Sex-related differences in GH secretion in rat using reverse hemolytic plaque assay.
It is not known whether enhanced growth hormone (GH)-releasing factor (GRF)-stimulated GH release observed in the male reflects differences in somatotrope numbers and/or secretory response to GRF. We addressed this question by using the hemolytic plaque assay which allows quantification of hormone secretion by single pituitary cells. Time-course studies and GRF-GH concentration-response relationships (0.01, 0.1, 1, 10, 100, 1,000 nM GRF) in age-matched male and diestrous day 2 female rats were compared by quantitating the percent of GH plaque-forming cells, and measuring the plaque areas. The male pituitary contained a greater percent (P less than 0.05) of somatotropes (% of plaque-forming cells 45 +/- 2 vs. 27 +/- 4% in the female; mean +/- SE). GRF induced a greater concentration-dependent increase in plaque areas in the male. Maximal responses were attained at 10 nM GRF in both sexes. However, mean maximal plaque area was significantly greater (P less than 0.001) and the EC50 was significantly lower (P less than 0.05) in the male (0.25 +/- 0.09 vs. 1.78 +/- 0.64 nM in the female). The data suggest that the greater percent of somatotropes in the male and greater secretory capacity and sensitivity to GRF may contribute to sex-related differences in GH secretion in the rat.


Leadership by fragmented destruction after a merger: an example from a facility of acute psychiatry.
Hospitals are labor intensive facilities based on highly skilled employees. A merger of hospitals is an effort to increase and rationalize this production. Decisions behind a merger are made at the top leadership level. How this might be done is demonstrated by examples from a 36 bed acute psychiatric facility. The aim of the study was to calculate the hidden costs of fragmented destruction of parts of a total hospital supply to patients after a merger. Fragmented destruction is the deliberate stopping of activities deemed not part of the core activities of the hospital without due consideration of the impact on core activities. The proposed changes to operational expenses at a single acute psychiatric hospital were materials for the study. The changes included activities as a reduction in local laboratory service, cleaning services, closure of physiotherapy unit, closing of cultural activities and reduced productivity. The selected activities are calculated as giving an imputed gain of € 630,000 as indicated by the leadership. The not calculated costs of reducing or removing the selected activities are estimated at € 1,955,640. The cost of staff disappointment after a merger is difficult to assess, but is probably higher than assumed in the present calculations. Imputed cost containment is not attained. The calculations indicate that implemented changes may increase cost, contrary to the belief of the leadership at both the hospital level and further up in the hospital trust. Arguments in favor of a merger have to be scrutinized thoroughly for optimistic neglect of uncalculated costs of mergers. Future hospital mergers and selected fragmentation of productive tasks at ward or hospital levels should include calculations of unavoidable costs as shown in the present paper.


Surface and Bulk Charge Density Wave Structure in 1 T-TaS2.
Incommensurate charge density waves (CDWs) in some materials form domains within which the CDWs may be commensurate. However, two questions have remained controversial: What is the geometrical structure of these domains, and are they or are they not identical on the surface and in the bulk? To address these issues in the triclinic (T) phase of tantalum disuffide (1T-TaS(2)) the CDW domain structure has been accurately determined for both the crystal surface and the crystal bulk. By analyzing the bulk CDW wave vectors and associated satellites by x-ray diffraction, it is found that the bulk contains three dimensionally ordered striped domains that have previously been misidentified. Scanning tunneling microscope images show that the striped domain configuration propagates unaltered to the crystal surface, and their Fourier transforms yield the same satellite positions as the x-rays. These observations demonstrate that the surface and bulk CDW domain structures in 1T-TaS(2) are identical.


[Prevalence of hepatitis B infection in children under 5 years old on indigenous communities of the Peruvian Amazonia after immunization interventions].
To determine the prevalence of hepatitis B virus (HVB) infection and seroprotection in indigenous children living in three draining basins of the Peruvian Amazon. A cross sectional study was conducted in three draining basins (Pastaza, Morona and Lower Urubamba rivers), home to six indigenous populations: shapra, candoshi, machiguenga, yine, ashaninka and nanti. Children under 5 years were included. A survey and review of the vaccination card was applied, and a serum sample was processed with ELISA for HBsAg, total anti-HBc and anti-HBs was obtained, defining "seroprotected children" if they were reactive to anti-HBs and non-reactive to anti-HBc. A total of 742 children under 5 years old residents in six indigenous communities were included in the study. 380 (51.2%) were male and 169 (22.9%) one year old or younger. Regarding recorded HVB vaccination on the card, only 434 (58.5%) had received three doses; 208 (38.8%) received the first dose within 24 hours. No cases were detected with HBsAg. 88.8% of children had seroconverted, varying between 67-100% across the populations. No cases of chronic HBV infection were detected. The vaccination coverage by card revision is low, contrasting with high seroconversion rates.


Lifetime health effects and medical costs of integrated stroke services - a non-randomized controlled cluster-trial based life table approach.
Economic evaluation of stroke services indicates that such services may lead to improved quality of life at affordable cost. The present study assesses lifetime health impact and cost consequences of stroke in an integrated service setting. The EDISSE study is a prospective non-randomized controlled cluster trial that compared stroke services (n = 151 patients) to usual care (n = 187 patients). Health status and cost trial-data were entered in multi-dimensional stroke life-tables. The tables distinguish four levels of disability which are defined by the modified Rankin scale. Quality-of-life scores (EuroQoL-5D), transition and survival probabilities are based on concurrent Dutch follow-up studies. Outcomes are quality-adjusted life years lived and lifetime medical cost by disability category. An economic analysis compares outcomes from a successful stroke service to usual care, by bootstrapping individual costs and effects data from patients in each arm. Lifetime costs and QALYs after stroke depend on age-of-onset of first-ever stroke. Lifetime QALYs after stroke are 2.42 (90% CI - 0.49 - 2.75) for male patients in usual care and 2.75 (-0.61; 6.26) for females. Lifetime costs for men in the usual care setting are €39,335 (15,951; 79,837) and €42,944 (14,081; 95,944) for women. A comparison with the stroke service results in an ICER of €11,685 saved per QALY gained (€14,211 and €7,745 for men and women respectively). This stroke service is with 90% certainty cost-effective. Our analysis shows the potential of large health benefits and cost savings of stroke services, taking a lifetime perspective, also in other European settings.


Risk of cancer in patients with scleroderma: a population based cohort study.
Previous studies have suggested an increased risk of cancer among patients with scleroderma. To study a population based cohort of patients with scleroderma in South Australia. Subjects with scleroderma were identified from the South Australian Scleroderma Registry established in 1993. All subjects on the scleroderma registry were linked to the South Australian Cancer Registry to identify all cases of cancer until 31 December 2000. Standardised incidence ratios (SIRs) for cancer for subjects with scleroderma were determined using the age- and sex-specific rates for South Australia. In 441 patients with scleroderma, 90 cases of cancer were identified, 47 of which developed after inclusion on the scleroderma registry. The SIRs for all cancers among these patients were significantly increased (SIR=1.99; 95% confidence interval (95% CI) 1.46 to 2.65) compared with the cancer incidence rates for South Australia. The SIRs for lung cancer (SIR=5.9; 95% CI 3.05 to 10.31) were also significantly increased. The SIRs for all cancers among the subgroups with diffuse scleroderma (SIR=2.73; 95% CI 1.31 to 5.02) and limited scleroderma (SIR=1.85; 95% CI 1.23 to 2.68) were significantly increased. This population based cohort study provides evidence that scleroderma is associated with cancer, and in particular, lung cancer. In addition, both diffuse and limited forms of scleroderma are associated with a similarly increased risk of cancer.


Effect of laboratory or clerical error on presymptomatic risk calculations for Huntington disease: a simulation study.
Linked markers are useful in prenatal diagnosis as well as presymptomatic diagnosis in late age-of-onset diseases such as Huntington disease (HD). It is widely assumed that most laboratory or clerical errors will be detected because of incompatibility of marker haplotypes within the family. However, errors in marker phenotypes that are compatible but wrong may result in a consultand being given an incorrect risk estimate. We have addressed this issue using simulated marker data in pedigrees similar to those seen in our HD testing program. In Family Structure I (an 11-member, 3-generation family), a particular family was more likely to be detected as inconsistent than incorrectly assigned. In a small nuclear family (Family Structure IV), fewer errors would be detected, and more would appear consistent but give incorrect risk estimates (e.g., low risk misclassified as noninformative or high). Given the presence of tight linkage, risk estimates are often calculated based on a small number of relatives. However, these computer simulations demonstrated that increasing the number of relatives typed decreases the probability that the family will remain consistent with an error present, and, therefore, decreases the probability of an incorrect assignment of risk. It is important to decrease the level of such errors by duplicated readings of raw marker data and validation of computer input.


Association between comorbidities and disease activity in axial spondyloarthritis: results from the BSRBR-AS.
Whether comorbidities influence disease activity assessment in axial SpA (axSpA) is unclear. Comorbidities inflate DAS28 in rheumatoid arthritis through the patient global score. We examined whether axSpA disease activity measures are differentially affected, and whether comorbidities inflate the AS disease activity score (ASDAS) through the patient global component. We used baseline data from the British Society for Rheumatology Biologics Register for AS, including 14 physician diagnosed comorbidities. Linear models were used to compare disease activity (BASDAI, spinal pain, ASDAS) and ESR/CRP according to comorbidity count, adjusted for age, gender, BMI, smoking, socioeconomic status, and education. The same models were used to examine whether the patient global score was associated with comorbidities, additionally adjusting for other ASDAS components. The number of participants eligible for analysis was 2043 (67% male, mean age 49 years); 44% had at least one comorbidity. Each additional comorbidity was associated with higher BASDAI by 0.40 units (95% CI: 0.27, 0.52) and spinal pain by 0.53 (95% CI: 0.37, 0.68). Effect size for ASDAS (0.09 units; 95% CI: 0.03, 0.15) was not clinically significant. ESR and CRP were not associated with comorbidity count. Depression, heart failure and peptic ulcer were consistently associated with higher disease activity measures, but not CRP/ESR. Patient global was associated with comorbidity count, but not independently of other ASDAS components (P = 0.75). Comorbidities were associated with higher patient reported disease activity in axSpA. Clinicians should be mindful of the potential impact of comorbidities on patient reported outcome measures and consider additionally collecting ASDAS when comorbidities are present.


