Organizational commitment of military physicians.
An individual's loyalty or bond to his or her employing organization, referred to as organizational commitment, influences various organizational outcomes such as employee motivation, job satisfaction, performance, accomplishment of organizational goals, employee turnover, and absenteeism. Therefore, as in other sectors, employee commitment is crucial also in the healthcare market. This study investigates the effects of organizational factors and personal characteristics on organizational commitment of military physicians using structural equation modeling (SEM) on a self-report, cross-sectional survey that consisted of 635 physicians working in the 2 biggest military hospitals in Turkey. The results of this study indicate that professional commitment and organizational incentives contribute positively to organizational commitment, whereas conflict with organizational goals makes a significantly negative contribution to it. These results might help develop strategies to increase employee commitment, especially in healthcare organizations, because job-related factors have been found to possess greater impact on organizational commitment than personal characteristics.


Polymerase chain reaction detection of human papillomavirus: quantitation may improve clinical utility.
A case-control study compared detection by polymerase chain reaction (PCR) specific for human papillomavirus (HPV) type 16 with restriction enzyme analysis and Southern blot hybridization detection of HPV type 16. Cervicovaginal lavage samples from 64 women with histopathologic evidence of a cervical squamous intraepithelial lesion and 55 samples from cytologically healthy women were studied. Several methods of PCR product analysis, including radioactive and nonradioactive probing, were compared. The sensitivity of HPV detection by PCR when the amplified DNA fragment was visualized on a gel was equivalent to those of detection by restriction enzyme and Southern blot analyses. Hybridization of the PCR product with radioactively or nonradioactively labeled oligonucleotide probes increased the sensitivity of HPV detection by 100-fold. However, an increase in the sensitivity of the assay preferentially identified low levels of the virus in cytologically healthy women. Therefore, the value of HPV detection in identifying women with cervical neoplastic disease was greater, and the odds ratio for the presence of a cervical squamous intraepithelial lesion was higher when the less sensitive modalities were used. These results suggest that quantitation of HPV by PCR may maximize the clinical significance of a positive test result. Further studies will be needed to determine the optimal level of virus detection which has the highest positive predictive value of clinical disease.


Fusarium spp., Cylindrocarpon spp., and Environmental Stress in the Etiology of a Canker Disease of Cold-Stored Fruit and Nut Tree Seedlings in California.
The principal objective of this study was to determine the etiology of a canker disease in dormant stone fruit and apple tree seedlings maintained in refrigerated storage that has significantly impacted California fruit and nut tree nurseries. Signs and symptoms of the disease develop during storage or soon after planting, with subsequent decline and death of young trees. Isolations from both diseased and healthy almond and apple trees and Koch's postulates using stem segments of desiccation-stressed almond trees as hosts implicated Fusarium avenaceum and F. acuminatum as the primary causal agents. F. solani, Ilyonectria robusta, and Cylindrocarpon obtusiusculum were also capable of causing similar symptoms but were less frequently encountered in isolations of diseased tissue. Loss of bark turgidity in excised almond stem segments, as can occur in cold-stored seedlings, correlated with increased susceptibility to F. acuminatum, with maximum canker development occurring after relative bark turgidity dropped below a threshold of approximately 86%. Healthy almond trees, almond scion budwood, and a wheat cover crop used in fields where tree seedlings were grown and maintained until cold storage all possessed asymptomatic infections of F. acuminatum, F. avenaceum, and C. obtusiusculum as determined by activation following overnight freezing, cold storage, or desiccation.


Type I and type III interferons drive redundant amplification loops to induce a transcriptional signature in influenza-infected airway epithelia.
Interferons (IFNs) are a group of cytokines with a well-established antiviral function. They can be induced by viral infection, are secreted and bind to specific receptors on the same or neighbouring cells to activate the expression of hundreds of IFN stimulated genes (ISGs) with antiviral function. Type I IFN has been known for more than half a century. However, more recently, type III IFN (IFNλ, IL-28/29) was shown to play a similar role and to be particularly important at epithelial surfaces. Here we show that airway epithelia, the primary target of influenza A virus, produce both IFN I and III upon infection, and that induction of both depends on the RIG-I/MAVS pathway. While IRF3 is generally regarded as the transcription factor required for initiation of IFN transcription and the so-called "priming loop", we find that IRF3 deficiency has little impact on IFN expression. In contrast, lack of IRF7 reduced IFN production significantly, and only IRF3(-/-)IRF7(-/-) double deficiency completely abolished it. The transcriptional response to influenza infection was largely dependent on IFNs, as it was reduced to a few upregulated genes in epithelia lacking receptors for both type I and III IFN (IFNAR1(-/-)IL-28Rα(-/-)). Wild-type epithelia and epithelia deficient in either the type I IFN receptor or the type III IFN receptor exhibit similar transcriptional profiles in response to virus, indicating that none of the induced genes depends selectively on only one IFN system. In chimeric mice, the lack of both IFN I and III signalling in the stromal compartment alone significantly increased the susceptibility to influenza infection. In conclusion, virus infection of airway epithelia induces, via a RIG-I/MAVS/IRF7 dependent pathway, both type I and III IFNs which drive two completely overlapping and redundant amplification loops to upregulate ISGs and protect from influenza infection.


A survey of patient satisfaction in a metropolitan Emergency Department: comparing nurse practitioners and emergency physicians.
The Emergency Nurse Practitioner role was introduced to an Emergency Department, Melbourne in 2004 as an alternative health-care model to provide accessible and efficient patient care. The aim of the study was to explore patient satisfaction using a questionnaire from their emergency department experience comparing Emergency Nurse Practitioners and emergency department doctors. Patients who received care from either Emergency Nurse Practitioners or emergency department doctors were given a self-administered questionnaire to complete. Descriptive statistics and non-parametric tests were used for data analysis. A total of 202 patients completed the survey with 103 seen by the Emergency Nurse Practitioners and 99 seen by emergency department doctors. Significant differences were reported in 12 of the 16 questions comparing patient satisfaction with either Emergency Nurse Practitioners or emergency department doctors with greater patient satisfaction demonstrated with the Emergency Nurse Practitioners. The Emergency Nurse Practitioner model demonstrates consistent levels of patient satisfaction with patients reporting more favourable satisfaction with the Emergency Nurse Practitioners compared with emergency department doctors.


Middle Cerebral Artery Calcification: Association With Ischemic Stroke.
Calcification of the middle cerebral artery (MCA) is uncommon in the healthy elderly. Whether calcification of the MCA is associated with cerebral ischemic stroke remains undetermined. We intended to investigate the association using Agatston calcium scoring of the MCA. This study retrospectively included 354 subjects with ischemic stroke in the MCA territory and 1518 control subjects who underwent computed tomography (CT) of the brain. We recorded major known risk factors for ischemic stroke, including age, gender, hypertension, diabetes mellitus, smoking, hyperlipidemia, and obesity, along with the MCA calcium burden, measured with the Agatston calcium scoring method. Univariate and modified logistic regression analyses were performed to examine the association between the MCA calcification and ischemic stroke. The univariate analyses showed significant associations of ischemic stroke with age, hypertension, diabetes mellitus, smoking, total MCA Agatston score, and the presence of calcification on both or either side of the MCA. Subjects with the presence of MCA calcification on both or either side of the MCA were 8.46 times (95% confidence interval, 4.93-14.53; P < 0.001) more likely to have a cerebral infarct than subjects without MCA calcification after adjustment for the major known risk factors, including age, hypertension, diabetes mellitus, and smoking. However, a higher degree of MCA calcification reflected by the Agatston score was not associated with higher risk of MCA ischemic stroke after adjustment for the confounding factors and presence of MCA calcification. These results suggest that MCA calcification is associated with ischemic stroke in the MCA territory. Further prospective studies are required to verify the clinical implications of the MCA calcification.


Lessons learned from HIV-1 vaccine trials: new priorities and directions.
A vaccine against human immunodeficiency virus (HIV) seems to be on the horizon. Correlates of risk of infection for [corrected] the RV144 vaccine trial have been found. There is understanding of what makes HIV envelope-specific antibodies broadly neutralizing and new T cell vaccine approaches can overcome virus variability.


Design rules for light-emitting electrochemical cells delivering bright luminance at 27.5 percent external quantum efficiency.
The light-emitting electrochemical cell promises cost-efficient, large-area emissive applications, as its characteristic in-situ doping enables use of air-stabile electrodes and a solution-processed single-layer active material. However, mutual exclusion of high efficiency and high brightness has proven a seemingly fundamental problem. Here we present a generic approach that overcomes this critical issue, and report on devices equipped with air-stabile electrodes and outcoupling structure that deliver a record-high efficiency of 99.2 cd A-1 at a bright luminance of 1910 cd m-2. This device significantly outperforms the corresponding optimized organic light-emitting diode despite the latter employing calcium as the cathode. The key to this achievement is the design of the host-guest active material, in which tailored traps suppress exciton diffusion and quenching in the central recombination zone, allowing efficient triplet emission. Simultaneously, the traps do not significantly hamper electron and hole transport, as essentially all traps in the transport regions are filled by doping.


Transient stimulated Raman scattering in gas mixtures.
We studied, for the first time to our knowledge, transient stimulated Raman scattering in H(2)- D(2) and H(2)- CH(4) gas mixtures excited with 200-fs and 2-ps, 390-nm pulses. Depending on the composition and partial pressure, of those gases, we observed approximately 40 output lines simultaneously. Under optimum conditions a conversion efficiency of as much as 8% in the combination Raman line at 556 nm was obtained in the H(2)- CH(4) gas mixtures. The difference between picosecond and femtosecond pumping is due to self-phase modulation and to generation of a spectral continuum in the latter. Our calculation of the ratio of the partial pressures is in a good agreement with the experimental results. A new method for measuring the unknown Raman scattering cross section of gas molecules is suggested.


