Video-rate fluorescence lifetime imaging camera with CMOS single-photon avalanche diode arrays and high-speed imaging algorithm.
A high-speed and hardware-only algorithm using a center of mass method has been proposed for single-detector fluorescence lifetime sensing applications. This algorithm is now implemented on a field programmable gate array to provide fast lifetime estimates from a 32 × 32 low dark count 0.13 μm complementary metal-oxide-semiconductor single-photon avalanche diode (SPAD) plus time-to-digital converter array. A simple look-up table is included to enhance the lifetime resolvability range and photon economics, making it comparable to the commonly used least-square method and maximum-likelihood estimation based software. To demonstrate its performance, a widefield microscope was adapted to accommodate the SPAD array and image different test samples. Fluorescence lifetime imaging microscopy on fluorescent beads in Rhodamine 6G at a frame rate of 50 fps is also shown.


Effect of surface carbon coating on sintering of silver nanoparticles: in situ TEM observations.
In this work, in situ transmission electron microscopy heating has been used to investigate the effects of a carbon capping layer on sintering of silver nanoparticles. For the first time, we make direct and real-time measurements of surface diffusivity of silver in nanoparticles coated with carbon. We observe that the carbon surface coatings may significantly inhibit sintering in silver nanoparticles.


Comparison of fesoterodine and tolterodine extended release for the treatment of overactive bladder: a head-to-head placebo-controlled trial.
To compare the efficacy and tolerability of fesoterodine 8 mg with tolterodine extended-release (ER) 4 mg and placebo in a randomized clinical trial of patients with an overactive bladder (OAB). In this 12-week double-blind, double-dummy, placebo-controlled, randomized clinical trial, eligible patients reported OAB symptoms for > or = 3 months and recorded > or = 8 voids and > or = 1 urgency urinary incontinence (UUI) episode per 24 h in 3-day bladder diaries at baseline. Patients were randomized in a 2:2:1 ratio to fesoterodine (4 mg for 1 week then 8 mg for 11 weeks); tolterodine ER 4 mg; or placebo (with sham dose escalation for tolterodine ER and placebo). Endpoints were changes from baseline to week 12 in UUI episodes (primary endpoint), total and nocturnal voids, urgency episodes, severe urgency episodes, and frequency-urgency sum per 24 h; mean voided volume per void (MVV); and the OAB questionnaire (OAB-q), Patient Perception of Bladder Condition (PPBC), and Urgency Perception Scale (UPS). Safety and tolerability were assessed and summarized over the 12-week study period. Fesoterodine (636 patients) significantly improved UUI episodes at week 12 (primary endpoint) compared with tolterodine ER (641 patients; P = 0.017) and placebo (313 patients; P < 0.001). Fesoterodine also produced significantly greater improvements than tolterodine ER in MVV (P = 0.005). Fesoterodine significantly improved all diary endpoints compared with placebo (P < 0.001), except for nocturnal voids (P = 0.327). Tolterodine ER significantly improved all diary endpoints vs placebo (P < 0.001), except for nocturnal voids (P = 0.506) and MVV (P = 0.103). Diary dry rates (the proportion of patients reporting no UUI episodes at endpoint among those with one or more UUI episodes at baseline) were significantly higher with fesoterodine (64%) than with tolterodine ER (57%; P = 0.015) and placebo (45%; P < 0.001). Improvements in PPBC, UPS and OAB-q scale and domain scores at week 12 were all significantly better with fesoterodine than placebo (all P < 0.001) and tolterodine ER (all P < 0.05) except for the OAB-q Sleep domain vs tolterodine ER (P = 0.081). Dry mouth and constipation rates were 28% and 5% in the fesoterodine group, 16% and 4% in the tolterodine ER group, and 6% and 3% with placebo, respectively. Discontinuations due to treatment-emergent adverse events were 6%, 4% and 2% in the fesoterodine, tolterodine ER, and placebo groups, respectively. In patients with OAB, fesoterodine 8 mg showed superior efficacy over tolterodine ER 4 mg and placebo in reducing UUI episodes (primary endpoint) and in improving most patient-reported outcome measures. Both active treatments were well tolerated.


Influence of the UV Environment on the Synthesis of Prebiotic Molecules.
Ultraviolet radiation is common to most planetary environments and could play a key role in the chemistry of molecules relevant to abiogenesis (prebiotic chemistry). In this work, we explore the impact of UV light on prebiotic chemistry that might occur in liquid water on the surface of a planet with an atmosphere. We consider effects including atmospheric absorption, attenuation by water, and stellar variability to constrain the UV input as a function of wavelength. We conclude that the UV environment would be characterized by broadband input, and wavelengths below 204 nm and 168 nm would be shielded out by atmospheric CO2 and water, respectively. We compare this broadband prebiotic UV input to the narrowband UV sources (e.g., mercury lamps) often used in laboratory studies of prebiotic chemistry and explore the implications for the conclusions drawn from these experiments. We consider as case studies the ribonucleotide synthesis pathway of Powner et al. (2009) and the sugar synthesis pathway of Ritson and Sutherland (2012). Irradiation by narrowband UV light from a mercury lamp formed an integral component of these studies; we quantitatively explore the impact of more realistic UV input on the conclusions that can be drawn from these experiments. Finally, we explore the constraints solar UV input places on the buildup of prebiotically important feedstock gasses like CH4 and HCN. Our results demonstrate the importance of characterizing the wavelength dependence (action spectra) of prebiotic synthesis pathways to determine how pathways derived under laboratory irradiation conditions will function under planetary prebiotic conditions.


A comparison study of repetitive transcranial magnetic stimulation for tinnitus treatment in an Asian population.
Tinnitus, a subjective auditory perception of sounds or noise not triggered by external auditory stimuli, carries considerable morbidity. To date, pharmacological, physical or behavioral therapy is the mainstay of management. We compared repetitive transcranial magnetic stimulation (rTMS) of 1000 or 2000 stimulations/day at 1 Hz and 110% of the motor threshold for 5 consecutive days over the left auditory cortex. Ratings based on the Tinnitus Handicap Inventory (THI) rating scale were completed weekly for 4 weeks. None of the patients had significant hearing impairment. All 28 patients (age range 21-72; 18 men) tolerated rTMS well and no adverse effects were observed. Analysis of variance (ANOVA) showed significant decrease in THI scores over the entire time period (F (1,26)=11.33, p=0.002). At every weekly time point of evaluation, ANOVA with repeated measures demonstrated significantly lower THI score compared to baseline (p<0.02 for all). RTMS treatment had resulted in tinnitus reduction in the range of 15-25% over the 4 week period. Separately, ANOVA also demonstrated significantly reducing THI for both the 1000 pulse (F (1,14)=4.8, p=0.04) and 2000 pulse (F (1,14)=6.56, p=0.02) rTMS treatment arms. Comparison of THI ratings between the 2 treatment arms did not result in significant difference (F (1,26)=1.48, p=0.24). The present study has revalidated the efficacy and safety of rTMS for improving tinnitus up to 4 weeks post-treatment in Asians. However, there was no significant difference with THI evaluation between the 1000 pulse and 2000 pulse treatment arms.


Prevalence of amblyopia and strabismus in Eastern China: results from screening of preschool children aged 36-72 months.
To determine the prevalence of amblyopia and strabismus in preschool children in Eastern China. The Nanjing Pediatric Vision Project was a cross-sectional, population-based cohort study conducted in preschool children aged 36-72 months from 2011 to 2012 in Yuhua District, Nanjing, China, using an age-stratified random sampling procedure. A questionnaire totalling 117 items was sent to be completed by the corresponding parents or legal guardians of each eligible child. Comprehensive eye examinations, including visual acuity, anterior segment examination, autorefraction, cover test and ocular motility, were conducted. Postcycloplegic refraction and fundus examinations were performed if necessary. Amblyopia was present in 68 children (1.20%, 95% CI 0.92% to 1.48%), with no statistical differences in gender (p=0.903) and age (p=0.835). Among these, 27 had bilateral amblyopia and 41 had unilateral amblyopia, including 26 anisometropic without strabismus, 26 binocular refractive, 7 strabismic, 6 combined strabismic/anisometropic and 3 deprivation amblyopia. Strabismus was found in 320 children (5.65%, 95% CI 5.05% to 6.25%), including 43 with concomitant esotropia, 259 with concomitant exotropia (1 esotropia at near but exotropia at distance), 8 with microtropia (3 esotropia and 5 exotropia), 10 with pure vertical strabismus (3 dissociated vertical deviation and 7 oblique muscle dysfunction) and 1 with Type I Duane syndrome. The prevalence of strabismus had no statistical differences in gender (p=0.952), but had significant statistical differences between different age groups (p=0.021). The prevalence of amblyopia and strabismus in preschool children in Eastern China were 1.20% and 5.65%, respectively. The refractive and strabismus are the main factors associated with amblyopia.


Impact of AB0-blood group incompatibility on the outcome of recipients of bone marrow transplants from unrelated donors in the Japan Marrow Donor Program.
Although the AB0 blood group is one of two major antigen systems of relevance for transplantation in humans, there are still conflicting data concerning the influence of AB0 incompatibility on transplant outcome. This study investigated the effect of AB0 incompatibility in recipients of bone marrow transplants from unrelated donors. We retrospectively analyzed data from 5,549 patients who underwent bone marrow transplantation from unrelated donors in the Japan Marrow Donor Program. Overall survival rates in the group with major and minor mismatches were significantly lower than the rate in the AB0-identical group (AB0-identical 63.0%; major mismatch, 56.9%; minor mismatch, 57.1% at 1 year). Treatment-related mortality was higher in the major and minor mismatch groups, but there was no significant difference in the rate of relapse. Cox proportional hazards modeling showed that both major and minor AB0 incompatibility were significant risk factors for transplant-related mortality, independently of disease, patients' age, and HLA incompatibility. Delayed engraftment of neutrophils, platelets, and erythrocytes was observed in transplants with major incompatibility. There was a high incidence of grade 3 and 4 acute graft-versus-host disease in the groups with major and minor mismatches, which was caused by a high incidence of stage 2 to 4 liver graft-versus-host disease. Interestingly, the risk of grade 2 to 4 graft-versus-host disease in the major mismatch group was higher in patients with early engraftment of erythrocytes. Among the patients receiving reduced-intensity conditioning, the transplant-related mortality was also increased in AB0-incompatible transplants. Major and minor AB0 incompatibility have specific effects on transplant-related mortality and acute graft-versus-host disease in recipients of bone marrow transplants from unrelated donors.


