Comparison of magnetic resonance spectroscopy and perfusion-weighted imaging in presurgical grading of oligodendroglial tumors.
Oligodendroglial tumors form an uncommon, but distinct, subgroup of gliomas with longer survival, better treatment response, and characteristic genetic alterations. Noninvasive grading of oligodendroglial tumors using functional and metabolic magnetic resonance imaging may be helpful in guiding the treatment approach and predicting malignant transformation of these tumors. We used perfusion-weighted magnetic resonance imaging and proton magnetic resonance spectroscopic imaging (MRSI) to predict the oligodendroglioma grade. Twenty-four patients with pathologically confirmed oligodendrogliomas underwent dynamic contrast-enhanced perfusion-weighted magnetic resonance imaging and/or proton MRSI before surgery. We assessed the ability of tumor contrast enhancement, normalized cerebral blood volume, normalized choline, and the presence of either lactate or lipid metabolites to correctly predict the World Health Organization tumor grade. The accuracy of tumor grading using each method was also compared. Tumor contrast enhancement (P = 0.069) and normalized cerebral blood volume (P = 0.181) were not significantly different between low and high-grade oligodendrogliomas. The MRSI measurement of normalized choline was significantly higher in high-grade (2.82 +/- 0.64) than in low-grade (1.62 +/- 0.46) oligodendrogliomas (P < 0.001), and the presence of lactate or lipid metabolites also correctly predicted high-grade tumors (P = 0.014). The maximum accuracy of contrast enhancement, normalized cerebral blood volume, normalized choline, and lactate or lipid metabolites in grading oligodendroglioma was 71, 83, 90, and 85%, respectively. MRSI measurements are more accurate than perfusion-weighted magnetic resonance imaging or conventional contrast enhancement in differentiating oligodendroglial tumor grade. In these inherently vascular tumors, metabolic measurements of mitosis and necrosis may be better than measures of neovascularity in presurgical grading.


JUUL use and reasons for initiation among adult tobacco users.
The electronic nicotine delivery system (ENDS) JUUL has quickly captured the ENDS market, representing 74.6% of the total dollar share for this category as of November 2018. Although JUUL is marketed as an alternative to cigarettes intended for current adult smokers, evidence suggests that a majority of ENDS users are concurrently current cigarette smokers. Little is known about the dual use of JUUL and cigarettes, as well as the reasons for trying JUUL among adult tobacco users. A survey fielded via web and phone of 1332 current cigarette, cigar, little cigar or cigarillo (CLCC), and ENDS users aged 18-54 years was conducted from March to May 2018. Weighted descriptive and bivariate analyses examined JUUL use and reasons for trying JUUL by demographics, combustible tobacco use, ENDS use and intention to quit. Approximately 81% of our sample reported current use of two or more tobacco products, including cigarettes, CLCCs or ENDS. Among current tobacco users, 15% ever used JUUL and 12% used JUUL in the past 30 days. Most individuals (74%) reporting ever JUUL use indicated using it for 5 days or fewer in the past 30 days. The most common reason for trying JUUL was trying to quit smoking cigarettes (37%), followed by family, friends or colleagues using the product (32%). Although some tobacco users may be initiating JUUL to decrease combustible use, most were using the product infrequently and concurrently with other products. Findings have significant implications for cessation intervention efforts and policy development to help smokers quit.


Early mixed farming of millet and rice 7800 years ago in the Middle Yellow River region, China.
The Peiligang Culture (9000-7000 cal. yr BP) in the Middle Yellow River region, North China, has long been considered representative of millet farming. It is still unclear, however, if broomcorn millet or foxtail millet was the first species domesticated during the Peiligang Culture. Furthermore, it is also unknown whether millet was cultivated singly or together with rice at the same period. In this study, phytolith analysis of samples from the Tanghu archaeological site reveals early crop information in the Middle Yellow River region, China. Our results show that broomcorn millet was the early dry farming species in the Peiligang Culture at 7800 cal. yr BP, while rice cultivation took place from 7800 to 4500 cal. yr BP. Our data provide new evidence of broomcorn millet and rice mixed farming at 7800 cal. yr BP in the Middle Yellow River region, which has implications for understanding the domestication process of the two crops, and the formation and continuance of the Ancient Yellow River Civilization.


Near-atomic resolution visualization of human transcription promoter opening.
In eukaryotic transcription initiation, a large multi-subunit pre-initiation complex (PIC) that assembles at the core promoter is required for the opening of the duplex DNA and identification of the start site for transcription by RNA polymerase II. Here we use cryo-electron microscropy (cryo-EM) to determine near-atomic resolution structures of the human PIC in a closed state (engaged with duplex DNA), an open state (engaged with a transcription bubble), and an initially transcribing complex (containing six base pairs of DNA-RNA hybrid). Our studies provide structures for previously uncharacterized components of the PIC, such as TFIIE and TFIIH, and segments of TFIIA, TFIIB and TFIIF. Comparison of the different structures reveals the sequential conformational changes that accompany the transition from each state to the next throughout the transcription initiation process. This analysis illustrates the key role of TFIIB in transcription bubble stabilization and provides strong structural support for a translocase activity of XPB.


A prospective, randomised trial of immediate exercise following lumbar microdiscectomy: a preliminary study.
To investigate whether immediate commencement of exercises after lumbar microdiscectomy enabled patients to become independently mobile more rapidly with no increase in risk of complications. Randomised, single-blind, controlled trial. One secondary care teaching hospital in the UK. Thirty patients were recruited. The follow-up rate was 28 (93%) at 4 weeks and 23 (77%) at 3 months. Patients were randomised to an intervention group commencing exercises within 2 hours after surgery, or a control group commencing exercises on the first postoperative day. Primary outcome measures included the time taken for the patient to become independently mobile and attain discharge criteria following surgery. Secondary outcome measures were disability and pain scores collected before surgery, and 4 weeks and 3 months after surgery; and return to work rates. The two groups were similar at baseline. The results indicated significantly reduced time to independent mobility [median 7 vs 19 hours, median difference 9 hours, 95% confidence interval (CI) 1.25 to 14.5, P=0.009] and return to work (median 6 vs 8 weeks, median difference 2 weeks, 95% CI 0 to 6, P=0.002) in the intervention group compared with the control group. At 15 hours after surgery, independent mobility was attained in 80 and 40% of the intervention and control groups, respectively. There were no significant differences in disability and pain scores at 4 weeks and 3 months. Immediate commencement of exercises following first-time single-level lumbar microdiscectomy enabled patients to become independently mobile more rapidly and return to work sooner. Immediate commencement of exercises may enable patients to be discharged earlier, with associated cost benefits to health care and no increase in the rate of revision surgery.


Factors involved in long- and short-term mandibular plate exposure.
To evaluate and to compare rates and timing of exposure of alloplastic mandibular plates by plate type and tissue reconstruction technique. A retrospective review series of 92 consecutive patients for 4 years (mean follow-up, 30 months). National Cancer Institute-designated comprehensive cancer center in a freestanding cancer hospital. Seventy-nine patients received alloplastic mandibular plates for segmental defects, and 13 patients received compression plates for mandibular osteotomies following ablative cancer surgery, including 21 titanium hollow osseointegrating reconstruction, 41 Storz, 16 Synthes, and 5 AO/ASIF (Arbeitsgemein schott fur Ostcosynthese fragen/Association for the Study of Internal Fixation) plates. Primary flap repair was provided by 71 pedicled soft tissue and 19 osseocutaneous free flaps, with primary closure in the remaining 2. Most of the reconstructions of the mandibular defect was with an alloplastic plate with musculocutaneous flap or revascularized bone graft. Clinically apparent intraoral or extraoral plate exposure. Plate exposure occurred in 25 cases. Nine plates were exposed extraorally, at a mean postoperative interval of 40 weeks. The remaining 16 plates were exposed intraorally at a mean postoperative interval of 16 weeks. There was no significant difference in the exposure rates of different plate types or methods of reconstruction. The titanium hollow osseointegrating reconstruction plate had a similar exposure rate compared with the other plates. Size and site of the defect were the only significant predictors of plate exposure Radiotherapy and postoperative complications did not affect the rate of exposure. Extraoral plate exposure occurs less commonly and later in the postoperative period than intraoral exposure, suggesting different causes. Plate type and type of flap reconstruction do not affect the rate of exposure. This may reflect long follow-up.


Effect of light/dark cycles on expression of nitrate assimilatory genes in maize shoots and roots.
The level of nitrate reductase (NR) and nitrite reductase (NiR) varied in both shoot and root tissue from nitrate-fed Zea mays L. grown under a 16-hour light/8-hour dark regime over a 10-day period postgermination, with peak activity occurring in days 5 to 6. To study the effect of different light regimes on NR and NiR enzyme activity and mRNA levels, 6-day-old plants were grown in the presence of continuous KNO(3) (10 millimolar). Both shoot NRA and mRNA varied considerably, peaking 4 to 8 hours into the light period. Upon transferring plants to continuous light, the amplitude of the peaks increased, and the peaks moved closer together. In continuous darkness, no NR mRNA or NR enzyme activity could be detected by 8 hours and 12 hours, respectively. In either a light/dark or continuous light regime, root NRA and mRNA did not vary substantially. However, when plants were placed in continuous darkness, both declined steadily in the roots, although some remained after 48 hours. Although there was no obvious cycling of NiR enzyme activity in shoot tissue, changes in mRNA mimicked those seen for NR mRNA. The expression of NR and NiR genes is affected by the light regime adopted, but light does not have a direct effect on the expression of these genes.


Effect of the Synchronization-Based Control of a Wearable Robot Having a Non-Exoskeletal Structure on the Hemiplegic Gait of Stroke Patients.
We have been developing the robotic wear curara as both a welfare device and rehabilitation robot that assists the elderly and disabled. curara is aimed at user friendliness. We have, thus, chosen a non-exoskeleton structure made of a plastic so that the robot is as light in weight as possible and to minimize the restraining stress against natural human movement. We verified the assistance effect of curara on 15 hemiplegic patients with stroke by comparing gait parameters (i.e., velocity, step length, cadence, and symmetry of joint angles) among three conditions. The conditions were "without assistance" (i.e., a control mode that cancels frictional resistances in actuators), Condition A (where joint angles are enlarged but there is no change in gait cycle), and Condition B (where there is no change in joint angles but the gait cycle is shortened). curara improved the walking velocity by 19% and 27% under Conditions A and B, respectively. Improvements in step length and cadence were, respectively, 11% and 7% under Condition A and 14% and 11% under Condition B. Moreover, the two assistance conditions reduced the difference in joint angles between unaffected and paralyzed legs. We consider that curara will facilitate the rehabilitation of stroke patients.