TGF alpha can act as a chemoattractant to perioptic mesenchymal cells in developing mouse eyes.
Growth factors are believed to play an important role in regulating cell fate and cell behavior during embryonic development. Transforming growth factor alpha (TGF alpha), a member of the epidermal growth factor (EGF) superfamily, is a small polypeptide growth factor. Upon binding to its receptor, the EGF receptor (EGFR), TGF alpha can exert diverse biological activities, such as induction of cell proliferation or differentiation. To explore the possibility that TGF alpha might regulate cell fate during murine eye development, we generated transgenic mice that express human TGF alpha in the lens under the control of the mouse alpha A-crystallin promoter. The transgenic mice displayed multiple eye defects, including corneal opacities, cataracts and microphthalmia. At early embryonic stages TGF alpha induced the perioptic mesenchymal cells to migrate abnormally into the eye and accumulate around the lens. In situ hybridization revealed that the EGFR mRNA is highly expressed in the perioptic mesenchyme, suggesting that the migratory response is mediated by receptor activation. In order to test this model, the TGF alpha transgenic mice were bred to EGFR mutant waved-2 (wa-2) mice. We found that the eye defects of the TGF alpha transgenic mice are significantly abated in the wa-2 homozygote background. Because the EGFR mutation in the wa-2 mice is located in the receptor kinase domain, this result indicates that the receptor tyrosine kinase activity is critical for signaling the migratory response. Taken together, our studies demonstrate that TGF alpha is capable of altering the migratory decisions and behavior of perioptic mesenchyme during eye development.


Cellular stresses and cytokines activate multiple mitogen-activated-protein kinase kinase homologues in PC12 and KB cells.
The identities of the upstream activators of the mitogen-activated protein (MAP) kinase homologues termed stress-activated-protein (SAP) kinase-1 (also known as JNK or SAPK) and SAP kinase-2 (also known as p38, RK and CSBP) were investigated in rat PC12 cells and human KB cells after exposure to cellular stresses and cytokines. In PC12 cells, the same two upstream activators, SAP kinase kinase-1 (SAPKK-1) and SAPKK-2 were activated after exposure to osmotic shock, ultraviolet irradiation or the protein synthesis inhibitor anisomycin, and more weakly in response to sodium arsenite. SAPKK-1 was capable of activating both SAP kinase-1 and SAP kinase-2 and was similar, if not identical, to the previously described MAP kinase kinase homologue MKK4, as judged by immunological criteria and by its ability to be activated by MEK kinase in vitro. In contrast, SAPKK-2 activated SAP kinase-2, but not SAP kinase-1 in vitro. In KB cells, five distinct upstream activators of SAP kinase-1 and SAP kinase-2 were induced, namely SAPKK-1, SAPKK-2, SAPKK-3, SAPKK-4 and SAPKK-5, whose appearance depended on the nature of the stimulus. SAPKK-3, which was strongly induced by every stimulus tested (osmotic shock, ultraviolet irradiation, anisomycin or IL-1), accounted for about 95% of the SAP kinase-2 activator activity in these cells, did not activate SAP kinase-1 and eluted from Mono S at a lower salt concentration than SAPKK-2. SAPKK-4 and SAPKK-5 were also eluted from Mono S at higher NaC1 concentrations than SAPKK-3 and these enzymes activated SAP kinase-1 but not SAP kinase-2. SAPKK-4 was the only SAP kinase-1 activator induced by interleukin-1 or ultraviolet irradiation, while two SAP kinase-1 activators, SAPKK-1 and SAPKK-5, were induced by osmotic shock or anisomycin. SAPKK-2, SAPKK-3, SAPKK-4 and SAPKK-5, were not activated by MEK kinase in vitro, were separable from the major activator(s) of p42 MAP kinase, and were not recognised by anti-MKK4 antibodies. At least two of these enzymes are likely to be novel MAP kinase kinase homologues. Our results demonstrate unexpected complexity in the upstream regulation of stress and cytokine-stimulated kinase cascades and indicate that the selection of the appropriate SAPKK varies with both the stimulus and the cell type.


Color-selective attention need not be mediated by spatial attention.
It is well-established that attention can select stimuli for preferential processing on the basis of non-spatial features such as color, orientation, or direction of motion. Evidence is mixed, however, as to whether feature-selective attention acts by increasing the signal strength of to-be-attended features irrespective of their spatial locations or whether it acts by guiding the spotlight of spatial attention to locations containing the relevant feature. To address this question, we designed a task in which feature-selective attention could not be mediated by spatial selection. Participants observed a display of intermingled dots of two colors, which rapidly and unpredictably changed positions, with the task of detecting brief intervals of reduced luminance of 20% of the dots of one or the other color. Both behavioral indices and electrophysiological measures of steady-state visual evoked potentials showed selectively enhanced processing of the attended-color items. The results demonstrate that feature-selective attention produces a sensory gain enhancement at early levels of the visual cortex that occurs without mediation by spatial attention.


Why our ethics curricula don't work.
The impact our ethics curricula have on students seems marginal at best. Students take the ethics courses we offer and pass the tests we give, but no one's behavior changes as a result. We fundamentally see ourselves teaching about ethics, which is slightly different than teaching ethics--and expecting behavior to change as a result of what is taught. The premise of this article is that our ethics courses are inadequate in content and form to the extent that they do not cultivate an introspective orientation to professional life. In some cases they amount to little more than a study of various state dental practice acts or the Code of Ethics of the American Dental Association. Three specific weaknesses are identified in a typical ethics curriculum: 1) failure to recognize that more education is not the answer to everything; 2) ethics is boring; and 3) course content is qualitatively inadequate because it does not foster an introspective basis for true behavioral change. A fourth element, an innovation, is directed to this third weakness and entails implementing a precurriculum very early in the dental educational experience to address the disconnect between knowledge and action.


Reduced functional performance in the lower extremity predicted radiographic knee osteoarthritis five years later.
Reduced quadriceps strength is an early finding in subjects with knee osteoarthritis, but it is not clear whether it is a cause or a consequence of knee osteoarthritis. To determine whether reduced functional performance in the lower extremity predicts the incidence or progression of radiographic knee osteoarthritis. Prospective, epidemiological, population based cohort study. 148 subjects (62 women), aged 35-54 (mean 44.8), with chronic knee pain from a population based cohort. Predictors analysed were age, sex, body mass index, baseline knee pain, and three tests of lower extremity functional performance: maximum number of one-leg rises from sitting, time spent walking 300 m, and timed standing on one leg. Weightbearing tibiofemoral knee radiographs were obtained at baseline and after 5 years (median 5.1, range 4.2-6.1), and classified according to Kellgren and Lawrence as no osteoarthritis (Kellgren and Lawrence = 0, n = 94) or prevalent osteoarthritis (Kellgren and Lawrence >/=1, n = 54). Fewer one-leg rises (median 17 v 25) predicted incident radiographic osteoarthritis five years later (OR 2.6, 95% CI 1.1 to 6.0). The association remained significant after controlling for age, sex, body mass index, and pain. No significant predictor of radiographic progression in the group with prevalent osteoarthritis was found. Reduced functional performance in the lower extremity predicted development of radiographic knee osteoarthritis 5 years later among people aged 35-55 with chronic knee pain and normal radiographs at baseline. These findings suggest that a test of one-leg rises may be useful, and interventions aimed at improving functional performance may be protective against development of knee osteoarthritis.


Cryopreserved microvascular conduits: current applications and future prospects.
Modern cryopreservation techniques have resulted in the clinical use of cryopreserved allograft heart valves and large diameter veins. In addition to these large vascular tissues, there is a need for small-diameter vascular allografts. We review our studies of cryopreserved rat microvenous allografts (1 mm diameter) in both the arterial and venous systems. The clinical utilization of cryopreserved allografts in reconstructive microsurgery and the potential problems associated with the use of allogeneic tissues will be addressed. Further applications for vascular cryopreservation in reconstructive surgery include small-diameter arteries, vascularized tissue flaps, and limbs.


Incidence and Clinical Impact of Anti-TNFα Treatment of Severe Immune Checkpoint Inhibitor-induced Colitis in Advanced Melanoma: The Mecolit Survey.
Immune checkpoint inhibitors (ICI) significantly improve overall survival (OS) in patients with advanced melanoma, but immune-related colitis may occur and warrant anti-tumor necrosis factor α (TNFα) treatment in severe forms. A nationwide, multicenter retrospective survey was conducted to assess both, the real-life incidence of grade 3/4 ICI-induced colitis treated with anti-TNFα, in patients with advanced melanoma, and the consequence of this therapeutic strategy on disease outcome. All patients with advanced melanoma treated with anti-TNFα agents for severe ICI-related colitis in the participating centers were included. Relative incidence was calculated according to the total number of patients treated with ICI in network centers during the period of inclusion. The possible impact of anti-TNFα treatment on disease outcome was evaluated through comparison of objective response rate, progression-free survival, and OS with pivotal literature data. Twenty-seven patients from 13 tertiary referral centers were included. Overall, severe ICI-related colitis treated with anti-TNFα occurred in 1% of patients with advanced melanoma, mostly with ipilimumab. Infliximab was successfully used in all patients but 1, mostly after 1 infusion. OS and progression-free survival of 12 and 3 months, respectively, were observed in these patients, along with an objective response rate of 41% at 12 months. This survey shows a low real-life incidence of severe colitis requiring anti-TNFα. Response rates to immunotherapy and survival data do not appear to significantly differ from those observed in pivotal studies. Severe ICI-induced colitis requiring anti-TNFα treatment appears to be a rare event in advanced melanoma, and infliximab does not seem to adversely affect disease outcome.