Improving the identification and treatment of depression in low-income primary care clinics: a qualitative study of providers in the VitalSign6 program.
Despite its global burden and prevalence, Major Depressive Disorder often goes undetected and untreated, and is particularly pervasive in the primary care setting. One in four Texans lack health insurance, and people with behavioral health disorders are disproportionately affected. It is possible to provide high-quality depression treatment in primary care settings with outcomes equal to those provided by specialty care. The Center for Depression Research and Clinical Care offered an opportunity to transform service delivery practices in underserved primary care practices to improve quality, health status, patient experience and coordination. A point-of-care, web-based, self-report based software program, VitalSign6, was developed to provide universal depression screening in primary care practices and assist providers in monitoring and treating patients' symptoms using principles of Measurement-Based Care. Implementation included a multi-faceted training program designed to build confidence and competence in participating clinics' medical providers and staff as well as ongoing performance improvement delivered by the VitalSign6 team. Primary care providers (N = 11) were interviewed, using a semi-structured interview guide, with a focus on barriers and challenges to full integration, perceptions of the most/least valuable aspects of the program, and the program's impact on knowledge, attitudes and behaviors about depression screening and treatment. More efficient technology is needed to reduce time wasted, as is training to reduce stigma and correct misconceptions about antidepressant medications. Provider buy-in is essential. Despite barriers, VitalSign6 increased knowledge, changed attitudes and enhanced providers' depression screening and treatment skills over time.


Medicare coverage for vision assistive equipment.
Vision loss that cannot be corrected medically, surgically, or by refractive means is considered low vision. Low vision often results in impairment of daily activities, loss of independence, increased risk of fractures, excess health care expense, and reduced physical functioning, quality of life, and life expectancy. Vision rehabilitation can enable more independent functioning for individuals with low vision. The Centers for Medicare and Medicaid Services recognizes the importance of rehabilitation for achieving medically necessary goals but has denied Medicare coverage for vision assistive equipment that is necessary to complete these goals, although they provide coverage for assistive equipment to provide compensation for other disabilities. We believe that this is discriminatory and does not comport with congressional intent. The Centers for Medicare and Medicaid Services should provide coverage for vision assistive equipment, allowing beneficiaries with vision loss to benefit fully from Medicare-covered rehabilitation to achieve the cost-effective results of these services.


Stories from the field: students' descriptions of gender discrimination and sexual harassment during medical school.
Previous studies have documented the prevalence of gender discrimination and sexual harassment during medical training, but very few have examined the behaviors that students perceive as discriminatory or harassing. The authors addressed this lack of information by examining graduating medical students' written descriptions of personal experiences with such behaviors during medical school. The authors reviewed the responses of graduating seniors at 12 U.S. medical schools to a questionnaire, administered in 2001-02, that asked them to provide written descriptions of their personal experiences with gender discrimination and sexual harassment. Seven response categories were created on the basis of recurring themes: educational inequalities; stereotypical comments; sexual overtures; offensive, embarrassing, or sexually explicit comments; inappropriate touching; sexist remarks; and not classifiable. The three authors examined the students' written accounts and placed each into one or more of the categories. Of the students' responses, 290 (36.6%) contained 313 written descriptions of personal experiences that the students perceived as either discriminatory or harassing. The most frequently reported experiences involved educational inequalities; experiences in this category were reported more frequently by men than by women. All other categories of experiences were reported more frequently by women. The results support earlier findings of the prevalence of gender discrimination and sexual harassment during undergraduate medical education. Perhaps formal antiharassment policies should provide examples of unacceptable behavior that are based on categories such as those revealed by this analysis. Perhaps, too, medical students' comments could be used to develop educational interventions for physicians in supervisory positions.


Cemented total hip replacement cable debris and acetabular construct durability.
Third-body wear can adversely affect the outcome of total hip arthroplasty by causing increased polyethylene wear, osteolysis, and component loosening. We hypothesized that there would be greater generation and migration of metal debris to the bearing surfaces in hips in which cobalt-chromium cables were used to reattach the osteotomized greater trochanter when compared with hips in which stainless steel wires were used. Between June 1981 and December 1983, 196 consecutive total hip arthroplasties were performed with use of an Iowa stem and a titanium-backed cemented acetabular component, with cobalt-chromium cable trochanteric reattachment. After nineteen to twenty years of follow-up, the patients were evaluated with regard to the depth of head penetration into the polyethylene (as a surrogate for wear), osteolysis, loosening, and the need for revision. The results were compared with those for a series of 304 total hip arthroplasties that were performed by the same surgeon from January 1984 to December 1985 with use of the same components and the same surgical technique, but with stainless steel wire trochanteric reattachment. The two groups had a comparable nineteen to twenty-year follow-up. All living patients (fifty-nine hips in the cable group and ninety-two hips in the wire group) had minimum ten-year follow-up radiographs. The polyethylene wear rate was 0.101 mm/yr for the cable group and 0.082 mm/yr for the wire group (p = 0.039). For the living patients, the rate of revision of the acetabular component because of aseptic loosening was 37.3% (twenty-two hips) for the cable group and 20.7% (nineteen hips) for the wire group (p = 0.025). The rate of acetabular osteolysis was 44% (twenty-six hips) for the cable group and 26% (twenty-four hips) for the wire group (p = 0.022). Kaplan-Meier analysis with revision of the acetabular component because of aseptic loosening as the end point demonstrated survival rates of 73.7% +/- 9% and 83% +/- 7% for the cable and wire groups, respectively, at twenty years (p = 0.03). Because cable trochanteric attachment led to significantly greater polyethylene wear, osteolysis, acetabular loosening, and acetabular revision, presumably due to third-body metallic debris generation in this cemented total hip replacement construct, surgeons should be aware of the deleterious effects of third-body debris and avoid the use of potential debris generators in the total hip arthroplasty construct. If cable is used and fretting is recognized, especially with intra-articular migration of metallic material or nonunion of the greater trochanter, consideration should be given to cable removal.


Computational strategies to address chromatin structure problems.
While the genetic information is contained in double helical DNA, gene expression is a complex multilevel process that involves various functional units, from nucleosomes to fully formed chromatin fibers accompanied by a host of various chromatin binding enzymes. The chromatin fiber is a polymer composed of histone protein complexes upon which DNA wraps, like yarn upon many spools. The nature of chromatin structure has been an open question since the beginning of modern molecular biology. Many experiments have shown that the chromatin fiber is a highly dynamic entity with pronounced structural diversity that includes properties of idealized zig-zag and solenoid models, as well as other motifs. This diversity can produce a high packing ratio and thus inhibit access to a majority of the wound DNA. Despite much research, chromatin's dynamic structure has not yet been fully described. Long stretches of chromatin fibers exhibit puzzling dynamic behavior that requires interpretation in the light of gene expression patterns in various tissue and organisms. The properties of chromatin fiber can be investigated with experimental techniques, like in vitro biochemistry, in vivo imagining, and high-throughput chromosome capture technology. Those techniques provide useful insights into the fiber's structure and dynamics, but they are limited in resolution and scope, especially regarding compact fibers and chromosomes in the cellular milieu. Complementary but specialized modeling techniques are needed to handle large floppy polymers such as the chromatin fiber. In this review, we discuss current approaches in the chromatin structure field with an emphasis on modeling, such as molecular dynamics and coarse-grained computational approaches. Combinations of these computational techniques complement experiments and address many relevant biological problems, as we will illustrate with special focus on epigenetic modulation of chromatin structure.


Right-wing authoritarianism and stereotype-driven expectations interact in shaping intergroup trust in one-shot vs multiple-round social interactions.
Trust towards unrelated individuals is often conditioned by information about previous social interactions that can be derived from either personal or vicarious experience (e.g., reputation). Intergroup stereotypes can be operationalized as expectations about other groups' traits/attitudes/behaviors that heavily influence our behavioral predictions when interacting with them. In this study we investigated the role of perceived social dimensions of the Stereotype Content Model (SCM)-Warmth (W) and Competence (C)-in affecting trusting behavior towards different European national group members during the Trust Game. Given the well-known role of ideological attitudes in regulating stereotypes, we also measured individual differences in right-wing authoritarianism (RWA). In Experiment 1, we designed an online survey to study one-shot intergroup trust decisions by employing putative members of the European Union states which were also rated along SCM dimensions. We found that low-RWA participants' trusting behavior was driven by perceived warmth (i.e., the dimension signaling the benevolence of social intentions) when interacting with low-C groups. In Experiment 2, we investigated the dynamics of trust in a multiple-round version of the European Trust Game. We found that in low-RWA participants trusting behavior decreased over time when interacting with high-W groups (i.e., expected to reciprocate trust), but did not change when interacting with low-W groups (i.e., expected not to reciprocate trust). Moreover, we found that high-RWA participants' trusting behavior decreased when facing low-W groups but not high-W ones. This suggests that low-RWA individuals employ reputational priors but are also permeable to external evidence when learning about others' trustworthiness. In contrast, high-RWA individuals kept relying on stereotypes despite contextual information. These results confirm the pivotal role played by reputational priors triggered by perceived warmth in shaping social interactions.


Identification of two lysosomal membrane glycoproteins.
Two murine lysosome-associated membrane proteins, LAMP-1 of 105,000-115,000 D and LAMP-2 of 100,000-110,000 D, have been identified by monoclonal antibodies that bind specifically to lysosomal membranes. Both glycoproteins were distinguished as integral membrane components solubilized by detergent solutions but not by various chaotropic agents. The lysosome localization was demonstrated by indirect immunofluorescent staining, co-localization of the antigen to sites of acridine orange uptake, and immunoelectron microscopy. Antibody binding was predominantly located at the limiting lysosomal membrane, distinctly separated from colloidal gold-labeled alpha-2-macroglobulin accumulated in the lumen during prolonged incubation. LAMP-1 and LAMP-2 also appeared to be present in low concentrations on Golgi trans-elements but were not detected in receptosomes marked by the presence of newly endocytosed alpha-2-macroglobulin, or in other cellular structures. LAMP-1 and LAMP-2 were distinguished as different molecules by two-dimensional gel analysis, 125I-tryptic peptide mapping, and sequential immunoprecipitations of 125I-labeled cell extracts. Both glycoproteins were synthesized as a precursor protein of approximately 90,000 D, and showed a marked heterogeneity of apparent molecular weight expression in different cell lines. LAMP-2 was closely related or identical to the macrophage antigen, MAC-3, as indicated by antibody adsorption and tryptic peptide mapping. It is postulated that these glycoproteins, as major protein constituents of the lysosomal membrane, have important roles in lysosomal structure and function.


