed system was subsequently confirmed by experimental results, based on an FCS analysis of microspheres in distilled water. We expect that the proposed multipoint site-specific fluorescence measurement system can be used as an inexpensive fluorescence measurement tool to study many intracellular and molecular dynamics in cell biology.


Structural insight into an atomic layer deposition (ALD) grown Al2O3 layer on Ni/SiO2: impact on catalytic activity and stability in dry reforming of methane.
The development of stable Ni-based dry reforming of methane (DRM) catalysts is a key challenge owing to the high operating temperatures of the process and the propensity of Ni for promoting carbon deposition. In this work, Al2O3-coated Ni/SiO2 catalysts have been developed by employing atomic layer deposition (ALD). The structure of the catalyst at each individual preparation step was characterized in detail through a combination of in situ XAS-XRD, ex situ 27Al NMR and Raman spectroscopy. Specifically, in the calcination step, the ALD-grown Al2O3 layer reacts with the SiO2 support and Ni, forming aluminosilicate and NiAl2O4. The Al2O3-coated Ni/SiO2 catalyst exhibits an improved stability for DRM when compared to the benchmark Ni/SiO2 and Ni/Al2O3 catalysts. In situ XAS-XRD during DRM together with ex situ Raman spectroscopy and TEM of the spent catalysts confirm that the ALD-grown Al2O3 layer suppresses the sintering of Ni, in turn reducing also coke formation significantly. In addition, the formation of an amorphous aluminosilicate phase by the reaction of the ALD-grown Al2O3 layer with the SiO2 support inhibited catalysts deactivation via NiAl2O4 formation, in contrast to the reference Ni/Al2O3 system. The in-depth structural characterization of the catalysts provided an insight into the structural dynamics of the ALD-grown Al2O3 layer, which reacts both with the support and the active metal, allowing to rationalize the high stability of the catalyst under the harsh DRM conditions.


Analysing the Unequal Effects of Positive and Negative Information on the Behaviour of Users of a Taiwanese On-Line Bulletin Board.
The impact of social influence causes people to adopt the behaviour of others when interacting with other individuals. The effects of social influence can be direct or indirect. Direct social influence is the result of an individual directly influencing the opinion of another, while indirect social influence is a process taking place when an individual's opinion and behaviour is affected by the availability of information about others' actions. Such indirect effect may exhibit a more significant impact in the on-line community because the internet records not only positive but also negative information, for example on-line written text comments. This study focuses on indirect social influence and examines the effect of preceding information on subsequent users' opinions by fitting statistical models to data collected from an on-line bulletin board. Specifically, the different impacts of information on approval and disapproval comments on subsequent opinions were investigated. Although in an anonymous situation where social influence is assumed to be at minimum, our results demonstrate the tendency of on-line users to adopt both positive and negative information to conform to the neighbouring trend when expressing opinions. Moreover, our results suggest unequal effects of the local approval and disapproval comments in affecting the likelihood of expressing opinions. The impact of neighbouring disapproval densities was stronger than that of neighbouring approval densities on inducing subsequent disapproval relative to approval comments. However, our results suggest no effects of global social influence on subsequent opinion expression.


Mitochondrial network genes in the skeletal muscle of amyotrophic lateral sclerosis patients.
Recent evidence suggested that muscle degeneration might lead and/or contribute to neurodegeneration, thus it possibly play a key role in the etiopathogenesis and progression of amyotrophic lateral sclerosis (ALS). To test this hypothesis, this study attempted to categorize functionally relevant genes within the genome-wide expression profile of human ALS skeletal muscle, using microarray technology and gene regulatory network analysis. The correlation network structures significantly change between patients and controls, indicating an increased inter-gene connection in patients compared to controls. The gene network observed in the ALS group seems to reflect the perturbation of muscle homeostasis and metabolic balance occurring in affected individuals. In particular, the network observed in the ALS muscles includes genes (PRKR1A, FOXO1, TRIM32, ACTN3, among others), whose functions connect the sarcomere integrity to mitochondrial oxidative metabolism. Overall, the analytical approach used in this study offer the possibility to observe higher levels of correlation (i.e. common expression trends) among genes, whose function seems to be aberrantly activated during the progression of muscle atrophy.