Executive dysfunction in subcortical ischaemic vascular disease.
Executive dysfunction has been reported in patients with subcortical-frontal pathology, even in the absence of dementia. This study was undertaken to determine if impairments in executive functioning could be found in non-demented patients with subcortical lacunes. Cross sectional comparison between older control subjects (n=27) and non-demented patients with one or more subcortical lacunes (n=12). All participants were administered a neuropsychological test battery incorporating three measures of executive functioning, the Stroop interference test, California card sorting test, and the initiation-perseveration subtest of the Mattis dementia rating scale. No group differences were found on measures of recent verbal memory, language, or spatial ability. Normal controls performed better than patients with lacunes in visual memory. On the Stroop interference test, patients with lacunes performed as well as controls on the colour naming condition but slower on the interference condition. Patients with lacunes also generated fewer correct sorts on the California card sort test and achieved lower scores on the initiation-perseveration subtest. Executive measures were correlated with extent of white matter signal hyperintensity but not number of lacunes. Subcortical ischaemic vascular disease is associated with subtle declines in executive functioning and visual memory, even in non-demented patients. The pattern of cognitive impairment after subcortical lacunes is consistent with models of subcortical-frontal circuits.


Hyperhydration-Induced Decrease in Urinary Luteinizing Hormone Concentrations of Male Athletes in Doping Control Analysis.
Low urinary luteinizing hormone (LH) values have been discussed as a marker to detect steroid abuse. However, suppressed LH concentrations related to highly diluted urine samples could be a misleading indication of anabolic steroid abuse. One aim of the present study was to examine the effect of hyperhydration on the interpretation of LH findings during doping control analysis and to investigate different possibilities to correct volume-related changes in urinary LH concentrations. Seven healthy, physically active, nonsmoking White males were examined for a 72-hr period, using water and a commercial sports drink as hyperhydration agents (20 ml/kg body weight). Urine samples were collected and analyzed according to the World Anti-Doping Agency's technical documents. Baseline urinary LH concentrations, expressed as the mean ± SD for each individual, were within the acceptable physiological range (7.11 ± 5.42 IU/L). A comparison of the measured LH values for both hyperhydration phases (Phase A: 4.24 ± 5.60 IU/L and Phase B: 4.74 ± 4.72 IU/L) with the baseline ("normal") values showed significant differences (Phase A: p < .001 and Phase B: p < .001), suggesting the clear effect of urine dilution due to hyperhydration. However, an adjustment of urinary LH concentrations by specific gravity based on a reference value of 1.020 seems to adequately correct the hyperhydration-induced decrease on the LH levels.


A Redescription of Serrasentis sagittifer (Rhadinorhynchidae: Serrasentinae) from Rachycentron canadum (Rachycentridae) with Comments on its Biology and its Relationship to Other Species of Serrasentis.
Adult and cystacanth forms of the acanthocephalan Serrasentis sagittifer from Australian coastal waters are redescribed and verified as the same species using both molecular and morphological data. This study provides the baseline 18S rDNA, 28S rDNA, and cox1 sequence data to serve as genetic barcode for S. sagittifer. The validity of the currently recognized species of Serrasentis is discussed. The most recently described species are junior synonyms of either Serrasentis nadakali or S. sagittifer, and a number of species are species inquirenda. When using morphological characters to distinguish the species of Serrasentis, consideration needs to be given to the maturity of the specimens, since the trunk elongates and the number and distribution of the ventral combs changes as worms mature, although the proboscis armature itself does not change. A simple key to assist in the identification of species of Serrasentis is provided. Adult S. sagittifer appear to be highly host specific to the cobia, Rachycentron canadum, in northern Australian waters, whereas cystacanths have been reported from a wide range of fish species. The relationship between host length and number of cystacanths shows that most paratenic infections are acquired as young fish, most likely via a crustacean intermediate host.


Out-of-hospital coronary death in an urban population--validation of death certificate diagnosis. The Minnesota Heart Survey.
To assess the validity of death certificate diagnoses of out-of-hospital coronary heart disease deaths, the authors studied a one-third random sample of out-of-hospital deaths occurring in 1979 in Minneapolis-St. Paul, Minnesota, residents. Death certificates with diagnoses possibly containing coronary heart disease deaths were enumerated, and cause of death was recorded from the certificate in two ways: as the first listed ("immediate") cause and as the "underlying cause" assigned by a trained nosologist. Validation was performed by standardized physician review of information obtained about the death, which included one or more of the following: an interview with a relative or friend, physician report, autopsy report, medical record, and/or nursing home record. Missing information was frequent, but cases with at least an informant interview and/or autopsy report (82%) were representative and could be used for validation. The sensitivity and specificity of the underlying cause of coronary heart disease (International Classification of Diseases, Ninth Revision, codes 410-414, 427) on the death certificate were 90.3% and 82.7%, respectively, compared with the physician-assigned diagnosis. For the immediate cause, sensitivity and specificity were 90.3% and 67.9%, respectively. These findings suggest that the validity of death certificates for out-of-hospital coronary heart disease death is high, as assessed by this method of retrospective physician review.


Pneumothoraces Prevented With Use of Electromagnetic Device to Place Feeding Tubes.
A US Food and Drug Administration safety letter warned about the risk for pneumothoraces during feeding tube insertion despite the use of electromagnetic placement devices that provide real-time visualization of feeding tube position. To systematically assess pulmonary placement and pneumothoraces in CORTRAK-assisted feeding tube insertions. CINAHL, MEDLINE, and Cochrane databases were searched for studies of CORTRAK-assisted feeding tube insertion. Thirty-two studies documenting pulmonary placement and/or complications of feeding tube insertion were found. Operators recognized pulmonary placement on insertion tracings during 202 CORTRAK-assisted feeding tube insertion procedures, resulting in the immediate withdrawal of 199 feeding tubes. One pneumothorax was identified later by radiography. Seven pulmonary placements were not recognized by CORTRAK operators at the time of feeding tube insertion, resulting in 2 pneumothoraces. The incidence of pneumothorax for CORTRAK-assisted feeding tube insertions was 0.02% (3 of 17039). Of the feeding tubes inserted into the pulmonary system - either found during or after the procedure -1.4% (3 of 209) resulted in pneumothoraces (as opposed to the 19% to 28% incidence of pneumothorax for blind feeding tube insertions. Operators recognizing pulmonary placement on CORTRAK insertion tracings may have prevented 97% (202 of 209) of feeding tubes from being inserted farther into the respiratory tract. Feeding tube insertion with an electromagnetic placement device is advantageous over blind feeding tube insertion because the operator can recognize pulmonary placement early and withdraw the feeding tube, thus decreasing the risk of pulmonary complications.


Hemi-epiphysiodesis for unclassified congenital scoliosis: immediate results and mid-term follow-up.
Complex spinal anomalies involved in congenital scoliosis consist of a jumble of vertebral defects. Progressive scoliotic curves are frequently encountered in these patients. We evaluated the results of hemi-epiphysiodesis for this patient population. A series of ten patients with unclassified scoliosis, involving multiple hemivertebrae and/or bars, was reviewed retrospectively. Hemi-epiphysiodesis was the primary treatment in all patients. Mortality, complications and wound healing problems did not occur. The average Cobb angle for all patients changed from 54 degrees (range: 40 degrees -74 degrees), preoperatively, to 58 degrees (range: 30 degrees -104 degrees), postoperatively. Applying the criterion of a minimum change of 20 degrees, to take into account measurement variability, an epiphysiodesis effect was achieved in two procedures, progression was arrested in six procedures and failure occurred in two procedures. The mean rate of Cobb angle change per annum decreased from 2.9 degrees (range: -35 degrees to 14 degrees), preoperatively, to 2.4 degrees (range: -4 degrees to 13 degrees), following surgery. Repeat surgery was necessitated by coexisting progressive kyphosis and pseudoarthrosis in one patient, and involved extension of primary epiphysiodesis in two patients. A mean intervention-free period of 58 months was established. These results suggest that hemi-epiphysiodesis stabilized the unclassified congenital scoliosis. Being under 5 years of age, having a thoracolumbar curve location, and the absence of coexisting kyphosis were found to be associated with a more favorable outcome.


Plasmonic caged gold nanorods for near-infrared light controlled drug delivery.
A new near-infrared light-controlled drug delivery system based on caged gold nanorods (CGNRs) is demonstrated. The loading and release process of drug payloads into/from CGNR nanocarriers were systematically investigated. The drug-loaded CGNR constructs could enable combined chemotherapy and photo-thermal effects in killing tumor cells upon light irradiation, therefore, enhance the killing efficiency. In conjunction with visibility under quenching-free dark-field imaging, CGNRs may serve as multifunctional theranostic reagents towards cancer diagnostics and therapeutics.


Plasma growth hormone is a potential biomarker of response to atezolizumab and bevacizumab in advanced hepatocellular carcinoma patients.
Hepatocellular carcinoma (HCC) has limited systemic therapy options when discovered at an advanced stage. Thus, there is a need for accessible and minimally invasive biomarkers of response to guide the selection of patients for treatment. This study investigated the biomarker value of plasma growth hormone (GH) level as a potential biomarker to predict outcome in unresectable HCC patients treated with current standard therapy, atezolizumab plus bevacizumab (Atezo/Bev). Study included unresectable HCC patients scheduled to receive Atezo/Bev. Patients were followed to determine progression-free survival (PFS) and overall survival (OS). Plasma GH levels were measured by ELISA and used to stratify the HCC patients into GH-high and GH-low groups (the cutoff normal GH levels in women and men are ≤3.7 μg/L and ≤0.9 μg/L, respectively). Kaplan-Meier method was used to calculate median OS and PFS and Log rank test was used to compare survival outcomes between GH-high and -low groups. Thirty-seven patients were included in this analysis, of whom 31 were males and 6 females, with a median age of 67 years (range: 37-80). At the time of the analysis, the one-year survival rate was 70% (95% CI: 0.51, 0.96) among GH low patients and 33% (95% CI: 0.16, 0.67) among GH high patients. OS was significantly superior in GH-low compared to GH-high patients (median OS: 18.9 vs. 9.3 months; p = 0.014). PFS showed a non-significant trend in favor of GH-low patients compared to the GH-high group (median PFS: 6.6 vs. 2.9 months; p = 0.053). Plasma GH is a biomarker candidate for predicting treatment outcomes in advanced HCC patients treated with Atezo/Bev. This finding should be further validated in larger randomized clinical trials in advanced HCC patients.