Hypertonic saline solution (NaCl 7.2%) enhances renal excretion of acids in cattle with acute ruminal lactic acidosis.
The present study aimed to evaluate the efficiency of hypertonic saline solution (HSS) as a novel treatment of acute ruminal lactic acidosis (ARLA) in cattle, focusing on urinary excretion of acids. Twelve cannulated steers were submitted to experimentally induced ARLA by adminis- tering sucrose into the rumen. Twenty hours later, the cattle were randomly divided into two equal groups. The first group was treated with 7.5% HSS (5 mL/kg) over 15 min, and isotonic saline solution (ISS; 20 mL/kg) for the subsequent 165 minutes. The control group was administered ISS instead of HSS. Rumen and urine samples were collected at different times during the experiment from the baseline to 64 h post-induction. The induction caused a medium-to-moderate ruminal acidosis, and a moderate degree of systemic acidosis and dehydration. Steers treated with HSS increased by 50% its glomerular filtration rate (1.61 mL/min) compared to ISS group (1.06 mL/ min; p⟨0.03). The overall volume of urine excreted by HSS group was higher than that in ISS group (1.62 L vs 0.7 L; p⟨0.02). This increase in total volume of urine provided by HSS favored a greater excretion of H+ ions in urine, which was 3.39-fold higher in HSS group (64.3*10-7 vs 18.9*10-7 Mol) as well as lactate (241.7 vs 181.8 mMol) and P urinary excretion (3.8 vs 1.1 mMol) that reduced the urine pH (5.3 vs 5.7). Only the HSS group decreased significantly blood total lactic acid concentration (20.3 %) throughout the treatment. A positive relationship was found between the excretion of urinary phosphorus and urinary pH (r2=0.562). The results showed that this novel treatment with HSS enhanced renal excretion of acids and may be recommended as an additional treatment for cattle with lactic acidosis.


Wi-Fi decreases melatonin protective effect and increases hippocampal neuronal damage in pentylenetetrazole induced model seizures in rats.
Epilepsy is a common brain disorder in which the seizures could cause a neuronal loss in the hippocampus. Oxidative stress has an important role in the pathology of epilepsy. Some studies indicate that Wi-Fi increases oxidative stress and suppresses antioxidant systems. The aim of this study is to investigate the effect of Wi-Fi on melatonin anticonvulsive effect and oxidative damage in pentylenetetrazole-induced epileptic seizures in rats. In our study, we used 30 male Wistar Albino rats, 230-250 grams of the body weight. The animals were divided into five groups as control, saline (1 ml/kg/day olive oil for 30 days), Wi-Fi (12 h/day for 30 days), melatonin (10 mg/kg/day for 30 days) and melatonin + Wi-Fi (10 mg/kg/day +12 h/day for 30 days). In the thirtieth day, thirty minutes after the last drugs administration at the indicated doses, PTZ in 45 mg/kg was administered to induce epileptic seizure. The animals were observed for 30 min during the seizure stages (according to the Racine Scale) and first myoclonic jerk times (FMJ). Twenty-four hours after PTZ injection, brain tissues were removed for biochemical and histopathological evaluation. The hippocampal Cornu Ammonis (CA) 1, CA3 and DG (dentate gyrus) regions were histopathologically evaluated in terms of a neuronal damage in addition that oxidative stress markers (total antioxidant status (TAS), total oxidant status (TOS) and oxidative stress index (OSI)) were measured in brain tissues. Wi-Fi was not found to affect behavioral changes associated with epilepsy (p > 0.05). However, Wi-Fi reduced anticonvulsive and antioxidant effect of melatonin (p < 0.05). Moreover, Wi-Fi increased neuronal damage in hippocampus (p < 0.05). Wi-Fi did not directly affect epileptic seizures. Nevertheless, it inhibits the positive effects of melatonin on epilepsy and it also has negative effects on hippocampal neuronal damage. These effects of Wi-Fi may occur via oxidative pathways.


Cutaneous stimulation evokes long-lasting excitation of spinal interneurons in the turtle.
1. We demonstrated multisecond increases in the excitability of the rostral-scratch reflex in the turtle by electrically stimulating the shell at sites within the rostral-scratch receptive field. To examine the cellular mechanisms for these multisecond increases in scratch excitability, we recorded from single cutaneous afferents and sensory interneurons that responded to stimulation of the shell within the rostral-scratch receptive field. A single segment of the midbody spinal cord (D4, the 4th postcervical segment) was isolated in situ by transecting the spinal cord at the segment's anterior and posterior borders. The isolated segment was left attached to its peripheral nerve that innervates part of the rostral-scratch receptive field. A microsuction electrode (4-5 microns ID) was used to record extracellularly from the descending axons of cutaneous afferents and interneurons in the spinal white matter at the posterior end of the D4 segment. 2. The turtle shell is innervated by slowly and rapidly adapting cutaneous afferents. All cutaneous afferents responded to a single electrical stimulus to the shell with a single action potential. Maintained mechanical stimulation applied to the receptive field of some slowly adapting afferents produced several seconds of afterdischarge at stimulus offset. We refer to the cutaneous afferent afterdischarge caused by mechanical stimulation of the shell as "peripheral afterdischarge." 3. Within the D4 spinal segment there were some interneurons that responded to a brief mechanical stimulus within their receptive fields on the shell with short afterdischarge and others that responded with long afterdischarge. Short-afterdischarge interneurons responded to a single electrical pulse to a site in their receptive fields either with a brief train of action potentials or with a single action potential. Long-afterdischarge interneurons responded to a single electrical shell stimulus with up to 30 s of afterdischarge. Long-afterdischarge interneurons also exhibited strong temporal summation in response to a pair of electrical shell stimuli delivered up to several seconds apart. Because all cutaneous afferents responded to an electrical shell stimulus with a single action potential, we conclude that electrically evoked afterdischarge in interneurons was produced by neural mechanisms in the spinal cord; we refer to this type of afterdischarge as "central afterdischarge." 4. These results demonstrate that neural mechanisms for long-lasting excitability changes in response to cutaneous stimulation reside in a single segment of the spinal cord. Cutaneous interneurons with long afterdischarge may serve as cellular loci for multise


Increase in neurokinin-1 receptor-mediated colonic motor response in a rat model of irritable bowel syndrome.
Irritable bowel syndrome (IBS) is a functional bowel disorder. Its major symptom is bowel dysmotility, yet the mechanism of the symptom is poorly understood. Since the neurokinin-1 receptor (NK1R)-mediated signaling in the gut is important in the control of normal bowel motor function, we aimed to investigate whether the NK1R-mediated bowel motor function was altered in IBS, using a rat IBS model that was previously reported to show colonic dysmotility in response to restraint stress. IBS symptoms were produced in male Sprague-Dawley rats by inducing colitis with acetic acid. Rats were left to recover from colitis for 6 d, and used for experiments 7 d post-induction of colitis. Motor activities of distal colon were recorded in vitro. The contractile sensitivity of isolated colon to a NK1R agonist (Sar9,Met(O2)11)-substance P (1-30 nmol/L) was higher in IBS rats than that in normal rats. After the enteric neurotransmission was blocked by tetrodotoxin (TTX, 1 micromol/L), the contractile sensitivity to the NK1R agonist was increased in normal colon but not in IBS rat colon. The NK1R agonist-induced contraction was not different between the two groups when the agonist was challenged to the TTX-treated colon or the isolated colonic myocytes. A nitric oxide synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME, 100 micromol/L) augmented the NK1R agonist-induced contraction only in normal rat colon. These results suggest that the NK1R-meidated colonic motor response is increased in IBS rats, due to the decrease in the nitrergic inhibitory neural component.


[A case of corticobasal degeneration that progressed from primary progressive aphasia].
A 65-year-old woman began to experience slowly progressive speech disturbance from 2001. She was admitted to our hospital for examination on May 2003. She had dysprosody, paragraphia, and mild disturbance in comprehension of spoken language. Repetition was preserved. No clear paraphasia was found. Calculation was disturbed, but there were no other neurological abnormalities including apraxia and agnosia. Brain MRI showed atrophy of the left frontal and parietal lobes. 99mTc-ECD SPECT showed decreased blood flow in the left frontal lobe and parietal lobe, especially in the Broca area and supplemental motor cortex. Primary progressive aphasia was diagnosed, because aphasia lasted for 2 years without other neurological deficits, and her daily activity was well preserved. During follow up, facial apraxia appeared from December 2003, and limb apraxia appeared from May 2004, followed by rigidity predominantly in the right upper limb and dementia. She was diagnosed as having corticobasal degeneration (CBD). The second SPECT on December 2004 showed progression of the decrease in cerebral flow at the same area showed by the first SPECT. SPECT is useful examination to predict the progress of the disease because the decrease of blood flow was recognized before the progression to CBD.


Combined association of occupational and leisure-time physical activity with all-cause and coronary heart disease mortality among a cohort of men followed-up for 22 years.
Leisure-time physical activity (LTPA) is a well-established protective factor for all-cause mortality and cardiovascular mortality while occupational physical activity (OPA) has shown contradictory results. We examined the association between OPA and all-cause and coronary heart disease (CHD) mortality, and tested its combined effect with LTPA. The CORDIS Study (Cardiovascular Occupational Risk Factor Determination in Israel Study) is a prospective cohort study of industrial workers examined during 1985-1989 and followed-up for 22 years. Data on self-reported OPA and LTPA among 4819 males (20-70 years old) were merged with data on all-cause and CHD mortality obtained from the National Death Registry. A higher incidence rate of all-cause mortality and CHD mortality was observed among men who performed moderate-hard OPA compared with those who performed none-mild OPA. Multiple regression analysis based on the Cox proportional hazards model showed that moderate-hard OPA was associated with increased risk of all-cause mortality (HR=1.42, 95% CI 1.16 to 1.74, p<0.001), while LTPA (30 min at least twice a week vs less or none) was associated with reduced risk for all-cause mortality (HR=0.61, 95% CI 0.48 to 0.79, p<0.001), after adjusting for potential confounders, including sociodemographic variables, body mass index, comorbidity and lifestyle habits. Employees who performed moderate-hard OPA and no LTPA had the greatest risk for all-cause mortality and employees who performed none-light OPA and LTPA had the lowest risk. Similar but non-significant trends were observed for the association with CHD mortality. Moderate-hard OPA among industrial male workers may be deleterious to health and should not be a substitute to LTPA.