A Mobilising Concept? Unpacking Academic Representations of Responsible Research and Innovation.
This paper makes a plea for more reflexive attempts to develop and anchor the emerging concept of responsible research and innovation (RRI). RRI has recently emerged as a buzzword in science policy, becoming a focus of concerted experimentation in many academic circles. Its performative capacity means that it is able to mobilise resources and spaces despite no common understanding of what it is or should be 'made of'. In order to support reflection and practice amongst those who are interested in and using the concept, this paper unpacks understandings of RRI across a multi-disciplinary body of peer-reviewed literature. Our analysis focuses on three key dimensions of RRI (motivations, theoretical conceptualisations and translations into practice) that remain particularly opaque. A total of 48 publications were selected through a systematic literature search and their content was qualitatively analysed. Across the literature, RRI is portrayed as a concept that embeds numerous features of existing approaches to govern and assess emerging technologies. Our analysis suggests that its greatest potential may be in its ability to unify and provide political momentum to a wide range of long-articulated ethical and policy issues. At the same time, RRI's dynamism and resulting complexity may represent its greatest challenge. Further clarification on what RRI has to offer in practice-beyond what has been offered to date-is still needed, as well as more explicit engagement with research and institutional cultures of responsibility. Such work may help to realise the high political expectations that are attached to nascent RRI.


PDE8A genetic variation, polycystic ovary syndrome and androgen levels in women.
Polycystic ovary syndrome (PCOS) is characterized by excessive theca cell androgen secretion, dependent upon LH, which acts through the intermediacy of 3',5'-cyclic adenosine monophosphate (cAMP). cAMP signaling pathways are controlled through regulation of its synthesis by adenylyl cyclases, and cAMP degradation by phosphodiesterases (PDEs). PDE8A, a high-affinity cAMP-specific PDE is expressed in the ovary and testis. Leydig cells from mice with a targeted mutation in the Pde8a gene are sensitized to the action of LH in terms of testosterone production. These observations led us to evaluate the human PDE8A gene as a PCOS candidate gene, and the hypothesis that reduced PDE8A activity or expression would contribute to excessive ovarian androgen production. We identified a rare variant (R136Q; NM_002605.2 c.407G > A) and studied another known single nucleotide polymorphism (SNP) (rs62019510, N401S) in the PDE8A coding sequence causing non-synonymous amino acid substitutions, and a new SNP in the promoter region (NT_010274.16:g.490155G > A). Although PDE8A kinetics were consistent with reduced activity in theca cell lysates, study of the expressed variants did not confirm reduced activity in cell-free assays. Sub-cellular localization of the enzyme was also not different among the coding sequence variants. The PDE8A promoter SNP and a previously described promoter SNP did not affect promoter activity in in vitro assays. The more common coding sequence SNP (N401S), and the promoter SNPs were not associated with PCOS in our transmission/disequilibrium test-based analysis, nor where they associated with total testosterone or dehydroepiandrosterone sulfate levels. These findings exclude a significant role for PDE8A as a PCOS candidate gene, and as a Las major determinant of androgen levels in women.


Primary pulmonary artery sarcoma resembling chronic thromboembolic pulmonary disease.
Two cases of primary pulmonary artery sarcoma resembling chronic thromboembolic disease features are presented. Tumour identification was achieved after pulmonarv thromboendarterectomy, which was indicated by documentation of a prothrombotic state in both patients. A doubtful history of pulmonary emboli or deep venous thrombosis should alert medical personnel to the possible presence of a primary pulmonary artery sarcoma. Advanced imaging methods such as gadolinium-enhanced magnetic resonance imaging could be useful in considering pulmonary thromboendarterectomy. If a tumour is detected, its surgical resection should be considered with caution, taking into account the poor survival results. Invasion of the adventitia or the right ventricle, as documented in the present cases, is unusual. As far as the present authors know, this is the first report of this kind of tumour and its coexistence with an activated protein C resistance state and type II heparin-induced thrombocytopenia.


The carboxylesterase/cholinesterase gene family in invertebrate deuterostomes.
Carboxylesterase/cholinesterase family members are responsible for controlling the nerve impulse, detoxification and various developmental functions, and are a major target of pesticides and chemical warfare agents. Comparative structural analysis of these enzymes is thus important. The invertebrate deuterostomes (phyla Echinodermata and Hemichordata and subphyla Urochordata and Cephalochordata) lie in the transition zone between invertebrates and vertebrates, and are thus of interest to the study of evolution. Here we have investigated the carboxylesterase/cholinesterase gene family in the sequenced genomes of Strongylocentrotus purpuratus (Echinodermata), Saccoglossus kowalevskii (Hemichordata), Ciona intestinalis (Urochordata) and Branchiostoma floridae (Cephalochordata), using sequence analysis of the catalytic apparatus and oligomerisation domains, and phylogenetic analysis. All four genomes show blurring of structural boundaries between cholinesterases and carboxylesterases, with many intermediate enzymes. Non-enzymatic proteins are well represented. The Saccoglossus and Branchiostoma genomes show evidence of extensive gene duplication and retention. There is also evidence of domain shuffling, resulting in multidomain proteins consisting either of multiple carboxylesterase domains, or of carboxylesterase/cholinesterase domains linked to other domains, including RING finger, chitin-binding, immunoglobulin, fibronectin type 3, CUB, cysteine-rich-Frizzled, caspase activation and 7tm-1, amongst others. Such gene duplication and domain shuffling in the carboxylesterase/cholinesterase family appears to be unique to the invertebrate deuterostomes, and we hypothesise that these factors may have contributed to the evolution of the morphological complexity, particularly of the nervous system and neural crest, of the vertebrates.


Endogenous kisspeptin tone is a critical excitatory component of spontaneous GnRH activity and the GnRH response to NPY and CART.
Kisspeptin is the major excitatory regulator of gonadotropin-releasing hormone (GnRH) neurons and is responsible for basal GnRH/LH release and the GnRH/LH surge. Although it is widely assumed, based on mutations in kisspeptin and Kiss1R, that kisspeptin acts to sustain basal GnRH neuronal activity, there have been no studies to investigate whether endogenous basal kisspeptin tone plays a direct role in basal spontaneous GnRH neuronal excitability. It is also of interest to examine possible interactions between endogenous kisspeptin tone and other neuropeptides that have direct effects on GnRH neurons, such as neuropeptide Y (NPY) or cocaine- and amphetamine-regulated transcript (CART), since the activity of all these neuropeptides changes during states of negative energy balance. Loose cell-attached and whole-cell current patch-clamp recordings were made from GnRH-GFP neurons in hypothalamic slices from female and male rats. Kisspeptin activated GnRH neurons in a concentration-dependent manner with an EC50 of 3.32 ± 0.02 nM. Surprisingly, a kisspeptin antagonist, Peptide 347, suppressed spontaneous activity in GnRH neurons, demonstrating the essential nature of the endogenous kisspeptin tone. Furthermore, inhibition of endogenous kisspeptin tone blocked the direct activation of GnRH cells that occurs in response to antagonism of NPY Y5 receptor or by CART. Our electrophysiology studies suggest that basal endogenous kisspeptin tone is not only essential for spontaneous GnRH neuronal firing, but it is also required for the net excitatory effects of other neuropeptides, such as CART or NPY antagonism, on GnRH neurons. Therefore, endogenous kisspeptin tone could serve as the linchpin in GnRH activation or inhibition.


Immersion diuresis in dogs.
The mechanism of diuresis during the 1st h of immersion was investigated using anesthetized dogs. Four different experiments were carried out. First, left atrial transmural pressure was measured before, during, and after immersion. The data suggest that, although the left atrium may or may not be stretched depending on the conditions of immersion, the amount of diuresis is independent of the amount of left atrial stretch, and therefore a causal relationship between diuresis and left atrial stretch could not be established. Second, bilateral cervical vagotomy was carried out. Immersion diuresis sometimes occurred despite this vagotomy, suggesting that the left atrial stretch reflex was not participating in those cases. Third, negative-pressure breathing was carried out to simulate the negative transthoracic pressure associated with uncompensated immersion. The average left atrial transmural pressure did not change. A slight hemodilution and a moderate diuresis occurred. There was no correlation between changes in left atrial transmural pressure and changes in urine ouput. Fourth, blood studies were done on splenectomized dogs subjected to immersion. Hemodilution occurred and was most marked in dogs which had had their kidneys removed. The hemodilution is sufficient to explain the early phase of the immersion diuresis. The data suggest that, in anesthetized dogs, hemodilution is the probable initiator of diuresis upon immersion and that, in dogs, left atrial stretch is unrelated to diuresis during immersion or negative-pressure breathing.


Inhibition of aldose reductase prevents experimental allergic airway inflammation in mice.
The bronchial asthma, a clinical complication of persistent inflammation of the airway and subsequent airway hyper-responsiveness, is a leading cause of morbidity and mortality in critically ill patients. Several studies have shown that oxidative stress plays a key role in initiation as well as amplification of inflammation in airways. However, still there are no good anti-oxidant strategies available for therapeutic intervention in asthma pathogenesis. Most recent studies suggest that polyol pathway enzyme, aldose reductase (AR), contributes to the pathogenesis of oxidative stress-induced inflammation by affecting the NF-kappaB-dependent expression of cytokines and chemokines and therefore inhibitors of AR could be anti-inflammatory. Since inhibitors of AR have already gone through phase-III clinical studies for diabetic complications and found to be safe, our hypothesis is that AR inhibitors could be novel therapeutic drugs for the prevention and treatment of asthma. Hence, we investigated the efficacy of AR inhibition in the prevention of allergic responses to a common natural airborne allergen, ragweed pollen that leads to airway inflammation and hyper-responsiveness in a murine model of asthma. Primary Human Small Airway Epithelial Cells (SAEC) were used to investigate the in vitro effects of AR inhibition on ragweed pollen extract (RWE)-induced cytotoxic and inflammatory signals. Our results indicate that inhibition of AR prevents RWE -induced apoptotic cell death as measured by annexin-v staining, increase in the activation of NF-kappaB and expression of inflammatory markers such as inducible nitric oxide synthase (iNOS), cycloxygenase (COX)-2, Prostaglandin (PG) E(2), IL-6 and IL-8. Further, BALB/c mice were sensitized with endotoxin-free RWE in the absence and presence of AR inhibitor and followed by evaluation of perivascular and peribronchial inflammation, mucin production, eosinophils infiltration and airway hyperresponsiveness. Our results indicate that inhibition of AR prevents airway inflammation and production of inflammatory cytokines, accumulation of eosinophils in airways and sub-epithelial regions, mucin production in the bronchoalveolar lavage fluid and airway hyperresponsiveness in mice. These results suggest that airway inflammation due to allergic response to RWE, which subsequently activates oxidative stress-induced expression of inflammatory cytokines via NF-kappaB-dependent mechanism, could be prevented by AR inhibitors. Therefore, inhibition of AR could have clinical implications, especially for the treatment of airway inflammation, a major cause of asthma pathogenesis.