[11C]Choline PET/CT detection of bone metastases in patients with PSA progression after primary treatment for prostate cancer: comparison with bone scintigraphy.
The aim of this study was to evaluate the clinical usefulness of [(11)C]choline positron emission tomography (PET)/CT in comparison with bone scintigraphy (BS) in detecting bone metastases (BM) of patients with biochemical progression after radical treatment for prostate cancer (PCa). Seventy-eight consecutive patients with biochemical progression of PCa (mean prostate-specific antigen 21.1 ng/ml, range 0.2-500.0 ng/ml) referred for both [(11)C]choline PET/CT and BS for restaging purposes were retrospectively analysed. The diagnostic accuracy of [(11)C]choline PET/CT and BS was assessed by using morphological imaging and/or follow-up as standards of reference. As equivocal findings were found, the accuracy analysis was performed twice, once including them as positive and once as negative. A separate analysis was also performed in hormone-resistant patients and data compared with those of patients who did not receive anti-androgenic treatment. Equivocal findings occurred in 1 of 78 (1%) cases in [(11)C]choline PET/CT and in 21 of 78 (27%) cases in BS. Depending on their attribution as either positive or negative, the ranges of sensitivity, specificity, positive predictive value, negative predictive value and accuracy for [(11)C]choline PET/CT were 89-89%, 98-100%, 96-100%, 94-96% and 95-96%, respectively. For BS they were 100-70%, 75-100%, 68--100%, 100-86% and 83-90%, respectively. Concordant findings between [(11)C]choline PET/CT and BS occurred in 55 of 78 (71%) cases. The accuracy of [(11)C]choline PET/CT did not significantly (p = 0.30) differ between hormone-resistant patients (97%) and those who did not receive anti-androgenic treatment (95%). In clinical practice, [(11)C]choline PET/CT may not replace BS because of its lower sensitivity. However, for its high specificity, [(11)C]choline PET/CT positive findings may accurately predict the presence of BM. Equivocal findings are more frequent in BS than [(11)C]choline PET/CT.


Activating mutations in BRAF characterize a spectrum of pediatric low-grade gliomas.
In the present study, DNA from 27 grade I and grade II pediatric gliomas, including ganglioglioma, desmoplastic infantile ganglioglioma, dysembryoplastic neuroepithelial tumor, and pleomorphic xanthoastrocytoma was analyzed using the Illumina 610K Beadchip SNP-based oligonucleotide array. Several consistent abnormalities, including gain of chromosome 7 and loss of 9p21 were observed. Based on our previous studies, in which we demonstrated BRAF mutations in 3 gangliogliomas, 31 tumors were screened for activating mutations in exons 11 and 15 of the BRAF oncogene or a KIAA1549-BRAF fusion product. There were no cases with a KIAA1549-BRAF fusion. A BRAF V600E mutation was detected in 14 of 31 tumors, which was not correlated with any consistent pattern of aberrations detected by the SNP array analysis. Tumors were also screened for mutations in codon 132 in exon 4 of IDH1, exons 2 and 3 of KRAS, and exons 2-9 of TP53. No mutations in KRAS or TP53 were identified in any of the samples, and there was only 1 IDH1 R132H mutation detected among the sample set. BRAF mutations constitute a major genetic alteration in this histologic group of pediatric brain tumors and may serve as a molecular target for biologically based inhibitors.


A reexamination of the folding mechanism of dihydrofolate reductase from Escherichia coli: verification and refinement of a four-channel model.
The mechanism of folding of dihydrofolate reductase from Escherichia coli was reinvestigated by studying the unfolding and refolding kinetics using absorbance and fluorescence spectroscopies. The original kinetic model proposed that folding involved a series of native, intermediate, and unfolded forms which interconverted through four independent channels linked by slow cis/trans isomerization reactions at Xaa-Pro peptide bonds [Touchette, N. A., Perry, K. M., & Matthews, C. R. (1986) Biochemistry 25, 5445]. Recently, alternative sequential models have been proposed [Frieden, C. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 4413; Kuwajima et al. (1991) Biochemistry 30, 7693] which challenge the original proposal. Stopped-flow studies of the intrinsic tryptophan fluorescence demonstrated the presence of three (and tentatively four) kinetic phases in unfolding which correlated well with four phases previously observed in refolding experiments. By monitoring the binding of the inhibitor methotrexate during folding at varying relative concentrations of inhibitor to protein, it was found that the selective loss of the slow-folding phases at substoichiometric levels could only be explained by a four-channel folding model. Double-jump experiments (native-->unfolded-->native) showed that the four refolding channels are populated within 20 s at 15 degrees C and are not likely to be due to proline isomerization. Reverse double-jump experiments (unfolded-->native-->unfolded) demonstrated that interconversions between native conformers are more rapid than originally proposed. Interestingly, the majority of the protein folds through a channel to a native conformer that is minimally populated at equilibrium. This implies that although the folding of dihydrofolate reductase is ultimately under thermodynamic control, kinetic factors contribute to the transient populations of native species during folding.