Genome-Wide Association Study for Anthracycline-Induced Congestive Heart Failure.
Anthracycline-induced congestive heart failure (CHF) is a rare but serious toxicity associated with this commonly employed anticancer therapy. The ability to predict which patients might be at increased risk prior to exposure would be valuable to optimally counsel risk-to-benefit ratio for each patient. Herein, we present a genome-wide approach for biomarker discovery with two validation cohorts to predict CHF from adult patients planning to receive anthracycline. We performed a genome-wide association study in 3,431 patients from the randomized phase III adjuvant breast cancer trial E5103 to identify single nucleotide polymorphism (SNP) genotypes associated with an increased risk of anthracycline-induced CHF. We further attempted candidate validation in two independent phase III adjuvant trials, E1199 and BEATRICE. When evaluating for cardiologist-adjudicated CHF, 11 SNPs had a P value <10-5, of which nine independent chromosomal regions were associated with increased risk. Validation of the top two SNPs in E1199 revealed one SNP rs28714259 that demonstrated a borderline increased CHF risk (P = 0.04, OR = 1.9). rs28714259 was subsequently tested in BEATRICE and was significantly associated with a decreased left ventricular ejection fraction (P = 0.018, OR = 4.2). rs28714259 represents a validated SNP that is associated with anthracycline-induced CHF in three independent, phase III adjuvant breast cancer clinical trials. Clin Cancer Res; 23(1); 43-51. ©2016 AACR.


Supravalvular aortic stenosis associated with a deletion disrupting the elastin gene.
Supravalvular aortic stenosis (SVAS) is an inherited vascular disease that can cause heart failure and death. SVAS can be inherited as an autosomal dominant trait or as part of a developmental disorder, Williams syndrome (WS). In recent studies we presented evidence suggesting that a translocation disrupting the elastin gene caused SVAS in one family while deletions involving the entire elastin locus caused WS. In this study, pulsed-field, PCR, and Southern analyses showed that a 100-kb deletion of the 3' end of the elastin gene cosegregated with the disease in another SVAS family. DNA sequence analysis localized the breakpoint between elastin exons 27 and 28, the same region disrupted by the SVAS-associated translocation. These data indicate that mutations in the elastin gene cause SVAS and suggest that elastin exons 28-36 may encode critical domains for vascular development.


Towards patient safety: assessment of medication errors in the intensive care unit in a developing country's tertiary care teaching hospital.
To determine the incidence, causes, patterns and outcomes of medication errors (MEs) in the intensive care unit. The ME reporting system was established using the principles based on prospective, voluntary, open, anonymous and stand-alone surveillance in a tertiary care teaching hospital located in southern India. MEs involving patients of either sex were included in the study, and the reporters were given the choice to remain anonymous. The analysis was carried out to determine the patterns, causes and outcomes of the reported errors and was discussed with healthcare professionals (HCPs) to minimise the recurrence of MEs. A total of 292 MEs were reported voluntarily among 5137 admitted patients and the incidence of MEs was 5.6%. Administration errors (n=143, 49%) were the most common type of MEs reported followed by prescription errors (n=56, 19%) and dispensing errors (n=43, 15%). Factors responsible for MEs were related to performance deficit of HCPs due to excessive workload, fatigue, unclear interpersonnel communications and patient-related factors, which accounted for 37.6%, 13.1%, 9.6% and 7.7%, respectively. The majority of the reported MEs had an outcome of category C and A, based on the National Coordinating Council for ME Reporting and Prevention (NCC MERP) outcome category scale, amounting to 42.2% and 41.7%, respectively. Although the majority of MEs that reached the patients did not cause any harm, providing continuous education and awareness of MEs to HCPs and patients may minimise the scope of the factors that may contribute to MEs and improve overall patient safety.


Stable oxygen isotope composition of plant tissue: a review.
With the development of rapid measurement techniques, stable oxygen isotope analysis of plant tissue is poised to become an important tool in plant physiological, ecological, paleoclimatic and forensic studies. Recent advances in mechanistic understanding have led to the improvement of process-based models that accurately predict variability in the oxygen isotope composition of plant organic material (δ18Op). δ18Op has been shown to reflect the isotope composition of soil water, evaporative enrichment in transpiring leaves, and isotopic exchange between oxygen atoms in organic molecules and local water in the cells in which organic molecules are formed. This review presents current theoretical models describing the influences on δ18Op, using recently published experimental work to outline strengths and weaknesses in the models. The potential and realised applications of the technique are described.


Hepatocyte growth factor signaling in intrapancreatic ductal cells drives pancreatic morphogenesis.
In a forward genetic screen for regulators of pancreas development in zebrafish, we identified donut(s908) , a mutant which exhibits failed outgrowth of the exocrine pancreas. The s908 mutation leads to a leucine to arginine substitution in the ectodomain of the hepatocyte growth factor (HGF) tyrosine kinase receptor, Met. This missense mutation impedes the proteolytic maturation of the receptor, its trafficking to the plasma membrane, and diminishes the phospho-activation of its kinase domain. Interestingly, during pancreatogenesis, met and its hgf ligands are expressed in pancreatic epithelia and mesenchyme, respectively. Although Met signaling elicits mitogenic and migratory responses in varied contexts, normal proliferation rates in donut mutant pancreata together with dysmorphic, mislocalized ductal cells suggest that met primarily functions motogenically in pancreatic tail formation. Treatment with PI3K and STAT3 inhibitors, but not with MAPK inhibitors, phenocopies the donut pancreatic defect, further indicating that Met signals through migratory pathways during pancreas development. Chimera analyses showed that Met-deficient cells were excluded from the duct, but not acinar, compartment in the pancreatic tail. Conversely, wild-type intrapancreatic duct and "tip cells" at the leading edge of the growing pancreas rescued the donut phenotype. Altogether, these results reveal a novel and essential role for HGF signaling in the intrapancreatic ducts during exocrine morphogenesis.


Insight into proteomic investigations of Neisseria meningitidis serogroup C strain L91543 from analysis of its genome sequence.
Here, we describe the draft sequence of a virulent isolate of Neisseria meningitidis strain L91543, belonging to serogroup C. The findings from previous proteomic and metabolomic studies of this strain can now be further interpreted with genomic analysis. Comparative analysis of the genome sequence revealed close similarity and localized synteny with the genome sequence of N. meningitidis serogroup C strain, FAM18. Polymorphisms were identified in the signal peptide sequence of factor H binding protein, a target for new meningococcal vaccines, which may result in its inefficient translocation across the cytoplasmic membrane affecting its processing (lipidation and cleavage of the signal peptide) and transportation to the outer membrane in strain L91543. This would explain the unusual proteomic data for factor H binding protein for this strain. NadA, another target for new vaccines, and the MtrR regulator, which controls expression of NadA, both contain SNPs between strains L91543 and FAM18. The genome sequence data were generated using Ion Torrent PGM sequencing, assembled into 50 contigs with 64× coverage and annotated with 2262 genes, 14 rRNAs and 56 tRNAs. The availability of the genome of N. meningitidis strain L91543 will aid our understanding of the proteome of this organism, importantly its vaccine antigens.


Rats and humans can optimally accumulate evidence for decision-making.
The gradual and noisy accumulation of evidence is a fundamental component of decision-making, with noise playing a key role as the source of variability and errors. However, the origins of this noise have never been determined. We developed decision-making tasks in which sensory evidence is delivered in randomly timed pulses, and analyzed the resulting data with models that use the richly detailed information of each trial's pulse timing to distinguish between different decision-making mechanisms. This analysis allowed measurement of the magnitude of noise in the accumulator's memory, separately from noise associated with incoming sensory evidence. In our tasks, the accumulator's memory was noiseless, for both rats and humans. In contrast, the addition of new sensory evidence was the primary source of variability. We suggest our task and modeling approach as a powerful method for revealing internal properties of decision-making processes.


The functionally active IE2 immediate-early regulatory protein of human cytomegalovirus is an 80-kilodalton polypeptide that contains two distinct activator domains and a duplicated nuclear localization signal.
The IE2 region of the human cytomegalovirus (CMV) strain Towne major immediate-early (MIE) gene encodes a transcriptional transactivator that stimulates expression from a variety of heterologous target promoters but specifically down-regulates its own promoter. By immunofluorescence and Western immunoblot analysis with monospecific peptide antisera, we found that human CMV MIE exon 5 encodes four overlapping polypeptides, two present at immediate-early times (80 and 55 kDa) and two others detected only at late times after infection (55 and 40 kDa). However, only the 80-kDa version (579 amino acids), which is derived from the small upstream exons 2 and 3 fused to the intact exon 5 region, was functionally active in both transactivation and autoregulation as assessed by cotransfection experiments. These results confirm the corrected assignment of the coding capacity of the exon 5 region based on amino acid homology with the equivalent IE2 protein from simian CMV (Colburn). In transient DNA transfection assays, IE2 expression plasmids also produced a predominant full-length 80-kDa protein, which was localized in a distinctive reticular pattern in the nucleus. Two short basic nuclear localization signals in IE2 were identified by deletion analysis and by conversion of a test cytoplasmic herpes simplex virus protein into a form that localized in the nucleus after insertion of either of these two human CMV motifs. Functional assays with MIE region plasmids containing deletions or truncations in exon 5 revealed that both transactivation and autoregulation required several distinct domains within the COOH half of the IE2 protein, whereas a region between codons 99 and 194 could be discarded. Three segments at the NH2 end of the protein between codons 1 to 85, 86 to 98, and 195 to 290 were also essential for transactivation but played no role in autoregulation. Finally, in domain swap experiments, GAL4-fusion proteins containing either an NH2-terminal 51-amino-acid domain from exon 3 (codons 25 to 85) or the COOH-terminal 33-amino-acid domain from exon 5 (codons 544 to 579) identified two distinct activator domains from IE2, both of which have acidic characteristics.