Headache in the elderly.
Aging is accompanied by a decline in the incidence of most primary headache disorders and an increase in the incidence of organic causes of headache. Headaches that begin after age 55 are more often due to serious conditions, or are exacerbated by comorbid disorders. When evaluating the older patient with new onset headache or a change in headache pattern, one must rule out serious secondary causes of headache, such as tumor, subdural hematoma, stroke, transient ischemic attack, and temporal arteritis. When older patients present with headache, a lowered threshold for ordering tests is justified, particularly if the headaches are of recent onset, are atypical, or are associated with neurologic findings. This article discusses the secondary causes of headache in the elderly and the symptoms and appropriate testing and treatment of these headaches. The epidemiology and treatment of the primary headache disorders in the elderly are reviewed.


Impact of different organic acids on solubility enhancement of cefpodxime proxetil immediate release tablet and its stability studies.
Cefpodoxime proxetil is a third generation cephalosporin antibiotic demonstrates pH dependent solubility and is highly soluble only in acidic pH. The purpose of this investigation was to design and develop immediate release tablets of cefpodoxime proxetil by direct compression method and determine the effect of different solid buffers (organic acids) such as fumaric acid (formulations F1-F4), maleic acid (formulations M1-M4) and citric acid (formulations C1-C4) by using cefpodoxime and acid in the ratios of 4:1, 2:1, 1:1 and 1:2 to achieve pH-independent release of the drug. Physical parameters and assay were found to be within the acceptable range as prescribed in USP 36 / NF 31. In vitro dissolution studies of each formulation were performed in distilled water, USP dissolution medium, HCl buffer solution of pH 1.2, phosphate buffer solutions of pH 4.5 and 6.8 to observe the drug release. The formulations F3, F4, M4 were selected for film coating on the basis of better drug release profile, to protect the drug from chemical degradation through hydrolysis. Film coated formulation F3, F4 and M4 showed a remarkable in vitro release of the drug (72.88±0.43 to 92.67±0.71%) within 30min of observation in all dissolution media and further evaluated by model independent and model dependent approaches. The drug release was found to be best fit to Weibull model as highest r2adjusted (0.924-0.998) and lowest AIC (18.416-54.710) values were obtained in all dissolution media. R Gui® applied for stability studies of F3 and F4 formulations, showing shelf lives of 28 & 27months at ambient and 33 months at accelerated temperatures. Formulation F4 was chosen as best formulation on the basis of physical properties, highest dissolution rate and stability studies.


Gene expression profiling of patient-derived pancreatic cancer xenografts predicts sensitivity to the BET bromodomain inhibitor JQ1: implications for individualized medicine efforts.
c-MYC controls more than 15% of genes responsible for proliferation, differentiation, and cellular metabolism in pancreatic as well as other cancers making this transcription factor a prime target for treating patients. The transcriptome of 55 patient-derived xenografts show that 30% of them share an exacerbated expression profile of MYC transcriptional targets (MYC-high). This cohort is characterized by a high level of Ki67 staining, a lower differentiation state, and a shorter survival time compared to the MYC-low subgroup. To define classifier expression signature, we selected a group of 10 MYC target transcripts which expression is increased in the MYC-high group and six transcripts increased in the MYC-low group. We validated the ability of these markers panel to identify MYC-high patient-derived xenografts from both: discovery and validation cohorts as well as primary cell cultures from the same patients. We then showed that cells from MYC-high patients are more sensitive to JQ1 treatment compared to MYC-low cells, in monolayer, 3D cultured spheroids and in vivo xenografted tumors, due to cell cycle arrest followed by apoptosis. Therefore, these results provide new markers and potentially novel therapeutic modalities for distinct subgroups of pancreatic tumors and may find application to the future management of these patients within the setting of individualized medicine clinics.


Thermal treatment of eggplant (Solanum melongena L.) increases the antioxidant content and the inhibitory effect on human neutrophil burst.
The aim of this study was to compare the amount and activity of phytonutrients in raw, grilled, and boiled eggplant fruit using chemical measures and a biological assay of oxidative bursts in human neutrophils. The thermally treated samples showed various changes in their chemical composition (dry matter, soluble solids, acidity, and the amount of alcohol insoluble substances) due to the cooking processes and were much richer in the main phenolic compounds such as chlorogenic and caffeic acids, which are known to be antioxidants. Consequently, their free radical scavenging activity was significantly higher, especially that of superoxide anion. The biological assay of oxidative bursts from human neutrophils in the presence of N-formyl-methionyl-leucyl-phenylalanine confirmed the greater activity of extracts of the cooked eggplants with respect to raw eggplants. Successive extract dilutions showed a significant activity up to 1.25 microg/mL after cooking, while raw fruits resulted in an activity up to 10.00 microg/mL. These results showed that the thermal treatment commonly used before consumption can increase the content and biological activity of antioxidant compounds of eggplants.


Early Decompression (< 8 h) after Traumatic Cervical Spinal Cord Injury Improves Functional Outcome as Assessed by Spinal Cord Independence Measure after One Year.
There is an ongoing controversy about the optimal timing for surgical decompression after acute traumatic cervical spinal cord injury (SCI). For this reason, we performed a retrospective study of patients who were operated on after traumatic cervical SCI at the Trauma Center Murnau, Germany, and who met inclusion as well as exclusion criteria (n = 70 patients). Follow-up data were collected prospectively according to the European Multicenter Study about Spinal Cord Injury (EMSCI) protocol over a period of 1 year. Early decompression was defined as within the first 8 h after the insult (n = 35 patients). Primary outcome was the difference in the SCIM (Spinal Cord Independence Measure) 1 year after the trauma. After the follow-up period, patients who were decompressed earlier had a significantly higher SCIM difference (45.8 vs. 27.1, p < 0.005). A regression analysis showed that timing of decompression, age, as well as basal AIS (American Spinal Injury Association Impairment Scale) and basal SCIM scores were independent predictors for a better functional outcome (SCIM). Further, patients from the early decompression group had better AIS grades (p < 0.006) and a higher AIS conversion rate (p < 0.029). Additionally, this cohort also had a better total motor performance as well as upper extremity motor function after 1 year (p < 0.025 and p < 0.002). The motor and neurological levels of patients who were operated on within 8 h were significantly more caudal (p < 0.003 and p < 0.014) after 1 year. The present study suggests that early decompression after traumatic cervical SCI might have a positive impact on the functional and neurological outcome of affected individuals.


Creeping edgework: carnivalesque consumption and the social experience of health risk.
This article contributes to an understanding of voluntary health risk based on the regular, excessive intake of food and alcohol in the micro-cultural setting. By drawing on and extending edgework theory our aim is to conceptualise the riskiness of carnivalesque consumption as a medium for expression and performance in two separate community contexts. Using ethnographic research methods, we explore the consumption of calorie-dense, low nutrient food for gamers and the use of alcohol for hipsters. Our findings are reported over four key themes. The first and last consider how carnivalesque consumption provides sensations for multi-sensory loss of self and a shared emancipation from day-to-day moderation. The second and third explore how community members prepare and exercise control over their consumption to manage risks related to an 'immediate edge'. We discuss how carnivalesque behaviour, when ritualised, establishes a trajectory that creeps towards a more 'distant edge' characterised by longer term health consequences. We argue that the transcendental experiences that are part and parcel of edgework can be enacted by products that are traditionally conceptualised as mundane and that the risks of consuming them are largely accumulative rather than instantaneous. Implications for health interventions are included.


Over- and under-expressed microRNAs in human colorectal cancer.
MicroRNAs (miRNAs) constitute a class of small (21-23 nucleotides) noncoding RNAs that function as post-transcriptional gene regulators. It is becoming increasingly clear that altered miRNA expression correlates with the pathogenesis of cancers. The purpose of this study was to determine the up-regulated miRNAs in human colorectal cancer. Total RNA was isolated from cancer tissues and corresponding noncancerous tissues from surgically resected colorectal cancers. The expression profiles of miRNAs were determined using a miRNA microarray containing 455 human miRNA probes. The expression status of selected miRNAs in paired clinical samples was then investigated by real-time RT-PCR. Twenty-one miRNAs were identified by miRNA array analysis as overexpressed in colorectal cancer tissues compared to normal epithelial tissues. Among them, the expression of miR-31, miR-183, miR-17-5p, miR-18a, miR-20a and miR-92 were confirmed to be significantly higher in cancer tissues than in normal tissues (P<0.05). In contrast, the expression of miR-143 and miR-145 in cancer tissues were significantly lower than in normal tissues (P<0.05). The miR-18a overexpression group tended to have a poorer clinical prognosis than the low expression group (P=0.07). We identified miRNAs that were overexpressed or under-expressed in colorectal cancers and which may be correlated with colorectal carcinogenesis.


A feasibility study of colorectal cancer diagnosis via circulating tumor DNA derived CNV detection.
Circulating tumor DNA (ctDNA) has shown great promise as a biomarker for early detection of cancer. However, due to the low abundance of ctDNA, especially at early stages, it is hard to detect at high accuracies while keeping sequencing costs low. Here we present a pilot stage study to detect large scale somatic copy numbers variations (CNVs), which contribute more molecules to ctDNA signal compared to point mutations, via cell free DNA sequencing. We show that it is possible to detect somatic CNVs in early stage colorectal cancer (CRC) patients and subsequently discriminate them from normal patients. With 25 normal and 24 CRC samples, we achieve 100% specificity (lower bound confidence interval: 86%) and ~79% sensitivity (95% confidence interval: 63% - 95%,), though the performance should be considered with caution given the limited sample size. We report a lack of concordance between the CNVs detected via cfDNA sequencing and CNVs identified in parent tissue samples. However, recent findings suggest that a lack of concordance is expected for CNVs in CRC because of their sub-clonal nature. Finally, the CNVs we detect very likely contribute to cancer progression as they lie in functionally important regions, and have been shown to be associated with CRC specifically. This study paves the path for a larger scale exploration of the potential of CNV detection for both diagnoses and prognoses of cancer.