p53 mutations are common and early events that precede tumor invasion in squamous cell neoplasia of the skin.
Mutations of the p53 gene are the most common genetic abnormality described in human cancer; p53 mutations have recently been reported in more than half of the cases of squamous cell carcinomas of the skin. We have previously reported positive p53 immunostaining in Bowen's disease and actinic keratosis. To determine if this abnormal immunostaining reflects p53 mutation or alternative pathways of p53 protein inactivation we have performed direct sequencing of p53 in 20 further cases of Bowen's disease. We found eight mutations in 20 cases, seven of which would produce alterations in the p53 protein product. Our results suggest that p53 mutation is an early event in malignant conversion, frequently preceding invasion in squamous cell neoplasia of the skin. The type and site of the observed mutations reflect known mutational hotspots and support the role of ultraviolet radiation in the pathogenesis of these tumors.


Duration of lactation and incidence of type 2 diabetes.
Lactation is associated with improved glucose and insulin homeostasis, independent of weight change. To evaluate the association between lactation history and incidence of type 2 diabetes. Prospective observational cohort study of 83,585 parous women in the Nurses' Health Study (NHS) and retrospective observational cohort study of 73,418 parous women in the Nurses' Health Study II (NHS II). Incident cases of type 2 diabetes mellitus. In the NHS, 5145 cases of type 2 diabetes were diagnosed during 1,239,709 person-years of follow-up between 1986 and 2002, and in the NHS II, 1132 cases were diagnosed during 778,876 person-years of follow-up between 1989 and 2001. Among parous women, increasing duration of lactation was associated with a reduced risk of type 2 diabetes. For each additional year of lactation, women with a birth in the prior 15 years had a decrease in the risk of diabetes of 15% (95% confidence interval, 1%-27%) among NHS participants and of 14% (95% confidence interval, 7%-21%) among NHS II participants, controlling for current body mass index and other relevant risk factors for type 2 diabetes. Longer duration of breastfeeding was associated with reduced incidence of type 2 diabetes in 2 large US cohorts of women. Lactation may reduce risk of type 2 diabetes in young and middle-aged women by improving glucose homeostasis.


Hydrogen: Its Future Role in the Nation's Energy Economy.
In examining the potential role of hydrogen in the energy economy of the future, we take an optimistic view. All the technology required for implementation is feasible but a great deal of development and refinement is necessary. A pessimistic approach would obviously discourage further thinking about an important and perhaps the most reasonable alternative for the future. We have considered a limited number of alternative energy systems involving hydrogen and have shown that hydrogen could be a viable secondary source of energy derived from nuclear power; for the immediate future, hydrogen could be derived from coal. A hydrogen supply system could have greater flexibility and be competitive with a more conventional all-electric delivery system. Technological improvements could make hydrogen as an energy source an economic reality. The systems examined in this article show how hydrogen can serve as a general-purpose fuel for residential and automotive applications. Aside from being a source of heat and motive power, hydrogen could also supply the electrical needs of the household via fuel cells (19), turbines, or conventional "total energy systems." The total cost of energy to a residence supplied with hydrogen fuel depends on the ratio of the requirements for direct fuel use to the requirements for electrical use. A greater direct use of hydrogen as a fuel without conversion to electricity reduces the overall cost of energy supplied to the household because of the greater expense of electrical transmission and distribution. Hydrogen fuel is especially attractive for use in domestic residential applications where the bulk of the energy requirement is for thermal energy. Although a considerable amount of research is required before any hydrogen energy delivery system can be implemented, the necessary developments are within the capability of present-day technology and the system could be made attractive economically .Techniques for producing hydrogen from water by electrolysis, from coal, and directly from thermal energy could be found that are less expensive than those now available; inexpensive fuel cells could be developed, and high-temperature turbines could be used for the efficient conversion of hydrogen (and oxygen) to electricity. The use of hydrogen as an automotive fuel would be a key factor in the development of a hydrogen energy economy, and safe storage techniques for carrying sufficient quantities of hydrogen in automotive systems can certainly be developed. The use of hydrogen in automobiles would significantly reduce urban pollution because the dispersed fossil fuel emissions would be replaced by radioactive wastes generated at large central stations. The conversion of internal or external combustion engines for combustion of hydrogen fuel would probably have less economic impact on the automotive industry than the mass introduction of electric automobiles. However, this is a subject that requires more detailed study. All of the safety aspects of hydrogen utilization will have to be examined, especially the problems of safety in the domestic use and the long distance transport of hydrogen in pipelines at high pressures. It is our opinion that the various energy planning agencies should now begin to outline the mode of implementing hydrogen energy delivery systems in the energy economy. The initial transition to hydrogen energy derived from available fossil fuels such as coal should be considered together with the long range view of all the hydrogen being derived eventually from nuclear energy. By the year 1985 when petroleum imports may be in excess of the domestic supply, these plans could set the stage for the transition period from fossil to a predominantly nuclear energy economy able to supply abundant synthetic fuels such as hydrogen. Synthetic fuels will obviously be more expensive than fuels now derived from petroleum; however, there may be no other viable choice. Thus, it is essential that the analysis and technological feasibility of a hydrogen energy system be considered now. It is of vital importance to the nation to develop some general-purpose fuel that can be Produced from a variety of domestic energy sources and reduce our dependence on imported oil.


