Intentional Staged Clipping Following Previous Coil Embolization of Ruptured Aneurysm with a Branch Arising from the Neck: Technical Note.
To present a substitute strategy for clipping: coil embolization of the ruptured aneurysm followed by intentional, staged clipping. We treated five cases of ruptured intracranial aneurysms with branches arising from the neck. The mean aneurysm diameter was 4.4 mm. In the acute rupture phase, coiling was performed without adjunctive endovascular techniques with intentional preservation of the neck in all cases. To treat recurrence during follow-up, the previously coiled aneurysm was clipped, which did not occasion any complications. Postoperative imaging showed completely obliterated aneurysms and preserved branches. No rebleeding occurred during the interval between coiling and clipping, and no rebleeding or recurrence occurred after clipping. No treatment-related complications occurred after coiling and clipping. This two-stage strategy may be effective for hemostasis and branch preservation for small- and medium-sized ruptured aneurysms with a branch arising from the neck. This intentional two-stage strategy can be a substitute strategy for clipping in the acute rupture phase with an acceptable outcome if the patient cannot undergo clipping as the first-line treatment.


Association study of molecular polymorphisms in candidate genes related to stress responses with production and meat quality traits in pigs.
The hypothalamic-pituitary-adrenal (HPA) axis exerts a large range of effects on metabolism, the immune system, inflammatory processes, and brain functions. Together with the sympathetic nervous system, it is also the most important stress-responsive neuroendocrine system. Both systems influence production traits, carcass composition, and meat quality. The HPA axis may be a critical target for genetic selection of more robust animals. Indeed, numerous studies in various species have demonstrated the importance of genetic factors in shaping the individual HPA axis phenotype, and genetic polymorphism can be found at each level of the axis, including hormone production by the adrenal cortices under stimulation by adrenocorticotropic hormone (ACTH), hormone bioavailability, or receptor and postreceptor mechanisms. The aim of the present experiment was to extend these findings to the brain neurochemical systems involved in stress responses. To this end, a number of candidate genes were sequenced for molecular polymorphisms and their association was studied with stress neuroendocrine and production traits in a genetically diverse population consisting of 100 female pigs from an advanced intercross (F10-F12) between 2 highly divergent breeds, Large White (LW) and Meishan (MS). The LW breed has a high production potential for lean meat and a low HPA axis activity, and the MS breed has low growth rate, fat carcasses-but large litters of highly viable piglets-and a high HPA axis activity. Candidate genes were chosen in the catecholaminergic and serotonergic pathways, in the pituitary control of cortisol production, among genes previously demonstrated to be differentially expressed in ACTH-stimulated adrenal glands from LW and MS pigs, and in cortisol receptors. Sixty new polymorphisms were found. The association study with carcass and meat quality traits and with endocrine traits showed a number of significant results, such as monoamine oxidase (MAOA) polymorphisms with growth rate (P = 0.01); lean content and intramuscular fat (P < 0.01), which are the most important traits for carcass value; dopamine receptor D3 (DRD3) with carcass composition (P < 0.05); and vasopressin receptor 1B (AVPR1B) with meat quality traits (P ≤ 0.05). The effect of these polymorphisms on neuroendocrine parameters (eg DRD3 and HPA axis or AVPR1B and catecholamines) indicates information regarding their biological mechanism of action.


Real-time dynamics of string breaking in quantum spin chains.
String breaking is a central dynamical process in theories featuring confinement, where a string connecting two charges decays at the expense of the creation of new particle-antiparticle pairs. Here, we show that this process can also be observed in quantum Ising chains where domain walls get confined either by a symmetry-breaking field or by long-range interactions. We find that string breaking occurs, in general, as a two-stage process. First, the initial charges remain essentially static and stable. The connecting string, however, can become a dynamical object. We develop an effective description of this motion, which we find is strongly constrained. In the second stage, which can be severely delayed due to these dynamical constraints, the string finally breaks. We observe that the associated timescale can depend crucially on the initial separation between domain walls and can grow by orders of magnitude by changing the distance by just a few lattice sites. We discuss how our results generalize to one-dimensional confining gauge theories and how they can be made accessible in quantum simulator experiments such as Rydberg atoms or trapped ions.


Haemoglobin Q-Thailand and hereditary spherocytosis in a Chinese family.
A Chinese family with concurrent hereditary spherocytosis (HS) and haemoglobin (Hb) Q-Thailand is described. The Hb Q-Thailand mutation was found on the remaining alpha1 globin gene on a chromosome 16 containing the (-alpha 4.2) deletion. Active haemolysis in members of this family is segregated with the HS phenotype, and the Hb Q-Thailand in the heterozygous state does not seem to show any modulating effect on HS.


Postpartum depression risk factors among recent refugee, asylum-seeking, non-refugee immigrant, and Canadian-born women: results from a prospective cohort study.
The objective was to examine and compare risk factors for postpartum depression among: (1) recent (≤5 years) migrant and Canadian-born women, and (2) refugee, asylum-seeking, and non-refugee immigrant women. A sample of 1536 women (1024 migrant and 512 Canadian-born) were recruited from 12 hospitals. Women completed questionnaires at 1-2 and 16 weeks postpartum including questions on socio-demographics, biomedical history, health services, and migration and resettlement factors. Bivariate analyses and multivariate logistic regression were performed to examine and compare risk factors for postpartum depressive symptoms at 16 weeks postpartum. Recent migrant women had significantly higher rates (6%) of depressive symptoms at 16 weeks postpartum than Canadian-born women (2.9%). Asylum-seekers had the highest rate (14.3%), followed by refugee (11.5%) and non-refugee immigrant women (5.1%). Migrant women at greatest risk to develop depressive symptoms were those who experienced abuse, had pain post-birth, worried about family members left behind, had food insecurity, and had reduced access to healthcare (limited insurance and/or no regular care-provider). Conversely, those with higher levels of social support and who felt they belonged to a community had a lower risk of developing depressive symptoms. All childbearing recent migrant women should be considered at risk for postpartum depression. To prevent and support migrant women suffering postpartum depressive symptoms, barriers to healthcare need to be addressed and interventions should include assessments and support/programmes for abuse/violence, lack of social support, food insecurity, and stress/poor mental health. Treatment of pain during the postpartum period is also critical.


Visualization of focal sites of transcription within human nuclei.
HeLa cells were encapsulated in agarose microbeads, permeabilized and incubated with Br-UTP in a 'physiological' buffer; then sites of RNA synthesis were immunolabelled using an antibody that reacts with Br-RNA. After extending nascent RNA chains by < 400 nucleotides in vitro, approximately 300-500 focal synthetic sites can be seen in each nucleus by fluorescence microscopy. Most foci also contain a component of the splicing apparatus detected by an anti-Sm antibody. alpha-amanitin, an inhibitor of RNA polymerase II, prevents incorporation into these foci; then, using a slightly higher salt concentration, approximately 25 nucleolar foci became clearly visible. Both nucleolar and extra-nucleolar foci remain after nucleolytic removal of approximately 90% chromatin. An underlying structure probably organizes groups of transcription units into 'factories' where transcripts are both synthesized and processed.


Comparison of diagnostic accuracy of Doppler USG and contrast-enhanced magnetic resonance angiography and selective renal arteriography in patients with atherosclerotic renal artery stenosis.
There are many systemic complications of conventional selective renal arteriography (SRA), such as contrast-mediated nephropathy. Contrast-enhanced magnetic resonance angiography (CE-MRA) and renal artery Doppler ultrasonography (DUSG) have been used increasingly for renal artery stenosis (RAS). The aim of this study was to evaluate the accuracy of CE-MRA and DUSG as used for diagnosis of RAS. We divided 130 consecutive patients investigated for resistant hypertension into 2 groups based on age: group 1 was patients <60 years old and group 2 was patients >60 year. DUSG, CE-MRA, and SRA were performed in group 1 and group 2 patients. Seventy-two patients (24 males [M], 48 females [F]) in group 1, and 58 patients (26 M, 32 F) in group 2 were included in the study. In the evaluation of clinically significant renal artery stenosis with DUSG, in group 1 the overall sensitivity was 83.33% and overall specificity was 81.82%, and in group 2 they were 69.23% and 0%, respectively, when compared with SRA. In the evaluation of clinically significant renal artery stenosis with CE-MRA, the overall sensitivity and specificity were 92.31% and 36.36%, respectively, in group 1 and 100.00% and 73.33%, respectively in group 2, when compared with SRA. CE-MRA is an accurate, non-invasive method for the diagnosis of RAS in patients above 60 years of age and DUSG may be the choice of diagnostic method for RAS in patients under 60 years of age.


Dietary fluoride restriction does not alter femoral biomechanical strength in col1a2-deficient (oim) mice with type I collagen glomerulopathy.
Osteogenesis imperfecta (OI) is a clinically and genetically heterogeneous disease due primarily to mutations in the type I procollagen genes, COL1A1 and COL1A2, causing bone deformity and numerous lifetime fractures. OI murine (oim) model mice carry a mutation in the col1a2 gene causing aberrant production of homotrimeric type I collagen [α1(I)(3)], leading to bone fragility and glomerular accumulation of type I collagen. Previous studies demonstrated that heterozygous (+/oim) and homozygous (oim/oim) mice have elevated tibiae fluoride concentrations but reduced femoral biomechanics. However, it is unclear whether these 2 variables are causally related, because impaired renal function could reduce urinary fluoride excretion, thus elevating bone fluoride concentrations regardless of disease status. Our goal in this study was to determine whether dietary fluoride restriction would improve femoral biomechanics in oim mice. Wild-type, +/oim, and oim/oim mice were fed a control (5 mg/kg fluoride) or fluoride-restricted diet (0 mg/kg fluoride) for ∼13 wk, at which time plasma and femora were analyzed for fluoride concentrations and bone biomechanical properties. In wild-type, +/oim, and oim/oim mice, dietary fluoride restriction reduced femoral fluoride burden by 54-74%, respectively (P < 0.05), without affecting glomerular collagen deposition. Oim/oim mice fed the fluoride-restricted diet had reduced material tensile strength (P < 0.05) compared with oim/oim mice fed the control diet. However, dietary fluoride restriction did not affect stiffness or whole bone femoral breaking strength, regardless of genotype. These data suggest that oim mice have reduced bone strength due to homotrimeric type I collagen, independent of bone fluoride content.