Noninvasive electroanatomic mapping of human ventricular arrhythmias with electrocardiographic imaging.
The rapid heartbeat of ventricular tachycardia (VT) can lead to sudden cardiac death and is a major health issue worldwide. Efforts to identify patients at risk, determine mechanisms of VT, and effectively prevent and treat VT through a mechanism-based approach would all be facilitated by continuous, noninvasive imaging of the arrhythmia over the entire heart. Here, we present noninvasive real-time images of human ventricular arrhythmias using electrocardiographic imaging (ECGI). Our results reveal diverse activation patterns, mechanisms, and sites of initiation of human VT. The spatial resolution of ECGI is superior to that of the routinely used 12-lead electrocardiogram, which provides only global information, and ECGI has distinct advantages over the currently used method of mapping with invasive catheter-applied electrodes. The spatial resolution of this method and its ability to image electrical activation sequences over the entire ventricular surfaces in a single heartbeat allowed us to determine VT initiation sites and continuation pathways, as well as VT relationships to ventricular substrates, including anatomical scars and abnormal electrophysiological substrate. Thus, ECGI can map the VT activation sequence and identify the location and depth of VT origin in individual patients, allowing personalized treatment of patients with ventricular arrhythmias.


Audit of outcomes in motor neuron disease (MND) patients treated with riluzole.
A review of riluzole in MND by NICE concluded that riluzole was modestly effective. NICE however suggested that further studies were needed to gain further insights into its clinical effectiveness. As these studies have not been carried out we audited outcomes in our use of riluzole. Median survival of 148 MND patients receiving riluzole was 3.07 years (2.25 years for 327 patients not given riluzole), hazard ratio 1.66, 95% confidence interval (CI) 1.32-2.12; 3.61 years in 103 limb onset patients given riluzole (2.62 years for 124 limb onset patients not given riluzole), hazard ratio 1.50, 95% CI 1.09-2.05; 2.19 years in 43 bulbar onset patients receiving riluzole (1.84 years for 103 bulbar onset patients not given riluzole), hazard ratio 1.34, 95% CI 0.89-2.07 and 3.80 years in 87 patients with a PEG given riluzole (2.21 years for 261 patients with a PEG not given riluzole), hazard ratio 1.86, 95% CI 1.44-2.39. These findings are comparable with the results of similar analyses from other groups. The uncertainties they raise emphasize the importance of adequate randomized studies being completed prior to licensing of new drugs. They further illustrate the desirability of controlled introduction of new drugs into clinical practice when RCTs have only suggested a modest level of efficacy.


Health-related quality of life as predictor for mortality in patients treated with long-term mechanical ventilation.
The Severe Respiratory Insufficiency (SRI) questionnaire is a specific measure of health-related quality of life (HRQoL) in patients treated with long-term mechanical ventilation (LTMV). The aim of the present study was to examine whether SRI sum scores and related subscales are associated with mortality in LTMV patients. The study included 112 LTMV patients (non-invasive and invasive) from the Norwegian LTMV registry in Western Norway from 2008 with follow-up in August 2014. SRI data were obtained through a postal questionnaire, whereas mortality data were obtained from the Norwegian Cause of Death Registry. The SRI questionnaire contains 49 items and seven subscales added into a summary score (range 0-100) with higher scores indicating a better HRQoL. The association between the SRI score and mortality was estimated as hazard ratios (HRs) with 95% confidence intervals (95% CI) using Cox regression models and HRs were estimated per one unit change in the SRI score. Of the 112 participating patients in 2008, 52 (46%) had died by August 2014. The mortality rate was the highest in patients with chronic obstructive pulmonary disease (75%), followed by patients with neuromuscular disease (46%), obesity hypoventilation syndrome (31%) and chest wall disease (25%) (p < 0.001). Higher SRI sum scores in 2008 were associated with a lower mortality risk after adjustment for age, education, hours a day on LTMV, time since initiation of LTMV, disease category and comorbidity (HR 0.98, 95% CI: 0.96-0.99). In addition, SRI-Physical Functioning (HR 0.98, 95% CI: 0.96-0.99), SRI-Psychological Well-Being (HR 0.98, 95% CI: 0.97-0.99), and SRI-Social Functioning (HR 0.98, 95% CI: 0.97-0.99) remained significant risk factors for mortality after covariate adjustment. In the subgroup analyses of patient with neuromuscular diseases we found significant inverse associations between some of the SRI subscales and mortality. SRI score is associated with mortality in LTMV-treated patients. We propose the use of SRI in the daily clinic with repeated measurements as part of individual follow-up. Randomized clinical trials with interventions aimed to improve HRQoL in LTMV patients should consider the SRI questionnaire as the standard HRQoL measurement.


Seroprevalence of Human Immunodeficiency Virus Among Antenatal Women in One of the Institute of Northern India.
As per NACO publication there are an estimated 2.08 Million HIV/AIDS cases in India as of February 2014. The estimated adult HIV prevalence in India is 0.27%, with a gender spit of 0.32% males and 0.22% females. Estimating the seroprevalence of HIV in a low risk population such as pregnant women provides essential information for an effective implementation of AIDS control programmes. In spite of an estimated adult HIV prevalence rate of 0.18% in Punjab, there are no previous studies among antenatal women to assess seroprevalence. To assess the prevalence of HIV infection, among otherwise healthy pregnant females. This retrospective study reviewed treatment records of 3686 antenatal women who underwent HIV testing. Demographic, obstetric and laboratory data was recorded into a preset proforma. Available data was analysed using SPSS version 13. Total of 38 subjects tested positive for HIV infection. Seroprevalence rate was 1.03%. A 25 -30 years age group had most number of infected individuals with 22 (57.89%) subjects. Seropositivity was common among rural domicile. Only 9 (23.68%) was aware of their seropositive status. A total of 20 (52.63%) subjects were illiterate, 14 (36.84%) had received primary school education. Seroprevalence rate of 1.03% reported in this study has been the highest among the contemporary articles published in last decade. Efficient surveillance along with proper implementation of Prevention of Parent to Child Transmission (PPTCT) is the need of the day. A prompt action at the hands of the authority will enable us to safeguard our present and future alike.


Protective Effects of AGE and Its Components on Neuroinflammation and Neurodegeneration.
Garlic (Allium sativum) is used for culinary and medicinal purposes in diverse cultures worldwide. When fresh garlic is soaked in aqueous ethanol under ambient environment over 4 months or longer, the majority of irritating taste and odor is eliminated and the antioxidant profile in the resulting aged garlic extract (AGE) changes significantly. Recently, AGE and its components have been demonstrated to exert neuroprotective effects in neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, Huntington's disease, and cerebral ischemia. Because of its health supporting potential, there is increasing interest in understanding the antioxidant and anti-inflammatory properties and the underlying mechanisms for its protective effects in heath and disease. There is evidence for AGE to exert its action on distinct signaling pathways associated with oxidative stress and neuroinflammation, although the primary molecular mechanisms remain unclear. By utilizing quantitative proteomic approaches, we demonstrated that AGE and two of its major ingredients, S-allyl-L-cysteine and N (α)-(1-deoxy-D-fructos-1-yl)-L-arginine, can attenuate neuroinflammatory responses in microglial cells through modulation of Nrf2-mediated signaling as well as other oxidative stress-related pathways. These experimental data provide information for the molecular targets of AGE and its components to mitigate neurodegeneration and neuroinflammation and show a promising potential of these compounds as dietary supplements for health maintenance.


Cyclic olefin homopolymer-based microfluidics for protein crystallization and in situ X-ray diffraction.
Microfluidics is a promising technology for the rapid identification of protein crystallization conditions. However, most of the existing systems utilize silicone elastomers as the chip material which, despite its many benefits, is highly permeable to water vapour. This limits the time available for protein crystallization to less than a week. Here, the use of a cyclic olefin homopolymer-based microfluidics system for protein crystallization and in situ X-ray diffraction is described. Liquid handling in this system is performed in 2 mm thin transparent cards which contain 500 chambers, each with a volume of 320 nl. Microbatch, vapour-diffusion and free-interface diffusion protocols for protein crystallization were implemented and crystals were obtained of a number of proteins, including chicken lysozyme, bovine trypsin, a human p53 protein containing both the DNA-binding and oligomerization domains bound to DNA and a functionally important domain of Arabidopsis Morpheus' molecule 1 (MOM1). The latter two polypeptides have not been crystallized previously. For X-ray diffraction analysis, either the cards were opened to allow mounting of the crystals on loops or the crystals were exposed to X-rays in situ. For lysozyme, an entire X-ray diffraction data set at 1.5 A resolution was collected without removing the crystal from the card. Thus, cyclic olefin homopolymer-based microfluidics systems have the potential to further automate protein crystallization and structural genomics efforts.