Topical steroids for nasal polyps.
Chronic rhinosinusitis with nasal polyps (CRSwNP) represents inflammatory changes throughout the nose and sinuses from a group of disorders which all lead to swelling and overgrowth of the nasal mucosa. Topical corticosteroids have been the most widely used treatment, with each clinician using different regimes, at different doses, in different settings and with or without sinus surgery. CRSwNP requires ongoing medical management to prevent recurrence. To assess the effects of topical corticosteroids on CRSwNP and to analyse various subgroups, including patients who had sinus surgery immediately prior to the delivery of the corticosteroids, surgery any time prior to the topical corticosteroids or patients who had never had previous surgery. Also to assess the most effective dose and delivery methods for topical corticosteroids. We searched the Cochrane Ear, Nose and Throat Disorders Group Trials Register; the Cochrane Central Register of Controlled Trials (CENTRAL); PubMed; EMBASE; CINAHL; Web of Science; BIOSIS Previews; Cambridge Scientific Abstracts; ICTRP and additional sources for published and unpublished trials. The date of the search was 10 April 2012. Randomised controlled trials studying topical corticosteroids for patients with CRSwNP. At least two authors reviewed the search results and selected trials meeting the eligibility criteria, obtaining full texts and contacting authors. We documented our justification for the exclusion of studies. At least two authors extracted data using a pre-determined, standardised data form. Forty studies (3624 patients) met the inclusion criteria. The trials were at low (21 trials), medium (13 trials) and high (six trials) risk of bias. The primary outcomes were sino-nasal symptoms, polyp size and polyp recurrence after surgery. When compared to placebo, topical corticosteroids improved overall symptom scores (standardised mean difference (SMD) -0.46; 95% confidence interval (CI) -0.65 to -0.27, P < 0.00001; seven trials, n = 445) and had a higher proportion of patients whose symptoms improved (responders) (risk ratio (RR) 1.71; 95% CI 1.29 to 2.26, P = 0.0002; four trials, n = 234). Topical corticosteroids also decreased the polyp score (SMD -0.73; 95% CI -1.00 to -0.46, P < 0.00001; three trials, n = 237) and had a greater proportion of patients with a reduction in polyp size (responders) (RR 2.09; 95% CI 1.65 to 2.64, P < 0.00001; eight trials, n = 785) when compared to placebo. Topical corticosteroids also prevented polyp recurrence after surgery (RR 0.59; 95% CI 0.45 to 0.79, P = 0.0004; six trials, n = 437). Subgroup analyses by sinus surgery status revealed a greater benefit in reduction of polyp score when topical steroid was administered any time after sinus surgery (SMD -1.19; 95% CI -1.54 to -0.83) compared to patients who had never had surgery (SMD -0.13; 95% CI -0.53 to 0.28, P < 0.00001). There was no difference between groups in terms of adverse events. Topical corticosteroids are a beneficial treatment for CRSwNP and the adverse effects are minor, with benefits outweighing the risks. They improve symptoms, reduce polyp size and prevent polyp recurrence after surgery. Patients having sinus surgery may have a greater response to topical corticosteroids but further research is required.


Cytomegalovirus and Epstein-Barr virus in breast cancer.
Findings of polymerase chain reaction (PCR) studies of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) and breast cancer vary, making it difficult to determine whether either, both, or neither virus is causally associated with breast cancer. We investigated CMV and EBV in paired samples of breast cancer and normal breast tissue from 70 women using quantitative PCR. A serum sample from each woman was tested for CMV and EBV IgG. To place our results in context, we reviewed the existing literature and performed a meta-analysis of our results together with previous PCR studies of EBV, CMV, and breast cancer. Of the serology samples, 67 of 70 (96%) were EBV IgG positive and 49 of 70 (70%) were CMV IgG positive. QPCR detected EBV in 24 (34%) of the tumour and 9 (13%) of the paired normal specimens and CMV in 0 (0%) of the tumour and 2 (3%) of the paired normal specimens. Our findings, together with earlier results summarised in the meta-analysis, suggest several possibilities: variable findings may be due to limitations of molecular analyses; 'hit and run' oncogenesis may lead to inconsistent results; one or both viruses has a role at a later stage in breast cancer development; infection with multiple viruses increases breast cancer risk; or neither virus has a role. Future studies should focus on ways to investigate these possibilities, and should include comparisons of breast cancer tissue samples with appropriate normal tissue samples.


Big data or bust: realizing the microbial genomics revolution.
Pathogen genomics has the potential to transform the clinical and public health management of infectious diseases through improved diagnosis, detection and tracking of antimicrobial resistance and outbreak control. However, the wide-ranging benefits of this technology can only fully be realized through the timely collation, integration and sharing of genomic and clinical/epidemiological metadata by all those involved in the delivery of genomic-informed services. As part of our review on bringing pathogen genomics into 'health-service' practice, we undertook extensive stakeholder consultation to examine the factors integral to achieving effective data sharing and integration. Infrastructure tailored to the needs of clinical users, as well as practical support and policies to facilitate the timely and responsible sharing of data with relevant health authorities and beyond, are all essential. We propose a tiered data sharing and integration model to maximize the immediate and longer term utility of microbial genomics in healthcare. Realizing this model at the scale and sophistication necessary to support national and international infection management services is not uncomplicated. Yet the establishment of a clear data strategy is paramount if failures in containing disease spread due to inadequate knowledge sharing are to be averted, and substantial progress made in tackling the dangers posed by infectious diseases.


Mice with an N-Ethyl-N-Nitrosourea (ENU) Induced Tyr209Asn Mutation in Natriuretic Peptide Receptor 3 (NPR3) Provide a Model for Kyphosis Associated with Activation of the MAPK Signaling Pathway.
Non-syndromic kyphosis is a common disorder that is associated with significant morbidity and has a strong genetic involvement; however, the causative genes remain to be identified, as such studies are hampered by genetic heterogeneity, small families and various modes of inheritance. To overcome these limitations, we investigated 12 week old progeny of mice treated with the chemical mutagen N-ethyl-N-nitrosourea (ENU) using phenotypic assessments including dysmorphology, radiography, and dual-energy X-ray absorptiometry. This identified a mouse with autosomal recessive kyphosis (KYLB). KYLB mice, when compared to unaffected littermates, had: thoraco-lumbar kyphosis, larger vertebrae, and increased body length and increased bone area. In addition, female KYLB mice had increases in bone mineral content and plasma alkaline phosphatase activity. Recombination mapping localized the Kylb locus to a 5.5Mb region on chromosome 15A1, which contained 51 genes, including the natriuretic peptide receptor 3 (Npr3) gene. DNA sequence analysis of Npr3 identified a missense mutation, Tyr209Asn, which introduced an N-linked glycosylation consensus sequence. Expression of wild-type NPR3 and the KYLB-associated Tyr209Asn NPR3 mutant in COS-7 cells demonstrated the mutant to be associated with abnormal N-linked glycosylation and retention in the endoplasmic reticulum that resulted in its absence from the plasma membrane. NPR3 is a decoy receptor for C-type natriuretic peptide (CNP), which also binds to NPR2 and stimulates mitogen-activated protein kinase (MAPK) signaling, thereby increasing the number and size of hypertrophic chondrocytes. Histomorphometric analysis of KYLB vertebrae and tibiae showed delayed endochondral ossification and expansion of the hypertrophic zones of the growth plates, and immunohistochemistry revealed increased p38 MAPK phosphorylation throughout the growth plates of KYLB vertebrae. Thus, we established a model of kyphosis due to a novel NPR3 mutation, in which loss of plasma membrane NPR3 expression results in increased MAPK pathway activation, causing elongation of the vertebrae and resulting in kyphosis.


A Proposed Role for Pro-Inflammatory Cytokines in Damaging Behavior in Pigs.
Sickness can change our mood for the worse, leaving us sad, lethargic, grumpy and less socially inclined. This mood change is part of a set of behavioral symptoms called sickness behavior and has features in common with core symptoms of depression. Therefore, the physiological changes induced by immune activation, for example following infection, are in the spotlight for explaining mechanisms behind mental health challenges such as depression. While humans may take a day off and isolate themselves until they feel better, farm animals housed in groups have only limited possibilities for social withdrawal. We suggest that immune activation could be a major factor influencing social interactions in pigs, with outbreaks of damaging behavior such as tail biting as a possible result. The hypothesis presented here is that the effects of several known risk factors for tail biting are mediated by pro-inflammatory cytokines, proteins produced by the immune system, and their effect on neurotransmitter systems. We describe the background for and implications of this hypothesis.


Contrast-Enhanced Ultrasound Tumor Therapy With Abdominal Imaging Transducer.
A diagnostic ultrasound machine add-on module (AOM) was created to enable an off-the-shelf abdominal imaging transducer to perform contrast-enhanced therapeutic ultrasound. The AOM creates plane-wave ultrasound through an abdominal imaging transducer targeting intravascular microbubbles within tumors. This therapeutic antivascular ultrasound (AVUS) causes heating and cavitation effects that destroy tumor vasculature and starves it of nutrients. The AOM can switch between therapeutic and imaging modes for monitoring AVUS treatment. The therapeutic capability of the AOM was validated in murine hepatocellular carcinomas (HCC) grown in adult mice. Contrast-enhanced ultrasound imaging performed before and after the therapeutic treatment evaluated the AVUS response to the treatment. The peak enhancement (PE), perfusion index (PI), and area under the curve (AUC) were measured for the control and AOM treatment groups. The AOM group showed a substantial decrease in these parameters compared to the control group. The difference between the pre- and post-therapy was significant, (p < 0.001) for the AOM group and not significant (p > 0.5) for the control group. Tumor temperatures increased markedly for the AOM group with a thermal dose (CEM43) of 124.8 (±2.5). Histochemical analysis of the excised HCC samples revealed several hemorrhagic pools in tumors from the AOM group, absent in the tumors of the control group. These results demonstrate the theranostic potential of the AOM to induce and monitor vascular disruption within murine tumors.


Speaking to the yet unknowing world: Hamlet, Horatio and the problem of imperfect witness.
Every day doctors bear witness to others about the experiences, needs and feelings of their patients, drawing on what they have learnt from clinical consultations. This paper considers the medical task of bearing honourable and truthful witness through an examination of the role and actions of Horatio in Hamlet. Horatio is simultaneously located among the background machinery of the play, separate from the lives of the protagonists, and in the foreground, where his authoritative witness is repeatedly called upon by the play's characters. Horatio is invited to watch an unfolding disaster, his warnings are not heard, and at its conclusion he stands apart from the drama to give its account. The tensions between engagement and observation, and between partial and impartial accounting echo those faced by doctors in everyday clinical practice. The act of bearing witness, Shakespeare suggests, even for those who are tasked with being objective, is necessarily imperfect, and not dispassionate. Those people, including doctors, who are expected to construct authoritative accounts of the motives and actions of others may find in Hamlet a small lesson on the need to approach their summary narratives about others more humbly, aware of the narrowness and partiality of their perspective.