Molecular characterization of the vanD gene cluster and a novel insertion element in a vancomycin-resistant enterococcus isolated in Canada.
A single vanD-containing Enterococcus faecium strain (N97-330) was isolated in Canada. The vanD-containing region was cloned and sequenced. Although the proteins have more than 96% identity to a previously described vanD region in BM4339, the vanS(D) gene contains a frameshift mutation that leads to a predicted truncated protein. Furthermore, sequence analysis of the ddl gene revealed the presence of an IS982-like element (ISEfm1) which interrupted the D-Ala-D-Ala ligase. This suggested the constitutive expression of the vanD operon, which was confirmed. Pulsed-field gel electrophoresis fingerprinting demonstrated that BM4339 was not related to N97-330 (>15 band differences). Both strains contained multiple copies of the IS982-like element.


Supramedullary inputs to cardiovascular neurons of rostral ventrolateral medulla in rats.
Experiments were done to test the hypothesis that selective activation of cell bodies in different nuclei known to be involved in central cardiovascular control could excite or inhibit the discharge of neurons in the rostral ventrolateral medulla (RVLM). It is known that chemical stimulation of the lateral parabrachial nucleus (LPBN), locus ceruleus (LC), and lateral hypothalamic area (LHA) in anesthetized animals elicits increases (LPBN) or decreases (LC and LHA) in arterial pressure. We therefore recorded extracellularly spontaneous activity from RVLM units in urethan-anesthetized rats and monitored the changes in firing frequency of these neurons during chemical stimulation of one of LPBN, LC, and LHA. Thirty-two units were classified as cardiovascular neurons because their activity was inhibited by baroreceptor activation (1-3 micrograms phenylephrine iv) and displayed a cardiac cycle-related rhythmicity. Chemical stimulation with sodium glutamate of arterial pressor sites in the ipsilateral LPBN increased the firing frequency (40.3 +/- 1.3%) of 11 cardiovascular neurons. Activation of cell bodies in arterial depressor sites in the ipsilateral LC inhibited the firing rate (59.1 +/- 7.1%) of 10 cardiovascular neurons and excited 1 unit. Activation of cell bodies in arterial depressor sites in the ipsilateral LHA inhibited the discharge rate (25.4 +/- 4.7%) of six cardiovascular neurons, excited one unit, and did not alter the rate of the remaining three units. These results provide direct evidence for the existence of excitatory and inhibitory pathways from neurons located in the LPBN, LC, and LHA to cardiovascular neurons in the RVLM.


Socioeconomic inequalities in placental vascular resistance: a prospective cohort study.
To examine the association between socioeconomic position (SEP) and umbilical and uterine placental resistance indices in the second and third trimester, and to what extent this could be explained by lifestyle-related behaviors. Prospective cohort study. Rotterdam, the Netherlands. 7,033 pregnant women of mean age (± standard deviation) 29.9 (±5.2) years. None. Uterine artery resistance index (UARI) and umbilical artery pulsatility index (UAPI) in second and third trimester measured with Doppler ultrasound. Third-trimester UARI and both second- and third-trimester UAPI were statistically significantly higher for women with lower educational levels as compared with those with higher educational levels. Educational level was strongly associated with the risk of continuously high levels of UARI and UAPI from second to third trimester of pregnancy. Notching was not associated with SEP. Smoking was a significant contributor to the association of SEP and increased placental resistance indices; body mass index, folic acid supplementation use, and alcohol use were not. Women from low socioeconomic subgroups have higher placental resistance indices, which may cause a higher prevalence of pregnancy complications. This was mainly explained by maternal smoking during pregnancy.


The effect of histone gene deletions on chromatin structure in Saccharomyces cerevisiae.
As a way of studying nucleosome assembly and maintenance in Saccharomyces cerevisiae, mutants bearing deletions or duplications of the genes encoding histones H2A and H2B were analyzed. Previous genetic analysis had shown that only one of these mutants exhibited dramatic and pleiotropic phenotypes. This mutant was also the only one that contained disrupted chromatin, suggesting that the original phenotypes were attributable to alterations in chromosome structure. The chromatin disruption in the mutant, however, did not extend over the entire genome, but rather was localized to specific regions. Thus, while the arrangement of nucleosomes over the HIS4 and GAL1 genes, the telomeres, and the long terminal repeats (delta sequences) of Ty retrotransposons appeared essentially normal, nucleosomes over the CYH2 and UBI4 genes and the centromere of chromosome III were dramatically disrupted. The observation that the mutant exhibited localized chromatin disruptions implies that the assembly or maintenance of nucleosomes differs over different parts of the yeast genome.


Dynamic postural stability in blind athletes using the biodex stability system.
Three systems affect the upright standing posture in humans - visual, vestibular, and somatosensory. It is well known that the visually impaired individuals have bad postural balance. On the other hand, it is a well documented fact that some sports can improve postural balance. Therefore, it is aimed in this study to evaluate the dynamic postural stability in goal-ball athletes. Twenty blind goal-ball players, 20 sighted and 20 sedentary blind controls were evaluated using the Biodex Stability System. Three adaptation trials and three test evaluations (a 20-second balance test at a platform stability of 8) were applied to the blind people, and to the sighted with eyes open and closed. Dynamic postural stability was measured on the basis of three indices: overall, anteroposterior, and mediolateral. Means of each test score were calculated. The tests results were compared for the blind athletes, sighted (with eyes open and closed) subjects, and sedentary blind people. There were significant differences between the results of the blind people and the sighted subjects with regards to all of the three indices. Although the stability of goal-ball players was better than sedentary blinds', only ML index values were statistically different (4.47 +/- 1.24 in the goal-ball players; 6.46 +/- 3.42 in the sedentary blind, p = 0.04). Dynamic postural stability was demonstrated to be affected by vision; and it was found that blind people playing goal-ball 1 - 2 days per week have higher ML stability than the sedentary sighted people.


Platelet-Rich Plasma for Patellar Tendinopathy: A Randomized Controlled Trial of Leukocyte-Rich PRP or Leukocyte-Poor PRP Versus Saline.
A small number of randomized controlled trials have found ultrasound-guided injection of platelet-rich plasma (PRP) to be no more effective than saline for several tendinopathies; limited information exists for patellar tendinopathy. In addition, different PRP formulations that produce varying concentrations of leukocytes have not been directly compared for patellar tendinopathy. To determine if a single ultrasound-guided PRP injection, either leukocyte-rich PRP (LR-PRP) or leukocyte-poor PRP (LP-PRP), was superior to saline injection for the treatment of patellar tendinopathy. The null hypothesis was that no treatment would be superior to another for the treatment of patellar tendinopathy. Randomized controlled trial; Level of evidence, 1. Athletes with patellar tendinopathy for ≥6 months (Blazina stage IIIB) were assessed for eligibility in a multisite single-blind controlled trial. There were 3 injection arms: LR-PRP, LP-PRP, and saline. Patients received a single ultrasound-guided injection, followed by 6 weeks of supervised rehabilitation (heavy slow resistance training, concentric and eccentric, 3 times per week). Outcome measures-Victorian Institute of Sport Assessment (patellar; VISA-P), pain during activity, and global rating of change-were assessed at 6 and 12 weeks and 6 and 12 months. VISA-P score at 12 weeks was the primary outcome. Fifty-seven patients (19 in each group) were included in an intention-to-treat analysis. Secondary outcome measures included pain during activity and patients' global rating of change. Study retention was 93% at 12 weeks and 79% after 1 year. There was no significant difference in mean change in VISA-P score, pain, or global rating of change among the 3 treatment groups at 12 weeks or any other time point. After 1 year, the mean (SD) outcomes for the LR-PRP, LP-PRP, and saline groups were as follows, respectively: VISA-P-58 (29), 71 (20), and 80 (18); pain-4.0 (2.4), 2.4 (2.3), and 2.0 (1.9); global rating of change-4.7 (1.6), 5.6 (1.0), and 5.7 (1.2) ( P > .05 for all outcomes). Combined with an exercise-based rehabilitation program, a single injection of LR-PRP or LP-PRP was no more effective than saline for the improvement of patellar tendinopathy symptoms. NCT02116946 (ClinicalTrials.gov identifier).


Long non-coding RNA H19 regulates endothelial cell aging via inhibition of STAT3 signalling.
Long non-coding RNAs (lncRNAs) have been shown to regulate numerous processes in the human genome, but the function of these transcripts in vascular aging is largely unknown. We aim to characterize the expression of lncRNAs in endothelial aging and analyse the function of the highly conserved lncRNA H19. H19 was downregulated in endothelium of aged mice. In human, atherosclerotic plaques H19 was mainly expressed by endothelial cells and H19 was significantly reduced in comparison to healthy carotid artery biopsies. Loss of H19 led to an upregulation of p16 and p21, reduced proliferation and increased senescence in vitro. Depletion of H19 in aortic rings of young mice inhibited sprouting capacity. We generated endothelial-specific inducible H19 deficient mice (H19iEC-KO), resulting in increased systolic blood pressure compared with control littermates (Ctrl). These H19iEC-KO and Ctrl mice were subjected to hindlimb ischaemia, which showed reduced capillary density in H19iEC-KO mice. Mechanistically, exon array analysis revealed an involvement of H19 in IL-6 signalling. Accordingly, intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 were upregulated upon H19 depletion. A luciferase reporter screen for differential transcription factor activity revealed STAT3 as being induced upon H19 depletion and repressed after H19 overexpression. Furthermore, depletion of H19 increased the phosphorylation of STAT3 at TYR705 and pharmacological inhibition of STAT3 activation abolished the effects of H19 silencing on p21 and vascular cell adhesion molecule 1 expression as well as proliferation. These data reveal a pivotal role for the lncRNA H19 in controlling endothelial cell aging.


The Tucson Children's Respiratory Study. I. Design and implementation of a prospective study of acute and chronic respiratory illness in children.
The Tucson Children's Respiratory Study, Tucson, Arizona, has been established as a long-term, longitudinal, prospective study of the risk factors for acute lower respiratory tract illnesses in early childhood and for chronic obstructive airways disease in later life. A total of 1,246 newborns were enrolled into the study between May 1980 and January 1984, representing 78% of eligible infants. Cord blood for immunologic studies, neonatal blood specimens for blood counts and differentials, and blood specimens at nine to 15 months of age for immunologic studies, blood counts, and differentials have been obtained on the majority of enrolled children. Pre-illness physiologic and more detailed immunologic studies have also been done on large subgroups of subjects. The majority of lower respiratory tract illnesses suffered by these children in the first three years of life have been assessed in detail for etiologic agents by means of culture and serologic techniques; 1,052 illnesses have been evaluated thus far. The type of illness and nature of etiologic agents are very similar to those reported in other epidemiologic studies. Thus, this group of enrolled infants and their family members constitute an appropriate population for the long-term study of risk factors for acute and chronic respiratory disorders.