A novel cardiovascular risk stratification model incorporating ECG and heart rate variability for patients presenting to the emergency department with chest pain.
Risk stratification models can be employed at the emergency department (ED) to evaluate patient prognosis and guide choice of treatment. We derived and validated a new cardiovascular risk stratification model comprising vital signs, heart rate variability (HRV) parameters, and demographic and electrocardiogram (ECG) variables. We conducted a single-center, observational cohort study of patients presenting to the ED with chest pain. All patients above 21 years of age and in sinus rhythm were eligible. ECGs were collected and evaluated for 12-lead ECG abnormalities. Routine monitoring ECG data were processed to obtain HRV parameters. Vital signs and demographic data were obtained from electronic medical records. Thirty-day major adverse cardiac events (MACE) were the primary endpoint, including death, acute myocardial infarction, and revascularization. Candidate variables were identified using univariate analysis; the model for the final risk score was derived by multivariable logistic regression. We compared the performance of the new model with that of the thrombolysis in myocardial infarct (TIMI) score using receiver operating characteristic (ROC) analysis. In total, 763 patients were included in this study; 254 (33 %) met the primary endpoint, the mean age was 60 (σ = 13) years, and the majority was male (65 %). Nineteen candidate predictors were entered into the multivariable model for backward variable elimination. The final model contained 10 clinical variables, including age, gender, heart rate, three HRV parameters (average R-to-R interval (RR), triangular interpolation of normal-to-normal (NN) intervals, and high-frequency power), and four 12-lead ECG variables (ST elevation, ST depression, Q wave, and QT prolongation). Our proposed model outperformed the TIMI score for prediction of MACE (area under the ROC curve 0.780 versus 0.653). At the cutoff score of 9 (range 0-37), our model had sensitivity of 0.709 (95 % CI 0.653, 0.765), specificity of 0.674 (95 % CI 0.633, 0.715), positive predictive value of 0.520 (95 % CI 0.468, 0.573), and negative predictive value of 0.823 (95 % CI 0.786, 0.859). A non-invasive and objective ECG- and HRV-based risk stratification tool performed well against the TIMI score, but future research warrants use of an external validation cohort.


Solution conformation of the (-)-trans-anti-[BP]dG adduct opposite a deletion site in a DNA duplex: intercalation of the covalently attached benzo[a]pyrene into the helix with base displacement of the modified deoxyguanosine into the minor groove.
A combined NMR-computational approach was employed to determine the solution structure of the (-)-trans-anti-[BP]dG adduct positioned opposite a -1 deletion site in the d(C1-C2-A3-T4-C5- [BP]G6-C7-T8-A9-C10-C11).d(G12-G13-T14-A15-G1 6-G17-A18-T19-G20-G21) sequence context. The (-)-trans-anti-[BP]dG moiety is derived from the binding of the (-)-anti-benzo[a]pyrene diol epoxide [(-)-anti-BPDE] to N2 of dG6 and has a 10R absolute configuration at the [BP]dG linkage site. The exchangeable and non-exchangeable protons of the benzo[a]pyrenyl moiety and the nucleic acid were assigned following analysis of two-dimensional NMR data sets in H2O and D2O solution. The solution conformation has been determined by incorporating intramolecular and intermolecular proton-proton distances defined by lower and upper bounds deduced from NOESY spectra as restraints in molecular mechanics computations in torsion angle space followed by restrained molecular dynamics calculations based on a NOE distance and intensity refinement protocol. Our structural studies establish that the aromatic BP ring system intercalates into the helix opposite the deletion site, while the modified deoxyguanosine residue is displaced into the minor groove with its face parallel to the helix axis. The intercalation site is wedge-shaped and the BP aromatic ring system stacks over intact flanking Watson-Crick dG.dC base pairs. The modified deoxyguanosine stacks over the minor groove face of the sugar ring of the 5'-flanking dC5 residue. The BP moiety is positioned with the benzylic ring oriented toward the minor groove and the distal pyrenyl aromatic ring directed toward the major groove. This conformation strikingly contrasts with the corresponding structure in the full duplex with the same 10R (-)-trans-anti-[BP]dG lesion positioned opposite a complementary dC residue [de los Santos et al. (1992) Biochemistry 31, 5245-5252); in this case the aromatic BP ring system is located in the minor groove, and there is no disruption of the [BP]dG.dC Watson-Crick base pairing alignment. The intercalation-base displacement features of the 10R (-)-trans-anti-[BP]dG adduct opposite a deletion site have features in common to those of the 10S (+)-trans-anti-[BP]dG adduct opposite a deletion site previously reported by Cosman et al. [(1994)(Biochemistry 33, 11507-11517], except that there is a nearly 180 degrees rotation of the BP residue about the axis of the helix at the base-displaced intercalation site and the modified deoxyguanosine is positioned in the opposite groove. In the 10S adduct, the benzylic ring is in the major groove and the aromatic ring systems point toward the minor groove. This work extends the theme of opposite orientations of adducts derived from chiral pairs of (+)- and (-)-anti-BPDE enantiomers; both 10S and 10R adducts can be positioned with opposite orientations either in the minor groove or at base displaced intercalation sites, depending on the presence or absence of the partner dC base in the complementary strand.


Compulsive sexual behavior disorder in 42 countries: Insights from the International Sex Survey and introduction of standardized assessment tools.
Despite its inclusion in the 11th revision of the International Classification of Diseases, there is a virtual paucity of high-quality scientific evidence about compulsive sexual behavior disorder (CSBD), especially in underrepresented and underserved populations. Therefore, we comprehensively examined CSBD across 42 countries, genders, and sexual orientations, and validated the original (CSBD-19) and short (CSBD-7) versions of the Compulsive Sexual Behavior Disorder Scale to provide standardized, state-of-the-art screening tools for research and clinical practice. Using data from the International Sex Survey (N = 82,243; Mage = 32.39 years, SD = 12.52), we evaluated the psychometric properties of the CSBD-19 and CSBD-7 and compared CSBD across 42 countries, three genders, eight sexual orientations, and individuals with low vs. high risk of experiencing CSBD. A total of 4.8% of the participants were at high risk of experiencing CSBD. Country- and gender-based differences were observed, while no sexual-orientation-based differences were present in CSBD levels. Only 14% of individuals with CSBD have ever sought treatment for this disorder, with an additional 33% not having sought treatment because of various reasons. Both versions of the scale demonstrated excellent validity and reliability. This study contributes to a better understanding of CSBD in underrepresented and underserved populations and facilitates its identification in diverse populations by providing freely accessible ICD-11-based screening tools in 26 languages. The findings may also serve as a crucial building block to stimulate research into evidence-based, culturally sensitive prevention and intervention strategies for CSBD that are currently missing from the literature.


Therapy of collagen VI-related myopathies (Bethlem and Ullrich).
The collagen VI-related myopathies comprise two major forms, Bethlem myopathy (BM) and Ullrich congenital muscular dystrophy (UCMD), which show a variable combination of muscle wasting and weakness, joint contractures, distal laxity, and respiratory compromise. Specific diagnosis requires molecular genetic testing showing mutation in one of the three genes involved. This review summarizes current treatments, in particular indication for physiotherapy, orthopedic treatment for correction of foot deformity, scoliosis, and flexion contractures of elbows, and treatment of respiratory failure. The turning point in basic research on collagen VI myopathies was the discovery of an unexpected mitochondrial dysfunction as a pathogenetic mechanism underlying the myopathic syndrome seen in Col6a1 null mice. Treatment of Col6a1(-/-) mice with cyclosporin A (CsA) rescued the mitochondrial dysfunction and decreased apoptosis. Similar mitochondrial defects were revealed in cultures of UCMD patients. The results of an open pilot trial with CsA in five patients with collagen VI-related myopathies are summarized and discussed. With the availability of new potential effective treatments, several challenges must be addressed in conducting trials in orphan diseases and in neuromuscular disorders in particular. Outcome measures are discussed in the context of the expected effect of the cure. Randomized clinical trials often are not feasible for rare diseases, and sometimes would be ethically inappropriate. The need to develop alternative outcome measures or biomarkers using platforms such as genomics and proteomics is stressed in this context.


Association Between Employment Status and Objectively Measured Physical Activity and Sedentary Behavior-The Maastricht Study.
To examine the association between employment status and physical activity and sedentary behavior. We included 2045 participants from The Maastricht Study, who used a thigh-worn accelerometer. We compared time spent sedentary, standing, stepping, and higher intensity physical activity between participants with different employment status (non-employed or low-, intermediate- or high-level occupation) with analysis of variance. Participants in low-level occupations were less sedentary and standing and stepping more than those in other occupational categories and non-employed participants. Among the employed, the differences were mostly observed on weekdays, whereas the differences in sedentary time and standing between those in low-level occupations and non-employed participants were evident both on weekdays and weekend days. Those in low-level occupational category were less sedentary and more active than non-employed and those in other occupational categories, especially on weekdays.


The microRNA signature of mouse spermatozoa is substantially modified during epididymal maturation.
In recent years considerable effort has been devoted to understanding the epigenetic control of sperm development, leading to an increased appreciation of the importance of RNA interference pathways, and in particular miRNAs, as key regulators of spermatogenesis and epididymal maturation. It has also been shown that sperm are endowed with an impressive array of miRNA that have been implicated in various aspects of fertilization and embryo development. However, to date there have been no reports on whether the sperm miRNA signature is static or whether it is influenced by their prolonged maturation within the male reproductive tract. To investigate this phenomenon, we employed next-generation sequencing to systematically profile the miRNA signature of maturing mouse spermatozoa. In so doing we have provided the first evidence for the posttesticular modification of the sperm miRNA profile under normal physiological conditions. Such modifications include the apparent loss and acquisition of an impressive cohort of some 113 and 115 miRNAs, respectively, between the proximal and distal epididymal segments. Interestingly, the majority of these changes occur late in maturation and include the uptake of novel miRNA species in addition to a significant increase in many miRNAs natively expressed in immature sperm. Because sperm are not capable of de novo transcription, these findings identify the epididymis as an important site in establishing the sperm epigenome with the potential to influence the peri-conceptual environment of the female reproductive tract, contribute to the inheritance of acquired characteristics, and/or alter the developmental trajectory of the resulting offspring.


Referential adjustment during discourse production in Alzheimer's disease.
Several studies have shown that people with Alzheimer's disease (AD) tend to use more pronouns than healthy aged adults when referring to entities during discourse. This referential behavior has been associated with the decrease of cognitive abilities, such as lexical retrieval difficulties or reduced abilities in working memory. However, the influence of certain important discourse factors on the referential choices made by people with AD has yet to be established. This study examines referential choices made at three discourse stages during narrative discourse (the introduction of a referent, the maintaining of the referent in focus, and the shift from one referent in focus to another). These referential choices are examined in increasingly complex referential contexts. In addition, this study investigates the relationships between referential choices and various cognitive abilities. To do so, the narrative discourses of 21 people with AD and 21 healthy adults were elicited using a newly developed storytelling in sequence task. The analyses focused on the production of three major referential expressions (indefinite expressions, definite expressions and pronouns) which are expected to vary according to discourse stage and the referential complexity of the stories. The results show that AD participants produce significantly fewer of the referential expressions expected at the introduction and shift stages than healthy aged adults produce. Nevertheless, the variation in the categories of referential expressions produced by the AD participants between the discourse stages is similar to that produced by the healthy aged adults, suggesting a preserved sensitivity to the factors manipulated in the task (i.e., discourse stages and referential complexity). This study also highlights the fact that different cognitive competences, especially executive abilities, are greatly involved in referential choices. The results add further evidence that referential choices rely on a variety of cognitive skills, depending on the discourse context in which they are made.