Free radicals and extrinsic skin aging.
Human skin is constantly directly exposed to the air, solar radiation, environmental pollutants, or other mechanical and chemical insults, which are capable of inducing the generation of free radicals as well as reactive oxygen species (ROS) of our own metabolism. Extrinsic skin damage develops due to several factors: ionizing radiation, severe physical and psychological stress, alcohol intake, poor nutrition, overeating, environmental pollution, and exposure to UV radiation (UVR). It is estimated that among all these environmental factors, UVR contributes up to 80%. UV-induced generation of ROS in the skin develops oxidative stress, when their formation exceeds the antioxidant defence ability of the target cell. The primary mechanism by which UVR initiates molecular responses in human skin is via photochemical generation of ROS mainly formation of superoxide anion (O(2) (-) (∙)), hydrogen peroxide (H(2)O(2)), hydroxyl radical (OH(∙)), and singlet oxygen ((1)O(2)). The only protection of our skin is in its endogenous protection (melanin and enzymatic antioxidants) and antioxidants we consume from the food (vitamin A, C, E, etc.). The most important strategy to reduce the risk of sun UVR damage is to avoid the sun exposure and the use of sunscreens. The next step is the use of exogenous antioxidants orally or by topical application and interventions in preventing oxidative stress and in enhanced DNA repair.


Symbiont community diversity is more variable in corals that respond poorly to stress.
Coral reefs are declining globally as climate change and local water quality press environmental conditions beyond the physiological tolerances of holobionts-the collective of the host and its microbial symbionts. To assess the relationship between symbiont composition and holobiont stress tolerance, community diversity metrics were quantified for dinoflagellate endosymbionts (Family: Symbiodiniaceae) from eight Acropora millepora genets that thrived under or responded poorly to various stressors. These eight selected genets represent the upper and lower tails of the response distribution of 40 coral genets that were exposed to four stress treatments (and control conditions) in a 10-day experiment. Specifically, four 'best performer' coral genets were analyzed at the end of the experiment because they survived high temperature, high pCO2 , bacterial exposure, or combined stressors, whereas four 'worst performer' genets were characterized because they experienced substantial mortality under these stressors. At the end of the experiment, seven of eight coral genets mainly hosted Cladocopium symbionts, whereas the eighth genet was dominated by both Cladocopium and Durusdinium symbionts. Symbiodiniaceae alpha and beta diversity were higher in worst performing genets than in best performing genets. Symbiont communities in worst performers also differed more after stress exposure relative to their controls (based on normalized proportional differences in beta diversity), than did best performers. A generalized joint attribute model estimated the influence of host genet and treatment on Symbiodiniaceae community composition and identified strong associations among particular symbionts and host genet performance, as well as weaker associations with treatment. Although dominant symbiont physiology and function contribute to host performance, these findings emphasize the importance of symbiont community diversity and stochasticity as components of host performance. Our findings also suggest that symbiont community diversity metrics may function as indicators of resilience and have potential applications in diverse disciplines from climate change adaptation to agriculture and medicine.


A phase II, randomized study on an investigational DTPw-HBV/Hib-MenAC conjugate vaccine administered to infants in Northern Ghana.
Combining meningococcal vaccination with routine immunization in infancy may reduce the burden of meningococcal meningitis, especially in the meningitis belt of Africa. We have evaluated the immunogenicity, persistence of immune response, immune memory and safety of an investigational DTPw-HBV/Hib-MenAC conjugate vaccine given to infants in Northern Ghana. In this phase II, double blind, randomized, controlled study, 280 infants were primed with DTPw-HBV/Hib-MenAC or DTPw-HBV/Hib vaccines at 6, 10 and 14 weeks of age. At 12 months of age, children in each group received a challenge dose of serogroup A+C polysaccharides. Antibody responses were assessed pre, and one month-post dose 3 of the priming schedule and pre and 1 month after administration of the challenge dose. One month post-dose 3, 87.8% and 88.2% of subjects in the study group had bactericidal meningococcal serogroup A (SBA-MenA) and meningococcal serogroup C (SBA-MenC) antibody titres > or = 1:8 respectively. Seroprotection/seropositivity rates to the 5 antigens administered in the routine EPI schedule were non-inferior in children in the study group compared to those in the control group. The percentages of subjects in the study group with persisting SBA-MenA titres > or = 1:8 or SBA-MenC titres > or = 1:8 at the age of 12 months prior to challenge were significantly higher than in control group (47.7% vs 25.7% and 56.4% vs 5.1% respectively). The administration of 10 microg of serogroup A polysaccharide increased the SBA-MenA GMT by 14.0-fold in the DTPW-HBV/HibMenAC-group compared to a 3.8 fold increase in the control-group. Corresponding fold-increases in SBA-MenC titres following challenge with 10 microg of group C polysaccharide were 18.8 and 1.9 respectively. Reactogenicity following primary vaccination or the administration of the challenge dose was similar in both groups, except for swelling (Grade 3) after primary vaccination which was more frequent in children in the vaccine than in the control group (23.7%; 95%CI [19.6-28.1] of doses vs 14.1%; 95% CI [10.9-17.8] of doses). Fifty-nine SAEs (including 8 deaths), none of them related to vaccination, were reported during the entire study. Three dose primary vaccination with DTPw-HBV/Hib-MenAC was non-inferior to DTPw-HBV/Hib for the 5 common antigens used in the routine EPI schedule and induced bactericidal antibodies against Neisseria meningitidis of serogroups A and C in the majority of infants. Serogroup A and C bactericidal antibody levels had fallen below titres associated with protection in nearly half of the infants by the age of 12 months confirming that a booster dose is required at about that age. An enhanced memory response was shown after polysaccharide challenge. This vaccine could provide protection against 7 important childhood diseases (including meningococcal A and C) and be of particular value in countries of the African meningitis belt. Controlled-Trials.com ISRCTN35754083.


Host life history and the evolution of parasite virulence.
We present a general epidemiological model of host-parasite interactions that includes various forms of superinfection. We use this model to study the effects of different host life-history traits on the evolution of parasite virulence. In particular, we analyze the effects of natural host death rate on the evolutionarily stable parasite virulence. We show that, contrary to classical predictions, an increase in the natural host death rate may select for lower parasite virulence if some form of superinfection occurs. This result is in agreement with the experimental results and the verbal argument presented by Ebert and Mangin (1997). This experiment is discussed in the light of the present model. We also point out the importance of superinfections for the effect of nonspecific immunity on the evolution of virulence. In a broader perspective, this model demonstrates that the occurrence of multiple infections may qualitatively alter classical predictions concerning the effects of various host life-history traits on the evolution of parasite virulence.


The expression ratio of Map7/B2M is prognostic for survival in patients with stage II colon cancer.
Colorectal cancer (CRC) is the second most frequent cause of cancer-related death in the United States. To determine whether certain molecular markers might be prognostic for survival, we measured by quantitative real-time RT-PCR the expression levels of 15 previously studied genes that are known to be up-regulated or down-regulated in the progression of epithelial cancers. The tumor samples were extracted from formalin-fixed paraffin-embedded primary tissues derived from patients with Stage II CRC who developed disease recurrence within two years (n=10), or were disease-free for at least 4 years (n=12). We were able to determine, by AUC curve analysis, that the ratio of microtubule associated protein 7 (Map7)/B2M was predictive of outcome in our sample set. Further, using Kaplan-Meier survival analysis, we observed significantly different curves as a function of marker positivity for the Map7/B2M (p=0.0001; HR=11) expression ratio. This suggests that the expression ratio of Map7/B2M may serve as a valuable prognostic marker in patients with Stage II colon cancer, and potentially guide therapeutic decision making.


High-Content RNAi Phenotypic Screening Unveils the Involvement of Human Ubiquitin-Related Enzymes in Late Cytokinesis.
CEP55 is a central regulator of late cytokinesis and is overexpressed in numerous cancers. Its post-translationally controlled recruitment to the midbody is crucial to the structural coordination of the abscission sequence. Our recent evidence that CEP55 contains two ubiquitin-binding domains was the first structural and functional link between ubiquitin signaling and ESCRT-mediated severing of the intercellular bridge. So far, high-content screens focusing on cytokinesis have used multinucleation as the endpoint readout. Here, we report an automated image-based detection method of intercellular bridges, which we applied to further our understanding of late cytokinetic signaling by performing an RNAi screen of ubiquitin ligases and deubiquitinases. A secondary validation confirmed four candidate genes, i.e., LNX2, NEURL, UCHL1 and RNF157, whose downregulation variably affects interconnected phenotypes related to CEP55 and its UBDs, as follows: decreased recruitment of CEP55 to the midbody, increased number of midbody remnants per cell, and increased frequency of intercellular bridges or multinucleation events. This brings into question the Notch-dependent or independent contributions of LNX2 and NEURL proteins to late cytokinesis. Similarly, the role of UCHL1 in autophagy could link its function with the fate of midbody remnants. Beyond the biological interest, this high-content screening approach could also be used to isolate anticancer drugs that act by impairing cytokinesis and CEP55 functions.


Optical Super-Resolution Imaging of β-Amyloid Aggregation In Vitro and In Vivo: Method and Techniques.
Super-resolution microscopy has emerged as a powerful and non-invasive tool for the study of molecular processes both in vitro and in live cells. In particular, super-resolution microscopy has proven valuable for research studies in protein aggregation. In this chapter we present details of recent advances in this method and the specific techniques, enabling the study of amyloid beta aggregation optically, both in vitro and in cells. First, we show that variants of optical super-resolution microscopy provide a capability to visualize oligomeric and fibrillar structures directly, providing detailed information on species morphology in vitro and even in situ, in the cellular environment. We focus on direct Stochastic Optical Reconstruction Microscopy, dSTORM, which provides morphological detail on spatial scales below 20 nm, and provide detailed protocols for its implementation in the context of amyloid beta research. Secondly, we present a range of optical techniques that offer super-resolution indirectly, which we call multi-parametric microscopy. The latter offers molecular scale information on self-assembly reactions via changes in protein or fluorophore spectral signatures. These techniques are empowered by our recent discovery that disease related amyloid proteins adopt intrinsic energy states upon fibrilisation. We show that fluorescence lifetime imaging provides a particularly sensitive readout to report on the aggregation state, which is robustly quantifiable for experiments performed either in vitro or in vivo.