Birth prevalence of malformations in members of different ethnic groups and in the offspring of matings between them, in Birmingham, England.
The aims were: (1) to compare the birth prevalence of malformations in different ethnic groups and (2) to explore the reasons for the ethnic variations found by examining birth prevalence in the offspring of matings between ethnic groups. Analysis of data from a register of malformations and register of births. Birmingham, England. A total of 432,778 infants (including stillbirths) born in 1960-84. Significant differences (p < 0.01) between ethnic groups were exhibited by the birth prevalence of neural tube defects (NTD), cleft palate, cleft lip, oesophageal atresia/fistula, hypospadias, hip dislocation, clubfoot, polydactyly, and syndactyly. In the offspring of matings between parents of European and Caribbean origin, the birth prevalence of NTD, cleft lip, hypospadias, hip dislocation, polydactyly, and syndactyly seemed more likely to be influenced by the ethnicity of both parents than by that of the mother alone. The reverse was true for the birth prevalence of NTD in subjects with one parent of Irish origin and on of British. Genetic differences may be responsible for Europeans being at lower risk of polydactyly and at higher risk of NTD, cleft lip, hypospadias, hip dislocation, and syndactyly than Caribbeans. Variations in the intrauterine environment are more likely to account for NTD being more common in Irish than in British subjects.


Role of epigenetics in the etiology of germ cell cancer.
Embryonic development is strictly controlled by functionality of genes in which the existing networks can act both on transcription and translation regulation. Germ cell cancers (GCC) are unique because of a number of characteristics. In spite of their clinical presentation, i.e., predominantly after puberty, they arise from primordial germ cells/gonocytes that have failed appropriate maturation to either pre-spermatogonia or oogonia. GCC mimic embryonal development to a certain extent, including capacity for totipotency. This knowledge has allowed the identification of informative diagnostic markers, including OCT3/4 (POU5F1), SOX2 and SOX17. An additional marker is the overall demethylated status of the genome. Genetic mutations in GCC are rare, which is exceptional for solid cancers. Our hypothesis is that a disturbed epigenetic regulation (through combined interaction of genetic or environmental parameters; referred to as genvironment) affect embryonic germ cell development, resulting in delayed or blocked maturation, and potentially progression to GCC. In this respect, studies of patients with Disorders of Sex Development (DSD) have increased our knowledge significantly. Genvironmental influences can lead to retention of existence of embryonic germ cells, the first step in the pathogenesis of GCC, resulting into the precursor lesions gonadoblastoma or carcinoma in situ. Identification of epigenetic alterations could lead to better understanding these processes and development of specific markers for early detection, eventually leading to development of targeted treatment. This review describes an interactive model related to the role of epigenetics in GCC pathogenesis, focusing on DNA methylation, histone modifications, epigenetic memory and inheritance, as well as environmental factors.


Experiences of longing in daily life and associations to well-being among frail older adults receiving home care: a qualitative study.
Purpose: All over the world, communities face the challenge of maintaining well-being among older adults. More specifically, understanding the resources required to establish and maintain well-being among community-dwelling older adults is an essential issue. Although longing from a caring science perspective is considered a driver for well-being, it has not yet been investigated among frail older adults. The aim of this study was to explore frail older adults' experiences of longing in daily life and the relation between longing and well-being from a caring science perspective. Method: The study uses a hermeneutical approach and follows a qualitative explorative design. The data comprises texts from 17 interviews with frail older adults and was analysed by content analysis. Results: The results uncovered three themes: Longing for social contacts; Longing for nature and Longing creativity, aesthetics, and music. Longing was positively related to well-being when the older adults were able to fulfil their longings. Conclusion: This study provides an understanding of the mechanisms of longing among frail older adults. Longing, here, is an inner resource for setting into motion the transition towards well-being. Further studies could focus on how frail older adults can be supported to combat the negative forms of longing in daily life.