Susceptibility of low-density lipoprotein particles to aggregate depends on particle lipidome, is modifiable, and associates with future cardiovascular deaths.
Low-density lipoprotein (LDL) particles cause atherosclerotic cardiovascular disease (ASCVD) through their retention, modification, and accumulation within the arterial intima. High plasma concentrations of LDL drive this disease, but LDL quality may also contribute. Here, we focused on the intrinsic propensity of LDL to aggregate upon modification. We examined whether inter-individual differences in this quality are linked with LDL lipid composition and coronary artery disease (CAD) death, and basic mechanisms for plaque growth and destabilization. We developed a novel, reproducible method to assess the susceptibility of LDL particles to aggregate during lipolysis induced ex vivo by human recombinant secretory sphingomyelinase. Among patients with an established CAD, we found that the presence of aggregation-prone LDL was predictive of future cardiovascular deaths, independently of conventional risk factors. Aggregation-prone LDL contained more sphingolipids and less phosphatidylcholines than did aggregation-resistant LDL. Three interventions in animal models to rationally alter LDL composition lowered its susceptibility to aggregate and slowed atherosclerosis. Similar compositional changes induced in humans by PCSK9 inhibition or healthy diet also lowered LDL aggregation susceptibility. Aggregated LDL in vitro activated macrophages and T cells, two key cell types involved in plaque progression and rupture. Our results identify the susceptibility of LDL to aggregate as a novel measurable and modifiable factor in the progression of human ASCVD.


Biolimus-eluting stent with biodegradable polymer improves clinical outcomes in patients with acute myocardial infarction.
To investigate clinical outcomes of coronary intervention using a biolimus-eluting stent (BES) compared with a sirolimus-eluting stent (SES) in patients with acute myocardial infarction (AMI) in the Limus Eluted from A Durable versus ERodable Stent (LEADERS) coating trial at the final 5-year follow-up. The LEADERS trial is a multicentre all-comer study, where patients (n=1707) were randomised to percutaneous intervention with either BES containing biodegradable polymer or SES containing durable polymer. Out of 1707 patients enrolled in this trial, 573 patients had percutaneous coronary intervention for AMI (BES=280, SES=293) and were included in the current analysis. Patient-oriented composite endpoint (POCE, including all death, all myocardial infarction (MI) and all revascularisations), major adverse cardiac events (MACE, including cardiac death, MI and clinically indicated target vessel revascularisation) and stent thrombosis were assessed at 5-year follow-up. The baseline clinical, angiographic and procedural characteristics were well matched between BES and SES groups. In all patients with AMI, coronary intervention with a BES, compared with SES, significantly reduced POCE (28.9% vs 42.3%; relative risk (RR) 0.61, 95% CI 0.47 to 0.82, p=0.001) at 5-year follow-up. There was also a reduction in MACE rate in the BES group (18.2% vs 25.9%; RR 0.67, 95% CI 0.47 to 0.95, p=0.025); however, there was no difference in cardiac death and stent thrombosis. In patients with ST-elevation MI (STEMI), coronary intervention with BES significantly reduced POCE (24.4% vs 39.3%; RR 0.55, 95% CI 0.36 to 0.85, p=0.006), MACE (12.6% vs 25.0%; RR 0.47, 95% CI 0.26 to 0.83, p=0.008) and cardiac death (3.0% vs 11.4%; RR 0.25, 95% CI 0.08 to 0.75, p=0.007), along with a trend towards reduction in definite stent thrombosis (3.7% vs 8.6%; RR 0.41, 95% CI 0.15 to 1.18, p=0.088), compared with SES. BES, compared with SES, significantly improved safety and efficacy outcomes in patients with AMI, especially those with STEMI, at 5-year follow-up. NCT 00389220.


Insulin gene region-encoded susceptibility to IDDM maps upstream of the insulin gene.
The gene region on chromosome 11p15.5 known to be involved in insulin-dependent diabetes mellitus (IDDM) susceptibility was recently mapped to a 4.1-kilobase region including the insulin gene. The region contains 10 candidate polymorphisms that are in strong linkage disequilibrium. By genotyping 7 of these 10 polymorphisms and the tyrosine hydroxylase microsatellite in Finnish Caucasoid IDDM patients and control subjects, we demonstrate that many of the polymorphisms found to be associated with IDDM in other Caucasoid populations do not show any association in this Finnish population. Of the polymorphisms typed, only those at -23 Hph I and the variable number of tandem repeats (VNTR) sites confer significant relative risk. Furthermore, we have demonstrated that the -23 Hph I polymorphism cannot explain the association. Comparison of the genotypic patterns observed here and previously suggests that the VNTR is the most likely candidate for IDDM2. The VNTR is located adjacent to defined regulatory DNA sequences affecting insulin gene expression, which suggests a possible effect on expression of insulin or one of the neighboring genes, tyrosine hydroxylase or insulin-like growth factor 2.


The influence of prior synaptic activity on the induction of long-term potentiation.
Long-term potentiation (LTP) is an extensively studied model of synaptic plasticity, in part because it is a plausible biological correlate for the Hebbian synaptic modification that forms the basis for theoretical models of neural development, learning, and memory. Although these models must incorporate algorithms that constrain synaptic weight changes, physiological evidence for such mechanisms is limited. Examination of LTP in area CA1 of the hippocampus revealed that the threshold for LTP induction was not fixed but could be strongly influenced by the recent history of synaptic activity. This effect was transient, synapse-specific, and dependent on postsynaptic N-methyl-D-aspartate (NMDA) receptor activation. These results suggest that the threshold for LTP induction may be continually adjusted according to the recent history of NMDA receptor activation and provide a physiological mechanism by which LTP can be transiently inhibited.


Bone involvement predicts poor outcome in atypical meningioma.
The authors identified clinical features associated with progression and death in atypical meningioma (AM). Forty-seven cases of primary AM treated at Massachusetts General Hospital were retrospectively evaluated for clinical features. Associations with progression-free survival (PFS) and overall survival were assessed. The estimated median PFS was 56 months (95% CI 35 months-not estimable). The overall 3- and 5-year PFS rates were 65% (95% CI 44-80%) and 48% (95% CI 26-67%), respectively. The median survival time and 5- and 10-year survival rates were 158 months (95% CI 103 months-not estimable), and 86% (95% CI 69-94%) and 61% (95% CI 35-79%), respectively. Subtotal resection was associated with increased rate of progression compared to gross-total resection (p=0.05) and trended toward an association with decreased survival (p=0.09). Bone involvement was associated with an increased rate of disease progression (p=0.001) and decreased survival (p=0.04). Bone involvement remained significantly associated with progression after Bonferroni adjustment for multiple comparisons (p=0.008) and in bivariate Cox regression models. Seventy-eight percent of patients with bone involvement at primary diagnosis had tumor recurrence within bone, whereas only 25% of patients without evidence of bone invasion at primary diagnosis experienced osseous recurrence. Osseous involvement is associated with a poor outcome in patients with AMs; bone assessment is therefore extremely important. Further investigation is warranted to assess the effectiveness of bone resection and/or bone-directed radiation therapy in improving outcome.


Hydrodynamic Effects on Drug Dissolution and Deaggregation in the Small Intestine-A Study with Felodipine as a Model Drug.
The aim of this study was to understand and predict the influence of hydrodynamic effects in the small intestine on dissolution of primary and aggregated drug particles. Dissolution tests of suspensions with a low-solubility drug, felodipine, were performed in a Couette cell under hydrodynamic test conditions corresponding to the fed small intestine. Dissolution was also performed in the USP II apparatus at two paddle speeds of 25 and 200 rpm and at different surfactant concentrations below critical micelle concentration. The experimental dissolution rates were compared with theoretical calculations. The different levels of shear stress in the in vitro tests did not influence the dissolution of primary or aggregated particles and experimental dissolution rates corresponded very well to calculations. The dissolution rate for the aggregated drug particles increased after addition of surfactant because of deaggregation, but there were still no effect of hydrodynamics. In conclusion, hydrodynamics do not influence dissolution and deaggregation of micronized drug particles in the small intestine of this model drug. Surface tension has a strong effect on the deaggregation and subsequent dissolution. Addition of surfactants at in vivo relevant surface tension levels is thus critical for in vivo predictive in vitro dissolution testing.


Reconstituting donor T cells increase their biomass following hematopoietic stem cell transplantation.
In this study, we used a rapid, highly-sensitive, single-cell biomass measurement method, Live Cell Interferometry (LCI), to measure biomass in populations of CD3 + T cells isolated from hematopoietic stem cell transplant (SCT) patients at various times pre- and post-transplant (days 0-100). CD3 + T cell 'mass spectra' were obtained from five autologous and 20 allogenic transplant recipients. We found a pronounced rise in median T cell biomass (+25%; p <0.001) shortly after transplant (day 14), which moderated by day 60. Further, the inter-patient and intra-patient cell masses were most variable at days 14 and 30 post-transplant. T cell biomass trends were similar in both autologous and allogenic transplant recipients. These data suggest that T cell biomass changes are associated with immune reconstitution occurring in the first few weeks post-transplant. To our knowledge, this is the first time single-cell biomass measurements have been studied in human clinical trials. With refinement, these data may prove useful in guiding the withdrawal of immunosuppression following SCT, reducing the likelihood of Graft-Versus-Host Disease or cancer relapse occurring.


In vivo bioimaging with tissue-specific transcription factor activated luciferase reporters.
The application of transcription factor activated luciferase reporter cassettes in vitro is widespread but potential for in vivo application has not yet been realized. Bioluminescence imaging enables non-invasive tracking of gene expression in transfected tissues of living rodents. However the mature immune response limits luciferase expression when delivered in adulthood. We present a novel approach of tissue-targeted delivery of transcription factor activated luciferase reporter lentiviruses to neonatal rodents as an alternative to the existing technology of generating germline transgenic light producing rodents. At this age, neonates acquire immune tolerance to the conditionally responsive luciferase reporter. This simple and transferrable procedure permits surrogate quantitation of transcription factor activity over the lifetime of the animal. We show principal efficacy by temporally quantifying NFκB activity in the brain, liver and lungs of somatotransgenic reporter mice subjected to lipopolysaccharide (LPS)-induced inflammation. This response is ablated in Tlr4(-/-) mice or when co-administered with the anti-inflammatory glucocorticoid analogue dexamethasone. Furthermore, we show the malleability of this technology by quantifying NFκB-mediated luciferase expression in outbred rats. Finally, we use somatotransgenic bioimaging to longitudinally quantify LPS- and ActivinA-induced upregulation of liver specific glucocorticoid receptor and Smad2/3 reporter constructs in somatotransgenic mice, respectively.