Research on the characteristics of the water quality of rainwater runoff from green roofs.
This paper investigates the water quality characteristics of rainwater runoff from dual-substrate-layer green roofs in Tianjin, China. The data were collected from four different assemblies and three types of simulated rains. The storm-water runoff quality was monitored from early June through late October 2012 and from July through late November 2013. The results revealed that the runoff water quality would be improved to some extent with the ageing of green roofs and that the quality retention rate better reflected the pollutant retention capacity of the green roof than the pollutant concentration in the runoff water. The investigation clearly demonstrated that green roofs also effectively reduced the chemical oxygen demand and turbidity value and neutralised acid rain to stabilise the pH of the runoff.


A multi-level typology of abstract visualization tasks.
The considerable previous work characterizing visualization usage has focused on low-level tasks or interactions and high-level tasks, leaving a gap between them that is not addressed. This gap leads to a lack of distinction between the ends and means of a task, limiting the potential for rigorous analysis. We contribute a multi-level typology of visualization tasks to address this gap, distinguishing why and how a visualization task is performed, as well as what the task inputs and outputs are. Our typology allows complex tasks to be expressed as sequences of interdependent simpler tasks, resulting in concise and flexible descriptions for tasks of varying complexity and scope. It provides abstract rather than domain-specific descriptions of tasks, so that useful comparisons can be made between visualization systems targeted at different application domains. This descriptive power supports a level of analysis required for the generation of new designs, by guiding the translation of domain-specific problems into abstract tasks, and for the qualitative evaluation of visualization usage. We demonstrate the benefits of our approach in a detailed case study, comparing task descriptions from our typology to those derived from related work. We also discuss the similarities and differences between our typology and over two dozen extant classification systems and theoretical frameworks from the literatures of visualization, human-computer interaction, information retrieval, communications, and cartography.


The role of social support in family socio-economic disparities in depressive symptoms during early pregnancy: Evidence from a Chinese birth cohort.
Few studies have examined the association between a composite measure of family socio-economic position (SEP)2 and depressive symptoms among Chinese pregnant women, nor any potential underlying mechanisms which may be amenable to preventative interventions. We investigated the association between a composite SEP measure and depressive symptoms during early pregnancy, and tested for mediation by social support and moderation by parity in the Born in Guangzhou Cohort Study (n = 12,382) using adjusted logistic regression and causal mediation analysis. In this population, 18.4% of women experienced depressive symptoms before the 20th gestational week. Compared with the highest quartile, the lowest SEP score quartile was associated with a higher risk of depressive symptoms (OR 1.50, 95% CI 1.31-1.71), which was more pronounced among multiparous women than nulliparous women (P for interaction <0.001). Social support mediated the association between SEP and depressive symptoms, with greater proportion mediated in nulliparous women (73.4% for the lowest SEP score quartile) than multiparous women (30.5%). Depressive symptoms were measured by Self-rated Depression Scale, which is not designed as a clinical diagnosis tool for depression. We only had information on perceived social support but not actual social support, although these two parameters were modestly correlated. Lower SEP was associated with higher risk of depressive symptoms in pregnant women, driven by social support. And the association between SEP and depressive symptoms and mediation by social support were modified by parity. Whether this association extends to the post pregnancy period or is amenable to cost-effective interventions should be investigated in further studies.


Effects of alpha-adrenergic blockade on coronary autoregulation in dogs.
In alpha-chloralose-anesthetized, open-chest dogs, left coronary blood flow and oxygen extraction were measured at perfusion pressures from 25 to 175 mmHg. Pressure-flow relationships were obtained before and after alpha-blockade with prazosin or alpha 2-blockade with yohimbine in the presence of beta-blockade with propranolol. The efficiency of flow autoregulation with increments in pressure was calculated as the closed-loop gain and as the slope of flow to pressure. In all animals the highest gain and the lowest slope were seen when pressure increased from 75 to 125 mmHg. In the prazosin group before alpha-blockade the gain with this pressure increase was 0.76 +/- 0.05, whereas in the yohimbine group before alpha-blockade this gain was 0.75 +/- 0.03. Prazosin reduced the gain within the pressure range of 75-125 mmHg by 41%, and this reduction was statistically significant (P less than 0.05). Prazosin also significantly increased the flow to pressure slope within this range. In addition, at all pressures greater than or equal to 50 mmHg, the level of coronary flow was significantly increased by prazosin. The increase in flow and the reduction in autoregulatory efficiency with prazosin were not attributable to an increase in myocardial oxygen consumption. Yohimbine had no effects on the level of coronary flow or on the autoregulatory efficiency. These data indicate that an alpha 1-adrenoceptor mediated coronary constriction modulates coronary autoregulation.


Band structure evolution during the ultrafast ferromagnetic-paramagnetic phase transition in cobalt.
The evolution of the electronic band structure of the simple ferromagnets Fe, Co, and Ni during their well-known ferromagnetic-paramagnetic phase transition has been under debate for decades, with no clear and even contradicting experimental observations so far. Using time- and spin-resolved photoelectron spectroscopy, we can make a movie on how the electronic properties change in real time after excitation with an ultrashort laser pulse. This allows us to monitor large transient changes in the spin-resolved electronic band structure of cobalt for the first time. We show that the loss of magnetization is not only found around the Fermi level, where the states are affected by the laser excitation, but also reaches much deeper into the electronic bands. We find that the ferromagnetic-paramagnetic phase transition cannot be explained by a loss of the exchange splitting of the spin-polarized bands but instead shows rapid band mirroring after the excitation, which is a clear signature of extremely efficient ultrafast magnon generation. Our result helps to understand band structure formation in these seemingly simple ferromagnetic systems and gives first clear evidence of the transient processes relevant to femtosecond demagnetization.


Interactions of human hMSH2 with hMSH3 and hMSH2 with hMSH6: examination of mutations found in hereditary nonpolyposis colorectal cancer.
Mutations in the human mismatch repair protein hMSH2 have been found to cosegregate with hereditary nonpolyposis colorectal cancer (HNPCC). Previous biochemical and physical studies have shown that hMSH2 forms specific mispair binding complexes with hMSH3 and hMSH6. We have further characterized these protein interactions by mapping the contact regions within the hMSH2-hMSH3 and the hMSH2-hMSH6 heterodimers. We demonstrate that there are at least two distinct interaction regions of hMSH2 with hMSH3 and hMSH2 with hMSH6. Interestingly, the interaction regions of hMSH2 with either hMSH3 or hMSH6 are identical and there is a coordinated linear orientation of these regions. We examined several missense alterations of hMSH2 found in HNPCC kindreds that are contained within the consensus interaction regions. None of these missense mutations displayed a defect in protein-protein interaction. These data support the notion that these HNPCC-associated mutations may affect some other function of the heterodimeric complexes than simply the static interaction of hMSH2 with hMSH3 or hMSH2 with hMSH6.


[Pharmaceutical Care at the primary care clinic in Garðabær - number and type of drug therapy problems identified among elderly clients].
Elderly people are a rising population in Iceland. With higher age the likelihood of drug consumption increases and thus drug therapy problems. Pharmaceutical care has been established abroad, where the pharmacist works in collaboration with other healthcare professionals to reduce patients' drug therapy problems. The aim of this research was to study the number and types of drug therapy problems of older individuals in primary care in Garðabær, by providing pharmacist-led pharmaceutical care in collaboration with general practitioners. Five general practitioners selected patients, 65 years and older, and asked the pharmacist to provide them with pharmaceutical care service. The pharmacist provided pharmaceutical care using a well-defined process. A total of 100 patients participated in the research, 44 men and 56 women. On average the pharmacist identified two drug therapy problems per patient. The most frequent drug therapy problem was related to noncompliance (30.1%), next was adverse drug reaction (26.7%) and the third was unnecessary drug therapy (18.2%). Almost all pharmacist comments were accepted by the general practitioners (90.3%). Our results reveal that a pharmacist providing pharmaceutical care makes, on average, two comments regarding each drug therapy. In almost all cases the general practitioners accept the comments.


Evaluation for regional difference of skin-gas ethanol and sweat rate using alcohol dehydrogenase-mediated fluorometric gas-imaging system (sniff-cam).
Skin gas that contains volatile metabolites (volatilome) is emanated continuously and is thus expected to be suitable for non-invasive monitoring. The aim of this study was to investigate the relationship between the regional difference of sweat rate and skin volatilome distribution to identify the suitable site to monitor metabolisms. In this study, we developed a biofluorometric gas-imaging system (sniff-cam) based on nicotinamide adenine dinucleotide (NAD)-dependent alcohol dehydrogenase (ADH) to visualize transcutaneous ethanol (EtOH) distribution. The EtOH distribution was converted to a fluorescence distribution of reduced NAD with autofluorescence property. First, we optimized the solution volume and concentration of the oxidized NAD, which was a coenzyme of ADH. Owing to the optimization, a two-dimensional distribution of EtOH could be visualized from 0.05-10 ppm with good sensitivity and selectivity. Subsequently, transcutaneous EtOH imaging and measurement of sweat rate were performed at the palm, dorsum of hand, and wrist of participants who consumed alcohol. Transcutaneous EtOH from all skin parts was imaged using the sniff-cam; the concentrations initially increased until 30 min after drinking, followed by a gradual decrease. Although the determined peak EtOH concentrations of typical subjects were approximately 1100 ± 35 ppb (palm), which were higher than 720 ± 18 ppb (dorsum) and 620 ± 13 ppb (wrist), the results of sweat rate suggested that the dorsum of hand and the wrist were appropriate sites. Finally, the sniff-cam could visualize the individual difference of alcohol metabolism capacity originating from aldehyde dehydrogenase phenotype by imaging transcutaneous EtOH.