Inverse association between body mass and frequency of milk consumption in children.
Recent studies have shown an inverse association between the level of dietary Ca, particularly from dairy sources, and body weight in adults; there is, however, a paucity of data regarding this relationship in children. We therefore investigated this issue in 1087 children who underwent body weight and height measurement during a survey on childhood obesity. Lifestyle and dietary habits were investigated by a questionnaire. After excluding children who were following a dietary regimen for any reason, 884 children (M:F 451:433; age 7.5 (sd 2.1) years) were selected. Milk consumption was pooled into four frequency categories: poor (< or =1/week; n 125), moderate (>1 but < or =5-6/week; n 133), regular (1/d; n 408) and high (> or =2/d; n 218). The frequency of consumption of milk was inversely and significantly associated (t=-2.964, P=0.003) with age- and sex-specific BMI z-scores by linear regression analysis, controlling for sex, age, physical activity, birth weight and parental overweight and education. The statistical association remained significant (t=-2.831, P=0.005) after the inclusion of children consuming only skimmed milk (n 91). Milk consumption was still significantly and inversely associated with BMI z score (t=-2.791, P=0.005) in the whole-milk consumers when controlling for age and the frequency of consumption of various foods; this association was no longer significant (P=0.21) when children consuming skimmed milk were included in the analysis. This is the first report showing a significant inverse association between frequency of milk consumption and body mass in children. Regardless of the mechanisms involved, our results might encourage further research on this issue and might have important implications for the prevention of obesity.


High levels of variation in Salix lignocellulose genes revealed using poplar genomic resources.
Little is known about the levels of variation in lignin or other wood related genes in Salix, a genus that is being increasingly used for biomass and biofuel production. The lignin biosynthesis pathway is well characterized in a number of species, including the model tree Populus. We aimed to transfer the genomic resources already available in Populus to its sister genus Salix to assess levels of variation within genes involved in wood formation. Amplification trials for 27 gene regions were undertaken in 40 Salix taxa. Twelve of these regions were sequenced. Alignment searches of the resulting sequences against reference databases, combined with phylogenetic analyses, showed the close similarity of these Salix sequences to Populus, confirming homology of the primer regions and indicating a high level of conservation within the wood formation genes. However, all sequences were found to vary considerably among Salix species, mainly as SNPs with a smaller number of insertions-deletions. Between 25 and 176 SNPs per kbp per gene region (in predicted exons) were discovered within Salix. The variation found is sizeable but not unexpected as it is based on interspecific and not intraspecific comparison; it is comparable to interspecific variation in Populus. The characterisation of genetic variation is a key process in pre-breeding and for the conservation and exploitation of genetic resources in Salix. This study characterises the variation in several lignocellulose gene markers for such purposes.


Antioxidative and anti-inflammatory activities of polyhydroxyflavonoids of Scutellaria baicalensis GEORGI.
The active ingredients of 'golden root' of Scutellaria baicalensis GEORGI (Huang-Qin), a valuable traditional Chinese medicine, are polyhydroxyflavonoids, namely baicalein, oroxylin A and wogonin. With the objective of overcoming their poor solubility and to investigate their structure and activity relationships, baicaleinyl 7-O-sulfate was prepared, and extensive comparative antioxidative and anti-inflammatory tests were conducted. All the polyhydroxyflavonoids exhibited significant antioxidative and free-radical scavenging activities. In respect of their nitric oxide (NO) inhibition, wogonin was superior to all the other flavonoids, while oroxylin A was most potent in the inhibition of lipid peroxidation. Wogonin proved to be the most potent (82.9% inhibition, p<0.05) in its anti-inflammatory activity against carrageenan-induced rat hind paw edema. There was a correlation between the in-vivo anti-inflammatory activity and the in-vitro antioxidative activities.


Reading a suspenseful literary text activates brain areas related to social cognition and predictive inference.
Stories can elicit powerful emotions. A key emotional response to narrative plots (e.g., novels, movies, etc.) is suspense. Suspense appears to build on basic aspects of human cognition such as processes of expectation, anticipation, and prediction. However, the neural processes underlying emotional experiences of suspense have not been previously investigated. We acquired functional magnetic resonance imaging (fMRI) data while participants read a suspenseful literary text (E.T.A. Hoffmann's "The Sandman") subdivided into short text passages. Individual ratings of experienced suspense obtained after each text passage were found to be related to activation in the medial frontal cortex, bilateral frontal regions (along the inferior frontal sulcus), lateral premotor cortex, as well as posterior temporal and temporo-parietal areas. The results indicate that the emotional experience of suspense depends on brain areas associated with social cognition and predictive inference.


Capillary imbibition in open-cell monodisperse foams.
Although capillary imbibition of solid foams is involved in many industrial applications, general theory for capillary imbibition has never been proved to apply for this specific class of porous materials. In order to compare accurately experiment and theory we produce solid foam samples with monodisperse pore size distributions and tunable pore volume fraction, and we measure their permeability (Darcy), their capillary pressure and their imbibition rate. Our findings reveal that the imbibition velocity is qualitatively compatible with the Washburn theory but it is one order of magnitude smaller than the predicted value. This deviation is attributed to the excess time spent by the liquid-gas interface through connections between pores, for which an empirical expression is provided as a function of pore size and solid volume fraction. Our results provide the first step to understand deeply the imbibition process in foams and to predict imbibition rates for various foamed materials.


A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes.
Electron microscopy-based connectomics aims to comprehensively map synaptic connections in neural tissue. However, current approaches are limited in their capacity to directly assign molecular identities to neurons. Here, we use serial multiplex immunogold labeling (siGOLD) and serial-section transmission electron microscopy (ssTEM) to identify multiple peptidergic neurons in a connectome. The high immunogenicity of neuropeptides and their broad distribution along axons, allowed us to identify distinct neurons by immunolabeling small subsets of sections within larger series. We demonstrate the scalability of siGOLD by using 11 neuropeptide antibodies on a full-body larval ssTEM dataset of the annelid Platynereis. We also reconstruct a peptidergic circuitry comprising the sensory nuchal organs, found by siGOLD to express pigment-dispersing factor, a circadian neuropeptide. Our approach enables the direct overlaying of chemical neuromodulatory maps onto synaptic connectomic maps in the study of nervous systems.


Nurses' perceptions of climate and environmental issues: a qualitative study.
The aim of this study was to explore nurses' perceptions of climate and environmental issues and examine how nurses perceive their role in contributing to the process of sustainable development. Climate change and its implications for human health represent an increasingly important issue for the healthcare sector. According to the International Council of Nurses Code of Ethics, nurses have a responsibility to be involved and support climate change mitigation and adaptation to protect human health. This is a descriptive, explorative qualitative study. Nurses (n = 18) were recruited from hospitals, primary care and emergency medical services; eight participated in semi-structured, in-depth individual interviews and 10 participated in two focus groups. Data were collected from April-October 2013 in Sweden; interviews were transcribed verbatim and analysed using content analysis. Two main themes were identified from the interviews: (i) an incongruence between climate and environmental issues and nurses' daily work; and (ii) public health work is regarded as a health co-benefit of climate change mitigation. While being green is not the primary task in a lifesaving, hectic and economically challenging context, nurses' perceived their profession as entailing responsibility, opportunities and a sense of individual commitment to influence the environment in a positive direction. This study argues there is a need for increased awareness of issues and methods that are crucial for the healthcare sector to respond to climate change. Efforts to develop interventions should explore how nurses should be able to contribute to the healthcare sector's preparedness for and contributions to sustainable development.


Structure-activity relationships for flow cytometric data of smaller polycyclic aromatic hydrocarbons.
Using flow cytometry, select polycyclic aromatic hydrocarbons (PAHs) were evaluated for induction of apoptosis in human monocytic THP-1 cells. Based on structure, the PAHs were divided into linear and bay-region-containing compounds. Except for fluorene, the linear PAHs failed to induce apoptosis; all of the bay-region-containing PAHs induce apoptosis. The relationship that a bay-region is required to induce apoptosis is supported by results for benzo[a]pyrene (positive) and 2-methylanthracene (negative). The data for bay-region containing, four-ringed PAH compounds reveal that possessing a linear-region of more than two rings diminishes the ability of a PAH to induce apoptosis. Owing to the steric interactions of the hydrogen atoms of the methyl group and those on the ring carbons, 1-methylanthracene does not have a true bay-region. However, the methyl group substituted in the 1-position does confer a bay-like conformation, which may explain its activity in contrast to its parent derivative anthracene and its 2-postion homologue.


Autoantibody profile and other immunological parameters in recurrent spontaneous abortion patients.
An autoimmune cause and related immunological alterations resulting in recurrent spontaneous abortion (RSA) have been suggested in patients with unknown etiology. This study evaluated the autoantibody profile and other immunological parameters among RSA patients and normal pregnant women from Mumbai western India. Fifty RSA patients with unknown cause and greater than three consecutive abortions along with 50 normal pregnant women were studied for various auto antibodies such as ANA, anti-dsDNA, ANCA, AECA, 2 micro globulin, anti-HLA antibodies and ACLA using immunofluorescence microlymphocytotoxicity and ELISA. Immunological parameters such as HLA class I monoclonal antibody expression, CD3 (T cell), CD19 (B cell), and CD56 (NK cell) were estimated by flow cytometry. The results revealed 34% positivity of all auto antibodies tested among patients. ANA(12%), ANCA (20%), AECA (24%), ACLA (8%), anti-dsDNA(0%), β2 microglobulin (14%), and anti-HLA antibodies(10%) among RSA patients were identified. An increased expression of HLA class I specific monoclonal antibody (10%) with HLA A3 (16%) specificity were found to correlate with shared HLA alleles among the RSA couples. Among normal pregnant (control) group ANA (2%), ANCA (2%), AECA (3%), ACLA (4%) and increased expression of CD56 with reduced HLA class I monoclonal were observed. Our findings suggest a possible role of various autoantibodies along with the related immunological parameters underlying RSA.