Iron deficiency anaemia and delayed diagnosis of colorectal cancer: a retrospective cohort study.
The extent to which different referral pathways following a primary care diagnosis of iron deficiency anaemia (IDA) are associated with delay in diagnosis of colorectal cancer (CRC) was determined. Eligible patients aged 40 or more years, with IDA diagnosed in primary care, and a subsequent diagnosis of CRC, were studied retrospectively. Referral pathways were identified using the specialty of first recorded GP referral following IDA diagnosis. Differences in time to diagnosis of CRC were assessed by referral specialty. Differences in the proportion of cases referred before and after the re-issue of the NICE urgent referral guidelines for suspected lower gastrointestinal (GI) cancer were also assessed. Of 628,882 eligible patients, 3.1% (n = 19,349) were diagnosed with IDA during the study period; 3.0% (n = 578) were subsequently diagnosed with CRC. Two hundred and fifty-nine (44.8%) patients had no recorded referral or a referral unrelated to anaemia or the GI tract. Only 35% (n = 201) of patients were referred to a relevant specialty. Median time to CRC diagnosis ranged from 2.5 months (referral to a relevant surgical specialty) to 31.9 months (haematology). Time to diagnosis was longer in patients referred to a medical compared with a relevant surgical specialty (P = 0.024). There was no significant difference in time to CRC diagnosis before and after the NICE guidelines were re-issued in 2005. Significant differences exist between referral specialties in time to CRC diagnosis following a primary care diagnosis of IDA. Despite NICE referral recommendations, a significant proportion of patients are still not managed within recommended care pathways to CRC diagnosis.


Threshold criterion for wetting at the triple point
Grand canonical simulations are used to calculate adsorption isotherms of various classical gases on alkali metal and Mg surfaces. Ab initio adsorption potentials and Lennard-Jones gas-gas interactions are used. Depending on the system, the resulting behavior can be nonwetting for all temperatures studied, complete wetting, or (in the intermediate case) exhibit a wetting transition. An unusual variety of wetting transitions at the triple point is found in the case of a specific adsorption potential of intermediate strength. The general threshold for wetting near the triple point is found to be close to that predicted with a heuristic model of Cheng et al. This same conclusion was drawn in a recent experimental and simulation study of Ar on CO2 by Mistura et al. These results imply that a dimensionless wetting parameter w is useful for predicting whether wetting behavior is present at and above the triple temperature. The nonwetting/wetting crossover value found here is w approximately 3.3.


Sequence variants of the Phytophthora sojae RXLR effector Avr3a/5 are differentially recognized by Rps3a and Rps5 in soybean.
The perception of Phytophthora sojae avirulence (Avr) gene products by corresponding soybean resistance (Rps) gene products causes effector triggered immunity. Past studies have shown that the Avr3a and Avr5 genes of P. sojae are genetically linked, and the Avr3a gene encoding a secreted RXLR effector protein was recently identified. We now provide evidence that Avr3a and Avr5 are allelic. Genetic mapping data from F(2) progeny indicates that Avr3a and Avr5 co-segregate, and haplotype analysis of P. sojae strain collections reveal sequence and transcriptional polymorphisms that are consistent with a single genetic locus encoding Avr3a/5. Transformation of P. sojae and transient expression in soybean were performed to test how Avr3a/5 alleles interact with soybean Rps3a and Rps5. Over-expression of Avr3a/5 in a P. sojae strain that is normally virulent on Rps3a and Rps5 results in avirulence to Rps3a and Rps5; whereas silencing of Avr3a/5 causes gain of virulence in a P. sojae strain that is normally avirulent on Rps3a and Rps5 soybean lines. Transient expression and co-bombardment with a reporter gene confirms that Avr3a/5 triggers cell death in Rps5 soybean leaves in an appropriate allele-specific manner. Sequence analysis of the Avr3a/5 gene identifies crucial residues in the effector domain that distinguish recognition by Rps3a and Rps5.


Starch-to-sugar conversion in wood parenchyma of field-growing Laurus nobilis plants: a component of the signal pathway for embolism repair?
The ability of stems of Laurus nobilis (L.) to refill embolised xylem conduits was studied in plants both at optimal water supply (W) and under conditions of soil drought inducing xylem pressures (Px) of -1.54 (S1) and -2.35 MPa (S2). Starch depolymerisation in wood parenchyma was measured as percentage of cells 'with high starch content' (HSC-cells) counted under a microscope. HSC-cells decreased during embolism and increased again in refilled stems. A direct relationship was found between percentage of HSC-cells and Px, with HSC-cells between 65 and 75% of the total at Px ≥ -0.6 MPa, at which recovery from PLC was recorded. At low transpiration, starch re-appeared in wood parenchyma cells but only in plants that showed diurnal stomatal opening (W- and S1-plants). In S2-plants showing diurnal stomatal closure and nocturnal opening with Px between -1.2 to -2.4 MPa, HSC-cells were only 25% and plants did not recover from PLC. This finding suggests that (i) the Px threshold for embolism repair was ≥ -0.6 MPa, and (ii) impeded phloem loading limits starch content in wood parenchyma and embolism repair. We conclude that starch depolymerisation acts as a signal to phloem unloading sugars to embolised conduits thus generating the necessary osmotic gradients driving refilling.


Methanobacterium paludis sp. nov. and a novel strain of Methanobacterium lacus isolated from northern peatlands.
Two mesophilic, hydrogenotrophic methanogens, designated strains SWAN1T and AL-21, were isolated from two contrasting peatlands: a near circumneutral temperate minerotrophic fen in New York State, USA, and an acidic boreal poor fen site in Alaska, USA, respectively. Cells of the two strains were rod-shaped, non-motile, stained Gram-negative and resisted lysis with 0.1% SDS. Cell size was 0.6×1.5-2.8 µm for strain SWAN1T and 0.45-0.85×1.5-35 µm for strain AL-21. The strains used H2/CO2 but not formate or other substrates for methanogenesis, grew optimally around 32-37 °C, and their growth spanned through a slightly low to neutral pH range (4.7-7.1). Strain AL-21 grew optimally closer to neutrality at pH 6.2, whereas strain SWAN1T showed a lower optimal pH at 5.4-5.7. The two strains were sensitive to NaCl with a maximal tolerance at 160 mM for strain SWAN1T and 50 mM for strain AL-21. Na2S was toxic at very low concentrations (0.01-0.8 mM), resulting in growth inhibition above these values. The DNA G+C content of the genomes was 35.7 mol% for strain SWAN1T and 35.8 mol% for strain AL-21. Phylogenetic analysis of the 16S rRNA gene sequences showed that the strains are members of the genus Methanobacterium. Strain SWAN1T shared 94-97% similarity with the type strains of recognized species of the genus Methanobacterium, whereas strain AL-21 shared 99% similarity with Methanobacterium lacus 17A1T. On the basis of phenotypic, genomic and phylogenetic characteristics, strain SWAN1T (=DSM 25820T=JCM 18151T) is proposed as the type strain of a novel species, Methanobacterium paludis sp. nov., while strain AL-21 is proposed as a second strain of Methanobacterium lacus.


Increased secretion of atrial and brain natriuretic peptides during acute myocardial ischaemia induced by dynamic exercise in patients with angina pectoris.
1. This study was conducted to assess the role of artrial and brain natriuretic peptides during acute myocardial ischaemia associated with dynamic exercise. 2. Study subjects consisted of 35 angiographically proven patients with angina pectoris and 35 angiographically normal control subjects. All subjects underwent 201Tl dynamic exercise testing. The presence and localization of the exercise-induced acute myocardial perfusion defect were assessed by 201Tl single-photon emission computed tomography. The severity score was calculated using the early image for quantitative assessment of the acute myocardial perfusion defect. 3. Plasma levels of atrial natriuretic peptide increased from 21.3 +/- 3.8 to 72.2 +/- 26.7 pg/ml (P < 0.01) in the angina pectoris group, and increased from 19.4 +/- 2.4 to 36.4 +/- 17.4 pg/ml (P < 0.01) in the control group during dynamic exercise. Plasma levels of brain natriuretic peptide increased from 2.8 +/- 0.8 to 6.9 +/- 2.6 pg/ml (P < 0.01) in the angina pectoris group, but did not change significantly in the control group (from 2.7 +/- 0.7 to 2.9 +/- 1.0 pg/ml) during dynamic exercise. At peak exercise, plasma levels of these natriuretic peptides in the angina pectoris group were significantly higher than those in the control group (P < 0.01). 4. At peak exercise, there were correlations between the plasma level of atrial natriuretic peptide and heart rate in both the angina pectoris and control groups (P < 0.01, r = 0.46; P < 0.01, r = 0.51, respectively), but no significant correlations between the plasma level of brain natriuretic peptide and heart rate in either group. The plasma levels of these peptides at peak exercise correlated well with the severity score in the angina pectoris group (atrial natriuretic peptide, r = 0.71, P < 0.01; brain natriuretic peptide, r = 0.69, P < 0.01).


Obturator hernia, a rare cause of small bowel obstruction: case report.
Obturator hernia is a rare hernia in the world, diagnosed late since it has no specific symptoms and findings and generally occur in thin and old women with comorbidity.For this reason obturator hernia has high morbidity and mortality rates. In this study, we present an obturator hernia case that Howship-Romberg sign is positive and has typical appearance in computerized tomography. Laparotomy was performed on 89 years old female patient with body mass index 18.08 kg/m(2) by low middle line incision. Following the segmentectomy to the strangulated small bowel loop, obturator canal is repaired by retroperitoneal application. No complication occurred in the postoperative period. Obturator hernia should be taken into consideration in old and thin female patients with intestinal obstruction. Computerized tomography should be performed for early diagnosis of the obturator hernia.


Synthetic protein interactions reveal a functional map of the cell.
To understand the function of eukaryotic cells, it is critical to understand the role of protein-protein interactions and protein localization. Currently, we do not know the importance of global protein localization nor do we understand to what extent the cell is permissive for new protein associations - a key requirement for the evolution of new protein functions. To answer this question, we fused every protein in the yeast Saccharomyces cerevisiae with a partner from each of the major cellular compartments and quantitatively assessed the effects upon growth. This analysis reveals that cells have a remarkable and unanticipated tolerance for forced protein associations, even if these associations lead to a proportion of the protein moving compartments within the cell. Furthermore, the interactions that do perturb growth provide a functional map of spatial protein regulation, identifying key regulatory complexes for the normal homeostasis of eukaryotic cells.