Developing and validating a cardiovascular risk score for patients in the community with prior cardiovascular disease.
Patients with atherosclerotic cardiovascular disease (CVD) vary significantly in their risk of future CVD events; yet few clinical scores are available to aid assessment of risk. We sought to develop a score for use in primary care that estimates short-term CVD risk in these patients. Adults aged <80 years with prior CVD were identified from a New Zealand primary care cohort study (PREDICT), and linked to national mortality, hospitalisation and dispensing databases. A Cox model with an outcome of myocardial infarction, stroke or CVD death within 2 years was developed. External validation was performed in a cohort from the UK. 24 927 patients, 63% men, 63% European, median age 65 years (IQR 58-72 years), experienced 1480 CVD events within 2 years after a CVD risk assessment. A risk score including ethnicity, comorbidities, body mass index, creatine creatinine and treatment, in addition to established risk factors used in primary prevention, predicted a median 2-year CVD risk of 5.0% (IQR 3.5%-8.3%). A plot of actual against predicted event rates showed very good calibration throughout the risk range. The score performed well in the UK cohort but overestimated risk for those at highest risk, who were predominantly patients defined as having heart failure. The PREDICT-CVD secondary prevention score uses routine measurements from clinical practice that enable it to be implemented in a primary care setting. The score will facilitate risk communication between primary care practitioners and patients with prior CVD, particularly as a resource to show the benefit of risk factor modification.


The evaluation of an evidence-based clinical answer format for pediatricians.
Clinicians are increasingly using electronic sources of evidence to support clinical decision-making; however, there are multiple demands on clinician time, and summarised and synthesised evidence is needed. Clinical Answers (CA) have been developed to address this need; the CA is a synthesised evidence-based summary that supports point-of-care clinical decision-making. The aim of this paper is to report on a survey used to test and improve the CA format. An online survey was sent to pediatricians via e-mail and posted on a child health clinical standards website. Quantitative data analysis consisted primarily of descriptive statistics; qualitative data analysis consisted of content analysis. Eighty-three pediatricians responded to the survey. Most respondents found the CA useful or very useful (93%) and agreed or strongly agreed that the layout was effective and allowed them to quickly locate critical information (82%). Quantitative and qualitative data suggested that respondents thought there should be less detail in the linked figures and tables (p = 0.0002), but overall respondents seemed to think there was an appropriate level of detail in most sections of the CA. Based on the quantitative and qualitative survey responses, major and minor modifications to the CA format were implemented, such as removing forest plots, adding links in each addendum to bring the user back to the front page, and adding an 'Implications for practice' section to the CA. Findings suggest that CAs will be a useful tool for pediatricians; thus, the research team has now begun creating CAs to assist busy clinicians in their day-to-day clinical practice by providing high-quality information for decision-making at the point-of-care.


Tetracycline fiber used alone or with scaling and root planing in periodontal maintenance patients: clinical results.
This study evaluated the 6-month clinical response to sustained-release tetracycline fibers used alone or with scaling and root planing in 25 adult periodontal maintenance patients. All subjects had at least one pocket > or = 4.0 mm that bled on probing and required therapy. Thirty-six teeth were treated with tetracycline fibers for 7 to 12 days; twelve of the 36 teeth also received scaling and root planing. The selection of teeth for scaling and root planing was based on the condition of the teeth. Therapeutic results were evaluated by changes in probing depth and frequency of bleeding on probing. Use of tetracycline fibers and fibers with scaling produced 1.8- and 1.7-mm reductions in probing depth, respectively, 1 month after treatment; reductions declined to 1.3 and 0.8 mm at 3 months, but rebounded to 1.5 and 1.3 mm at 6 months. The percentage of teeth exhibiting bleeding on probing decreased from 100% at baseline to 68% and 50% in the fiber and fiber plus scaling groups, respectively, at 6 months. None of the differences was statistically significant. Tetracycline fibers clearly decreased clinical signs of periodontal inflammation. Addition of scaling and root planing at the time of fiber placement further decreased, although not significantly, the degree of inflammation.


Development and implementation of a formalized geriatric surgery curriculum for general surgery residents.
Despite the growth of the elderly population, most surgical training programs lack formalized geriatric education. The authors' aim was to implement a formalized geriatric surgery curriculum at an academic medical center. Surgery residents were surveyed on attitudes toward the care of elderly patients and the importance of various geriatric topics to daily practice. A curriculum consisting of 16 didactic sessions was created with faculty experts moderating. After curriculum completion, residents were surveyed to assess curriculum impact. Residents expressed increased comfort in accessing community resources. A greater percentage of residents recognized the significance of delirium and acute renal failure in elderly patients. Implementing a geriatric surgery curriculum geared toward surgery residents is feasible and can increase resident comfort with multidisciplinary care and recognition of clinical conditions pertinent to elderly surgical patients. This initiative also provided valuable experience for geriatric surgery curriculum development.


An integrated approach to the taxonomic identification of prehistoric shell ornaments.
Shell beads appear to have been one of the earliest examples of personal adornments. Marine shells identified far from the shore evidence long-distance transport and imply networks of exchange and negotiation. However, worked beads lose taxonomic clues to identification, and this may be compounded by taphonomic alteration. Consequently, the significance of this key early artefact may be underestimated. We report the use of bulk amino acid composition of the stable intra-crystalline proteins preserved in shell biominerals and the application of pattern recognition methods to a large dataset (777 samples) to demonstrate that taxonomic identification can be achieved at genus level. Amino acid analyses are fast (<2 hours per sample) and micro-destructive (sample size <2 mg). Their integration with non-destructive techniques provides a valuable and affordable tool, which can be used by archaeologists and museum curators to gain insight into early exploitation of natural resources by humans. Here we combine amino acid analyses, macro- and microstructural observations (by light microscopy and scanning electron microscopy) and Raman spectroscopy to try to identify the raw material used for beads discovered at the Early Bronze Age site of Great Cornard (UK). Our results show that at least two shell taxa were used and we hypothesise that these were sourced locally.


Genome-wide DNA methylation meta-analysis in the brains of suicide completers.
Suicide is the second leading cause of death globally among young people representing a significant global health burden. Although the molecular correlates of suicide remains poorly understood, it has been hypothesised that epigenomic processes may play a role. The objective of this study was to identify suicide-associated DNA methylation changes in the human brain by utilising previously published and unpublished methylomic datasets. We analysed prefrontal cortex (PFC, n = 211) and cerebellum (CER, n = 114) DNA methylation profiles from suicide completers and non-psychiatric, sudden-death controls, meta-analysing data from independent cohorts for each brain region separately. We report evidence for altered DNA methylation at several genetic loci in suicide cases compared to controls in both brain regions with suicide-associated differentially methylated positions enriched among functional pathways relevant to psychiatric phenotypes and suicidality, including nervous system development (PFC) and regulation of long-term synaptic depression (CER). In addition, we examined the functional consequences of variable DNA methylation within a PFC suicide-associated differentially methylated region (PSORS1C3 DMR) using a dual luciferase assay and examined expression of nearby genes. DNA methylation within this region was associated with decreased expression of firefly luciferase but was not associated with expression of nearby genes, PSORS1C3 and POU5F1. Our data suggest that suicide is associated with DNA methylation, offering novel insights into the molecular pathology associated with suicidality.


A simple and versatile strategy for realizing bright multicolor mechanoluminescence.
Multicolor mechanoluminescence (ML) was first realized by using the stable organic blue ML emitter N-phenylcarbazole as the host matrix. It is claimed that a good ML host should have a moderate melting point and be able to dissolve or disperse organic dyes but maintain high ML activity and crystallinity. This strategy is versatile and can avoid difficult molecular design and troublesome chemical synthesis.


Measurement of residual renal function in patients treated with continuous ambulatory peritoneal dialysis.
Renal function contributes markedly to the adequacy of continuous ambulatory peritoneal dialysis (CAPD). The best way to measure it in clinical practice has not been established. Ten stable CAPD patients with residual renal function were investigated to compare the GFR measured as inulin clearance (Cli) with the creatinine clearance (Clc), the urea clearance (Clu), and with 0.5(Clc + Clu). Thereafter, an analysis of whether the administration of cimetidine could improve the accuracy of these clearances was performed. Two clearance periods (CP) of 24 h were investigated. During CP-2, patients received 400 mg cimetidine twice daily, for a total dose of 1200 mg. Two h before the urine and dialysate collection period, inulin was administered iv. Calculations were done for each CP for Cli, Clc, Clu, Clc-Cli, the Clc/Cli ratio, and the tubular secretion of creatinine (TSc). No differences between CP-1 and CP-2 were present for urinary excretion of volume and solutes, and clearance rates of inulin and urea. The median TSc decreased from 0.71 mumol/min (range, -0.24 to 5.90) in CP-1 to 0.30 mumol/min (range, -0.18 to 0.64) in CP-2 (P < 0.05). Therefore, the median ratio of Clc/Cli decreased from 1.23 (range, 0.87 to 2.20) in CP-1 to 1.11 (range, 0.95 to 1.51) in CP-2 (P < 0.05). The median overestimation of the Cli in CP-1 by the Clc was 0.90 mL/min (range, -0.28 to 3.80) and by the 0.5(Clc + Clu) was 0.30 (range, -0.67 to 1.52). The median overestimation of Cli during cimetidine treatment in CP-2 was 0.43 mL/min (range, -0.21 to 1.20). The range, in differences between Cli and Clc, in CP-2 was smaller than that between Cli and 0.5(Clc + Clu) in CP-1. The difference between the clearance rate of inulin and creatinine or the combined clearance rate of urea and creatinine was not influenced by the magnitude of the average GFR. It can be concluded that the administration of cimetidine improved the accuracy of measuring the GFR with the Clc in CAPD patients.