Quantification of circulating endothelial progenitor cells using the modified ISHAGE protocol.
Circulating endothelial progenitor cells (EPC), involved in endothelial regeneration, neovascularisation, and determination of prognosis in cardiovascular disease can be characterised with functional assays or using immunofluorescence and flow cytometry. Combinations of markers, including CD34+KDR+ or CD133+KDR+, are used. This approach, however may not consider all characteristics of EPC. The lack of a standardised protocol with regards to reagents and gating strategies may account for the widespread inter-laboratory variations in quantification of EPC. We, therefore developed a novel protocol adapted from the standardised so-called ISHAGE protocol for enumeration of haematopoietic stem cells to enable comparison of clinical and laboratory data. In 25 control subjects, 65 patients with coronary artery disease (CAD; 40 stable CAD, 25 acute coronary syndrome/acute myocardial infarction (ACS)), EPC were quantified using the following approach: Whole blood was incubated with CD45, KDR, and CD34. The ISHAGE sequential strategy was used, and finally, CD45(dim)CD34(+) cells were quantified for KDR. A minimum of 100 CD34(+) events were collected. For comparison, CD45(+)CD34(+) and CD45(-)CD34(+) were analysed simultaneously. The number of CD45(dim)CD34(+)KDR(+) cells only were significantly higher in healthy controls compared to patients with CAD or ACS (p = 0.005 each, p<0.001 for trend). An inverse correlation of CD45(dim)CD34(+)KDR(+) with disease activity (r = -0.475, p<0.001) was confirmed. Only CD45(dim)CD34(+)KDR(+) correlated inversely with the number of diseased coronaries (r = -0.344; p<0.005). In a second study, a 4-week de-novo treatment of atorvastatin in stable CAD evoked an increase only of CD45(dim)CD34(+)KDR(+) EPC (p<0.05). CD45(+)CD34(+)KDR(+) and CD45(-)CD34(+)KDR(+) were indifferent between the three groups. Our newly established protocol adopted from the standardised ISHAGE protocol achieved higher accuracy in EPC enumeration confirming previous findings with respect to the correlation of EPC with disease activity and the increase of EPC during statin therapy. The data of this study show the CD45(dim) fraction to harbour EPC.


Assignment of the genome segment coding for the neutralizing epitope(s) of orbiviruses in the Great Island subgroup (Kemerovo serogroup).
Reassortant viruses were produced from high-frequency recombination events between temperature-sensitive (ts) mutants of Broadhaven (BRD) (S. R. Moss and P. A. Nuttall, Virus Res. 4, 331-336, 1986) and Wexford (WEX) viruses, two serotypes in the Kemerovo serogroup of orbiviruses. The parental origin of each of the 10 genomic segments comprising each reassortant was determined by polyacrylamide gel electrophoresis. Comparison of neutralization titers with the results of genomic dsRNA analyses revealed that genome segment 5 codes for the neutralizing epitope(s). Kemerovo group viruses therefore differ from orbiviruses of the bluetongue serogroup in which segment 2 codes for the neutralizing epitopes (M. J. Grubman, J. A. Appleton, and G. J. Letchworth, Virology 131, 355-366, 1983; J. Kahlon, K. Sugiyama, and P. Roy, J. Virol. 48, 627-632, 1983). Results also indicated that the ts lesions in mutants of recombination groups II, III, and VI were in segments 1, 5, and 4, respectively. Reassortment of segments 2 and 10 appeared to be nonrandom, and evidence of possible "linkage" was obtained for segments 3 and 9.


Distinct classes of mitotic cyclins are differentially expressed in the soybean shoot apex during the cell cycle.
Protein phosphorylation by the complexes of cyclin and cyclin-dependent kinase plays a key role in cell cycle progression in all eukaryotes. The amplification by polymerase chain reaction of a cyclin box from developing root nodules and root apices of soybean showed the expression of a number of different molecular species of mitotic cyclins in plant meristems, and they were classified into five distinct groups based on their sequence similarities. The complete soybean cyclin cDNAs, cyc1Gm to cyc5Gm, corresponding to each group were isolated, and their predicted amino acid sequences showed clear similarities to mitotic cyclins identified from various organisms. These genes are expressed predominantly in such meristematic tissues as root and shoot apices and young developing nodules. Double-target in situ hybridization involving histone H4 as an S-phase marker allowed us to estimate the phases during which these cyclin genes are abundantly expressed. The results indicated that cyc5Gm is expressed in G2-to-M phases and cyc3Gm is expressed from late S-to-G2 phases. These expression patterns, together with the sequence criteria, strongly suggest that cyc3Gm and cyc5Gm encode the plant cognates for A- and B-type cyclins, respectively. In addition, the expression of cyc1Gm was restricted during a short period in S phase, suggesting that it belongs to a novel class of plant cyclins. Sequence comparison of 18 plant mitotic cyclins cloned thus far showed that they can be divided into four distinct structural groups with different functions in cell cycle progression.


Thermodynamics of aminoglycoside and acyl-coenzyme A binding to the Salmonella enterica AAC(6')-Iy aminoglycoside N-acetyltransferase.
Kinetic and mechanistic studies on the chromosomally encoded aminoglycoside 6'-N-acetyltransferase, AAC(6')-Iy, of Salmonella enterica that confers resistance toward aminoglycosides have been previously reported [Magnet et al. (2001) Biochemistry 40, 3700-3709]. In the present study, equilibrium binding and the thermodynamic parameters of binding of aminoglycosides and acyl-coenzyme A derivatives to AAC(6')-Iy and of two mutants, C109A and the C109A/C70A double mutant, have been studied using fluorescence spectroscopy and isothermal titration calorimetry (ITC). Association constants for different aminoglycosides varied greatly (4 x 10(4)-150 x 10(4)) while the association constants of several acyl-coenzyme A derivatives were similar (3.2 x 10(4)-4.5 x 10(4)). The association constants and van't Hoff enthalpy changes derived from intrinsic protein fluorescence changes were in agreement with independently measured values from isothermal titration calorimetry studies. Binding of both aminoglycosides and acyl-coenzyme A derivatives is strongly enthalpically driven and revealed opposing negative entropy changes, resulting in enthalpy-entropy compensation. The acetyltransferase exhibited a temperature-dependent binding of tobramycin with a negative heat capacity value of 410 cal mol(-1) K(-1). Isothermal titration studies of acetyl-coenzyme A and tobramycin binding to mutant forms of the enzyme indicated that completely conserved C109 does not play any direct role in the binding of either of the substrates, while C70 is directly involved in aminoglycoside binding. These results are discussed and compared with previous steady-state kinetic studies of the enzyme.


Design and Characterization of Superpotent Bivalent Ligands Targeting Oxytocin Receptor Dimers via a Channel-Like Structure.
Dimeric/oligomeric states of G-protein coupled receptors have been difficult to target. We report here bivalent ligands consisting of two identical oxytocin-mimetics that induce a three order magnitude boost in G-protein signaling of oxytocin receptors (OTRs) in vitro and a 100- and 40-fold gain in potency in vivo in the social behavior of mice and zebrafish. Through receptor mutagenesis and interference experiments with synthetic peptides mimicking transmembrane helices (TMH), we show that such superpotent behavior follows from the binding of the bivalent ligands to dimeric receptors based on a TMH1-TMH2 interface. Moreover, in this arrangement, only the analogues with a well-defined spacer length (∼25 Å) precisely fit inside a channel-like passage between the two protomers of the dimer. The newly discovered oxytocin bivalent ligands represent a powerful tool for targeting dimeric OTR in neurodevelopmental and psychiatric disorders and, in general, provide a framework to untangle specific arrangements of G-protein coupled receptor dimers.


In vitro study of the influence of the type of connection on the fracture load of zirconia abutments with internal and external implant-abutment connections.
To determine whether zirconia abutments with an internal connection exhibit similar fracture load as zirconia abutments with an external connection. The following zirconia abutments were divided into four groups of 20 each: StraumannCARES abutments on Straumann implants (group A), Procera abutments on Branemark implants (group B), Procera abutments on NobelReplace implants (group C), and Zirabut SynOcta prototype abutments on Straumann implants (group D). The abutments were fixed on their respective implants either internally via a secondary abutment (A) or a metallic coupling (C) (two-piece) or directly externally (B) and internally (D) (one-piece). In each group, 10 abutments were left unrestored (A1 to D1). Ten received glass-ceramic crowns (A2 to D2). Static loading was performed according to the ISO norm 14801 until failure. The bending moment was calculated for comparison of the groups and subjected to statistical analysis (Student t test). The mean bending moments of the unrestored abutments were 371.5 +/- 142.3 Ncm (A1), 276.5 +/- 47.6 Ncm (B1), 434.9 +/- 124.8 Ncm (C1), and 182.5 +/- 136.5 Ncm (D1). Two-piece internally connected abutments exhibited higher bending moments than one-piece internally (C1 versus D1 P = .003, A1 versus D1 P = .03) or externally (C1 versus B1 P = .004) connected abutments. The groups with restorations did not show different bending moments than those without restorations. The mean bending moments of the restored abutments were 283.3 +/- 44.8 Ncm (A2), 291.5 +/- 31.7 Ncm (B2), 351.5 +/- 58 Ncm (C2), and 184.3 +/- 77.7 Ncm (D2). Group C2 exhibited the highest bending moment (P < .05). Internally connected one-piece abutments (D2) were weaker than all other groups (D2 versus A2 P = .002; D2 versus B2 P = .001; D2 versus C2 P = .0003). The type of connection significantly influenced the strength of zirconia abutments. Superior strength was achieved by means of internal connection via a secondary metallic component.


Student nurses' experiences of the clinical learning environment in relation to the organization of supervision: a questionnaire survey.
The aim was to investigate student nurses' experiences of the clinical learning environment in relation to how the supervision was organized. The clinical environment plays an essential part in student nurses' learning. Even though different models for supervision have been previously set forth, it has been stressed that there is a need both of further empirical studies on the role of preceptorship in undergraduate nursing education and of studies comparing different models. A cross-sectional study with comparative design was carried out with a mixed method approach. Data were collected from student nurses in the final term of the nursing programme at three universities in Sweden by means of a questionnaire. In general the students had positive experiences of the clinical learning environment with respect to pedagogical atmosphere, leadership style of the ward manager, premises of nursing, supervisory relationship, and role of the nurse preceptor and nurse teacher. However, there were significant differences in their ratings of the supervisory relationship (p<0.001) and the pedagogical atmosphere (p 0.025) depending on how the supervision was organized. Students who had the same preceptor all the time were more satisfied with the supervisory relationship than were those who had different preceptors each day. Students' comments on the supervision confirmed the significance of the preceptor and the supervisory relationship. The organization of the supervision was of significance with regard to the pedagogical atmosphere and the students' relation to preceptors. Students with the same preceptor throughout were more positive concerning the supervisory relationship and the pedagogical atmosphere.