Distribution of fat, non-osseous lean and bone mineral mass in international Rugby Union and Rugby Sevens players.
Differences in the body composition of international Rugby Union and Rugby Sevens players, and between players of different positions are poorly understood. The purpose of this study was to examine differences in the quantity and regional distribution of fat, non-osseous lean and bone mineral mass between playing units in Rugby Union and Rugby Sevens. Male Rugby Union (n=21 forwards, 17 backs) and Rugby Sevens (n=11 forwards, 16 backs) players from the Australian national squads were measured using dual-energy X-ray absorptiometry. The digital image of each player was partitioned into anatomical regions including the arms, legs, trunk, and android and gynoid regions. Compared with backs, forwards in each squad were heavier and exhibited higher absolute regional fat (Union 43-67%; ±~17%, range of % differences; ±~95% confidence limits (CL); Sevens 20-26%; ±~29%), non-osseous lean (Union 14-22%; ±~5.8%; Sevens 6.9-8.4%; ±~6.6%) and bone mineral (Union 12-26%; ±~7.2%; Sevens 5.0-11%; ±~7.2%) mass. When tissue mass was expressed relative to regional mass, differences between Rugby Sevens forwards and backs were mostly unclear. Rugby Union forwards had higher relative fat mass (1.7-4.7%; ±~1.9%, range of differences; ±~95% CL) and lower relative non-osseous lean mass (-4.2 to -1.8%; ±~1.8%) than backs in all body regions. Competing in Rugby Union or Rugby Sevens characterized the distribution of fat and non-osseous lean mass to a greater extent than a player's positional group, whereas the distribution of bone mineral mass was associated more with a player's position. Differences in the quantity and distribution of tissues appear to be related to positional roles and specific demands of competition in Rugby Union and Rugby Sevens.


Increased sensitivity to mu opiate antinociception in the neonatal rat despite weaker receptor-guanyl nucleotide binding protein coupling.
The developmental changes in mu opiate receptor-mediated antinociception and coupling to guanyl nucleotide binding proteins were studied in the rat. ED50 values for morphine- and sufentanil-induced antinociception were determined in the paw-lift assay on days 10 and 27. The ED50 for morphine analgesia in 10-day-old pups was 0.35 mg/kg and increased with age to 5.3 mg/kg on day 27. Similarly, sufentanil was more potent in pups than in weanlings, the ED50 increasing from 1.7 to 7.6 micrograms/kg. Serum and brain morphine levels after 5 mg/kg of morphine were higher in neonates (day 10) than weanlings (day 27), largely due to a more rapid redistribution phase in weanlings (T1/2 = 26 min.) than in pups (T1/2 = 2.5 hr). Additionally, a substantial (70%) antinociceptive response was achieved in neonates at brain morphine levels that were one-half those producing an equal effect in weanlings. Radioligand binding studies indicated that the number of mu receptors increased 2-fold without change in affinity between days 10 and 27, whereas the GTP analog guanylylimidophosphate was nearly twice as effective in shifting the mu receptors from high to low affinity on day 27 than on day 10. These data indicate that neonatal rats are more sensitive to mu opiate antinociception despite apparently weaker receptor-guanyl nucleotide binding protein coupling. This greater sensitivity is enhanced by the pharmacokinetic differences between neonates and weanlings which result in higher drug levels in pups after a similar dose.


Osimertinib: A Review in T790M-Positive Advanced Non-Small Cell Lung Cancer.
Osimertinib (Tagrisso™) is an oral, CNS-active, third-generation epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) that targets EGFR TKI-sensitizing mutations and, crucially, the T790M mutation that often underlies acquired resistance to EGFR TKI therapy. Osimertinib has been approved in numerous countries for use in patients with T790M-positive advanced NSCLC. In the pivotal, international AURA3 trial in patients with T790M-positive advanced NSCLC who had disease progression after EGFR TKI therapy, osimertinib treatment significantly prolonged progression-free survival (PFS; primary endpoint) compared with platinum-pemetrexed therapy at the time of the primary analysis. PFS results were consistent across predefined subgroups of patients, including those with CNS metastases at baseline. There was no difference between treatment groups in overall survival at 26% maturity. Objective response rates (ORRs) and patient-reported outcomes for prespecified symptoms were also significantly improved with osimertinib relative to platinum-pemetrexed, with CNS ORRs in patients with CNS metastases more than twofold higher in the osimertinib than in the platinum-pemetrexed group. Osimertinib had a manageable tolerability profile, with relatively few patients permanently discontinuing treatment because of adverse events (AEs). With limited treatment options available in this setting, osimertinib is an important option in adult patients with advanced EGFR T790M-positive NSCLC.


Assessment of dose variation for accelerated partial-breast irradiation using rigid and deformable image registrations.
The aim of this study was to estimate the delivered dose to the target and organs at risk (OAR) for external beam accelerated partial-breast irradiation (APBI) accounting for day-to-day setup uncertainties, using rigid and deformable image registration. One planning computed tomography (CT) scan and 5 cone beam CT scans for each of 25 patients previously treated with tangential breast radiation therapy were used. All cone beam CT scans were registered to the planning CT scan using 3 techniques: (1) rigid registration based on bony anatomy only, (2) rigid registration based on soft-tissue only, and (3) deformable image registration. For each patient, 4 dose distributions were calculated for APBI. The first dose distribution was the original plan. The other 3 were "dose-of-the-day" for each of the registration approaches. The effects of image registrations on estimating delivered dose to targets and OAR were determined. The average reductions in V95 (percentage of the PTV that received 95% of the prescribed dose) were 6%, 7%, and 5% for bone, soft-tissue, and deformable registrations, respectively. The average increase in mean dose to the heart were 9%, 9%, and 18% for bone, soft-tissue, and deformable registrations, respectively, whereas the average increase in maximum dose to the contralateral breast were 19%, 20%, and 28%, respectively. The results of this study have shown that there are differences between the planned and estimated delivered dose for APBI because of day-to-day setup uncertainties that may need to be accounted for. Estimated dosimetric impact of setup variation and breast deformation assessed using deformable registration was greater for OARs and smaller for target volumes compared to rigid registration.


The longitudinal outcome of fibromyalgia: a study of 1555 patients.
To describe the diagnosis status and outcome of patients diagnosed with fibromyalgia (FM) by US rheumatologists. We assessed 1555 patients with FM with detailed outcome questionnaires during 11,006 semiannual observations for up to 11 years. At entry, all patients satisfied American College of Rheumatology preliminary 2010 FM criteria modified for survey research. We determined diagnosis status, rates of improvement, responder subgroups, and standardized mean differences (effect sizes) between start and study completion scores of global well-being, pain, sleep problems, and health related quality of life. (QOL) RESULTS: The 5-year improvement rates were pain 0.4 (95% CI 0.2, 0.5), fatigue 0.4 (95% CI 0.2, 0.05), and global 0.0 (95% CI -0.1, 0.1). The standardized mean differences were patient global 0.03 (95% CI -0.02, 0.08), pain 0.22 (95% CI 0.16, 0.28), sleep problems 0.20 (95% CI 0.14, 0.25), physical component summary of the Short-form 36 (SF-36) 0.11 (95% CI -0.14, -0.07), and SF-36 mental component summary 0.03 (95% CI -0.07, 0.02). Patients switched between criteria-positive and criteria-negative states, with 716 patients (44.0%) failing to meet criteria at least once during 4228.5 patient-years (7448 observations). About 10% of patients had substantial improvement and about 15% had moderate improvement of pain. Overall, FM severity worsened in 35.9% and pain in 38.6%. Although we found no average clinically meaningful improvement in symptom severity overall, 25% had at least moderate improvement of pain over time. The result that emerged from this longitudinal study was one of generally continuing high levels of self-reported symptoms and distress for most patients, but a slight trend toward improvement.


COPI activity coupled with fatty acid biosynthesis is required for viral replication.
During infection by diverse viral families, RNA replication occurs on the surface of virally induced cytoplasmic membranes of cellular origin. How this process is regulated, and which cellular factors are required, has been unclear. Moreover, the host-pathogen interactions that facilitate the formation of this new compartment might represent critical determinants of viral pathogenesis, and their elucidation may lead to novel insights into the coordination of vesicular trafficking events during infection. Here we show that in Drosophila cells, Drosophila C virus remodels the Golgi apparatus and forms a novel vesicular compartment, on the surface of which viral RNA replication takes place. Using genome-wide RNA interference screening, we found that this step in the viral lifecycle requires at least two host encoded pathways: the coat protein complex I (COPI) coatamer and fatty acid biosynthesis. Our results integrate, clarify, and extend numerous observations concerning the cell biology of viral replication, allowing us to conclude that the coupling of new cellular membrane formation with the budding of these vesicles from the Golgi apparatus allows for the regulated generation of this new virogenic organelle, which is essential for viral replication. Additionally, because these pathways are also limiting in flies and in human cells infected with the related RNA virus poliovirus, they may represent novel targets for antiviral therapies.


End-tidal carbon dioxide monitoring during procedural sedation.
To prospectively determine whether end-tidal carbon dioxide (ETCO2) monitors can detect respiratory depression (RD) and the level of sedation in emergency department (ED) patients undergoing procedural sedation (PS). This was a prospective observational study conducted in an urban county hospital of adult patients undergoing PS. Patients were monitored for vital signs, depth of sedation per the physician by the Observer's Assessment of Alertness/Sedation scale (OAA/S), pulse oximetry, and nasal-sample ETCO2 during PS. Respiratory depression was defined as an oxygen saturation <90%, an ETCO2 >50 mm Hg, or an absent ETCO2 waveform at any time during the procedure. The physician also determined whether protective airway reflexes were lost during the procedure and assisted ventilation was required, or whether there were any other complications. Rates of RD were compared with the physician assessment of airway loss and between agents using chi-square statistics. Spearman's rho analysis was used to determine whether there was a correlation between ETCO2 and the OAA/S score. Seventy-four patients were enrolled in the study. Forty (54.1%) received methohexital, 21 (28.4%) received propofol, ten (13.5%) received fentanyl and midazolam, and three (4.1%) received etomidate. Respiratory depression was seen in 33 (44.6%) patients, including 47.5% of patients receiving methohexital, 19% receiving propofol (p = 0.008), 80% receiving fentanyl and midazolam, and 66.6% receiving etomidate. No correlation between OAA/S and ETCO2 was detected. Eleven (14.9%) patients required assisted ventilation at some point during the procedure, all of whom met the criteria for RD. Pulse oximetry detected 11 of the 33 patients with RD. Post-hoc analysis revealed that all patients with RD had an ETCO2 >50 mm Hg, an absent waveform, or an absolute change from baseline in ETCO2 >10 mm Hg. There was no correlation between ETCO2 and the OAA/S score. Using the criteria of an ETCO2 >50 mm Hg, an absolute change >10 mm Hg, or an absent waveform may detect subclinical RD not detected by pulse oximetry alone. The ETCO2 may add to the safety of PS by quickly detecting hypoventilation during PS in the ED.