The liquidlike ordering of lipid A-diphosphate colloidal crystals: the influence of Ca2+, Mg2+, Na+, and K+ on the ordering of colloidal suspensions of lipid A-diphosphate in aqueous solutions.
A comprehensive study was performed on electrostatically stabilized aqueous dispersion of lipid A-diphosphate in the presence of bound Ca2+, Mg2+, K+, and Na+ ions at low ionic strength (0.10-10.0-mM NaCl, 25 degrees C) over a range of volume fraction of 1.0 x 10(-4)< or =phi< or =4.95 x 10(-4). These suspensions were characterized by light scattering (LS), quasielastic light scattering, small-angle x-ray scattering, transmission electron microscopy, scanning electron microscopy, conductivity measurements, and acid-base titrations. LS and electron microscopy yielded similar values for particle sizes, particle size distributions, and polydispersity. The measured static structure factor, S(Q), of lipid A-diphosphate was seen to be heavily dependent on the nature and concentration of the counterions, e.g., Ca2+ at 5.0 nM, Mg2+ at 15.0 microM, and K+ at 100.0 microM (25 degrees C). The magnitude and position of the S(Q) peaks depend not only on the divalent ion concentration (Ca2+ and Mg2+) but also on the order of addition of the counterions to the lipid A-diphosphate suspension in the presence of 0.1-microM NaCl. Significant changes in the rms radii of gyration (R2G) 1/2 of the lipid A-diphosphate particles were observed in the presence of Ca2+ (24.8+/-0.8 nm), Mg2+ (28.5+/-0.7 nm), and K+ (25.2+/-0.6 nm), whereas the Na+ salt (29.1+/-0.8 nm) has a value similar to the one found for the de-ionized lipid A-diphosphate suspensions (29.2+/-0.8 nm). Effective particle charges were determined by fits of the integral equation calculations of the polydisperse static structure factor, S(Q), to the light-scattering data and they were found to be in the range of Z*=700-750 for the lipid A-diphosphate salts under investigation. The light-scattering data indicated that only a small fraction of the ionizable surface sites (phosphate) of the lipid A-diphosphate was partly dissociated (approximately 30%). It was also discovered that a given amount of Ca2+ (1.0-5.0 nM) or K+ (100 microM) influenced the structure much more than Na+ (0.1-10.0-mM NaCl) or Mg2+ (50 microM). By comparing the heights and positions of the structure factor peaks S(Q) for lipid A-diphosphate-Na+ and lipid A-diphosphate-Ca2+, it was concluded that the structure factor does not depend simply on ionic strength but more importantly on the internal structural arrangements of the lipid A-diphosphate assembly in the presence of the bound cations. The liquidlike interactions revealed a considerable degree of ordering in solution accounting for the primary S(Q) peak and also the secondary minimum at large particle separation. The ordering of lipid A-diphosphate-Ca2+ colloidal crystals in suspension showed six to seven discrete diffraction peaks and revealed a face-centered-cubic (fcc) lattice type (a=56.3 nm) at a volume fraction of 3.2 x 10(-4)< or =phi< or =3.9 x 10(-4). The K+ salt also exhibited a fcc lattice (a=55.92 nm) at the same volume fractions, but reveals a different peak intensity distribution, as seen for the lipid A-diphosphate-Ca2+ salt. However, the Mg2+ and the Na+ salts of lipid A-diphosphate showed body-centered-cubic (bcc) lattices with a=45.50 nm and a=41.50 nm, respectively (3.2 x 10(-4)< or =phi< or =3.9 x 10(-4)), displaying the same intensity distribution with the exception of the (220) diffraction peaks, which differ in intensity for both salts of lipid A-diphosphate.