The evidence for antidepressant use in bipolar depression.
Mood elevation, which includes mania, hypomania, and mixed states, was previously considered the defining symptom of bipolar disorder, but bipolar depression by comparison is actually a much more substantial challenge to diagnose and treat. Recent studies, including research by the Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD), found that patients with bipolar disorder spend longer periods of time in depressive episodes and are more likely to relapse to depression compared with mania or hypomania. However, the treatment of bipolar depression is hampered by the limited number and varying quality of available studies of pharmacologic treatments to guide clinical decision making. Clinicians should rely on studies with the highest level of evidence (category A) when prescribing appropriate antidepressant treatments. The standard care pathways outlined by STEP-BD to aid clinicians in treating varying phases of bipolar disorder provide data on the use of various treatments for bipolar depression and their outcomes. While some treatments have the potential to induce mania, others appear to have some efficacy without inducing mania.


Global sensing of the antigenic structure of herpes simplex virus gD using high-throughput array-based SPR imaging.
While HSV-2 typically causes genital lesions, HSV-1 is increasingly the cause of genital herpes. In addition, neonatal HSV infections are associated with a high rate of mortality and HSV-2 may increase the risk for HIV or Zika infections, reinforcing the need to develop an effective vaccine. In the GSK Herpevac trial, doubly sero-negative women were vaccinated with a truncated form of gD2 [gD2(284t)], then examined for anti-gD serum titers and clinical manifestations of disease. Surprisingly, few vaccinees were protected against genital HSV-2 but 86% were protected from genital HSV-1. These observations suggest that subtle differences in gD structure might influence a protective response. To better understand the antigenic structure of gD and how it impacts a protective response, we previously utilized several key anti-gD monoclonal antibodies (mAbs) to dissect epitopes in vaccinee sera. Several correlations were observed but the methodology limited the number of sera and mAbs that could be tested. Here, we used array-based surface plasmon imaging (SPRi) to simultaneously measure a larger number of protein-protein interactions. We carried out cross-competition or "epitope binning" studies with 39 anti-gD mAbs and four soluble forms of gD, including a form [gD2(285t)] that resembles the Herpevac antigen. The results from these experiments allowed us to organize the mAbs into four epitope communities. Notably, relationships within and between communities differed depending on the form of gD, and off-rate analysis suggested differences in mAb-gD avidity depending on the gD serotype and length. Together, these results show that gD1 and gD2 differ in their structural topography. Consistent with the Herpevac results, several mAbs that bind both gD1 and gD2 neutralize only HSV-1. Thus, this technology provides new insights into the antigenic structure of gD and provides a rationale as to how vaccination with a gD2 subunit may lead to protection from HSV-1 infection.


Readily accessible CT scoring method to quantify fibrosis in IPF.
There is currently no readily accessible measure to specifically quantify the amount of fibrosis in idiopathic pulmonary fibrosis (IPF). Such a measure could isolate contribution of fibrosis from other comorbidities to lung function abnormality and deterioration of disease, and potentially help determine if there has been response to antifibrotic treatment. In a pilot study of 39 IPF patients, we used a CT-based visual scoring method to examine the correlation between the sum of all fibrotic features (all traction bronchiectasis, ground glass with traction bronchiectasis, honeycombing and reticulation; referred to as Total Fibrosis Score, TFS) or the individual fibrotic features, with lung function, Composite Physiologic Index (CPI) and time to death in the 5 years following CT measurement. TFS measurements were highly reproducible (r=0.982; p<0.001) and correlated significantly with TLCO, FVC and CPI. Traction bronchiectasis score was superior to others in its correlation to lung function and CPI, and as good as TFS. TFS and traction bronchiectasis score were also the best correlates (individually) to time to death (r=0.60 for both, and p=0.002 and p=0.004, respectively). We suggest that TFS and our 6-slices method of quantifying traction bronchiectasis on CT scans could be readily accessible and simple methods of quantifying lung fibrosis in IPF. These scores could assist in determining if clinical deterioration is due to worsening fibrosis, for correlation of research findings to amount of lung fibrosis, and to stratify patients for established drug treatment and clinical trials. Our findings also provide a basis for larger studies to validate these findings and determine if the scores could measure change in fibrosis.


Anesthesia timing for children undergoing therapeutic cardiac catheterization after upper respiratory infection: a prospective observational study.
We aimed to analyze anesthesia timing and perioperative respiratory adverse event (PRAE) risk factors in children undergoing therapeutic cardiac catheterization after upper respiratory tract infection (URI). We prospectively included children for elective therapeutic cardiac catheterization. Parents or legal guardians were asked to complete a questionnaire on the child's demographics, tobacco exposure, and URI symptoms. PRAEs (laryngospasm, bronchospasm, coughing, airway secretion, airway obstruction, and oxygen desaturation) as well as details of anesthesia management were recorded. Of 332 children, 201 had a history of URI in the preceding eight weeks. The occurrence rate of PRAEs in children with URI≤two weeks reached the highest proportion, which was higher than that in children without URI (66.3% vs. 46.6%, P=0.007). The overall incidence of PRAEs in children with URI in 3-8 weeks was significantly lower than that in children with URI in the recent ≤two weeks (49.0% vs. 66.3%, P=0.007), and similar to that in the control group (49.0% vs. 46.6%). Multivariate analysis showed association between PRAEs and type of congenital heart disease (CHD) (P<0.001), anesthesia timing (P=0.007), and age (P=0.021). Delayed schedule (two weeks after URI) minimized the risk of PRAEs to the level comparable to that observed in children without URI (OR, 1.11; 95% CI: 0.64-1.91; P=0.707). If treatment is not urgent, a pediatric patient at a high risk of PRAEs will be benefit from the postponement of an interventional operation by at least two weeks after URI.


Effect of Wnt3a on keratinocytes utilizing in vitro and bioinformatics analysis.
Wingless-type (Wnt) signaling proteins participate in various cell developmental processes. A suppressive role of Wnt5a on keratinocyte growth has already been observed. However, the role of other Wnt proteins in proliferation and differentiation of keratinocytes remains unknown. Here, we investigated the effects of the Wnt ligand, Wnt3a, on proliferation and differentiation of keratinocytes. Keratinocytes from normal human skin were cultured and treated with recombinant Wnt3a alone or in combination with the inflammatory cytokine, tumor necrosis factor α (TNFα). Furthermore, using bioinformatics, we analyzed the biochemical parameters, molecular evolution, and protein-protein interaction network for the Wnt family. Application of recombinant Wnt3a showed an anti-proliferative effect on keratinocytes in a dose-dependent manner. After treatment with TNFα, Wnt3a still demonstrated an anti-proliferative effect on human keratinocytes. Exogenous treatment of Wnt3a was unable to alter mRNA expression of differentiation markers of keratinocytes, whereas an altered expression was observed in TNFα-stimulated keratinocytes. In silico phylogenetic, biochemical, and protein-protein interaction analysis showed several close relationships among the family members of the Wnt family. Moreover, a close phylogenetic and biochemical similarity was observed between Wnt3a and Wnt5a. Finally, we proposed a hypothetical mechanism to illustrate how the Wnt3a protein may inhibit the process of proliferation in keratinocytes, which would be useful for future researchers.


Analysis of worm burden variation in human Schistosoma mansoni infections by determination of serum levels of circulating anodic antigen and circulating cathodic antigen.
Serum circulating anodic antigen (CAA) and circulating cathodic antigen (CCA) concentrations, as a possible direct measure of worm burden, were compared with fecal egg counts in a heavily Schistosoma mansoni-infected population from Zaire to allow differentiation between worm loads and worm fecundity in relation to age and intensity of infection. Of the 517 subjects, 95% excreted eggs and 97% demonstrated circulating antigens. Fecal egg counts showed an age-related pattern characteristic for an area in which schistosomiasis is endemic with intense transmission levels. Regression analysis showed that antigen concentrations were strongly associated with egg counts. For CAA, but not for CCA, this relation was found to be nonlinear, which would be consistent with density-dependent fecundity or crowding. The trend was uniform for all age groups, which for this particular population indicated a genuine reduction of worm loads rather than reduced worm fecundity with age of the host.


Analysis of relationship between Anopheles subpictus larval densities and environmental parameters using Remote Sensing (RS), a Global Positioning System (GPS) and a Geographic Information System (GIS).
Remote Sensing (RS), a Global Positioning System (GPS) and a Geographic Information System (GIS) were used to analyze relationship between Anopheles subpictus larval densities and environmental parameters in the Sekotong district on Lombok Island, Indonesia. Distance from the coast to larval habitats, season and surface water were considered as environmental parameters for determining An. subpictus larval densities. Japanese Earth Resources Satellite (JERS) Visible and Near Infrared Radiometer (VNIR) satellite imagery for the area acquired by National Space Development Agency of Japan (NASDA) were used to detect water, which could be used to characterize larval habitats. Data on larval sampling sites obtained from a GPS were entered into a GIS for mapping larval habitats to measure distance between the coast and the larval habitats. A GIS was used for overlaying of data coverages (i.e., water distribution from RS data and larval habitats coupled with data on larval densities) to identify factors that may explain the spatial distribution patterns of larval densities. An. subpictus larval densities were significantly associated with season and distance from the coast to larval habitats. The rainy season and the distance from the coast to larval habitats were critical environmental determinants for presence of An. subpictus larvae in the study. In this paper, we investigated relationship between An. subpictus larval densities and the environmental parameters using RS/GPS/GIS to determine if these tools could be used to predict larval densities.