Recovery of ambulation activity across the first six months post-stroke.
Stroke survivors commonly adopt sedentary activity behaviours by the chronic phase of recovery. However, the change in activity behaviours from the subacute to chronic phase of stroke is variable. This study explored the recovery of ambulation activity (volume and bouts) at one, three and six months after hospital discharge post-stroke. A total of 42 stroke survivors were recruited at hospital discharge and followed up one, three and six months later. At follow-up, ambulation activity was measured over four days using the ActivPAL™ accelerometer. Measures included volume of activity and frequency and intensity of ambulation activity bouts per day. Linear mixed effects modelling was used to determine changes over time. There was wide variation in activity. Total step counts across all time points were below required levels for health benefits (mean 4592 SD 3411). Most activity was spread across short bouts. While most number of bouts was of low intensity, most time was spent in moderate intensity ambulation across all time points. Daily step count and time spent walking and sitting/lying increased from one month to three and six months. The number of and time spent in short and medium duration bouts increased from one to six months. Time in long duration bouts increased at three months only. Time spent in moderate intensity ambulation increased over time. No change was observed for any other measures. In future, it would be valuable to identify strategies to increase engagement in activity behaviours to improve health outcomes after stroke.


Brain transcriptomic response of threespine sticklebacks to cues of a predator.
Predation pressure represents a strong selective force that influences the development and evolution of living organisms. An increasing number of studies have shown that both environmental and social factors, including exposure to predators, substantially shape the structure and function of the brain. However, our knowledge about the molecular mechanisms underlying the response of the brain to environmental stimuli is limited. In this study, we used whole-genome comparative oligonucleotide microarrays to investigate the brain transcriptomic response to cues of a predator in the threespine stickleback, Gasterosteus aculeatus. We found that repeated exposure to olfactory, visual and tactile cues of a predator (rainbow trout, Oncorrhynchus mykiss) for 6 days resulted in subtle but significant transcriptomic changes in the brain of sticklebacks. Gene functional analysis and gene ontology enrichment revealed that the majority of the transcripts differentially expressed between the fish exposed to cues of a predator and the control group were related to antigen processing and presentation involving the major histocompatibility complex, transmission of synaptic signals, brain metabolic processes, gene regulation and visual perception. The top four identified pathways were synaptic long-term depression, RAN signaling, relaxin signaling and phototransduction. Our study demonstrates that exposure of sticklebacks to cues of a predator results in the activation of a wide range of biological and molecular processes and lays the foundation for future investigations on the molecular factors that modulate the function and evolution of the brain in response to stressors.


Treatment patterns and prognostic indicators of response to therapy among patients with chronic myeloid leukemia in Australia, Canada, and South Korea.
Given the multiple options for treatment of chronic-phase chronic myeloid leukemia (CML) with tyrosine kinase inhibitors, our objective was to understand treatment patterns in routine practice and prognostic indicators of response. We conducted a retrospective medical record review of 681 patients with CML in Australia, Canada, and South Korea. Eligible patients had a diagnosis of chronic-phase CML, were Philadelphia chromosome and/or BCR-ABL positive, were aged 18 years or older, and had been treated with first-line imatinib therapy between January 2005 and September 2010. Data on patient demographics, medical history (e.g., comorbidities, Sokal score), and treatment characteristics (e.g., time to initiation, therapy duration) were abstracted. Descriptive analyses were stratified by country and therapy line. Prognostic indicators of response to imatinib were evaluated using multivariable logistic regression, adjusting for country, patient demographics, medical history, treatment characteristics, and side effects. Hematologic, cytogenetic, and molecular responses at 3, 6, 12, and 18 months following initiation of each therapy line. Patients' average age was 57 years, and 59% were male. Overall, imatinib was initiated approximately 4 months following CML diagnosis. Complete or major molecular response (C/MMR) at 6 months following imatinib initiation was 54% in Australia, 22% in Canada, and 38% in South Korea. At 18 months, over 60% of patients achieved C/MMR. Approximately 30% of patients discontinued imatinib primarily due to intolerance and lack of response. Among patients who received second-line treatment, dasatinib was used more frequently than nilotinib. Multivariable regression results indicated Sokal score was identified as a prognostic indicator of response to imatinib therapy at several time points. There are several limitations to this study. First, we selected a convenience sample of patients and physicians and therefore results may not be representative of the true population of patients with chronic-phase CML. Second, data were entered by the selected physician and could be subject to data entry errors or inaccuracies. Third, limited information was collected from the patient records, and it is possible that we did not capture additional prognostic or confounding factors related to the measured outcomes. Next, because this was an analysis of previously documented data (i.e., retrospective), we were unable to provide a priori definitions of response. Finally, multivariable analyses were limited to imatinib-related outcomes. Treatment patterns and prognostic indicators differed by country. Health care providers, payers, and patients can utilize these results to inform treatment and policies aimed at improving the effectiveness of care for patients with chronic-phase CML.


Macroporous nanofiber wraps promote axonal regeneration and functional recovery in nerve repair by limiting fibrosis.
Functional outcomes following nerve repair remain suboptimal. Scarring at the repair site is a major impediment to regeneration. A biomaterial scaffold applied around the coaptation site that decreases inflammation holds great potential in reducing scarring, enhancing axonal growth, and improving functional recovery. In this study, we evaluated the effect of a macroporous nanofiber wrap, comprised of nonwoven electrospun poly-ε-caprolactone (PCL), in improving axonal regeneration in a rat sciatic nerve cut and direct repair model. Controls consisted of conventional epineurial repair. We also evaluated our wrap against the commercially available AxoGuard wrap. At five weeks following repair, the nanofiber wrap group showed a significantly decreased intraneural macrophage invasion and collagen deposition at the repair site. This was associated with increased expression of the anti-inflammatory cytokine (IL-10), decreased expression of the pro-inflammatory cytokine (TNF-α), and a decrease in the M1:M2 macrophage phenotype ratio. These findings suggest that this nanofiber wrap, with its unique macroporosity, is modulating the inflammatory response at the repair site by polarizing macrophages towards a pro-regenerative M2 phenotype. Concomitantly, a higher number of regenerated axons was noted. At sixteen weeks, the nanofiber wrap resulted in enhanced functional recovery as demonstrated by electrophysiology, neuromuscular re-innervation, and muscle histology. When compared to the AxoGuard wrap, the nanofiber wrap showed similar inflammation at the repair site and similar nerve morphometric findings, but there was a trend towards a lower overall number of macrophages invading the wrap wall. These results demonstrate favorable outcomes of the macroporous nanofiber wrap in promoting neuroregeneration and functional recovery following nerve repair. STATEMENT OF SIGNIFICANCE: Electrospun nanofiber scaffolds, with specific fiber and pore sizes, were shown to modulate the immune response and create a regenerative environment. In this paper, we present a macroporous nanofiber wrap, made of poly-ε-caprolactone, to be applied at the coaptation site in primary nerve repair. We show that it regulates the inflammatory response at the repair site and decreases scarring/fibrosis. This results in enhanced axonal regeneration, allowing a higher number of axons to cross the suture line and reach the target muscle in a timely fashion. Functional outcomes are thus improved.


The integrin ligand c(RGDf(NMe)Nal) reduces neointimal hyperplasia in a polymer-free drug-eluting stent system.
The use of highly active and selective integrin ligands in combination with stent implantation is emerging as a promising alternative to the release of classical immunosuppressive drugs by current drug-eluting stents (DES), which has been associated with delayed vascular healing and late stent thrombosis. Herein we present the development and biological evaluation of the integrin ligand c(RGDf(NMe)Nal) as a potent anti-proliferative molecule that targets coronary artery smooth muscle cells (CASMCs). This peptide showed an antagonistic activity for αvβ3 and αvβ5 in the low-nanomolar range, and selectivity against the platelet receptor αIIbβ3. In vitro, it efficiently inhibited the proliferation of CASMCs, displaying higher potency than the anti-tumor drug candidate cilengitide. This peptide was then loaded into a polymer-free bare metal stent (BMS), and its release studied at different time points. Up to seven days of elution, the peptide-coated stents retained high anti-proliferative activity toward CASMCs. Finally, the peptide was examined in vivo in a polymer-free DES system in a rabbit iliac artery model. After 28 days of implantation, histopathological analysis revealed that the peptide clearly decreased neointimal growth and improved vessel healing and re-endothelialization compared with the FDA-approved Cypher DES. Our study shows that this type of lipophilic integrin ligand, when eluted from a polymer-free stent system, has the potential to successfully decrease in-stent restenosis in the absence of delayed vascular healing.


Persistent pediatric obstructive sleep apnea treated with skeletally anchored transpalatal distraction
The aim of this study was to evaluate the impact of nasomaxillary expansion using skeletally anchored transpalatal distraction (TPD) in children without transverse maxillary deficiency that were previously treated by rapid palatal expansion (RPE). Twenty-nine consecutive children were treated by TPD. Twenty-five children, aged 10-16 years completed pre- and post-operative clinical evaluations, questionnaires (OSA-18), cone beam computed tomography (CBCT), and polysomnography (PSG). The pre- and post-operative CBCT data were used to reconstruct the 3-dimensional shape of the upper airway. Two measures of airflow function (pressure and velocity) were simulated by using computational fluid dynamics (CFD) at four different airway segments (nasal, nasopharyngeal, oropharyngeal and hypopharyngeal). Twenty-three patients (92%) experienced improvement based on PSG. The apnea hypopnea index (AHI) improved from 6.72±4.34 to 3.59±5.11 (p<0.001) events per hour. Clinical symptoms based on OSA-18 scores were improved in all patients. Twenty-five patients (100%) had successful expansion defined as separation of the midpalatal suture at least 1 mm from anterior nasal spine (ANS) to posterior nasal spine (PNS). The nasal sidewall widening was 2.59±1.54 mm at canine, 2.91±1.23 mm at first molar and 2.30±1.29 mm at PNS. The ratio of dental expansion to nasal expansion was 1.12:1 (2.90 mm:2.59 mm) at canine and 1.37:1 (3.98 mm:2.91 mm) at first molar. The nasal airflow pressure reduced by 76% (-275.73 to -67.28 Pa) and the nasal airflow velocity reduced by over 50% (18.60 to 8.56 m/s). Nasomaxillary expansion by skeletally anchored TPD improves OSA in children without transverse maxillary deficiency that were previously treated by RPE. A nearly parallel anterior-posterior opening of the mid-palatal suture achieves enlargement of the entire nasal passage with improvement of the airflow characteristics in the nasal and pharyngeal airway. The improved airflow characteristic is significantly correlated with the improved polysomnographic findings, thus demonstrating that nasomaxillary expansion in previously expanded patients is a viable treatment option.