"I always walked out with an empty purse." Older adults' engagement with electronic gambling machines in Victoria, Australia.
Older adults are at an increased risk of experiencing gambling harm, which may be due to their use of high-intensity gambling products such as electronic gambling machines (EGMs). However, little research has explored the motivations behind older adults' engagement with EGMs, their understanding of the structural characteristics of EGMs, or their perceptions of risk associated with EGM gambling. This paper aims to address this gap in the literature. Focus groups were conducted in Melbourne, Australia with n = 126 adults aged 55+, who had attended a club or pub in the last 12 months. Topics included EGM attitudes and behaviours, structural characteristics of EGMs, and the potential risks associated with EGM gambling. Thematic analysis was used to interpret the data. For most participants, EGM gambling was secondary to their participation in other activities available within venues. Participants identified structural characteristics of EGMs; however, there were some misconceptions about how EGMs operated, including how or why machines paid out. Most participants perceived that they were not at risk of gambling harm because they engaged in "responsible" gambling practices such as setting limits. Older adults often engaged in EGM gambling because of its availability in community-based venues. Older adults' perception that they are implementing responsible gambling practices may be increasing their susceptibility to harm. SO WHAT?: There is a need to reduce the availability and accessibility of EGMs in community settings and develop public education programs that are tailored to the needs of older adults.


Angiotensin II-noradrenergic interactions in renovascular hypertensive rats.
This study tested the hypothesis that interactions of endogenous angiotensin II (AII) with the noradrenergic neuroeffector junction are important in renin-dependent hypertension. In the in situ blood-perfused rat mesentery, in normal rats exogenous AII potentiated mesenteric vascular responses to periarterial (sympathetic) nerve stimulation (PNS) more than vascular responses to exogenous norepinephrine (NE). In 2-kidney-1-clip (2K-1C) rats with renovascular hypertension mesenteric vascular responses to PNS and NE were greater than in sham-operated rats, and renovascular hypertension mimicked the effects of exogenous AII with respect to enhancing responses to PNS more than responses to NE. In 2K-1C rats, but not in sham-operated rats, 1-Sar-8-Ile-AII markedly suppressed vascular responses to PNS, without influencing responses to NE. Finally, 1-Sar-8-Ile-AII attenuated sympathetic nerve stimulation-induced neuronal spillover of NE in 2K-1C rats, but not in sham-operated rats. These data indicate that renovascular hypertension enhances noradrenergic neurotransmission, and that this enhancement is mediated in part by AII-induced facilitation of NE release.


Glutamate Excitotoxicity and Oxidative Stress in Epilepsy: Modulatory Role of Melatonin.
Epilepsy is thought to be associated with oxidative stress, glutamate excitotoxicity, and mitochondrial dysfunction. The enhanced synthesis and release of oxygen free radicals is linked to the low and oxidative potential of the central nervous system. Glutamate excitotoxicity also contributes significantly to the production of reactive nitrogen species that cause nitrosative stress. A decrease in adenosine triphosphate synthesis, which leads to free radical formation, is associated with mitochondrial dysfunction. The brain is very much susceptible to degeneration and oxidative stress because of its low antioxidant enzyme activity. Melatonin, a hormone secreted by the pineal gland, has remarkable antioxidant properties. Melatonin and its analogs that bind to melatonin receptors have a significant role in suppressing seizures. Melatonin scavenges oxygen free radicals such as hydroxyl radical, peroxy radical, peroxynitrite anion, and superoxide radical and stimulates synthesis of superoxide dismutase and glutathione peroxidase, which are potent antioxidant enzymes. Melatonin administration has been shown to be effective in both experimental models and patients suffering from epilepsy. In this review, we compile the literature supporting consequences of seizures and the protective role of melatonin during seizures.


Knockdown of zebrafish Lgi1a results in abnormal development, brain defects and a seizure-like behavioral phenotype.
Epilepsy is a common disorder, typified by recurrent seizures with underlying neurological disorders or disease. Approximately one-third of patients are unresponsive to currently available therapies. Thus, a deeper understanding of the genetics and etiology of epilepsy is needed to advance the development of new therapies. Previously, treatment of zebrafish with epilepsy-inducing pharmacological agents was shown to result in a seizure-like phenotype, suggesting that fish provide a tractable model to understand the function of epilepsy-predisposing genes. Here, we report the first model of genetically linked epilepsy in zebrafish and provide an initial characterization of the behavioral and neurological phenotypes associated with morpholino (MO) knockdown of leucine-rich, glioma-inactivated 1a (lgi1a) expression. Mutations in the LGI1 gene in humans have been shown to predispose to a subtype of autosomal dominant epilepsy. Low-dose Lgi1a MO knockdown fish (morphants) appear morphologically normal but are sensitized to epilepsy-inducing drugs. High-dose Lgi1a morphants have morphological defects which persist into adult stages that are typified by smaller brains and eyes and abnormalities in tail shape, and display hyperactive swimming behaviors. Increased apoptosis was observed throughout the central nervous system of high-dose morphant fish, accounting for the size reduction of neural tissues. These observations demonstrate that zebrafish can be exploited to dissect the embryonic function(s) of genes known to predispose to seizure-like behavior in humans, and offer potential insight into the relationship between developmental neurobiological abnormalities and seizure.


MicroRNA signatures in total peripheral blood as novel biomarkers for acute myocardial infarction.
MicroRNAs (miRNAs) are important regulators of adaptive and maladaptive responses in cardiovascular diseases and hence are considered to be potential therapeutical targets. However, their role as novel biomarkers for the diagnosis of cardiovascular diseases still needs to be systematically evaluated. We assessed here for the first time whole-genome miRNA expression in peripheral total blood samples of patients with acute myocardial infarction (AMI). We identified 121 miRNAs, which are significantly dysregulated in AMI patients in comparison to healthy controls. Among these, miR-1291 and miR-663b show the highest sensitivity and specificity for the discrimination of cases from controls. Using a novel self-learning pattern recognition algorithm, we identified a unique signature of 20 miRNAs that predicts AMI with even higher power (specificity 96%, sensitivity 90%, and accuracy 93%). In addition, we show that miR-30c and miR-145 levels correlate with infarct sizes estimated by Troponin T release. The here presented study shows that single miRNAs and especially miRNA signatures derived from peripheral blood, could be valuable novel biomarkers for cardiovascular diseases.


Surface texture limits transfer of S. aureus, T4 bacteriophage, influenza B virus and human coronavirus.
Spread of pathogens on contaminated surfaces plays a key role in disease transmission. Surface technologies that control pathogen transfer can help control fomite transmission and are of great interest to public health. Here, we report a novel bead transfer method for evaluating fomite transmission in common laboratory settings. We show that this method meets several important criteria for quantitative test methods, including reasonableness, relevancy, resemblance, responsiveness, and repeatability, and therefore may be adaptable for standardization. In addition, this method can be applied to a wide variety of pathogens including bacteria, phage, and human viruses. Using the bead transfer method, we demonstrate that an engineered micropattern limits transfer of Staphylococcus aureus by 97.8% and T4 bacteriophage by 93.0% on silicone surfaces. Furthermore, the micropattern significantly reduces transfer of influenza B virus and human coronavirus on silicone and polypropylene surfaces. Our results highlight the potential of using surface texture as a valuable new strategy in combating infectious diseases.


Analysis of bronchial reactivity in epidemiological studies.
The measurement of bronchial reactivity in epidemiological studies has the advantage of quantifying an objective physiological feature of asthma. Bronchial reactivity was developed in a clinical setting and has been conventionally expressed as the dose of agonist producing a 20% fall in FEV1 (PD20). As PD20 can be estimated for less than 20% of subjects in general community surveys with the doses of agonist that are usually given, data from most subjects must be censored. Thus PD20 alone is a poor index of bronchial reactivity for epidemiological studies. Data from 809 aluminium smelter workers were used to evaluate alternative methods of analysing bronchial reactivity. Dose-response relationships were analysed by four methods: (1) PD20 by the conventional method of interpolating the dose on a logarithmic scale between the last two measurements of FEV1; (2) PD20 (with allowance for extrapolation), estimated by fitting an exponential curve to the dose-response data; (3) the linear regression slope between dose and FEV1 when significant; (4) the dose-response slope obtained in all subjects as the % change in FEV1 from baseline in response to total dose. When each of these measures was related to symptoms, diagnosis, and treatment of asthma, all differentiated between "asthmatic" and "non-asthmatic" subjects. The dose-response slope (method 4) had the advantages of simplicity and no censored data, and was shown to be clinically relevant. It is suggested that the dose-response slope should be used for the analysis of bronchial reactivity in epidemiological studies.


Transcranial Near Infrared Light Stimulations Improve Cognition in Patients with Dementia.
Dementia is a complex syndrome with various presentations depending on the underlying pathologies. Low emission of transcranial near-infrared (tNIR) light can reach human brain parenchyma and be beneficial to a number of neurological and neurodegenerative disorders. We hereby examined the safety and potential therapeutic benefits of tNIR light stimulations in the treatment of dementia. Patients of mild to moderate dementia were randomized into active and sham treatment groups at 2:1 ratio. Active treatment consisted of low power tNIR light stimulations with an active photobiomodulation for 6 min twice daily during 8 consequent weeks. Sham treatment consisted of same treatment routine with a sham device. Neuropsychological battery was obtained before and after treatment. Analysis of variance (ANOVA) was used to analyze outcomes. Sixty subjects were enrolled. Fifty-seven subjects completed the study and had not reported health or adverse side effects during or after the treatment. Three subjects dropped out from trial for health issues unrelated to use of tNIR light treatment. Treatment with active device resulted in improvements of cognitive functions and changes were: an average increase of MMSE by 4.8 points; Logical Memory Tests I and II by ~3.0 points; Trail Making Tests A and B by ~24%; Boston Naming Test by ~9%; improvement of both Auditory Verbal Learning Tests in all subtest categories and overall time of performance. Many patients reported improved sleep after ~7 days of treatment. Caregivers noted that patients had less anxiety, improved mood, energy, and positive daily routine after ~14-21 days of treatment. The tNIR light treatments demonstrated safety and positive cognitive improvements in patients with dementia. Developed treatment protocol can be conveniently used at home. This study suggests that additional dementia treatment trials are warranted with a focus on mitigating caregivers' burden with tNIR light treatment of dementia patients.