Peak exposure concentrations of dust and flour aeroallergen in flour mills and bakeries.
As part of an epidemiological study amongst workers exposed to flour we measured peak exposure levels to total dust and flour aeroallergen with personal samplers in bakeries, flour mills and packing stations. Short-term tasks which were expected to give rise to high concentrations of exposure (peaks) were identified. The frequency and duration of these tasks were estimated and their levels of exposure to dust and flour aeroallergen measured. In total 209 samples were taken. The highest exposure concentrations both for dust (geometric mean > 30 mg m-3) and for flour aeroallergen (geometric mean > 500 micrograms m-3) were measured during certain operations. Exposure concentrations for the tasks were often much higher than the levels we had measured over a shift in a previous study. This might be important for sensitization and for the development of asthma. Peak exposure concentrations could be used to explore the exposure-response relationship more comprehensively. In general average flour aeroallergen concentrations increased linearly with average dust concentrations, although there were some exceptions.


[The experience of a serological examination of the rural population in a territory endemic for Lyme borreliosis].
Serological surveys have been performed in two communities (Kirovsk District, Leningrad Province) endemic for Lyme's borreliosis. 736 individuals (51% of the population) have been examined. Standard capillary blood samples (drops from the finger on a filter paper) have been investigated by indirect immunofluorescence. Blood sera of residents from nonendemic territories and patients with various rheumatic diseases served as a control. Serum dilution 1:80 was considered as a reliable screening titre. In general, the seroprevalence was shown to account for 9%.


Molecular and biochemical characterization of VR-EILs encoding mung bean ETHYLENE INSENSITIVE3-LIKE proteins.
ETHYLENE INSENSITIVE3 (EIN3) is a transcription factor involved in the ethylene signal transduction pathway in Arabidopsis. Two full-length cDNA clones, pVR-EIL1 and pVR-EIL2, encoding EIN3-LIKE proteins were isolated by reverse transcriptase-polymerase chain reaction and by screening the cDNA library of mung bean (Vigna radiata) hypocotyls. VR-EIL1 and VR-EIL2 share 70% identity and display varying degrees of sequence conservation (39%-65%) with previously isolated EIN3 homologs from Arabidopsis, tobacco (Nicotiana tabacum) and tomato (Lycopersicon esculentum) plants. Gel retardation assay revealed that both VR-EILs were able to interact specifically with optimal binding sequence-1, the recently identified optimal binding sequence for tobacco TEIL, with the binding of VR-EIL2 being more efficient than that of VR-EIL1. Transient expression analysis using a VR-EIL::smGFP fusion gene in onion (Allium cepa) epidermal cells indicated that the VR-EIL proteins were effectively targeted to the nucleus. The fusion protein of VR-EIL2 with GAL4 DNA-binding domain strongly activated transcription of a reporter gene in yeast cells, and an essential domain for transcription-stimulating activity was localized to the amino-terminal acidic region that consists of 50 amino acid residues. In contrast with what has been previously found in EIN3- and TEIL-overexpressing Arabidopsis plants, transgenic tobacco seedlings expressing the VR-EIL genes under the control of cauliflower mosaic virus 35S promoter did not exhibit a constitutive triple response. Instead, they displayed a markedly enhanced proliferation of root hairs, one of the typical ethylene response phenotypes, and increased sensitivity to exogenous ethylene. In addition, the pathogenesis-related (PR) genes encoding beta-1,3-glucanase, osmotin, and PR1 were constitutively expressed in 35S::VR-EIL lines without added ethylene, and were hyperinduced in response to ethylene treatment. These results indicate that VR-EILs are functional in tobacco cells, thereby effectively transactivating the GCC-box-containing PR genes and enhancing sensitivity to ethylene. The possible physiological role of VR-EILs is discussed in the light of the suggestion that they are active components of the ethylene-signaling pathway and their heterologous expressions constitutively turn on a subset of ethylene responses in tobacco plants.


A signal, from human mtDNA, of postglacial recolonization in Europe.
Mitochondrial HVS-I sequences from 10,365 subjects belonging to 56 populations/geographical regions of western Eurasia and northern Africa were first surveyed for the presence of the T-->C transition at nucleotide position 16298, a mutation which has previously been shown to characterize haplogroup V mtDNAs. All mtDNAs with this mutation were then screened for a number of diagnostic RFLP sites, revealing two major subsets of mtDNAs. One is haplogroup V proper, and the other has been termed "pre*V," since it predates V phylogenetically. The rather uncommon pre*V tends to be scattered throughout Europe (and northwestern Africa), whereas V attains two peaks of frequency: one situated in southwestern Europe and one in the Saami of northern Scandinavia. Geographical distributions and ages support the scenario that pre*V originated in Europe before the Last Glacial Maximum (LGM), whereas the more recently derived haplogroup V arose in a southwestern European refugium soon after the LGM. The arrival of V in eastern/central Europe, however, occurred much later, possibly with (post-)Neolithic contacts. The distribution of haplogroup V mtDNAs in modern European populations would thus, at least in part, reflect the pattern of postglacial human recolonization from that refugium, affecting even the Saami. Overall, the present study shows that the dissection of mtDNA variation into small and well-defined evolutionary units is an essential step in the identification of spatial frequency patterns. Mass screening of a few markers identified using complete mtDNA sequences promises to be an efficient strategy for inferring features of human prehistory.


Forced degradation of therapeutic proteins.
The scope of this paper is to review approaches used for forced degradation (synonym, stress testing) of therapeutic proteins. Forced degradation studies play a central role in the development of therapeutic proteins, for example, for candidate selection, molecule characterization, formulation development, assay development, and comparability studies. Typical stress methods are addressed within this review, such as exposure to elevated temperatures, freeze-thawing, mechanical stress, oxidation, light, as well as various materials and devices used in the clinics during final administration. Stability testing is briefly described as far as relevant to the discussion of forced degradation studies. Whereas stability-testing requirements are defined in regulatory guidelines, standard procedures for forced degradation of therapeutic proteins are largely unavailable, except for photostability. Possible selection criteria to identify appropriate stress conditions and recommendations for setting up forced degradation studies for the different phases of development of therapeutic proteins are presented.


Details in the catalytic mechanism of mammalian thioredoxin reductase 1 revealed using point mutations and juglone-coupled enzyme activities.
The mammalian selenoprotein thioredoxin reductase 1 (TrxR1) is a key enzyme in redox regulation, antioxidant defense, and cellular growth. TrxR1 can catalyze efficient reduction of juglone (5-hydroxy-1,4-naphthoquinone; walnut toxin) in a reaction which, in contrast to reduction of most other substrates of TrxR1, is not dependent upon an intact selenocysteine (Sec, U) residue of the enzyme. Using a number of TrxR1 mutant variants, we here found that a sole Cys residue at the C-terminal tail of TrxR1 is required for high-efficiency juglone-coupled NADPH oxidase activity of Sec-deficient enzyme, occurring with mixed one- and two-electron reactions producing superoxide. The activity also utilizes the FAD and the N-terminal redox active disulfide/dithiol motif of TrxR1. If a sole Cys residue at the C-terminal tail of TrxR1, in the absence of Sec, was moved further towards the C-terminal end of the protein compared to its natural position at residue 497, juglone reduction was, surprisingly, further increased. Ala substitutions of Trp407, Asn418 and Asn419 in a previously described "guiding bar", thought to mediate interactions of the C-terminal tail of TrxR1 with the FAD/dithiol site at the N-terminal domain of the other subunit in the dimeric enzyme, lowered turnover with juglone about 4.5-fold. Four residues of Sec-deficient TrxR1 were found to be easily arylated by juglone, including the Cys residue at position 497. Based upon our observations we suggest a model for involvement of the juglone-arylated C-terminal motif of TrxR1 to explain its high activity with juglone. This study thus provides novel insights into the catalytic mechanisms of TrxR1. One-electron juglone reduction by TrxR1 producing superoxide should furthermore contribute to the well-known prooxidant cytotoxicity of juglone.


Optical trapping and propulsion of red blood cells on waveguide surfaces.
We have studied optical trapping and propulsion of red blood cells in the evanescent field of optical waveguides. Cell propulsion is found to be highly dependent on the biological medium and serum proteins the cells are submerged in. Waveguides made of tantalum pentoxide are shown to be efficient for cell propulsion. An optical propulsion velocity of up to 6 µm/s on a waveguide with a width of ~6 µm is reported. Stable optical trapping and propulsion of cells during transverse flow is also reported.


Depressive disorders in primary care: prevalence, functional disability, and identification.
To assess the relative prevalence of subsyndromal depression (SubD) and major depression (MDD) in primary care patients and describe their associated functional impairments, and to define the operating characteristics of a short depression screen (SDS). Three primary care clinics: a university-affiliated Veterans Affairs clinic, a county general internal medicine clinic, and a community health center. Randomly selected adult patients (n = 221), aged > or = 30 years, with no history of psychiatric comorbidity, current substance abuse, major depressive disorder, chronic pain disorder, or dementia. The SDS and the Medical Outcomes Study Short Form 36 (SF-36) were interviewer-administered by an experienced bilingual research assistant to all subjects in the language of their choice. A physician administered independently the Structured Clinical Interview for DSM-III diagnosis (SCID) and the Hamilton Depression Rating Scale (HDRS) to all patients who exceeded a minimum threshold on the SDS and to a randomly selected sample of patients who had subthreshold scores. MDD was defined by DSM-III criteria and SubD was defined as two to four DSM-III criteria, of which one had to be depressed mood or anhedonia. Demographic and clinical characteristics of the patients were: Mexican American 53%, non-Hispanic white 38%, and African American 9%; men 68%; mean age 60 +/- 12.7 years; mean level of education 9.5 +/- 4.4 years; and hypertension 57%, diabetes mellitus 51%, and arthritis 45%. The prevalences of MDD and SubD (adjusted for sampling strategy) were 4% and 16%, respectively. For the patients who had MDD, the median HDRS score was 17 (interquartile range, 10-18), and for those who had SubD, the median HDRS score was 9 (interquartile range, 8-14). Compared with the patients who did not have depressive symptoms, those who had either MDD or SubD were significantly impaired in multiple domains of self-reported function. The sensitivity and specificity of the SDS for MDD were 100% (95% CI 57-100) and 72% (95% CI 63-81), respectively. For depressive disorders (MDD or SubD), the sensitivity was 66% (95% CI 49-83) and the specificity was 79% (95% CI 69-89). SubD was more prevalent than MDD in these primary care settings. Both MDD and SubD were associated with significant functional impairment. The sensitivity of the SDS was lower for identifying depressive disorders (MDD or SubD) than it was for identifying MD.