Nonoperative treatment compared with plate fixation of displaced midshaft clavicular fractures. A multicenter, randomized clinical trial.
Recent studies have shown a high prevalence of symptomatic malunion and nonunion after nonoperative treatment of displaced midshaft clavicular fractures. We sought to compare patient-oriented outcome and complication rates following nonoperative treatment and those after plate fixation of displaced midshaft clavicular fractures. In a multicenter, prospective clinical trial, 132 patients with a displaced midshaft fracture of the clavicle were randomized (by sealed envelope) to either operative treatment with plate fixation (sixty-seven patients) or nonoperative treatment with a sling (sixty-five patients). Outcome analysis included standard clinical follow-up and the Constant shoulder score, the Disability of the Arm, Shoulder and Hand (DASH) score, and plain radiographs. One hundred and eleven patients (sixty-two managed operatively and forty-nine managed nonoperatively) completed one year of follow-up. There were no differences between the two groups with respect to patient demographics, mechanism of injury, associated injuries, Injury Severity Score, or fracture pattern. Constant shoulder scores and DASH scores were significantly improved in the operative fixation group at all time-points (p = 0.001 and p < 0.01, respectively). The mean time to radiographic union was 28.4 weeks in the non-operative group compared with 16.4 weeks in the operative group (p = 0.001). There were two nonunions in the operative group compared with seven in the nonoperative group (p = 0.042). Symptomatic malunion developed in nine patients in the nonoperative group and in none in the operative group (p = 0.001). Most complications in the operative group were hardware-related (five patients had local irritation and/or prominence of the hardware, three had a wound infection, and one had mechanical failure). At one year after the injury, the patients in the operative group were more likely to be satisfied with the appearance of the shoulder (p = 0.001) and with the shoulder in general (p = 0.002) than were those in the nonoperative group. Operative fixation of a displaced fracture of the clavicular shaft results in improved functional outcome and a lower rate of malunion and nonunion compared with nonoperative treatment at one year of follow-up. Hardware removal remains the most common reason for repeat intervention in the operative group. This study supports primary plate fixation of completely displaced midshaft clavicular fractures in active adult patients.


Chronic osteoarthritis and adherence to exercise: a review of the literature.
Osteoarthritis (OA) is a chronic disease that disables many aging adults. People with OA are often asked to adhere to prescribed exercise regimens that must be undertaken in the presence of pain and other disease-related symptoms. We conducted a review of literature that focused on what is known about exercise adherence and the factors that influence exercise adherence among people with OA. Results revealed multiple determinants of exercise adherence; however, these determinants have not been carefully studied in the context of exercise adherence and OA. Almost all studies of exercise adherence among people with OA are short-term and do not use validated measures of adherence. Moreover, poor adherence is the most compelling explanation for the declining impact of the benefits of exercise over time. We conclude that interventions to enhance self-efficacy, social support, and skills in long-term monitoring of progress are necessary to foster exercise adherence among people with OA.


Dual and Specific Inhibition of NAMPT and PAK4 By KPT-9274 Decreases Kidney Cancer Growth.
Kidney cancer (or renal cell carcinoma, RCC) is the sixth most common malignancy in the United States and one of the relatively few whose incidence is increasing. Because of the near universal resistance which occurs with the use of current treatment regimens, reprogrammed metabolic pathways are being investigated as potential targets for novel therapies of this disease. Borrowing from studies on other malignancies, we have identified the PAK4 and NAD biosynthetic pathways as being essential for RCC growth. We now show, using the dual PAK4/NAMPT inhibitor KPT-9274, that interference with these signaling pathways results in reduction of G2-M transit as well as induction of apoptosis and decrease in cell invasion and migration in several human RCC cell lines. Mechanistic studies demonstrate that inhibition of the PAK4 pathway by KPT-9274 attenuates nuclear β-catenin as well as the Wnt/β-catenin targets cyclin D1 and c-Myc. Furthermore, NAPRT1 downregulation, which we show occurs in all RCC cell lines tested, makes this tumor highly dependent on NAMPT for its NAD requirements, such that inhibition of NAMPT by KPT-9274 leads to decreased survival of these rapidly proliferating cells. When KPT-9274 was administered in vivo to a 786-O (VHL-mut) human RCC xenograft model, there was dose-dependent inhibition of tumor growth with no apparent toxicity; KPT-9274 demonstrated the expected on-target effects in this mouse model. KPT-9274 is being evaluated in a phase I human clinical trial in solid tumors and lymphomas, which will allow this data to be rapidly translated into the clinic for the treatment of RCC. Mol Cancer Ther; 15(9); 2119-29. ©2016 AACR.


Nucleotide sequence of the 3'-terminal region of potato virus YN RNA.
The sequence of the 3'-terminal 1611 nucleotides of the genome of the tobacco veinal necrosis strain of potato virus Y (PVYN) was determined. The sequence revealed an open reading frame of 1285 nucleotides, of which the start was not identified, and an untranslated region of 316 nucleotides upstream of a poly(A) tract. Comparison of the open reading frame with the amino-terminal sequence of the viral coat protein enabled mapping of the start of the coat protein at amino acid -267, and indicated that maturation of this protein requires proteolytic processing from a larger polyprotein precursor at a glutamine/glycine dipeptide sequence. The coat protein of PVYN displayed significant (51 to 63%) sequence homology to the coat proteins of four other potyviruses, tobacco etch virus, tobacco vein mottling virus, plum pox virus and sugarcane mosaic virus. Even higher sequence homology (91%) was detected with the coat protein of a fifth potyvirus, pepper mottle virus (PeMV). This homology was of the same level as found between the coat proteins of PVYN and a second strain of this virus, PVYD. Since, moreover, PVYN and PeMV were the only potyviruses displaying homology in the 3'-terminal, non-translated regions of their genomes, we conclude that PeMV should be regarded as a strain of PVY.


Comparison of ophthalmic measurements obtained via high-frequency ultrasound imaging in four species of snakes.
To measure the dimensions of the eyes of living snakes by use of high-frequency ultrasound imaging and correlate those measurements with age, length, and weight. Animals-14 clinically normal snakes. Species, age, length, weight, and horizontal spectacle diameter were recorded, and each snake underwent physical and ophthalmic examinations; ultrasonographic examination of both eyes was performed by use of a commercially available ultrasound unit and a 50-MHz transducer. Ultrasonographic measurements included spectacle thickness, subspectacular space depth, corneal thickness, anterior chamber depth, lens thickness, vitreous cavity depth, and globe length. All measurements were made along the visual axis. 2 corn snakes, 5 California king snakes, 1 gopher snake, and 6 ball pythons were examined. There were no significant differences within or between the species with regard to mean spectacle thickness, corneal thickness, or subspectacular space depth. However, mean horizontal spectacle diameter, anterior chamber depth, and axial globe length differed among the 4 species; for each measurement, ball pythons had significantly larger values than California king snakes. Spectacle thickness, subspectacular space depth, and corneal thickness were similar among the species of snake examined and did not vary significantly with age, length, or weight. Measurements of these dimensions can potentially serve as baseline values to evaluate snakes of these species with a retained spectacle, subspectacular abscess, or subspectacular fluid accumulation. Anterior chamber depth and axial length appeared variable among species, but axial length did not vary with age, length, or weight in the species studied.


Lactic dehydrogenase and cancer: an overview.
Despite the intense scientific efforts made, there are still many tumors that are difficult to treat and the percentage of patient survival in the long-term is still too low. Thus, new approaches to the treatment of cancer are needed. Cancer is a highly heterogeneous and complex disease, whose development requires a reorganization of cell metabolism. Most tumor cells downregulate mitochondrial oxidative phosphorylation and increase the rate of glucose consumption and lactate release, independently of oxygen availability (Warburg effect). This metabolic rewiring is largely believed to favour tumor growth and survival, although the underlying molecular mechanisms are not completely understood. Importantly, the correlation between the aerobic glycolysis and cancer is widely regarded as a useful biochemical basis for the development of novel anticancer strategies. Among the enzymes involved in glycolysis, lactate dehydrogenase (LDH) is emerging as a very attractive target for possible pharmacological approaches in cancer therapy. This review addresses the state of the art and the perspectives concerning LDH both as a useful diagnostic marker and a relevant molecular target in cancer therapy and management.


Ordered and dynamic assembly of single spliceosomes.
The spliceosome is the complex macromolecular machine responsible for removing introns from precursors to messenger RNAs (pre-mRNAs). We combined yeast genetic engineering, chemical biology, and multiwavelength fluorescence microscopy to follow assembly of single spliceosomes in real time in whole-cell extracts. We find that individual spliceosomal subcomplexes associate with pre-mRNA sequentially via an ordered pathway to yield functional spliceosomes and that association of every subcomplex is reversible. Further, early subcomplex binding events do not fully commit a pre-mRNA to splicing; rather, commitment increases as assembly proceeds. These findings have important implications for the regulation of alternative splicing. This experimental strategy should prove widely useful for mechanistic analysis of other macromolecular machines in environments approaching the complexity of living cells.


Testing mechanistic models of growth in insects.
Insects are typified by their small size, large numbers, impressive reproductive output and rapid growth. However, insect growth is not simply rapid; rather, insects follow a qualitatively distinct trajectory to many other animals. Here we present a mechanistic growth model for insects and show that increasing specific assimilation during the growth phase can explain the near-exponential growth trajectory of insects. The presented model is tested against growth data on 50 insects, and compared against other mechanistic growth models. Unlike the other mechanistic models, our growth model predicts energy reserves per biomass to increase with age, which implies a higher production efficiency and energy density of biomass in later instars. These predictions are tested against data compiled from the literature whereby it is confirmed that insects increase their production efficiency (by 24 percentage points) and energy density (by 4 J mg(-1)) between hatching and the attainment of full size. The model suggests that insects achieve greater production efficiencies and enhanced growth rates by increasing specific assimilation and increasing energy reserves per biomass, which are less costly to maintain than structural biomass. Our findings illustrate how the explanatory and predictive power of mechanistic growth models comes from their grounding in underlying biological processes.


The role of Narrow Band Imaging (NBI) in the diagnosis of sinonasal diseases.
Narrow band imaging (NBI) endoscopy is an optical method that helps to characterise tissue vasculature. Its appli- cation in sinonasal pathology remains scarce and a systematic study of its application to rhinology is lacking. The aim of this study is to analyse and describe the normal sinonasal mucosa under NBI light and to characterise the microvascular features of various sinonasal pathologies. We also want to suggest a classification of the patterns, peculiar to this district, and to evaluate whether they can be indicative of a specific physiological or pathological condition. Digital videos and images under white light and NBI of 103 patients (82 evaluated) with 29 sinonasal pathologies and 55 controls (33 evaluated). were independently analysed by three otolaryngologists and the final pattern was then arranged for each image, reaching an agreement between the individual evaluations. Once the appearance of normal sinonasal (SN) mucosa was established (SN1), four patterns for the pathological mucosa were described and a working classification was proposed (SN2, SN3, SN4, SN5). We calculated specificity (80.6% vs 90.6%), sensi- tivity (20% vs 38.5%), PPV (46.1% vs 50%), NPV (54.7% vs 85.7%) and accuracy (53% vs 80.3%) of the ability of SN4 and SN5 pattern to discriminate between benign and malignant nasal neoformations. This is the first study to propose a systematic NBI description and a classification of the vasculature of healthy and pathological mucosa in the sinonasal tract. Our preliminary results show that this technique can help in the workup of several rhinologic conditions and especially in distinguishing benign from malignant tumors.