Modulation of locomotor activity in larval zebrafish during light adaptation.
The neural basis of behavioral choice in vertebrates remains largely unknown. Zebrafish larvae have a defined locomotor repertoire as well as a simple nervous system and are therefore an attractive vertebrate system in which to study this process. Here we describe a high-throughput system for quantifying the kinematics of motor events in zebrafish larvae in order to measure the initiation frequency of different maneuvers. We use this system to analyze responses to photic stimuli and find that larvae respond to changes in illumination with both acute responses and extended behavioral programs. Reductions in illumination elicit large angle turns, distinct from startle responses, which orient larvae toward the source of light. In continuing darkness, larvae are transiently hyperactive before adopting a quiescent state. Indeed, locomotor activity is controlled by the state of light or dark adaptation similar to masking phenomena in higher vertebrates where light directly regulates motor activity. We propose that regulation of motor activity by photic stimuli in zebrafish larvae serves a behavioral goal of maximizing exposure to well lit environments optimal for feeding.


The HPV-16 E6 and E6-AP complex functions as a ubiquitin-protein ligase in the ubiquitination of p53.
The ubiquitin-dependent proteolytic pathway plays a major role in selective protein degradation. Ubiquitination of proteins requires the sequential action of the ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzymes (E2), and in some cases ubiquitin-protein ligases (E3s). The oncogenic human papillomavirus (HPV) types 16 and 18 utilize this cellular proteolytic system to target the tumor suppressor protein p53. The HPV E6 oncoprotein binds to a cellular protein of 100 kd, termed E6-associated protein (E6-AP). The E6-E6-AP complex specifically interacts with p53, resulting in the rapid ubiquitin-dependent degradation of p53. Here we report the purification and identification of the factors necessary for the E6-E6-AP-mediated ubiquitination of p53. The ubiquitination of p53 requires the E1 enzyme and a novel E2 in mammalian cells, while E3 activity is conferred by the E6-E6-AP complex. Furthermore, E6-AP appears to have ubiquitin-protein ligase activity in the absence of E6.


Vascular risk factors and cognitive decline among elderly male twins.
Studies investigating the association between cardiovascular risk factors and cognitive decline report discrepant outcomes. The co-twin control method improves on traditional case-control approaches by controlling for within-twin pair similarities of genetic and early environmental influences. To examine the association of diabetes, hypertension, hypercholesterolemia, and elevated body mass index (BMI) (>30 kg/m(2)), individually and combined, with cognitive decline over a 12-year period in members of the National Academy of Sciences-National Research Council Twin Registry of World War II male veterans. Modified Telephone Interview for Cognitive Status (TICS-m) was administered four times at approximately 4-year intervals from 1990 to 2002 as part of an epidemiologic study of dementia. Self-report medical information was collected from 1996 to 2002. We examined the difference in cognitive decline within twin pairs discordant for the vascular risk factors while controlling for baseline TICS-m, education, smoking, and alcohol history. Among twin pairs discordant for diabetes (n = 177), the diabetic twins declined an average of almost 1 point more than their nondiabetic co-twins (p = 0.018) at the last screening time point. Further analyses showed that this was in large part due to greater decline among older men (age 76 to 84 years). Cognitive change was not significantly different between members of pairs discordant for hypertension (n = 326), hypercholesterolemia (n = 282), or elevated BMI (n = 166). Based on this study of twin pairs who share similar genetic and early environmental risks for cardiovascular risk factors, diabetes is associated with greater cognitive decline, particularly among the oldest individuals.


S100A9 Protein Aggregates Boost Hippocampal Glutamate Modifying Monoaminergic Neurochemistry: A Glutamate Antibody Sensitive Outcome on Alzheimer-like Memory Decline.
Alzheimer's disease (AD) involves dementia conceivably arising from integrated inflammatory processes, amyloidogenesis, and neuronal apoptosis. Glutamate can also cause neuronal death via excitotoxicity, and this is similarly implicated in some neurological diseases. The aim was to examine treatment with in vitro generated proinflammatory protein S100A9 aggregate species alone or with glutamate antibodies (Glu-Abs) on Morris water maze (MWM) spatial learning and memory performance in 12 month old mice. Amino acid and monoamine cerebral neurotransmitter metabolic changes were concurrently monitored. Initially, S100A9 fibrils were morphologically verified by atomic force microscopy and Thioflavin T assay. They were then administered intranasally alone or with Glu-Abs for 14 days followed by a 5 day MWM protocol before hippocampal and prefrontal cortical neurochemical analysis. S100A9 aggregates evoked spatial amnesia which correlated with disrupted glutamate and dopaminergic neurochemistry. Hippocampal glutamate release, elevation of DOPAC and HVA, as well as DOPAC/DA and HVA/DA ratios were subsequently reduced by Glu-Abs which simultaneously prevented the spatial memory deficit. The present outcomes emphasized the pathogenic nature of S100A9 fibrillar aggregates in causing spatial memory amnesia associated with enhanced hippocampal glutamate release and DA-ergic disruption in the aging brain. This finding might be exploited during dementia management through a neuroprotective strategy.


Self-assembly of peptide amphiphiles for drug delivery: the role of peptide primary and secondary structures.
Peptide amphiphiles (PAs), functionalized with alkyl chains, are capable of self-assembling into various nanostructures. Recently, PAs have been considered as ideal drug carriers due to their good biocompatibility, specific biological functions, and hypotoxicity to normal cells and tissues. Meanwhile, the nanocarriers formed by PAs are able to achieve controlled drug release and enhanced cell uptake in response to the stimulus of the physiological environment or specific biological factors in the location of the lesion. However, the underlying detailed drug delivery mechanism, especially from the aspect of primary and secondary structures of PAs, has not been systematically summarized or discussed. Focusing on the relationship between the primary and secondary structures of PAs and stimuli-responsive drug delivery applications, this review highlights the recent advances, challenges, and opportunities of PA-based functional drug nanocarriers, and their potential pharmaceutical applications are discussed.


Blood pressure of urban Native American school children.
In 307 Native American (NA), 1784 black (B), and 7777 white (W) children in grades 1, 2, and 3 in Minneapolis schools (99% overall response rate), blood pressure (BP) was measured supine in the right arm after 5 minutes' rest by trained technicians using a random zero BP device. In addition, height, weight, pulse rate, and triceps skinfold thickness were measured. Among children aged 6 through 9 years, NA children had slightly higher systolic BP (SBP) than B or W children overall (mean SBP: NA 106, B 104, W 105 mm Hg) and for nearly all age sex groups. In contrast, Phase 4 and 5 diastolic BP (DBP) were consistently lower in NA children ( mean DBP4: NA 64, B 69, W 67 mm Hg); NA children also had lower pulse rates, greater pulse pressures, similar or slightly lower mean BP, similar height, greater weight, body mass index, and triceps skinfold. Multiple regression analyses revealed that the slightly higher SBP in NA children was explained almost entirely by greater ponderosity. However, the lower DBP could not be explained statistically by any of the variables measured.


A multidisciplinary health care team's efforts to improve educational attainment in children with sickle-cell anemia and cerebral infarcts.
The primary objective of this study was to improve the educational success of children with sickle-cell disease (SCD) and cerebral infarcts. A prospective intervention trial was conducted; a multidisciplinary team was created to maximize educational resources for children with SCD and cerebral infarcts. Students were evaluated systematically before and after the intervention. A baseline evaluation was completed assessing the presence of an Individualized Education Plan (IEP), grade retention in school, and days absent from school in the year preceding the intervention. A postintervention assessment occurred 2 years later for these same measurements. At baseline, 74% (17 of 23) of the students were receiving IEPs. Two years later, 87% (20 of 23) students received IEPs (p = .34). Despite the intervention, the rate of children retained in their school grade increased from 0.6 per 100 years in school at baseline to 1.7 per 100 years, 95% CI (-3.86, 1.49). The school absenteeism rate did not significantly change after the intervention; the average days absent per student rose from 15.5 to 22.5, (p = .05). The multidisciplinary team effort alone was insufficient to decrease grade retention and absenteeism rate. Further support, from either the parents or school administration, is needed to increase education attainment of students with cerebral infarcts.


A Low-Cost Simulation Model for R-Wave Synchronized Atrial Pacing in Pediatric Patients with Postoperative Junctional Ectopic Tachycardia.
Postoperative junctional ectopic tachycardia (JET) occurs frequently after pediatric cardiac surgery. R-wave synchronized atrial (AVT) pacing is used to re-establish atrioventricular synchrony. AVT pacing is complex, with technical pitfalls. We sought to establish and to test a low-cost simulation model suitable for training and analysis in AVT pacing. A simulation model was developed based on a JET simulator, a simulation doll, a cardiac monitor, and a pacemaker. A computer program simulated electrocardiograms. Ten experienced pediatric cardiologists tested the model. Their performance was analyzed using a testing protocol with 10 working steps. Four testers found the simulation model realistic; 6 found it very realistic. Nine claimed that the trial had improved their skills. All testers considered the model useful in teaching AVT pacing. The simulation test identified 5 working steps in which major mistakes in performance test may impede safe and effective AVT pacing and thus permitted specific training. The components of the model (exclusive monitor and pacemaker) cost less than $50. Assembly and training-session expenses were trivial. A realistic, low-cost simulation model of AVT pacing is described. The model is suitable for teaching and analyzing AVT pacing technique.