Player-Tracking Technology: Half-Full or Half-Empty Glass?
With the ongoing development of microtechnology, player tracking has become one of the most important components of load monitoring in team sports. The 3 main objectives of player tracking are better understanding of practice (provide an objective, a posteriori evaluation of external load and locomotor demands of any given session or match), optimization of training-load patterns at the team level, and decision making on individual players' training programs to improve performance and prevent injuries (eg, top-up training vs unloading sequences, return to play progression). This paper discusses the basics of a simple tracking approach and the need to integrate multiple systems. The limitations of some of the most used variables in the field (including metabolic-power measures) are debated, and innovative and potentially new powerful variables are presented. The foundations of a successful player-monitoring system are probably laid on the pitch first, in the way practitioners collect their own tracking data, given the limitations of each variable, and how they report and use all this information, rather than in the technology and the variables per se. Overall, the decision to use any tracking technology or new variable should always be considered with a cost/benefit approach (ie, cost, ease of use, portability, manpower/ability to affect the training program).


Parasympathetic Tone Activity Evaluation to Discriminate Ketorolac and Ketorolac/Tramadol Analgesia Level in Swine.
Evaluation of nociceptive-antinociceptive balance during general anesthesia is still challenging and routinely based on clinical criteria. Analgesic drug delivered may be optimized with parasympathetic tone activity (PTA) monitor. This study compares ketorolac and ketorolac/tramadol balance analgesia using a PTA monitor. Pain intensity response was assessed using a 0-100 numerical state scale (PTA) after nociceptive stimuli in pigs under stable sevoflurane anesthesia. Bispectral index, heart rate, noninvasive blood pressure, and respiratory parameters were also measured. Animals were divided into 3 groups: without analgesia, ketorolac, and ketorolac/tramadol. Mean values or mean areas under the curve (AUC) in selected time periods were compared over time and between groups through a mixed-model repeated measures analysis of variance and nonparametric Kruskal-Wallis tests, followed by Bonferroni or Dunn's multiple comparisons. It was observed a significant decrease in the PTA AUC mean value after application of the stimulus in animals treated without analgesia and only with ketorolac. The PTA AUC mean value in the control group was significantly lower than the corresponding mean in ketorolac group. The ketorolac/tramadol group showed the highest PTA AUC mean values, significantly different from those obtained for the other 2 groups, with no significant differences detected over time. Bispectral index means showed no statistically significant differences either over time periods or between different treatment groups. Heart rate showed only a statistically significant increase in AUC mean between without analgesia and ketorolac/tramadol group, in the time period after the stimulus application. Noninvasive blood pressure means showed no statistically significant differences over time and between treatment groups. This study shows that a low dose combination of ketorolac and tramadol is sufficient to block the pain responses induced with a needle holder in pigs 20 minutes after its administration. The PTA monitor was able to clearly recognize the analgesic level between treatments and may be used to optimize analgesic drug delivered.


Statistical-mode sensor for fiber optic vibration sensing uses.
A method of sensing vibration using the detection of changes in the spatial distribution of energy in the output of a multimode optical fiber has been demonstrated. Two implementations of the sensor have been built and tested. The first implementation involved simple optical processing of the output fiber speckle pattern using spatial filtering. The second implementation involved projecting the pattern on a CCD array and digitally processing observed changes in the intensity distribution. A mathematical model has been developed which has shown good agreement with observed sensor behavior. The sensor technique has been used to detect induced structural vibration in laboratory test specimens. Simple field testing has also demonstrated the ability of the technique to detect personnel and vehicles passing over a buried and electrically undetectable sensing cable. The sensing technique is compatible with off-the-shelf components and fiber cable and even allows for simultaneous telecommunication and sensing using the same optical fiber cable. Near term application of this technology could provide significant benefits for vibration sensing, intrusion detection, and acoustic sensing.


Effects of insulin-like growth factor receptor inhibition on human melanomas in culture and in athymic mice.
The role of the insulin-like growth factor (IGF) receptor in regulating the growth of melanoma cells was evaluated by examining the effect of antibody-mediated IGF receptor inhibition on the growth of four human melanoma cell lines in culture and as xenotransplants in athymic mice. All four cell lines expressed typical type I IGF receptors and an antibody to this receptor (alpha IR-3) inhibited [125I]IGF-I binding. However, the cell lines varied widely in their in vitro responsiveness to IGF-I and alpha IR-3: in the WM 373 and WM 852 cell lines, IGF-I stimulated cell replication and alpha IR-3 inhibited this response, whereas in the WM 239-A and WM 266-4 cell lines neither the growth factor nor the antibody affected growth. A wide variation was also observed in the effect of the antibody on the growth of the different cell lines as xenotransplants but this qualitatively correlated with the responses observed in vitro: alpha IR-3 treatment significantly inhibited the growth of the WM 373 and WM 852 xenotransplants but did not inhibit the growth of the WM 239-A or WM 266-4 xenotransplants and may even have had a slight stimulatory effect. These results indicate that the IGF receptor pathway is a functional regulator of the in vivo growth of some melanomas and that this is reflected in the activity of this pathway as determined in vitro. These findings suggest that therapies aimed at inhibiting the IGF pathway may be beneficial in treating some melanomas.


Primary blastomycosis of oral cavity.
Blastomycosis is an uncommon male-predominant disease caused by the fungus Blastomyces dermatitidis. The lungs are most commonly affected, and other organs are usually involved by dissemination. Clinical feature and pathohistologic findings are similar to the appearance of squamous cell carcinoma. A 52-year-old male patient who has lived as a farmer on the countryside in Argentina for 35 years presented with an initial histopathologic diagnosis of a squamous cell carcinoma of the right lower jaw. There was no history of pulmonary disease, in particular fever, coughing, or hemoptysis. Final pathohistologic evaluation after resection revealed B. dermatitidis infection. This article presents the first described case of oral manifestation of B. dermatitidis infection in Switzerland. Manifestation of blastomycosis in oral tissue can mimic the feature of a squamous cell carcinoma and can therefore be a diagnostic pitfall that head and neck surgeons and a pathologist should be aware of.


Optimization of virulence functions through glucosylation of Shigella LPS.
Shigella, the leading cause of bacillary dysentery, uses a type III secretion system (TTSS) to inject proteins into human cells, leading to bacterial invasion and a vigorous inflammatory response. The bacterium is protected against the response by the O antigen of lipopolysaccharide (LPS) on its surface. We show that bacteriophage-encoded glucosylation of Shigella O antigen, the basis of different serotypes, shortens the LPS molecule by around half. This enhances TTSS function without compromising the protective properties of the LPS. Thus, LPS glucosylation promotes bacterial invasion and evasion of innate immunity, which may have contributed to the emergence of serotype diversity in Shigella.


Bacterial community analysis of activated sludge: an evaluation of four commonly used DNA extraction methods.
The effectiveness of three commercially available direct DNA isolation kits (Mobio, Fast, Qiagen) and one published direct DNA extraction protocol (Bead) for extracting bacterial DNA from different types of activated sludge was investigated and mutually compared. The DNA quantity and purity were determined using real-time PCR targeting the bacterial 16S rDNA gene. Microbial community fingerprints were assessed by automated ribosomal intergenic spacer analysis. The resulting community profiles were analyzed with canonical correspondence analysis. Our results clearly demonstrate that direct DNA extraction methods can significantly influence the DNA quantity, purity, and observed community patterns of microbiota in activated sludge. Fast and Mobio generated high amounts of good quality DNA compared to Bead and Qiagen. Mobio also resulted in the detection of the highest number of species while Fast scored the best in discriminating between the community patterns of different activated sludge types. With respect to the characterization of community profiles, our analyses demonstrated a strong sludge type dependent variability among methods. Taking into account our results, we recommend Fast as the most suitable DNA extraction method for activated sludge samples used for bacterial community studies.


Cytochrome c(y) of Rhodobacter capsulatus is attached to the cytoplasmic membrane by an uncleaved signal sequence-like anchor.
During the photosynthetic growth of Rhodobacter capsulatus, electrons are conveyed from the cytochrome (cyt) bc1 complex to the photochemical reaction center by either the periplasmic cyt c2 or the membrane-bound cyt c(y). Cyt c(y) is a member of a recently established subclass of bipartite c-type cytochromes consisting of an amino (N)-terminal domain functioning as a membrane anchor and a carboxyl (C)-terminal domain homologous to cyt c of various sources. Structural homologs of cyt c(y) have now been found in several bacterial species, including Rhodobacter sphaeroides. In this work, a C-terminally epitope-tagged and functional derivative of R. capsulatus cyt c(y) was purified from intracytoplasmic membranes to homogeneity. Analyses of isolated cyt c(y) indicated that its spectral and thermodynamic properties are very similar to those of other c-type cytochromes, in particular to those from bacterial and plant mitochondrial sources. Amino acid sequence determination for purified cyt c(y) revealed that its signal sequence-like N-terminal portion is uncleaved; hence, it is anchored to the membrane. To demonstrate that the N-terminal domain of cyt c(y) is indeed its membrane anchor, this sequence was fused to the N terminus of cyt c2. The resulting hybrid cyt c (MA-c2) remained membrane bound and was able to support photosynthetic growth of R. capsulatus in the absence of the cyt c(y) and c2. Therefore, cyt c2 can support cyclic electron transfer during photosynthetic growth in either a freely diffusible or a membrane-anchored form. These findings should now allow for the first time the comparison of electron transfer properties of a given electron carrier when it is anchored to the membrane or is freely diffusible in the periplasm.


To-Go medications for decreasing ED return visits.
The primary objective was to determine if providing patients with a complete course of antibiotics for select conditions would decrease the rate of return to the emergency department (ED) within 7 days of the initial visit. In an urban, academic medical center, we compared patients who received medications at discharge (To-Go medications) with patients who received standard care (a prescription at discharge). Emergency department patients were included if they were older than 18 years; had a discharge diagnosis International Classification of Diseases, Ninth Revision, code for urinary tract infection, pyelonephritis, cellulitis, or dental infection; and presented initially between January and December 2010. Candidates had limited health insurance or were discharged when nearby pharmacies were closed. Return visits were included if the condition was related to the initial diagnosis. Wound checks and scheduled revisits were excluded. Medications dispensed were penicillin, clindamycin, sulfamethoxazole-trimethoprim, and nitrofurantoin. A total of 4257 individuals were seen in initial ED visits for the included conditions. Comparing the 243 individuals given medications with the 4014 who were not given medications, the To-Go medications group was less likely to return than the comparison group (2.5% vs 5.9%; P = .026). The cellulitis subgroup also showed a significant reduction in return visits (1.6% vs 6.9%; P = .024). Three hundred eighteen courses of medication were given to the 243 individuals for a total cost of $1123. For a 1-year expense of $1123, we demonstrated a 50% reduction in ED return visits for patients who were given a free, complete course of antibiotics at discharge for select conditions.