Variability among strain variables derived from two-dimensional speckle tracking echocardiography in dogs by use of various software.
To determine variability of global longitudinal strain (GLS) and strain rate (SR) measurements in dogs with and without cardiac disease derived from 2-D speckle tracking echocardiography (STE) by use of various software. 2 cohorts comprising 44 dogs (23 cardiovascularly healthy and 21 with cardiac disease) and 40 dogs (18 cardiovascularly healthy and 22 with cardiac disease). Transthoracic echocardiographic images in each cohort were analyzed with vendor-independent software and vendor-specific 2-D STE software for each of 2 vendors. Values for GLS and SR obtained from the same left parasternal apical views with various software were compared. Intraobserver and interobserver variability was determined, and agreement among results for the various software was assessed. Strain analysis was not feasible with vendor-independent software for 20% of images obtained with the ultrasonography system of vendor 1. Intraobserver and interobserver coefficient of variation was < 10% for GLS values, whereas SR measurements had higher variance. There was a significant difference in GLS and SR obtained for each cohort with different software. Evaluation of Bland-Altman plots revealed wide limits of agreement, with variance for GLS of up to 6.3 units in a single dog. Results of longitudinal strain analysis were not uniform among software, and GLS was the most reproducible measurement. Significant variability in results among software warrants caution when referring to reference ranges or comparing serial measurements in the same patient because changes of < 6.5% in GLS might be within measurement error for different postprocessing software.


Spatial analysis of condyle position according to sagittal skeletal relationship, assessed by cone beam computed tomography.
The study aims to compare the condylar position in patients with different anteroposterior sagittal skeletal relationships through a cone beam computed generated tomography (CBCT) imaging generated space analysis. This was a retrospective study of clinically justified, previously taken CBCT images of 45 subjects. Based on a proper sample calculation, three groups of 15 CBCT images each were made according to their ANB angle and facial pattern: class I (normo facial pattern) and class II and III (long facial pattern). The CBCT images were of adult patients between 18 and 35 years old, with full permanent dentition at maximum occlusal intercuspidation. Anatomical references previously used by Ricketts for the condyle position inside the glenoid fossae were measured digitally through the EzImplant software. Analysis of variance, Tukey's, Kruskal-Wallis, and Mann-Whitney U statistical tests were used. The upper distance of the condyle to the glenoid fossa was smaller in the class II and class III compared with the class I group. The anterior distance of the condyle to the articular eminence showed significant differences when comparing the class I with the class II and class III groups. No statistically significant difference was noted in the posterior condylar distance between the groups. The angle of the eminence showed differences between the three groups, while the eminence height showed significant difference when comparing the class I with class III group. Spatial differences existed for the condylar position in relation to the glenoid fossa for skeletal class I, class II, and class III, but these spatial differences may not be clinically relevant.


Predicting trauma admissions: the effect of weather, weekday, and other variables.
One of the challenges all hospitals, especially designated trauma centers, face is how to make sure they have adequate staffing on various days of the week and at various times of the year. A number of studies have explored whether factors such as weather, temporal variation, holidays, and events that draw mass gatherings may be useful for predicting patient volume. This article looks at the effects of weather, mass gatherings, and calendar variables on daily trauma admissions at the three Level I trauma hospitals in the Minneapolis-St. Paul metropolitan area. Using ARIMA statistical modeling, we found that weekends, summer, lack of rain, and snowfall were all predictive of daily trauma admissions; holidays and mass gatherings such as sporting events were not. The forecasting model was successful in reflecting the pattern of trauma admissions; however, it's usefulness was limited in that the predicted range of daily trauma admissions was much narrower than the observed number of admissions. Nonetheless, the observed pattern of increased admission in the summer months and year-round on Saturdays should be helpful in resource planning.


Resection of symptomatic non-small cell lung cancer brain metastasis in the setting of multiple brain metastases.
Current guidelines primarily suggest resection of brain metastases (BMs) in patients with limited lesions. With a growing number of highly effective local and systemic treatment options, this view may be challenged. The purpose of this study was to evaluate the role of metastasectomy, disregarding BM count, in a comprehensive treatment setting. In this monocentric retrospective analysis, the authors included patients who underwent resection for at least 1 BM and collected demographic, clinical, and tumor-associated parameters. Prognostic factors for local control and overall survival (OS) were analyzed with the log-rank test and Cox proportional hazards analysis. The authors analyzed 216 patients. One hundred twenty-nine (59.7%) patients were diagnosed with a single/solitary BM, whereas 64 (29.6%) patients had 2-3 BMs and the remaining 23 (10.6%) had more than 3 BMs. With resection of symptomatic BMs, a significant improvement in Karnofsky Performance Scale (KPS) was achieved (p < 0.001), thereby enabling adjuvant radiotherapy for 199 (92.1%) patients and systemic treatment for 119 (55.1%) patients. During follow-up, 83 (38.4%) patients experienced local recurrence. BM count did not significantly influence local control rates. By the time of analysis, 120 (55.6%) patients had died; the leading cause of death was systemic tumor progression. The mean (range) OS after surgery was 12.7 (0-88) months. In univariate analysis, the BM count did not influence OS (p = 0.844), but age < 65 years (p = 0.007), preoperative and postoperative KPS ≥ 70 (p = 0.002 and p = 0.005, respectively), systemic metastases other than BM (p = 0.004), adjuvant radiation therapy (p < 0.001), and adjuvant systemic treatment (p < 0.001) were prognostic factors. In regression analysis, the presence of extracranial metastases (HR 2.30, 95% CI 1.53-3.48, p < 0.001), adjuvant radiation therapy (HR 0.97, 95% CI 0.23-0.86, p = 0.016), and adjuvant systemic treatment (HR 0.37, 95% CI 0.25-0.55, p < 0.001) remained as independent factors for survival. Surgery for symptomatic BM from non-small cell lung cancer may be indicated even for patients with multiple lesions in order to alleviate their neurological symptoms and to consequently facilitate further treatment.


Structure of the mammalian antimicrobial peptide Bac7(1-16) bound within the exit tunnel of a bacterial ribosome.
Proline-rich antimicrobial peptides (PrAMPs) produced as part of the innate immune response of animals, insects and plants represent a vast, untapped resource for the treatment of multidrug-resistant bacterial infections. PrAMPs such as oncocin or bactenecin-7 (Bac7) interact with the bacterial ribosome to inhibit translation, but their supposed specificity as inhibitors of bacterial rather than mammalian protein synthesis remains unclear, despite being key to developing drugs with low toxicity. Here, we present crystal structures of the Thermus thermophilus 70S ribosome in complex with the first 16 residues of mammalian Bac7, as well as the insect-derived PrAMPs metalnikowin I and pyrrhocoricin. The structures reveal that the mammalian Bac7 interacts with a similar region of the ribosome as insect-derived PrAMPs. Consistently, Bac7 and the oncocin derivative Onc112 compete effectively with antibiotics, such as erythromycin, which target the ribosomal exit tunnel. Moreover, we demonstrate that Bac7 allows initiation complex formation but prevents entry into the elongation phase of translation, and show that it inhibits translation on both mammalian and bacterial ribosomes, explaining why this peptide needs to be stored as an inactive pro-peptide. These findings highlight the need to consider the specificity of PrAMP derivatives for the bacterial ribosome in future drug development efforts.


How environmental and genetic factors combine to cause autism: A redox/methylation hypothesis.
Recently higher rates of autism diagnosis suggest involvement of environmental factors in causing this developmental disorder, in concert with genetic risk factors. Autistic children exhibit evidence of oxidative stress and impaired methylation, which may reflect effects of toxic exposure on sulfur metabolism. We review the metabolic relationship between oxidative stress and methylation, with particular emphasis on adaptive responses that limit activity of cobalamin and folate-dependent methionine synthase. Methionine synthase activity is required for dopamine-stimulated phospholipid methylation, a unique membrane-delimited signaling process mediated by the D4 dopamine receptor that promotes neuronal synchronization and attention, and synchrony is impaired in autism. Genetic polymorphisms adversely affecting sulfur metabolism, methylation, detoxification, dopamine signaling and the formation of neuronal networks occur more frequently in autistic subjects. On the basis of these observations, a "redox/methylation hypothesis of autism" is described, in which oxidative stress, initiated by environment factors in genetically vulnerable individuals, leads to impaired methylation and neurological deficits secondary to reductions in the capacity for synchronizing neural networks.


Molecular diversity of soil and marine 16S rRNA gene sequences related to beta-subgroup ammonia-oxidizing bacteria.
We have conducted a preliminary phylogenetic survey of ammonia-oxidizing beta-proteobacteria, using 16S rRNA gene libraries prepared by selective PCR and DNA from acid and neutral soils and polluted and nonpolluted marine sediments. Enrichment cultures were established from samples and analyzed by PCR. Analysis of 111 partial sequences of c. 300 bases revealed that the environmental sequences formed seven clusters, four of which are novel, within the phylogenetic radiation defined by cultured autotrophic ammonia oxidizers. Longer sequences from 13 cluster representatives support their phylogenetic positions relative to cultured taxa. These data suggest that known taxa may not be representative of the ammonia-oxidizing beta-proteobacteria in our samples. Our data provide further evidence that molecular and culture-based enrichment methods can select for different community members. Most enrichments contained novel Nitrosomonas-like sequences whereas novel Nitrosospira-like sequences were more common from gene libraries of soils and marine sediments. This is the first evidence for the occurrence of Nitrosospira-like strains in marine samples. Clear differences between the sequences of soil and marine sediment libraries were detected. Comparison of 16S rRNA sequences from polluted and nonpolluted sediments provided no strong evidence that the community composition was determined by the degree of pollution. Soil clone sequences fell into four clusters, each containing sequences from acid and neutral soils in varying proportions. Our data suggest that some related strains may be present in both samples, but further work is needed to resolve whether there is selection due to pH for particular sequence types.