Tle1 tumor suppressor negatively regulates inflammation in vivo and modulates NF-κB inflammatory pathway.
Tle1 (transducin-like enhancer of split 1) is a corepressor that interacts with a variety of DNA-binding transcription factors and has been implicated in many cellular functions; however, physiological studies are limited. Tle1-deficient (Tle1(Δ/Δ)) mice, although grossly normal at birth, exhibit skin defects, lung hypoplasia, severe runting, poor body condition, and early mortality. Tle1(Δ/Δ) mice display a chronic inflammatory phenotype with increased expression of inflammatory cytokines and chemokines in the skin, lung, and intestine and increased circulatory IL-6 and G-CSF, along with a hematopoietic shift toward granulocyte macrophage progenitor and myeloid cells. Tle1(Δ/Δ) macrophages produce increased inflammatory cytokines in response to Toll-like receptor (TLR) agonists and lipopolysaccharides (LPS), and Tle1(Δ/Δ) mice display an enhanced inflammatory response to ear skin 12-O-tetradecanoylphorbol-13-acetate treatment. Loss of Tle1 not only results in increased phosphorylation and activation of proinflammatory NF-κB but also results in decreased Hes1 (hairy and enhancer of split-1), a negative regulator of inflammation in macrophages. Furthermore, Tle1(Δ/Δ) mice exhibit accelerated growth of B6-F10 melanoma xenografts. Our work provides the first in vivo evidence, to our knowledge, that TLE1 is a major counterregulator of inflammation with potential roles in a variety of inflammatory diseases and in cancer progression.


Shared neural mechanisms between imagined and perceived egocentric motion - A combined GVS and fMRI study.
Many cognitive and social processes involve mental simulations of a change in perspective. Behavioral studies suggest that such egocentric mental rotations rely on brain areas that are also involved in processing actual self-motion, thus depending on vestibular input. In a combined galvanic vestibular stimulation (GVS) and functional Magnetic Resonance Imaging (fMRI) study, we investigated the brain areas that underlie both simulated changes in self-location and the processing of vestibular stimulation within the same individuals. Participants performed an egocentric mental rotation task, an object-based mental rotation task, or a pure lateralization task during GVS or sham stimulation. At the neural level, we expected an overlap between brain areas activated during vestibular processing and egocentric mental rotation (against object-based mental rotation) within area OP2 and the Posterior Insular Cortex (PIC), two core brain regions involved in vestibular processing. The fMRI data showed a small overlap within area OP2 and a larger overlap within the PIC for both egocentric mental rotation against object-based mental rotation and vestibular processing. GVS did not influence the ability to perform egocentric mental rotation. Our results provide evidence for shared neural mechanisms underlying perceived and simulated self-motion. We conclude that mental rotation of one's body involves neural activity in the PIC and area OP2, but the behavioral results also suggest that those mental simulations of one's body might be robust to modulatory input from vestibular stimulation.


The diet of the striped hyena in Nepal's lowland regions.
Striped hyenas (Hyaena hyaena) are extremely rare in Nepal, and only a few people have studied them in their natural forest and grassland habitat. Their rarity is due to anthropogenic pressures such as hunting, habitat modification, being killed on roads, and depletion of their natural prey. Here, we studied the feeding ecology of hyenas in lowland, Nepal. We employed an opportunistic sampling to collect hyena scats in a range of habitats and the line transect sampling to identify the prey of the hyena in the study site. We collected 68 hyena scats between 2015 and 2018. Most of the hyena scat (39.7%) was found in the Churia Hill forest followed by riverbed (26.4%), mixed forest (14.7%), Sal (Shorea robusta)-dominated forest (11.7%), and grassland area (7.3%). We found eleven mammalian prey species, plants, and some unidentified items in the hyena scats. The frequency of occurrence and relative biomass of the medium-sized wild boar (Sus scrofa) were higher than other smaller prey species such as hare (Lepus nigricollis) and rhesus macaque (Macaca mulatta). Similarly, the proportion of large prey species such as nilgai (Boselaphus tragocamelus) in the hyena diet was lower compared with wild boar, hares, and rhesus macaques indicating medium-sized wild boar is the most preferred prey species. Livestock contributed 17.3% of the total dietary biomass. Domesticated species such as goats, sheep, cows, and even dogs were found in the diet of hyenas. Predation of livestock by hyenas could cause conflict, especially if this ongoing issue continues in the future. Rather, more conservation effort is required in lowland areas of Nepal to protect the hyenas' natural prey species, particularly in wildlife habitats to reduce the lure of taking domestic livestock. Similarly, conservation education at the local level and active involvement of government authorities in the conservation of this species might be helpful to mitigate human-hyena conflict in the human-dominated landscape.


The evolution of transdermal/topical overactive bladder therapy and its benefits over oral therapy.
Multiple antimuscarinic agents are available for the treatment of overactive bladder. Many of the agents have undergone reformulation in an attempt to improve patient adherence and drug tolerability. Oxybutynin evolved from an immediate-release pill to a once-daily oral preparation, and is now available as a transdermal patch and gel. This article discusses the clinical impact of oxybutynin reformulation and reviews the evolution and benefits of transdermal therapy.


An integrated closed-tube 2-plex PCR amplification and hybridization assay with switchable lanthanide luminescence based spatial detection.
Switchable lanthanide luminescence is a binary probe technology that inherently enables a high signal modulation in separation-free detection of DNA targets. A luminescent lanthanide complex is formed only when the two probes hybridize adjacently to their target DNA. We have now further adapted this technology for the first time in the integration of a 2-plex polymerase chain reaction (PCR) amplification and hybridization-based solid-phase detection of the amplification products of the Staphylococcus aureus gyrB gene and an internal amplification control (IAC). The assay was performed in a sealed polypropylene PCR chip containing a flat-bottom reaction chamber with two immobilized capture probe spots. The surface of the reaction chamber was functionalized with NHS-PEG-azide and alkyne-modified capture probes for each amplicon, labeled with a light harvesting antenna ligand, and covalently attached as spots to the azide-modified reaction chamber using a copper(i)-catalyzed azide-alkyne cycloaddition. Asymmetric duplex-PCR was then performed with no template, one template or both templates present and with a europium ion carrier chelate labeled probe for each amplicon in the reaction. After amplification europium fluorescence was measured by scanning the reaction chamber as a 10 × 10 raster with 0.6 mm resolution in time-resolved mode. With this assay we were able to co-amplify and detect the amplification products of the gyrB target from 100, 1000 and 10,000 copies of isolated S. aureus DNA together with the amplification products from the initial 5000 copies of the synthetic IAC template in the same sealed reaction chamber. The addition of 10,000 copies of isolated non-target Escherichia coli DNA in the same reaction with 5000 copies of the synthetic IAC template did not interfere with the amplification or detection of the IAC. The dynamic range of the assay for the synthetic S. aureus gyrB target was three orders of magnitude and the limit of detection of 8 pM was obtained. This proof-of-concept study shows that the switchable lanthanide luminescent probes enable separation-free array-based multiplexed detection of the amplification products in a closed-tube PCR which can enable a higher degree of multiplexing than is currently feasible by using different spectrally separated fluorescent probes.


Combination of ERK and autophagy inhibition as a treatment approach for pancreatic cancer.
Pancreatic ductal adenocarcinoma (PDAC) is characterized by KRAS- and autophagy-dependent tumorigenic growth, but the role of KRAS in supporting autophagy has not been established. We show that, to our surprise, suppression of KRAS increased autophagic flux, as did pharmacological inhibition of its effector ERK MAPK. Furthermore, we demonstrate that either KRAS suppression or ERK inhibition decreased both glycolytic and mitochondrial functions. We speculated that ERK inhibition might thus enhance PDAC dependence on autophagy, in part by impairing other KRAS- or ERK-driven metabolic processes. Accordingly, we found that the autophagy inhibitor chloroquine and genetic or pharmacologic inhibition of specific autophagy regulators synergistically enhanced the ability of ERK inhibitors to mediate antitumor activity in KRAS-driven PDAC. We conclude that combinations of pharmacologic inhibitors that concurrently block both ERK MAPK and autophagic processes that are upregulated in response to ERK inhibition may be effective treatments for PDAC.


CRP subunit association and hinge conformation changes in response to cAMP binding: analysis of C-helix cysteine-substituted CRP.
We investigated the characteristics of 13 CRP variants having cysteine substituted at positions 113, 115, 116, 117, 118, 120, 122, 124, 126, 127, 129, 130, or 131, positions that span the length of the CRP C alpha-helix. Under reducing conditions, the WT and all Cys-substituted forms of CRP migrated as 23.5 kDa CRP monomer species on SDS-PAGE gels. In the absence of a reductant, 9 of 13 Cys-substituted forms of CRP including the L113C, S117C, M120C, L124C, V126C, T127C, E129C, K130C, and V131C CRP contained protein that migrated as 47 kDa CRP dimer species on SDS-PAGE gels. CNBr digestion of the protein preparations followed by MALDI-TOF MS analysis of the peptide fragments showed these 47 kDa species to be CRP dimers that originated from disulfide bonds formed between positional-pair C alpha-helix Cys residues. The ratio of monomer CRP and disulfide cross-linked CRP within a Cys-substituted CRP preparation was found to be independent of cAMP for Cys-substituted CRP preparations denatured and renatured in the presence of various cAMP concentrations. This finding suggests that there is no large-scale concerted motion (i.e., scissoring) of the CRP subunits in response to cAMP binding. In addition, we have identified three amino acid residues located along the CRP C alpha-helix that play a role in facilitating the conformation transition of the CRP hinge from that characteristic of apo-CRP to that characteristic of the CRP.cAMP complex.


M-CSF induces monocyte survival by activating NF-κB p65 phosphorylation at Ser276 via protein kinase C.
Macrophage colony-stimulating factor (M-CSF) promotes mononuclear phagocyte survival and proliferation. The transcription factor Nuclear Factor-kappaB (NF-κB) is a key regulator of genes involved in M-CSF-induced mononuclear phagocyte survival and this study focused at identifying the mechanism of NF-κB transcriptional activation. Here, we demonstrate that M-CSF stimulated NF-κB transcriptional activity in human monocyte-derived macrophages (MDMs) and the murine macrophage cell line RAW 264.7. The general protein kinase C (PKC) inhibitor Ro-31-8220, the conventional PKCα/β inhibitor Gö-6976, overexpression of dominant negative PKCα constructs and PKCα siRNA reduced NF-κB activity in response to M-CSF. Interestingly, Ro-31-8220 reduced Ser276 phosphorylation of NF-κBp65 leading to decreased M-CSF-induced monocyte survival. In this report, we identify conventional PKCs, including PKCα as important upstream kinases for M-CSF-induced NF-κB transcriptional activation, NF-κB-regulated gene expression, NF-κB p65 Ser276 phosphorylation, and macrophage survival. Lastly, we find that NF-κB p65 Ser276 plays an important role in basal and M-CSF-stimulated NF-κB activation in human mononuclear phagocytes.