Site-directed chemical conversion of serine to cysteine in penicillin acylase from Escherichia coli ATCC 11105. Effect on conformation and catalytic activity.
Penicillin acylase (EC 18.104.22.168) was completely inactivated with equimolar phenylmethane [35S]sulphonyl fluoride (PhMe35SO2F); the stability of the sulphonyl group in the modified protein was determined by measurement of the radioactivity in ultrafiltrates. In 8 M urea, the rate of loss of the sulphonyl group was similar to that observed in PhMeSO2F-inactivated chymotrypsin [Gold, A.M. & Fahrney, D. (1964) Biochemistry 3, 783-791]. Incubation of the PhMeSO2F-inactivated acylase with 0.7 M potassium thioacetate yielded an acetylthiol enzyme which was subsequently converted to a thiol-enzyme during incubation with 10 mM 6-aminopenicillanic acid. 4-Pyridyl-ethylcysteine was released by acid hydrolysis after reaction of the thiol-protein with 4-vinylpyridine. The rates of reaction of thiol-penicillin acylase with iodoacetic acid and 2,2'-dipyridyl disulphide were consistent with the presence of an incompletely accessible cysteinyl sidechain. After carboxymethylating the thiol-enzyme with iodo[2-3H]acetic acid, the label was shown by SDS/PAGE and sequencing analysis to be associated exclusively with the beta-chain NH2-terminal residue, indicating conversion of Ser290 to S-carboxymethyl-cysteine. Near-ultraviolet CD spectra showed the conformation of thiol-penicillin acylase to be indistinguishable from that of the native protein but the catalytic activity was less than 0.02% of that of the normal enzyme. The possibility that Ser290 acts as a nucleophile in catalysis is discussed.


First Report of Phytophthora nicotianae on Skimmia japonica in Italy.
Skimmia japonica, an evergreen flowering shrub, is becoming increasingly popular as a potted ornamental plant in northern Italy and represents 5% of acidophilous plant production; cv. Rubella accounts for 99% of production. During the spring of 2003, in many commercial nurseries located in northwestern Italy, plants of S. japonica cv. Rubella showed extensive chlorosis and root rot, and diseased plants eventually wilted and died without dropping leaves. The disease was widespread and severe, and in some nurseries, 40% of plants were affected. A Phytophthora-like organism was isolated consistently from infected lower stem and root pieces of S. japonica that had been disinfested for 1 min in 1% NaOCl and plated on a medium selective for oomycetes (2). The pathogen was identified based on morphological and physiological features as Phytophthora nicotianae (= P. parasitica [1]). The sporangia produced on V8 medium were ± spherical to obpyriform, obturbinate, papillate, and measured 33 to 94 × 25 to 48 μm (average 56.4 × 36.8 μm). Papillae measured 3.5 to 19 μm (average 7.8 μm). Chlamydospores were spherical with a diameter ranging from 26 to 32 μm (average 29.2 μm). Pathogenicity of four isolates obtained from infected plants was confirmed by inoculating 9-month-old plants of S. japonica cv. Rubella grown in 1-liter pots containing a substrate based on sphagnum peatmoss, pine bark, and clay (70-20-10% vol/vol/vol). Inocula, which consisted of 90-mm-diameter V8 agar disks per pot containing mycelium of each isolate, were introduced and mixed into the substrate in all pots before transplanting. One plant was transplanted into each pot and served as a replicate, and noninoculated plants served as controls. Eight replicates were used for each isolate and the control treatment, and the trial was repeated. All plants were kept outside at temperatures ranging from 16 to 38°C (average temperature 27°C). Inoculated plants developed symptoms of chlorosis, root rot, and wilt within 20 days, while control plants remained symptomless. P. nicotianae consistently was isolated from inoculated plants. Previously, P. nicotianae has been reported on S. japonica in Poland (3). To our knowledge, this is the first report of P. nicotianae on S. japonica in Italy. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St Paul, MN, 1996. (2) H. Masago et al. Phytopathology, 67:425, 1977 (3) G. Szkuta and L. B. Orlikowski. Prog. Plant Prot. 42:808, 2002.


Two-week administration of engineered Escherichia coli establishes persistent resistance to diet-induced obesity even without antibiotic pre-treatment.
Adverse alterations in the composition of the gut microbiota have been implicated in the development of obesity and a variety of chronic diseases. Re-engineering the gut microbiota to produce beneficial metabolites is a potential strategy for treating these chronic diseases. N-acyl-phosphatidylethanolamines (NAPEs) are a family of bioactive lipids with known anti-obesity properties. Previous studies showed that administration of Escherichia coli Nissle 1917 (EcN) engineered with Arabidopsis thaliana NAPE synthase to produce NAPEs imparted resistance to obesity induced by a high-fat diet that persisted after ending their administration. In prior studies, mice were pre-treated with ampicillin prior to administering engineered EcN for 8 weeks in drinking water. If use of antibiotics and long-term administration are required for beneficial effects, implementation of this strategy in humans might be problematic. Studies were therefore undertaken to determine if less onerous protocols could still impart persistent resistance and sustained NAPE biosynthesis. Administration of engineered EcN for only 2 weeks without pre-treatment with antibiotics sufficed to establish persistent resistance. Sustained NAPE biosynthesis by EcN was required as antibiotic treatment after administration of the engineered EcN markedly attenuated its effects. Finally, heterologous expression of human phospholipase A/acyltransferase-2 (PLAAT2) in EcN provided similar resistance to obesity as heterologous expression of A. thaliana NAPE synthase, confirming that NAPEs are the bioactive mediator of this resistance.


Endogenous testosterone and mortality due to all causes, cardiovascular disease, and cancer in men: European prospective investigation into cancer in Norfolk (EPIC-Norfolk) Prospective Population Study.
The relation between endogenous testosterone concentrations and health in men is controversial. We examined the prospective relationship between endogenous testosterone concentrations and mortality due to all causes, cardiovascular disease, and cancer in a nested case-control study based on 11 606 men aged 40 to 79 years surveyed in 1993 to 1997 and followed up to 2003. Among those without prevalent cancer or cardiovascular disease, 825 men who subsequently died were compared with a control group of 1489 men still alive, matched for age and date of baseline visit. Endogenous testosterone concentrations at baseline were inversely related to mortality due to all causes (825 deaths), cardiovascular disease (369 deaths), and cancer (304 deaths). Odds ratios (95% confidence intervals) for mortality for increasing quartiles of endogenous testosterone compared with the lowest quartile were 0.75 (0.55 to 1.00), 0.62 (0.45 to 0.84), and 0.59 (0.42 to 0.85), respectively (P<0.001 for trend after adjustment for age, date of visit, body mass index, systolic blood pressure, blood cholesterol, cigarette smoking, diabetes mellitus, alcohol intake, physical activity, social class, education, dehydroepiandrosterone sulfate, androstanediol glucuronide, and sex hormone binding globulin). An increase of 6 nmol/L serum testosterone ( approximately 1 SD) was associated with a 0.81 (95% confidence interval 0.71 to 0.92, P<0.01) multivariable-adjusted odds ratio for mortality. Inverse relationships were also observed for deaths due to cardiovascular causes and cancer and after the exclusion of deaths that occurred in the first 2 years. In men, endogenous testosterone concentrations are inversely related to mortality due to cardiovascular disease and all causes. Low testosterone may be a predictive marker for those at high risk of cardiovascular disease.


Potential of different species for use in removal of DDT from the contaminated soils.
Dichlorodiphenyltrichloroethane (DDT) and its main metabolites, p,p'-DDD and p,p'-DDE (DDTs in this study included DDT, DDD and DDE), are frequently detected in agricultural soils even though its usage in agriculture was banned in 1980s or earlier. In this study, eleven plants including eight maize (Zea mays) cultivars and three forage species (alfalfa, ryegrass and teosinte) widely cultivated in China were grown in the soils spiked with DDTs to investigate their potential for removal of DDT from the contaminated soils. The plants varied largely in their ability to accumulate and translocate DDTs, with the bioconcentration factor (BCF; DDT concentration ratio of the plant tissues to the soils) ranging from 0.014 to 0.25 and the translocation factor (TF; DDT concentration ratio of the shoots to the roots) varying from 0.35 (Zea mays cv Chaotian-23) to 0.76 (Zea mays spp. mexicana). The amount of DDT phytoextraction ranged from 3.89mug (ryegrass) to 27.0mug (teosinte) and accounted for <0.1% of the total initial DDTs spiked in the soils. After 70d, the removal rates reached 47.1-70.3% of the total initial DDTs spiked in the soils with plants while that was only 15.4% in the soils without plant. Moreover, the higher removal rates of DDTs occurred at the first 20d of experiment, and then the removal rate decreased with time. The highest amount of DDTs phytoextracted was observed in teosinte, followed by Zea mays spp. mexicana, but the highest removal rate of DDTs was found in maize (Zea mays cv Jinhai-6). Even though phytoextraction is not the main removal process for DDTs, the plant species especially Zea mays cv Jinhai-6 showed high potential for removing DDTs from the contaminated soils.


Brain mechanisms of serial and parallel processing during dual-task performance.
The psychological refractory period (PRP) refers to the fact that humans typically cannot perform two tasks at once. Behavioral experiments have led to the proposal that, in fact, peripheral perceptual and motor stages continue to operate in parallel, and that only a central decision stage imposes a serial bottleneck. We tested this model using neuroimaging methods combined with innovative time-sensitive analysis tools. Subjects performed a dual-task visual-auditory paradigm in which a delay of 300 ms was injected into the auditory task either within or outside of the dual-task interference period. Event-related potentials indicated that the first approximately 250 ms of processing were insensitive to dual-task interference, and that the PRP was mainly reflected in a delayed global component. By a clustering analysis based on time-resolved functional magnetic resonance imaging, we identified networks with qualitatively different timing properties: sensory areas tracked the objective time of stimulus presentation, a bilateral parietoprefrontal network correlated with the PRP delay, and an extended bilateral network that included bilateral posterior parietal cortex, premotor cortex, supplementary motor area, anterior part of the insula, and cerebellum was shared by both tasks during the extent of dual-task performance. The results provide physiological evidence for the coexistence of serial and parallel processes within a cognitive task.