Pharmacokinetics and Exposure Response Relationships of Ustekinumab in Patients With Crohn's Disease.
Ustekinumab is a monoclonal antibody that binds with high affinity to the p40 subunit of human interleukin 12 (IL12 and IL23) that has been approved for treatment of patients with moderate to severe Crohn's disease (CD). However, there are few data on its pharmacokinetic properties or the relationship between drug exposure levels and patient response. We collected data from 2 Phase 3 induction studies and 1 maintenance study to determine ustekinumab's pharmacokinetic features, relationship between exposure and response, and optimal serum concentrations for efficacy. We collected data on serum concentrations of ustekinumab and efficacy from induction studies of patients with moderate to severe CD given ustekinumab for 8 weeks following a single intravenous dose (either 130 mg or approximately 6 mg/kg). We collected the same data from a maintenance study of patients with a response to ustekinumab in the induction study who then received subcutaneous injections (90 mg) every 8 or 12 weeks for 44 weeks. At week 44 of the maintenance study (52 weeks after treatment began), patients were evaluated for the primary endpoint of clinical remission (defined as a CD activity index score below 150 points), endoscopic markers of efficacy, and serum level of C-reactive protein. Ustekinumab concentration data were categorized into quartiles and relationships between exposure and response were assessed. Optimal concentration cutoff values were evaluated using receiver operating characteristic curve analysis. Serum concentrations of ustekinumab over time were proportional to dose and did not differ significantly between the induction studies. In the maintenance study, ustekinumab concentration reached the steady state by the second maintenance dose; the median trough concentration was approximately threefold higher in patients given ustekinumab at 8-week intervals compared with 12-week intervals. Ustekinumab serum concentrations associated with rates of clinical remission and endoscopic efficacy endpoints, correlated inversely with level of C-reactive protein, and did not associate with use of immunomodulators. Trough concentrations of ustekinumab of 0.8 (or even up to 1.4 μg/mL) or greater were associated with maintenance of clinical remission in a higher proportion of patients than patients with lower trough concentrations. In an analysis of data from Phase 3 studies of patients with moderate to severe CD, we found serum concentrations of ustekinumab to be proportional to dose and associate with treatment efficacy. Concentrations of ustekinumab did not seem to be affected by cotreatment with immunomodulators. Clinicaltrials.gov no. NCT01369329 (UNITI 1), NCT01369342 (UNITI 2), and NCT01369355 (IM-UNITI).


Systematic review of multifaceted interventions to improve depression care.
Depression is a prevalent high-impact illness with poor outcomes in primary care settings. We performed a systematic review to determine to what extent multifaceted interventions improve depression outcomes in primary care and to define key elements, patients who are likely to benefit and resources required for these interventions. We searched Medline, HealthSTAR, CINAHL, PsycINFO and a specialized registry of depression trials from 1966 to February 2006; reviewed bibliographies of pertinent articles; and consulted experts. Searches were limited to the English language. We included 28 randomized controlled trials that: (a) involved primary care patients receiving acute-phase treatment; (b) tested a multicomponent intervention involving a patient-directed component; and (c) reported effects on depression severity. Pairs of investigators independently abstracted information regarding (a) setting and subjects, (b) components of the intervention and (c) outcomes. Twenty of 28 interventions improved depression outcomes over 3-12 months (an 18.4% median absolute increase in patients with 50% improvement in symptoms; range, 8.3-46%). Sustained improvements at 24-57 months were demonstrated in three studies addressing acute-phase and continuation-phase treatments. All interventions involved care management and required additional resources or staff reassignment to implement; interventions were delivered exclusively or predominantly by telephone in 16 studies. The most commonly used intervention features were: patient education and self-management, monitoring of depressive symptoms and treatment adherence, decision support for medication management, a patient registry and mental health supervision of care managers. Other intervention features were highly variable. There is strong evidence supporting the short-term benefits of care management for depression; critical elements for successful programs are emerging.


An assessment of the accuracy and usability of a novel optical wound measurement system.
Measurement of wound size can predict healing and provide information to guide treatment. This study assesses a novel optical wound imaging system that creates a three-dimensional image of the ulcer. Using a new camera-based digital system and traditional elliptical wound measurements, 36 foot ulcers from 31 patients (aged 44-94 years, median 70 years) were examined during a 12-week period at two centres. Median diabetes duration was 18 years (range 6-56 years). Seventeen percent had Type 1 diabetes, 93% had peripheral neuropathy and 57% had peripheral artery disease. Twenty-five were reviewed consecutively, resulting in 76 ulcer examinations. Median ulcer size was 94 mm(2), with size ranging from 3.1 to 2195 mm(2). Pearson, Spearman and Kendall rank coefficients showed a strong correlation (in all cases P < 0.001) between digital measurements of wounds against traditional hand-measured estimates. Intra-observer variation of wound length using digital elliptical measurement (DEM) gave a coefficient of variation of < 3.0%. Interobserver variation of wound length using DEM was < 6.5%. Variation from a standard known-size wound area was < 8.0% across 30 trials. This study shows a strong correlation between digital and traditional measurement techniques. The system can be easily deployed in routine clinical practice, providing an objective visual record, allowing remote in-depth analysis.


Prediction of SARS-CoV Interaction with Host Proteins during Lung Aging Reveals a Potential Role for TRIB3 in COVID-19.
COVID-19 is prevalent in the elderly. Old individuals are more likely to develop pneumonia and respiratory failure due to alveolar damage, suggesting that lung senescence may increase the susceptibility to SARS-CoV-2 infection and replication. Considering that human coronavirus (HCoVs; SARS-CoV-2 and SARS-CoV) require host cellular factors for infection and replication, we analyzed Genotype-Tissue Expression (GTEx) data to test whether lung aging is associated with transcriptional changes in human protein-coding genes that potentially interact with these viruses. We found decreased expression of the gene tribbles homolog 3 (TRIB3) during aging in male individuals, and its protein was predicted to interact with HCoVs nucleocapsid protein and RNA-dependent RNA polymerase. Using publicly available lung single-cell data, we found TRIB3 expressed mainly in alveolar epithelial cells that express SARS-CoV-2 receptor ACE2. Functional enrichment analysis of age-related genes, in common with SARS-CoV-induced perturbations, revealed genes associated with the mitotic cell cycle and surfactant metabolism. Given that TRIB3 was previously reported to decrease virus infection and replication, the decreased expression of TRIB3 in aged lungs may help explain why older male patients are related to more severe cases of the COVID-19. Thus, drugs that stimulate TRIB3 expression should be evaluated as a potential therapy for the disease.


Placoderm Assemblage from the Tetrapod-Bearing Locality of Strud (Belgium, Upper Famennian) Provides Evidence for a Fish Nursery.
The placoderm fauna of the upper Famennian tetrapod-bearing locality of Strud, Belgium, includes the antiarch Grossilepis rikiki, the arthrodire groenlandaspidid Turrisaspis strudensis and the phyllolepidid Phyllolepis undulata. Based on morphological and morphometric evidence, the placoderm specimens from Strud are predominantly recognised as immature specimens and this locality as representing a placoderm nursery. The Strud depositional environment corresponds to a channel in an alluvial plain, and the presence of a nursery in such environment could have provided nutrients and protection to the placoderm offspring. This represents one of the earliest pieces of evidence for this sort of habitat partitioning in vertebrate history, with adults living more distantly from the nursery and using the nursery only to spawn or give live birth.


In vitro and in vivo activities of HPi1, a selective antimicrobial against Helicobacter pylori.
A high-throughput screen (HTS) was performed to identify molecules specifically active against Helicobacter pylori, the causative agent of peptic ulcer and gastric carcinoma. Currently, treatment of H. pylori infection is suboptimal, with failure rates approaching 25%, despite triple therapy with two broad-spectrum antibiotics and a proton pump inhibitor or quadruple therapy with added bismuth. The HTS was performed in 384-well plates, and reduction of the metabolic indicator resazurin was used as a reporter for cell growth. Diverse molecules from commercial sources were identified as hits, and in vitro validations included measurements of MIC and time-dependent killing as well as anaerobic susceptibility testing against a panel of gut microbes. In vivo validation included testing in the mouse model of H. pylori infection. The small molecule HPi1 (3-hydrazinoquinoxaline-2-thiol) had excellent potency, with an MIC of 0.08 to 0.16 μg/ml and good selectivity for H. pylori compared to a panel of commensal bacteria. HPi1 was also effective in a mouse model of H. pylori infection, reducing colony counts to below the limit of detection after oral dosing of 25 mg/kg/day for 3 days. HPi1 is a promising lead in the search for more effective and specific H. pylori therapeutics.


Cofilin produces newly polymerized actin filaments that are preferred for dendritic nucleation by the Arp2/3 complex.
One of the earliest events in the process of cell motility is the massive generation of free actin barbed ends, which elongate to form filaments adjacent to the plasma membrane at the tip of the leading edge. Both cofilin and Arp2/3 complex have been proposed to contribute to barbed end formation during cell motility. Attempts to assess the functions of cofilin and Arp 2/3 complex in vivo indicate that both cofilin and Arp2/3 complex contribute to actin polymerization: cofilin by severing and Arp2/3 by nucleating and branching. In order to determine if the activities of cofilin and Arp2/3 complex interact, we employed a light microscope-based assay to visualize actin polymerization directly in the presence of both proteins. The results indicate that cofilin generates barbed ends to increase the mass of freshly polymerized F-actin but does not directly affect the activity of Arp2/3 complex. However, while ADP, ADP-Pi, and newly polymerized ATP-filaments are all capable of supporting Arp2/3-mediated branching, newly polymerized F-actin supports most of the Arp2/3-induced branch formation. The results suggest that, in vivo, cofilin contributes to barbed end formation by inducing the initial increase in the number of barbed ends leading to increased ATP-F-actin, which in turn supports higher levels of dendritic nucleation by active Arp2/3 complex.