Thermoelectric cooling and power generation
In a typical thermoelectric device, a junction is formed from two different conducting materials, one containing positive charge carriers (holes) and the other negative charge carriers (electrons). When an electric current is passed in the appropriate direction through the junction, both types of charge carriers move away from the junction and convey heat away, thus cooling the junction. Similarly, a heat source at the junction causes carriers to flow away from the junction, making an electrical generator. Such devices have the advantage of containing no moving parts, but low efficiencies have limited their use to specialty applications, such as cooling laser diodes. The principles of thermoelectric devices are reviewed and strategies for increasing the efficiency of novel materials are explored. Improved materials would not only help to cool advanced electronics but could also provide energy benefits in refrigeration and when using waste heat to generate electrical power.


Long-term effectiveness of azathioprine in IBD beyond 4 years: a European multicenter study in 1176 patients.
In Crohn's disease the optimal duration of azathioprine treatment is still controversial and for ulcerative colitis only limited data are available to support its efficacy. Charts of 1176 patients with IBD from 16 European centers were analyzed. Flare incidences and steroid dosages were assessed for the time before and during treatment and after discontinuation. Within the first 4 years, azathioprine suppressed flare incidence and steroid consumption in both diseases (P < 0.001). While in CD discontinuation after 3-4 years did not lead to reactivation, this was the case in UC. However, continuation beyond 4 years further improved clinical activity in CD and steroid requirement in both diseases (P < 0.001). Discontinuation of azathioprine may thus be considered after 3-4 years in CD patients in complete remission without steroid requirement. In all other CD patients and for UC patients in general, continuation seems beneficial. These results support a novel differential algorithm for long-term azathioprine therapy in IBD.


A genome-wide Drosophila screen for heat nociception identifies α2δ3 as an evolutionarily conserved pain gene.
Worldwide, acute, and chronic pain affects 20% of the adult population and represents an enormous financial and emotional burden. Using genome-wide neuronal-specific RNAi knockdown in Drosophila, we report a global screen for an innate behavior and identify hundreds of genes implicated in heat nociception, including the α2δ family calcium channel subunit straightjacket (stj). Mice mutant for the stj ortholog CACNA2D3 (α2δ3) also exhibit impaired behavioral heat pain sensitivity. In addition, in humans, α2δ3 SNP variants associate with reduced sensitivity to acute noxious heat and chronic back pain. Functional imaging in α2δ3 mutant mice revealed impaired transmission of thermal pain-evoked signals from the thalamus to higher-order pain centers. Intriguingly, in α2δ3 mutant mice, thermal pain and tactile stimulation triggered strong cross-activation, or synesthesia, of brain regions involved in vision, olfaction, and hearing.


Fabrication, chemical composition change and phase evolution of biomorphic hydroxyapatite.
Biomorphous, highly porous hydroxyapatite (HA) ceramics have been prepared by a combination of a novel biotemplating process and a sol-gel method, using natural plants like cane and pine as biotemplates. A HA sol was first synthesized from triethylphosphate and calcium nitrate used as the phosphorus and calcium precursors, respectively, and infiltrated into the biotemplates, and subsequently they were sintered at elevated temperatures to obtain porous HA ceramics. The microstructural changes, phase and chemical composition evolutions during the biotemplate-to-HA conversion were investigated by scanning electron microscopy (SEM), X-ray powder diffraction (XRD), and Fourier-transform infrared (FT-IR) spectroscopy. The XRD and FT-IR analysis revealed that the dominant phase of the product was HA, which contained a small amount of mixed A/B-type carbonated HA, closely resembling that of human bone apatite. Moreover, the appearance of a small amount of secondary phase CaCO(3) seemed unavoidable. The HA was not transformed to the other calcium phosphate phases up to 1400 degrees C, but it contained a trace amount of CaO when sintered at above 1100 degrees C. The possible transformation mechanism was proposed. The SEM observation and mechanical property test showed that as-produced HA ceramics retained the macro-/micro-porous structures of the biotemplates with high precision, and possessed acceptable mechanical strength, which is suggested to be potential scaffolds for bone tissue engineering.


Characterization of a New DGKE Intronic Mutation in Genetically Unsolved Cases of Familial Atypical Hemolytic Uremic Syndrome.
Genetic and acquired abnormalities causing dysregulation of the complement alternative pathway contribute to atypical hemolytic uremic syndrome (aHUS), a rare disorder characterized by thrombocytopenia, nonimmune microangiopathic hemolytic anemia, and acute kidney failure. However, in a substantial proportion of patients the disease-associated alterations are still unknown. Whole-exome and whole-genome sequencing were performed in two unrelated families with infantile recessive aHUS. Sequencing of cDNA from affected individuals was used to test for the presence of aberrant mRNA species. Expression of mutant diacylglycerol kinase epsilon (DGKE) protein was evaluated with western blotting. Whole-exome sequencing analysis with conventional variant filtering parameters did not reveal any obvious candidate mutation in the first family. The report of aHUS-associated mutations in DGKE, encoding DGKE, led to re-examination of the noncoding DGKE variants obtained from next-generation sequencing, allowing identification of a novel intronic DGKE mutation (c.888+40A>G) that segregated with disease. Sequencing of cDNA from affected individuals revealed aberrant forms of DGKE mRNA predicted to cause profound abnormalities in the protein catalytic site. By whole-genome sequencing, the same mutation was found in compound heterozygosity with a second nonsense DGKE mutation in all affected siblings of another unrelated family. Homozygous and compound heterozygous patients presented similar clinical features, including aHUS presentation in the first year of life, multiple relapsing episodes, and proteinuria, which are prototypical of DGKE-associated aHUS. This is the first report of a mutation located beyond the exon-intron boundaries in aHUS. Intronic mutations such as these are underreported because conventional filtering parameters used to process next-generation sequencing data routinely exclude these regions from downstream analyses in both research and clinical settings. The results suggest that analysis of noncoding regions of aHUS-associated genes coupled with mRNA sequencing might provide a tool to explain genetically unsolved aHUS cases.


Prognosis and prognostic factors of retinal infarction: a prospective cohort study.
To determine the prognosis and adverse prognostic factors in patients with retinal infarction due to presumed atheromatous thromboembolism or cardiogenic embolism. Prospective cohort study. University hospital departments of clinical neurology. 99 patients with retinal infarction, without prior stroke, referred to a single neurologist between 1976 and 1986 and evaluated and followed up prospectively until death or the end of 1986 (mean follow up 4.2 years). Cerebral angiography (55 patients), aspirin treatment (37), oral anticoagulant treatment (eight), carotid endarterectomy (13), cardiac surgery (six), and peripheral vascular surgery (two). Death, stroke, coronary events, contralateral retinal infarction; survival analysis confined to 98 patients with retinal infarction due to presumed artheromatous thromboembolism or cardiogenic embolism (one patient with giant cell arteries excluded), and Cox's proportional hazards regression analysis, including age as a prognostic factor. During follow up 29 patients died (21 of vascular causes and eight of non-vascular or unknown causes), 10 had a first ever stroke, 19 had a coronary event, and only one developed contralateral retinal infarction. A coronary event accounted for more than half (59%) of the deaths whereas stroke was the cause of only one death (3%). Over the first five years after retinal infarction the actuarial average absolute risk of death was 8% per year; of stroke 2.5% per year (7.4% in the first year); of coronary events 5.3% per year, exceeding that of stroke; and of stroke, myocardial infarction, or vascular death 7.4% per year. Prognostic factors associated with an increased risk of death were increasing age, peripheral vascular disease, cardiomegaly, and carotid bruit. Adverse prognostic factors for serious vascular events were increasing age and carotid bruit for stroke, and increasing age, cardiomegaly, and carotid bruit both for coronary events and for stroke, myocardial infarction, or vascular death. Patients who present with retinal infarction due to presumed atherothromboembolism or cardiogenic embolism are at considerable risk of a coronary event. The risk of stroke, although high, is not so great. Not all strokes occurring after retinal infarction relate directly to disease of the ipsilateral carotid system, although this is probably the most common cause. Few patients experience contralateral retinal infarction. Non-arteritic retinal infarction should be diagnosed or confirmed by an ophthalmologist, and the long term care of patients with the condition should involve a physician who has an active interest in managing vascular disease.


Statistical validity for testing associations between genetic markers and quantitative traits in family data.
In genetic analysis it is often of interest to analyze associations between traits of unknown genetic etiology and genetic markers from pedigree data. Statistical methods that assume independence of pedigree members cannot be used because they disregard the statistical dependencies of members in a pedigree. For quantitative traits, a regression model proposed by George and Elston [Genet Epidemiol 4:193-201, 1987] uses an asymptotic likelihood ratio test and incorporates a correlation structure that allows for statistical dependence among the pedigree members. The statistical validity of this test is assessed for finite samples by measuring the discrepancy between the empirical and theoretical chi-square distributions. The variance of the mean of the dependent variable is determined to be related to this discrepancy and can be used to determine whether a pedigree structure is large enough for making valid statistical inferences on the basis of the asymptotic test. A multi-generational pedigree of 200 or so individuals should in many cases be sufficient for valid results when using the asymptotic likelihood ratio test for the association between markers and continuous traits.


Comprehensive molecular portraits of human breast tumours.
We analysed primary breast cancers by genomic DNA copy number arrays, DNA methylation, exome sequencing, messenger RNA arrays, microRNA sequencing and reverse-phase protein arrays. Our ability to integrate information across platforms provided key insights into previously defined gene expression subtypes and demonstrated the existence of four main breast cancer classes when combining data from five platforms, each of which shows significant molecular heterogeneity. Somatic mutations in only three genes (TP53, PIK3CA and GATA3) occurred at >10% incidence across all breast cancers; however, there were numerous subtype-associated and novel gene mutations including the enrichment of specific mutations in GATA3, PIK3CA and MAP3K1 with the luminal A subtype. We identified two novel protein-expression-defined subgroups, possibly produced by stromal/microenvironmental elements, and integrated analyses identified specific signalling pathways dominant in each molecular subtype including a HER2/phosphorylated HER2/EGFR/phosphorylated EGFR signature within the HER2-enriched expression subtype. Comparison of basal-like breast tumours with high-grade serous ovarian tumours showed many molecular commonalities, indicating a related aetiology and similar therapeutic opportunities. The biological finding of the four main breast cancer subtypes caused by different subsets of genetic and epigenetic abnormalities raises the hypothesis that much of the clinically observable plasticity and heterogeneity occurs within, and not across, these major biological subtypes of breast cancer.