Efficacy of oxamyl against Heterodera schachtii on cabbage.
The efficacy of oxamyl in controlling Heterodera schachtii on cabbage was determined by applying various contbinations of soil drenches at 6.7 kg (a.i.)/ha and foliar sprays at 0.04 kg (a.i.)/100 liters of water to cabbage seedlings. Pretransplant drenches provided some control of H. schachtii over a 13-week period. A single foliar spray of oxamyl 1 week before transplanting apparently prevented penetration of H. schachtii larvae; post-transplant sprays were relatively ineffective. A pretransplant or transplant drench combined with a foliar application 2 weeks after transplanting provided the most effective control. The effectiveness of drenches plus post-transplant sprays is probably due to the spray augmenting the action of the drench in inhibiting the development of larvae after penelration.


Mice lacking histone deacetylase 6 have hyperacetylated tubulin but are viable and develop normally.
Posttranslational modifications play important roles in regulating protein structure and function. Histone deacetylase 6 (HDAC6) is a mostly cytoplasmic class II HDAC, which has a unique structure with two catalytic domains and a domain binding ubiquitin with high affinity. This enzyme was recently identified as a multisubstrate protein deacetylase that can act on acetylated histone tails, alpha-tubulin and Hsp90. To investigate the in vivo functions of HDAC6 and the relevance of tubulin acetylation/deacetylation, we targeted the HDAC6 gene by homologous recombination in embryonic stem cells and generated knockout mice. HDAC6-deficient mice are viable and fertile and show hyperacetylated tubulin in most tissues. The highest level of expression of HDAC6 is seen in the testis, yet development and function of this organ are normal in the absence of HDAC6. Likewise, lymphoid development is normal, but the immune response is moderately affected. Furthermore, the lack of HDAC6 results in a small increase in cancellous bone mineral density, indicating that this deacetylase plays a minor role in bone biology. HDAC6-deficient mouse embryonic fibroblasts show apparently normal microtubule organization and stability and also show increased Hsp90 acetylation correlating with impaired Hsp90 function. Collectively, these data demonstrate that mice survive well without HDAC6 and that tubulin hyperacetylation is not detrimental to normal mammalian development.


Autoimmune encephalitis: proposed recommendations for symptomatic and long-term management.
The objective of this paper is to evaluate available evidence for each step in autoimmune encephalitis management and provide expert opinion when evidence is lacking. The paper approaches autoimmune encephalitis as a broad category rather than focusing on individual antibody syndromes. Core authors from the Autoimmune Encephalitis Alliance Clinicians Network reviewed literature and developed the first draft. Where evidence was lacking or controversial, an electronic survey was distributed to all members to solicit individual responses. Sixty-eight members from 17 countries answered the survey. The most popular bridging therapy was oral prednisone taper chosen by 38% of responders while rituximab was the most popular maintenance therapy chosen by 46%. Most responders considered maintenance immunosuppression after a second relapse in patients with neuronal surface antibodies (70%) or seronegative autoimmune encephalitis (61%) as opposed to those with onconeuronal antibodies (29%). Most responders opted to cancer screening for 4 years in patients with neuronal surface antibodies (49%) or limbic encephalitis (46%) as opposed to non-limbic seronegative autoimmune encephalitis (36%). Detailed survey results are presented in the manuscript and a summary of the diagnostic and therapeutic recommendations is presented at the conclusion.


Vangl2 directs the posterior tilting and asymmetric localization of motile primary cilia.
Cilia are microtubule-based organelles that project into the extracellular space, function in the perception and integration of environmental cues, and regulate Hedgehog signal transduction. The emergent association of ciliary defects with diverse and pleiotropic human disorders has fuelled investigations into the molecular genetic regulation of ciliogenesis. Although recent studies implicate planar cell polarity (PCP) in cilia formation, this conclusion is based on analyses of proteins that are not specific to, or downstream effectors of PCP signal transduction. Here we characterize zebrafish embryos devoid of all Vangl2 function, a core and specific component of the PCP signalling pathway. Using Arl13b-GFP as a live marker of the ciliary axoneme, we demonstrate that Vangl2 is not required for ciliogenesis. Instead, Vangl2 controls the posterior tilting of primary motile cilia lining the neurocoel, Kupffer's vesicle and pronephric duct. Furthermore, we show that Vangl2 is required for asymmetric localization of cilia to the posterior apical membrane of neuroepithelial cells. Our results indicate a broad and essential role for PCP in the asymmetric localization and orientation of motile primary cilia, establishing directional fluid flow implicated in normal embryonic development and disease.


Major Co-localized QTL for Plant Height, Branch Initiation Height, Stem Diameter, and Flowering Time in an Alien Introgression Derived Brassica napus DH Population.
Plant height (PH), branch initiation height (BIH), and stem diameter (SD) are three stem-related traits that play crucial roles in plant architecture and lodging resistance. Herein, we show one doubled haploid (DH) population obtained from a cross between Y689 (one Capsella bursa-pastoris derived Brassica napus intertribal introgression) and Westar (B. napus cultivar) that these traits were significantly positively correlated with one another and with flowering time (FT). Based on a high-density SNP map, a total of 102 additive quantitative trait loci (QTL) were identified across six environments. Seventy-two consensus QTL and 49 unique QTL were identified using a two-round strategy of QTL meta-analysis. Notably, a total of 19 major QTL, including 11 novel ones, were detected for these traits, which comprised two QTL clusters on chromosomes A02 and A07. Conditional QTL mapping was performed to preliminarily evaluate the genetic basis (pleiotropy or tight linkage) of the co-localized QTL. In addition, QTL by environment interactions (QEI) mapping was performed to verify the additive QTL and estimate the QEI effect. In the genomic regions of all major QTL, orthologs of the genes involved in phytohormone biosynthesis, phytohormone signaling, flower development, and cell differentiation in Arabidopsis were proposed as candidate genes. Of these, BnaA02g02560, an ortholog of Arabidopsis GASA4, was suggested as a candidate gene for PH, SD, and FT; and BnaA02g08490, an ortholog of Arabidopsis GNL, was associated with PH, BIH and FT. These results provide useful information for further genetic studies on stem-related traits and plant growth adaptation.


Role of methylation in maintenance of positionally restricted transgene expression in developing muscle.
In transgenic mouse embryos, expression of a muscle-specific reporter, consisting of a chloramphenicol acetyltransferase gene linked to regulatory sequences from the rat myosin light chain 1/3 locus (MLC-CAT), is graded in developing axial muscles along the rostrocaudal axis and in cell cultures derived from these muscles. Here we demonstrate that maintenance of positional differences in MLC-CAT transgene expression cannot be attributed to differences in the transcriptional competence of corresponding muscles. Rather, patterns of transgene expression are reflected in the extent of CpG demethylation of both MLC1 promoter and MLC enhancer sequences. Variations in reporter gene expression can be reconstituted by in vitro methylation of specific CpGs in transfected MLC-CAT DNA. As the MLC-CAT transgene is activated during embryogenesis, demethylation of the MLC1 promoter lags behind that of the downstream MLC enhancer, which appears to be the initial target for epigenetic modification. In developing somites, demethylation of the transgenic MLC enhancer is not graded and therefore does not reflect early regional differences in MLC-CAT transgene expression patterns. These studies implicate selective methylation in the maintenance rather than in the establishment of transcriptional differences in developing muscles.


Male house mice evolving with post-copulatory sexual selection sire embryos with increased viability.
Although mating is costly, multiple mating by females is a taxonomically widespread phenomenon. Theory has suggested that polyandry may allow females to gain genetic benefits for their offspring, and thus offset the costs associated with this mating strategy. For example, the good sperm hypothesis posits that females benefit from mating multiply when genetically superior males have increased success in sperm competition and produce high quality offspring. We applied the powerful approach of experimental evolution to explore the potential for polyandry to drive evolutionary increases in female fitness in house mice, Mus domesticus. We maintained polygamously mated and monogamously mated selection lines of house mice for 14 generations, before determining whether selection history could account for divergence in embryo viability. We found that males from lineages evolving with post-copulatory sexual selection sire offspring with increased viability, suggesting that polyandry results in the production of higher quality offspring and thus provides long-term fitness benefits to females.


On the origin and nature of the grating interferometric dark-field contrast obtained with low-brilliance x-ray sources.
The x-ray dark-field contrast accessible via grating interferometry is sensitive to features at length scales well below what is resolvable by a detector system. It is commonly explained as arising from small-angle x-ray scattering (SAXS), and can be implemented both at synchrotron beamlines and with low-brilliance sources such as x-ray tubes. Here, we demonstrate that for tube based setups the underlying process of image formation can be fundamentally different. For focal spots or detector pixels that comprise multiple grating periods, we show that dark-field images contain a strong artificial and system-specific component not arising from SAXS. Based on experiments carried out with a nanofocus x-ray tube and the example of an excised rat lung, we demonstrate that the dark-field contrast observed for porous media transforms into a differential phase contrast for large geometric magnifications. Using a photon counting detector with an adjustable point spread function, we confirm that a dark-field image can indeed be formed by an intra-pixel differential phase contrast that cannot be resolved as such due to a dephasing between the periodicities of the absorption grating and the Talbot carpet. Our findings are further corroborated by a link between the strength of this pseudo-dark-field contrast and our x-ray tube's focal spot size in a three-grating setup. These results must not be ignored when measurements are intended to be reproducible across systems.


Sarcopenia, a strong determinant for prolonged feeding tube dependency after chemoradiotherapy for head and neck cancer.
Sarcopenia might be a relevant lead for optimization of the condition of patients with head and neck cancer (HNC) before chemoradiotherapy (CRT) to prevent long-term functional swallowing impairment, such as feeding tube dependency. Regression analyses were performed to assess the association between skeletal muscle mass index (SMI), as a measure of sarcopenia, and prolonged (>90 days) feeding tube dependency in 128 patients with HNC treated with primary CRT. Sixty-one patients (48%) became prolonged feeding tube-dependent. Lower SMI increased the risk of prolonged feeding tube dependency in multivariable analysis (risk ratio 1.08; 95% confidence interval 1.02-1.14, P = .01) adjusted for body mass index, abnormal diet, and socioeconomic status. Sarcopenia contributes to the risk of prolonged feeding tube dependency of patients with HNC treated with primary CRT. As sarcopenia might be a modifiable factor prior to treatment, it should be explored as a target for pretreatment patients' condition.