Filled carbon nanotubes in biomedical imaging and drug delivery.
Carbon nanotubes have been advocated as promising candidates in the biomedical field in the areas of diagnosis and therapy. In terms of drug delivery, the use of carbon nanotubes can overcome some limitations of 'free' drugs by improving the formulation of poorly water-soluble drugs, allowing targeted delivery and even enabling the co-delivery of two or more drugs for combination therapy. Two different approaches are currently being explored for the delivery of diagnostic and therapeutic agents by carbon nanotubes, namely attachment of the payload to the external sidewalls or encapsulation into the inner cavities. Although less explored, the latter confers additional stability to the chosen diagnostic or therapeutic agents, and leaves the backbone structure of the nanotubes available for its functionalization with dispersing and targeting moieties. Several drug delivery systems and diagnostic agents have been developed in the last years employing the inner tubular cavities of carbon nanotubes. The research discussed in this review focuses on the use of carbon nanotubes that contain in their interior drug molecules and diagnosis-related compounds. The approaches employed for the development of such nanoscale vehicles along with targeting and releasing strategies are discussed. The encapsulation of both biomedical contrast agents and drugs inside carbon nanotubes is further expanding the possibilities to allow an early diagnosis and treatment of diseases.


A single locus from the entomopathogenic bacterium Photorhabdus luminescens inhibits activated Manduca sexta phenoloxidase.
Insect blood (hemolymph) contains prophenoloxidase, a proenzyme that is activated to protective phenoloxidase when the insect is damaged or challenged with microorganisms. The Gram-negative bacterium Photorhabdus luminescens kills the lepidopteron insect Manduca sexta by using a variety of toxins. We screened P. luminescens and Photorhabdus asymbiotica cosmid libraries in an Escherichia coli host against previously activated M. sexta hemolymph phenoloxidase and identified three overlapping cosmid clones from P. luminescens and five from P. asymbiotica that suppressed the activity of the enzyme both in vitro and in vivo. Genome alignments of cosmid end sequences from both species confirmed that they contained orthologous loci. We examined one of the cosmids from P. luminescens in detail: it induced the formation of significantly fewer melanotic nodules, proliferated faster within the insect host and was significantly more virulent towards fifth-stage larvae than E. coli control bacteria. Insertional mutagenesis of this cosmid yielded 11 transposon mutants that were no longer inhibitory. All of these were insertions into a single 5.5-kb locus, which contained three ORFs and was homologous to the maltodextrin phosphorylase locus of E. coli. The implications of this novel inhibitory factor of insect phenoloxidase for Photorhabdus virulence are discussed.


Bioengineering of bacteria to assemble custom-made polyester affinity resins.
Proof of concept for the in vivo bacterial production of a polyester resin displaying various customizable affinity protein binding domains is provided. This was achieved by engineering various protein binding domains into a bacterial polyester-synthesizing enzyme. Affinity binding domains based on various structural folds and derived from molecular libraries were used to demonstrate the potential of this technique. Designed ankyrin repeat proteins (DARPins), engineered OB-fold domains (OBodies), and VHH domains from camelid antibodies (nanobodies) were employed. The respective resins were produced in a single bacterial fermentation step, and a simple purification protocol was developed. Purified resins were suitable for most lab-scale affinity chromatography purposes. All of the affinity domains tested produced polyester beads with specific affinity for the target protein. The binding capacity of these affinity resins ranged from 90 to 600 nmol of protein per wet gram of polyester affinity resin, enabling purification of a recombinant protein target from a complex bacterial cell lysate up to a purity level of 96% in one step. The polyester resin was efficiently produced by conventional lab-scale shake flask fermentation, resulting in bacteria accumulating up to 55% of their cellular dry weight as polyester. A further proof of concept demonstrating the practicality of this technique was obtained through the intracellular coproduction of a specific affinity resin and its target. This enables in vivo binding and purification of the coproduced "target protein." Overall, this study provides evidence for the use of molecular engineering of polyester synthases toward the microbial production of specific bioseparation resins implementing previously selected binding domains.


The predictive start of hunting archer fish: a flexible and precise motor pattern performed with the kinematics of an escape C-start.
Once their shots have successfully dislodged aerial prey, hunting archer fish monitor the initial values of their prey's ballistic motion and elicit an adapted rapid turning maneuver. This allows these fish to head straight towards the later point of catch with a speed matched to the distance to be covered. To make the catch despite severe competition the fish must quickly and yet precisely match their turn and take-off speed to the initial values of prey motion. However, the initial variables vary over broad ranges and can be determined only after prey is dislodged. Therefore, the underlying neuronal circuitry must be able to drive a maneuver that combines a high degree of precision and flexibility at top speed. To narrow down which neuronal substrate underlies the performance we characterized the kinematics of archer fish predictive starts using digital high-speed video. Strikingly, the predictive starts show all hallmarks of Mauthner-driven teleost C-type fast-starts, which have previously not been noted in feeding strikes and were not expected to provide the high angular accuracy required. The high demands on flexibility and precision of the predictive starts do not compromise their performance. On the contrary, archer fish predictive starts are among the fastest C-starts known so far among teleost fish, with peak linear speed beyond 20 body lengths s(-1), angular speed over 4500 deg. s(-1), maximum linear acceleration of up to 12 times gravitational acceleration and peak angular acceleration of more than 450 000 deg. s(-2). Moreover, they were not slower than archer fish escape C-starts, elicited in the same individuals. Rather, both escapes and predictive starts follow an identical temporal pattern and all kinematic variables of the two patterns overlap. This kinematic equivalence strongly suggests that archer fish recruit their C-start escape network of identified reticulospinal neurons, or elements of it, to drive their predictive starts. How the network drives such a rather complex behavior without compromising speed is a wide open question.


Maternal and perinatal outcomes in pregnant women with BMI >50: An international collaborative study.
To examine the association between maternal BMI>50kg/m2 during pregnancy and maternal and perinatal outcomes. An international cohort study was conducted using data from separate national studies in the UK and Australia. Outcomes of pregnant women with BMI>50 were compared to those of pregnant women with BMI<50. Multivariable logistic regression estimated the association between BMI>50 and perinatal and maternal outcomes. 932 pregnant women with BMI>50 were compared with 1232 pregnant women with BMI<50. Pregnant women with BMI>50 were slightly older, more likely to be multiparous, and have pre-existing comorbidities. There were no maternal deaths, however, extremely obese women had a nine-fold increase in the odds of thrombotic events compared to those with a BMI<50 (uOR: 9.39 (95%CI:1.15-76.43)). After adjustment, a BMI>50 during pregnancy had significantly raised odds of preeclampsia/eclampsia (aOR:4.88(95%CI: 3.11-7.65)), caesarean delivery (aOR: 2.77 (95%CI: 2.31-3.32)), induction of labour (aOR: 2.45(95% CI:2.00-2.99)) post caesarean wound infection (aOR:7.25(95%CI: 3.28-16.07)), macrosomia (aOR: 8.05(95%CI: 4.70-13.78)) compared a BMI<50. Twelve of the infants born to women in the extremely obese cohort died in the early neonatal period or were stillborn. Pregnant women with BMI>50 have a high risk of inferior maternal and perinatal outcomes.


Genetic spell-checking: gene editing using single-stranded DNA oligonucleotides.
Single-stranded oligonucleotides (ssODNs) can be used to direct the exchange of a single nucleotide or the repair of a single base within the coding region of a gene in a process that is known, generically, as gene editing. These molecules are composed of either all DNA residues or a mixture of RNA and DNA bases and utilize inherent metabolic functions to execute the genetic alteration within the context of a chromosome. The mechanism of action of gene editing is now being elucidated as well as an understanding of its regulatory circuitry, work that has been particularly important in establishing a foundation for designing effective gene editing strategies in plants. Double-strand DNA breakage and the activation of the DNA damage response pathway play key roles in determining the frequency with which gene editing activity takes place. Cellular regulators respond to such damage and their action impacts the success or failure of a particular nucleotide exchange reaction. A consequence of such activation is the natural slowing of replication fork progression, which naturally creates a more open chromatin configuration, thereby increasing access of the oligonucleotide to the DNA template. Herein, how critical reaction parameters influence the effectiveness of gene editing is discussed. Functional interrelationships between DNA damage, the activation of DNA response pathways and the stalling of replication forks are presented in detail as potential targets for increasing the frequency of gene editing by ssODNs in plants and plant cells.


Herpes simplex virus 1 infection activates poly(ADP-ribose) polymerase and triggers the degradation of poly(ADP-ribose) glycohydrolase.
Herpes simplex virus 1 infection triggers multiple changes in the metabolism of host cells, including a dramatic decrease in the levels of NAD(+). In addition to its role as a cofactor in reduction-oxidation reactions, NAD(+) is required for certain posttranslational modifications. Members of the poly(ADP-ribose) polymerase (PARP) family of enzymes are major consumers of NAD(+), which they utilize to form poly(ADP-ribose) (PAR) chains on protein substrates in response to DNA damage. PAR chains can subsequently be removed by the enzyme poly(ADP-ribose) glycohydrolase (PARG). We report here that the HSV-1 infection-induced drop in NAD(+) levels required viral DNA replication, was associated with an increase in protein poly(ADP-ribosyl)ation (PARylation), and was blocked by pharmacological inhibition of PARP-1/PARP-2 (PARP-1/2). Neither virus yield nor the cellular metabolic reprogramming observed during HSV-1 infection was altered by the rescue or further depletion of NAD(+) levels. Expression of the viral protein ICP0, which possesses E3 ubiquitin ligase activity, was both necessary and sufficient for the degradation of the 111-kDa PARG isoform. This work demonstrates that HSV-1 infection results in changes to NAD(+) metabolism by PARP-1/2 and PARG, and as PAR chain accumulation can induce caspase-independent apoptosis, we speculate that the decrease in PARG levels enhances the auto-PARylation-mediated inhibition of PARP, thereby avoiding premature death of the infected cell.