Interdependent self-construal predicts emotion suppression in Asian Americans: An electro-cortical investigation.
Although people with high interdependent self-construal (SC) are often assumed to be competent in emotion suppression, direct evidence is missing. We tested whether interdependent SC would predict the ability to down-regulate emotional arousal. Americans of both East Asian and European descent were shown a series of pictures. They were instructed to suppress or attend to their emotions. Their electroencephalogram was recorded and analyzed. The late positive potential (LPP) evoked by unpleasant (vs. neutral) pictures (a marker of emotional arousal) was reduced in the suppress (vs. attend) condition. This effect of emotion suppression was more pronounced for those high in interdependent SC than for those low in it. Curiously, the resulting valence x condition x interdependent SC interaction was robust among those of Asian descent, but not those of European descent. The 4-way interaction involving culture was statistically significant. Our work suggests that the purported link between interdependent SC and emotion suppression may be culture-bound.


Simvastatin prevents dopaminergic neurodegeneration in experimental parkinsonian models: the association with anti-inflammatory responses.
In addition to their original applications to lowering cholesterol, statins display multiple neuroprotective effects. N-methyl-D-aspartate (NMDA) receptors interact closely with the dopaminergic system and are strongly implicated in therapeutic paradigms of Parkinson's disease (PD). This study aims to investigate how simvastatin impacts on experimental parkinsonian models via regulating NMDA receptors. Regional changes in NMDA receptors in the rat brain and anxiolytic-like activity were examined after unilateral medial forebrain bundle lesion by 6-hydroxydopamine via a 3-week administration of simvastatin. NMDA receptor alterations in the post-mortem rat brain were detected by [³H]MK-801(Dizocilpine) binding autoradiography. 6-hydroxydopamine treated PC12 was applied to investigate the neuroprotection of simvastatin, the association with NMDA receptors, and the anti-inflammation. 6-hydroxydopamine induced anxiety and the downregulation of NMDA receptors in the hippocampus, CA1(Cornu Ammonis 1 Area), amygdala and caudate putamen was observed in 6-OHDA(6-hydroxydopamine) lesioned rats whereas simvastatin significantly ameliorated the anxiety-like activity and restored the expression of NMDA receptors in examined brain regions. Significant positive correlations were identified between anxiolytic-like activity and the restoration of expression of NMDA receptors in the hippocampus, amygdala and CA1 following simvastatin administration. Simvastatin exerted neuroprotection in 6-hydroxydopamine-lesioned rat brain and 6-hydroxydopamine treated PC12, partially by regulating NMDA receptors, MMP9 (matrix metalloproteinase-9), and TNF-a (tumour necrosis factor-alpha). Our results provide strong evidence that NMDA receptor modulation after simvastatin treatment could partially explain its anxiolytic-like activity and anti-inflammatory mechanisms in experimental parkinsonian models. These findings contribute to a better understanding of the critical roles of simvastatin in treating PD via NMDA receptors.


Myotonic dystrophy type 2: molecular, diagnostic and clinical spectrum.
Myotonic dystrophy types 1 (DM1) and 2 (DM2/proximal myotonic myopathy PROMM) are dominantly inherited disorders with unusual multisystemic clinical features. The authors have characterized the clinical and molecular features of DM2/PROMM, which is caused by a CCTG repeat expansion in intron 1 of the zinc finger protein 9 (ZNF9) gene. Three-hundred and seventy-nine individuals from 133 DM2/PROMM families were evaluated genetically, and in 234 individuals clinical and molecular features were compared. Among affected individuals 90% had electrical myotonia, 82% weakness, 61% cataracts, 23% diabetes, and 19% cardiac involvement. Because of the repeat tract's unprecedented size (mean approximately 5,000 CCTGs) and somatic instability, expansions were detectable by Southern analysis in only 80% of known carriers. The authors developed a repeat assay that increased the molecular detection rate to 99%. Only 30% of the positive samples had single sizeable expansions by Southern analysis, and 70% showed multiple bands or smears. Among the 101 individuals with single expansions, repeat size did not correlate with age at disease onset. Affected offspring had markedly shorter expansions than their affected parents, with a mean size difference of -17 kb (-4,250 CCTGs). DM2 is present in a large number of families of northern European ancestry. Clinically, DM2 resembles adult-onset DM1, with myotonia, muscular dystrophy, cataracts, diabetes, testicular failure, hypogammaglobulinemia, and cardiac conduction defects. An important distinction is the lack of a congenital form of DM2. The clinical and molecular parallels between DM1 and DM2 indicate that the multisystemic features common to both diseases are caused by CUG or CCUG expansions expressed at the RNA level.


Murine Wee1 plays a critical role in cell cycle regulation and pre-implantation stages of embryonic development.
Wee1 kinase regulates the G2/M cell cycle checkpoint by phosphorylating and inactivating the mitotic cyclin-dependent kinase 1 (Cdk1). Loss of Wee1 in many systems, including yeast and drosophila, leads to premature mitotic entry. However, the developmental role of Wee1 in mammals remains unclear. In this study, we established Wee1 knockout mice by gene targeting. We found that Wee-/- embryos were defective in the G2/M cell cycle checkpoint induced by gamma-irradiation and died of apoptosis before embryonic (E) day 3.5. To study the function of Wee1 further, we have developed MEF cells in which Wee1 is disrupted by a tamoxifen inducible Cre-LoxP approach. We found that acute deletion of Wee1 resulted in profound growth defects and cell death. Wee1 deficient cells displayed chromosome aneuploidy and DNA damage as revealed by gamma-H2AX foci formation and Chk2 activation. Further studies revealed a conserved mechanism of Wee1 in regulating mitotic entry and the G2/M checkpoint compared with other lower organisms. These data provide in vivo evidence that mammalian Wee1 plays a critical role in maintaining genome integrity and is essential for embryonic survival at the pre-implantation stage of mouse development.


Peptide receptor radionuclide therapy with 177Lu-DOTATATE for patients with somatostatin receptor-expressing neuroendocrine tumors: the first US phase 2 experience.
Peptide receptor radionuclide therapy with radiolabeled somatostatin analogs is a novel method of treatment in patients with metastatic neuroendocrine tumors (NETs). For the first time in the United States, we present preliminary results of the treatment with Lutetium (177)(Lu) DOTATATE in patients with progressive NETs. Thirty-seven patients with grade 1 and grade 2 disseminated and progressive gastroenteropancreatic NET were enrolled in a nonrandomized, phase 2 clinical trial. Repeated cycles of 200 mCi (7.4 GBq; ±10%) were administered up to the cumulative dose of 800 mCi (29.6 GBq; ±10%). Among 32 evaluable patients, partial response and minimal response to treatment were seen in 28% and 3%, respectively, and stable disease was seen in 41% of patients. A total of 28% had progressive disease. A response to treatment was significantly associated with lower burden of disease in the liver. No significant acute or delayed hematologic or kidney toxicity was observed. An impressive improvement of performance status and quality of life were seen after Lu-DOTATATE therapy. Treatment with multiple cycles of (177)Lu-DOTATATE peptide receptor radionuclide therapy is well tolerated. This treatment results in control of the disease in most patients, whereas systemic toxicities are limited and reversible. Quality of life is also improved.


Technical note: Evaluation of a scoring system for rumen fill in dairy cows.
Changes in feed intake are useful in early detection of disease in dairy cows. Cost and complexity limit our ability to monitor dry matter intake (DMI) of individual cows kept in loose-housing systems. A 5-point subjective scoring system has been developed to visually describe rumen fill, but no work to date has evaluated these scores as an indicator of feed intake. The objective of this study was to evaluate the performance of within-cow changes in visual rumen fill scores as estimates of changes of DMI and feed intake in dairy cows. Our results illustrate that rumen fill scored on a scale from 1 to 5 has substantial intra- (Cohen's kappa coefficient=0.69) and interobserver (Cohen's kappa coefficient=0.68) repeatability. Within-cow changes in visual rumen fill score are correlated with changes in DMI (Spearman's rank correlation=0.68). The depth of the paralumbar fossa (mean +/- SD; 5.6+/-0.9 cm) changes considerably (up to 4.8 cm) within 70+/-5 min. This more objective measure was also correlated with visual rumen fill scores (Spearman's rank correlation=-0.62). Our results indicate that subjective rumen fill scores are statistically associated with both an objective measure of paralumbar fossa indentation and feed intake. However, much of the variation in visual rumen fill scores is not associated with either measure, suggesting that caution is required in clinical usage of these scores.


Dietary Patterns and Blood Pressure in Adults: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.
Hypertension is a major risk factor for developing cardiovascular disease, stroke, and kidney disease. To lower blood pressure (BP), several lifestyle changes are recommended such as weight loss, exercise, and following a healthy diet. Investigating the effect of single nutrients may have positive results, but food is consumed as part of a whole diet, resulting in nutrient interactions. The aim of this systematic review and meta-analysis was to assess the effect of dietary patterns on BP in adults. Studies that were published between January 1999 and June 2014 were retrieved using Scopus, Web of Science, and the MEDLINE database. Seventeen randomized controlled trials were included in the meta-analysis. The results suggest that healthy dietary patterns such as the Dietary Approaches to Stop Hypertension diet, Nordic diet, and Mediterranean diet significantly lowered systolic BP and diastolic BP by 4.26 mm Hg and 2.38 mm Hg, respectively. These diets are rich in fruit, vegetables, whole grains, legumes, seeds, nuts, fish, and dairy and low in meat, sweets, and alcohol. Lifestyle factors such as exercise and weight loss in combination with dietary changes may also reduce BP. Further research is needed to establish the effect of dietary patterns on BP in different cultures other than those identified in this review. The review was registered on PROSPERO (International prospective register of systematic reviews) as CRD42015016272.