Shewanella oneidensis MR-1 sensory box protein involved in aerobic and anoxic growth.
Although little is known of potential function for conserved signaling proteins, it is hypothesized that such proteins play important roles to coordinate cellular responses to environmental stimuli. In order to elucidate the function of a putative sensory box protein (PAS domains) in Shewanella oneidensis MR-1, the physiological role of SO3389 was characterized. The predicted open reading frame (ORF) encodes a putative sensory box protein that has PAS, GGDEF, and EAL domains, and an in-frame deletion mutant was constructed (ΔSO3389) with approximately 95% of the ORF deleted. Under aerated conditions, wild-type and mutant cultures had similar growth rates, but the mutant culture had a lower growth rate under static, aerobic conditions. Oxygen consumption rates were lower for mutant cultures (1.5-fold), and wild-type cultures also maintained lower dissolved oxygen concentrations under aerated growth conditions. When transferred to anoxic conditions, the mutant did not grow with fumarate, iron(III), or dimethyl sulfoxide (DMSO) as electron acceptors. Biochemical assays demonstrated the expression of different c-type cytochromes as well as decreased fumarate reductase activity in the mutant transferred to anoxic growth conditions. Transcriptomic studies showed the inability of the mutant to up-express and down-express genes, including c-type cytochromes (e.g., SO4047/SO4048, SO3285/SO3286), reductases (e.g., SO0768, SO1427), and potential regulators (e.g., SO1329). The complemented strain was able to grow when transferred from aerobic to anoxic growth conditions with the tested electron acceptors. The modeled structure for the SO3389 PAS domains was highly similar to the crystal structures of FAD-binding PAS domains that are known O2/redox sensors. Based on physiological, genomic, and bioinformatic results, we suggest that the sensory box protein, SO3389, is an O2/redox sensor that is involved in optimization of aerobic growth and transitions to anoxia in S. oneidensis MR-1.


Ocular-specific ER stress reduction rescues glaucoma in murine glucocorticoid-induced glaucoma.
Administration of glucocorticoids induces ocular hypertension in some patients. If untreated, these patients can develop a secondary glaucoma that resembles primary open-angle glaucoma (POAG). The underlying pathology of glucocorticoid-induced glaucoma is not fully understood, due in part to lack of an appropriate animal model. Here, we developed a murine model of glucocorticoid-induced glaucoma that exhibits glaucoma features that are observed in patients. Treatment of WT mice with topical ocular 0.1% dexamethasone led to elevation of intraocular pressure (IOP), functional and structural loss of retinal ganglion cells, and axonal degeneration, resembling glucocorticoid-induced glaucoma in human patients. Furthermore, dexamethasone-induced ocular hypertension was associated with chronic ER stress of the trabecular meshwork (TM). Similar to patients, withdrawal of dexamethasone treatment reduced elevated IOP and ER stress in this animal model. Dexamethasone induced the transcriptional factor CHOP, a marker for chronic ER stress, in the anterior segment tissues, and Chop deletion reduced ER stress in these tissues and prevented dexamethasone-induced ocular hypertension. Furthermore, reduction of ER stress in the TM with sodium 4-phenylbutyrate prevented dexamethasone-induced ocular hypertension in WT mice. Our data indicate that ER stress contributes to glucocorticoid-induced ocular hypertension and suggest that reducing ER stress has potential as a therapeutic strategy for treating glucocorticoid-induced glaucoma.


Biogenesis of the crystalloid endoplasmic reticulum in UT-1 cells: evidence that newly formed endoplasmic reticulum emerges from the nuclear envelope.
The crystalloid endoplasmic reticulum (ER), a specialized smooth ER of the compactin-resistant UT-1 cell, is composed of multiple membrane tubules packed together in a hexagonal pattern. This membrane contains large amounts of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, an integral membrane protein that enzymatically regulates endogenous cholesterol biosynthesis. Using morphological and immunocytochemical techniques, we have traced the sequence of events in the biogenesis of this ER when compactin-withdrawn UT-1 cells, which do not have a crystalloid ER, are incubated in the presence of compactin. After 15 h of incubation in the presence of compactin, many cells had profiles of ER cisternae that were juxtaposed to the nuclear envelope and studded with ribosomes on their outer membrane. Both the outer nuclear membrane and the ER membrane contained HMG CoA reductase; however, there was little or no detectable enzyme in rough ER that was free in the cytoplasm. With longer times of incubation in the presence of compactin, these cells had lamellar stacks of smooth ER next to the nuclear envelope that contained HMG CoA reductase. Coordinate with the appearance of the smooth ER, crystalloid ER appeared in the same cell. Often regions of continuity were found between the membrane of the smooth ER and the membrane of the crystalloid ER tubules. These studies suggest that HMG CoA reductase is synthesized along the outer nuclear membrane and in response to increased enzyme synthesis, a membrane emerges from the outer nuclear membrane as smooth ER cisternae, which then transforms into crystalloid ER tubules.