Co-existing spinal intradural ependymal cyst and sacral Tarlov cyst in adult-onset tethered cord syndrome with syringomyelia: Case report and literature review.
Synchronous spinal intradural ependymal cysts and sacral Tarlov cysts in adult onset tethered cord syndrome are extremely rare. A 23-year-old male presented with back pain radiating into both lower extremities, accompanied by acute onset of gait difficulty and sphincter dysfunction. Magnetic resonance imaging identified a low lying conus medullaris, syringomyelia with septations extending from T12 to S1, a tethered cord, and a thickened filum terminale with a sacral Tarlov cyst. The patient underwent a L3-4 laminectomy for decompression of syringomyelia and excision/biopsy of a space occupying lesion along with S1-2 laminectomy for cord untethering and Tarlov cyst fenestration. Postoperative histopathology confirmed that the lesion was an ependymal cyst. Clinically, patient showed marked improvement in the neurological status. Simultaneous decompressive laminectomy of L3-4 and S1-2 effectively decompressed the syringomyelia while allowing for excision/biopsy of a space occupying lesion at the former and untethering and Tarlov cyst fenestration at the latter levels.


Inoculating Anopheles gambiae Mosquitoes with Beads to Induce and Measure the Melanization Immune Response.
The stimulation of immune responses is a common tool in invertebrate studies to examine the efficacy and the mechanisms of immunity. This stimulation is based on the injection of non-pathogenic particles into insects, as the particles will be detected by the immune system and will induce the production of immune effectors. We focus here on the stimulation of the melanization response in the mosquito Anopheles gambiae. The melanization response results in the encapsulation of foreign particles and parasites with a dark layer of melanin. To stimulate this response, mosquitoes are inoculated with beads in the thoracic cavity using microcapillary glass tubes. Then, after 24 hr, the mosquitoes are dissected to retrieve the beads. The degree of melanization of the bead is measured using image analysis software. Beads do not have the pathogenic effects of parasites, or their capacity to evade or suppress the immune response. These injections are a way to measure immune efficacy and the impact of immune stimulations on other life history traits, such as fecundity or longevity. It is not exactly the same as directly studying host-parasite interactions, but it is an interesting tool to study immunity and its evolutionary ecology.


Effects of pentoxifylline pretreatment on Kupffer cells in rat liver transplantation.
Previous research with pentoxifylline (PTX), a methylxanthine phosphodiesterase inhibitor, suggests that this drug may be capable of suppressing the activation of Kupffer cells and thereby help decrease liver injury after transplantation. To investigate this possibility, the current study sought to determine whether the release of O2- and tumor necrosis factor (TNF) from Kupffer cells in donor livers can be suppressed if the organs are exposed to PTX before preservation. In an in vitro experiment, rat livers were flushed with PTX (25 mg/kg body weight) in University of Washington (UW) solution or UW solution alone (control) and then and stored in UW solution for either 4 or 24 hours. Kupffer cells then were purified and their degree of activation determined by measuring O2- release and the production of TNF after lipopolysaccharide stimulation. In an in vivo experiment, a group of rats underwent orthotopic liver transplantation with grafts prepared in the same manner as in the in vitro study. TNF and aspartate transaminase (AST) were measured in blood samples taken 3 hours and 24 hours after transplantation. Compared with controls, the Kupffer cells from grafts pretreated with PTX produced significantly less O2- and TNF, and the recipients of PTX-pretreated grafts had lower levels of TNF and AST 3 hours after transplantation. The current data indicate that O2- and TNF production in liver grafts is suppressed by PTX pretreatment. Through its suppressive effect on Kupffer cells, PTX may help minimize preservation-reperfusion injury and improve graft survival.


Using in-clerkship tests to identify students with insufficient knowledge and assessing the effect of counseling on final examination performance.
To determine whether in-clerkship tests identify students with insufficient knowledge and whether counseling affects final examination pass rates. The authors reviewed students' mean scores from two internal medicine clerkship tests at the Wright State University School of Medicine from February 1993 to July 1996. To determine the sensitivity and specificity of the tests for identifying students with insufficient knowledge, they compared students in the lowest quartile of clerkship test results with those who scored 290 or less on the end-of-clerkship National Board of Medical Examiners' (NBME) subject examination in medicine. The authors also compared the final examination pass rates of counseled and non-counseled students. Twenty-five students scored 290 or less on the NBME subject examination. Of those, 17 had low mean clerkship test scores (sensitivity of 68%). The specificity of a low mean clerkship test score was 81%. Counseling did not improve final examination pass rates. In-clerkship tests can identify students who are at risk of failing an end-of-clerkship examination. Because counseling may be insufficient to raise final examination pass rates, further study into the appropriate clerkship intervention for low-achievement students is needed.


Femtosecond optical transfection of cells: viability and efficiency.
Photoporation is a rapidly expanding technique for the introduction of macromolecules into single cells. However, there remains no study into the true efficiency of this procedure. Here, we present a detailed analysis of transfection efficiency and cell viability for femtosecond optical transfection using a titanium sapphire laser at 800 nm. Photoporation of 4000 Chinese Hamster ovary cells was performed, representing the largest optical transfection study reported to date. We have investigated a range of laser fluences at the cell membrane and, at 1.2 microJ/cm(2), have found an average transfection efficiency of 50 +/- 10%. Contrary to recent literature, in which 100% efficiency is claimed, our measure of efficiency accounts for all irradiated cells, including those lost as a result of laser treatment, thereby providing a true biological measure of the technique.


Role of stress fiber-like structures in assembling nascent myofibrils in myosheets recovering from exposure to ethyl methanesulfonate.
When day 1 cultures of chick myogenic cells were exposed to the mutagenic alkylating agent ethyl methanesulfonate (EMS) for 3 d, 80% of the replicating cells were killed, but postmitotic myoblasts survived. The myoblasts fused to form unusual multinucleated "myosheets": extraordinarily wide, flattened structures that were devoid of myofibrils but displayed extensive, submembranous stress fiber-like structures (SFLS). Immunoblots of the myosheets indicated that the carcinogen blocked the synthesis and accumulation of the myofibrillar myosin isoforms but not that of the cytoplasmic myosin isoform. When removed from EMS, widely spaced nascent myofibrils gradually emerged in the myosheets after 3 d. Striking co-localization of fluorescent reagents that stained SFLS and those that specifically stained myofibrils was observed for the next 2 d. By both immunofluorescence and electron microscopy, individual nascent myofibrils appeared to be part of, or juxtaposed to, preexisting individual SFLS. By day 6, all SFLS had disappeared, and the definitive myofibrils were displaced from their submembranous site into the interior of the myosheet. Immunoblots from recovering myosheets demonstrated a temporal correlation between the appearance of the myofibrillar myosin isoforms and the assembly of thick filaments. The assembly of definitive myofibrils did not appear to involve desmin intermediate filaments, but a striking aggregation of sarcoplasmic reticulum elements was seen at the level of each I-Z-band. Our findings suggest that SFLS in the EMS myosheets function as early, transitory assembly sites for nascent myofibrils.


Compliance with results reporting at ClinicalTrials.gov.
The Food and Drug Administration Amendments Act (FDAAA) mandates timely reporting of results of applicable clinical trials to ClinicalTrials.gov. We characterized the proportion of applicable clinical trials with publicly available results and determined independent factors associated with the reporting of results. Using an algorithm based on input from the National Library of Medicine, we identified trials that were likely to be subject to FDAAA provisions (highly likely applicable clinical trials, or HLACTs) from 2008 through 2013. We determined the proportion of HLACTs that reported results within the 12-month interval mandated by the FDAAA or at any time during the 5-year study period. We used regression models to examine characteristics associated with reporting at 12 months and throughout the 5-year study period. From all the trials at ClinicalTrials.gov, we identified 13,327 HLACTs that were terminated or completed from January 1, 2008, through August 31, 2012. Of these trials, 77.4% were classified as drug trials. A total of 36.9% of the trials were phase 2 studies, and 23.4% were phase 3 studies; 65.6% were funded by industry. Only 13.4% of trials reported summary results within 12 months after trial completion, whereas 38.3% reported results at any time up to September 27, 2013. Timely reporting was independently associated with factors such as FDA oversight, a later trial phase, and industry funding. A sample review suggested that 45% of industry-funded trials were not required to report results, as compared with 6% of trials funded by the National Institutes of Health (NIH) and 9% of trials that were funded by other government or academic institutions. Despite ethical and legal obligations to disclose findings promptly, most HLACTs did not report results to ClinicalTrials.gov in a timely fashion during the study period. Industry-funded trials adhered to legal obligations more often than did trials funded by the NIH or other government or academic institutions. (Funded by the Clinical Trials Transformation Initiative and the NIH.).


Between thought and expression, a magnetoencephalography study of the "tip-of-the-tongue" phenomenon.
"Tip-of-the-tongue" (TOT) is the phenomenon associated with the inaccessibility of a known word from memory. It is universally experienced, increases in frequency with age, and is most common for proper nouns. It is a good model for the symptom of anomia experienced much more frequently by some aphasic patients following brain injury. Here, we induced the TOT state in older participants while they underwent brain scanning with magnetoencephalography to investigate the changes in oscillatory brain activity associated with failed retrieval of known words. Using confrontation naming of pictures of celebrities, we successfully induced the TOT state in 29% of trials and contrasted it with two other states: "Know" where the participants both correctly recognized the celebrity's face and retrieved their name and "Don't Know" when the participants did not recognize the celebrity. We wished to test Levelt's influential model of speech output by carrying out two analyses, one epoching the data to the point in time when the picture was displayed and the other looking back in time from when the participants first articulated their responses. Our main findings supported the components of Levelt's model, but not their serial activation over time as both semantic and motor areas were identified in both analyses. We also found enduring decreases in the alpha frequency band in the left ventral temporal region during the TOT state, suggesting ongoing semantic search. Finally, we identified reduced beta power in classical peri-sylvian language areas for the TOT condition, suggesting that brain regions that encode linguistic memories are also involved in their attempted retrieval.