The mechanism of fission yeast mating type interconversion: seal/replicate/cleave model of replication across the double-stranded break site at mat1.
The interconversion of cell type in the fission yeast, Schizosaccharomyces pombe, is initiated by a double-stranded break (DSB) found at the mating type locus (mat1). A heritable site- and strand-specific DNA "imprinting" event at mat1 was recently hypothesized to be required to make the mat1 locus cleavable, and the DSB was suggested to be produced one generation before the actual switching event. It is known that only one cell among four granddaughters of a cell ever switches, and the sister of the recently switched cell switches efficiently in consecutive cell divisions. The feature of consecutive switching creates a major difficulty of having to replicate chromosomes possessing the DSB. The mat1 cis-acting leaky mutation, called smt-s, reduces the level of the DSB required for switching and is shown here to be a 27-bp deletion located 50 bp away from the cut site. Determination of the pattern and frequency of switching of the mutant allele by cell lineage studies has allowed us to conclude the following: (1) the chromosome with the DSB is sealed and replicated, then one of the specific chromatids is cleaved again to generate switching-competent cells in consecutive cell divisions and (2) the smt-s mutation affects DNA cleavage and not the hypothesized DNA imprinting step.


Treatment of HeartMate II Short-to-Shield Patients With an Ungrounded Cable: Indications and Long-Term Outcomes.
Despite modifications and a procedure to externally replace the distal portion of the percutaneous lead, damage to the wiring insulation causing an electrical short to ground, referred to as a short to shield (STS), has become an important factor in the longevity of the HeartMate II left ventricular assist device (LVAD). Device exchange has been the suggested treatment option. The aim of this study was to evaluate the long-term clinical outcomes of patients with an STS supported on an ungrounded cable. A retrospective review of all patients (n = 479) implanted with a HeartMate II at our center between January 2008 and December 2017 was performed. Patients with a documented STS maintained on an ungrounded cable were examined. Patient characteristics, time from device implantation to STS, treatment strategies, and duration of support on an ungrounded cable were summarized. The association between support with an ungrounded cable and clinical outcomes was evaluated. A total of 53 (11% of 479) patients (83% males and 81% destination therapy) with an STS were supported on an ungrounded cable for a median duration of 195 days (range 2 days to 3.3 years). Patients were more active (New York Heart Association [NYHA] p < 0.001, 6 minute walk test [6MWT] p = 0.003) and had a trend toward increased weight gain (p = 0.055) from time of implant to STS. Duration of support before the STS was 1.9 years (range 165 days to 8.6 years). Twenty-two patients were treated directly with an ungrounded cable and 31 patients underwent an external driveline repair and still required an ungrounded cable within 2 days (range 0 days to 1.3 years). During the study period, 38 patients were maintained on an ungrounded cable: 21 patients were ongoing for 299 days (range 114 days to 2.8 years), 11 patients transplanted after 79 days (range 7-295 days), four patients died because of comorbid conditions after 1.6 years (range 141 days to 3.2 years), one patient exchanged for thrombosis after 229 days, and one patient explanted after 279 days. The other 15 patients developed a phase-to-phase electrical short after 51 days (range 2 days to 3.3 years): 14 patients underwent a successful pump exchange and one patient transplanted within 2 days. No patients died because of support with an ungrounded cable or worsening lead damage necessitating device exchange. With extended durations of support, some patients with a HeartMate II LVAD will experience device failure in the form of an STS. Select patients with an STS can be safely supported on an ungrounded cable for several years with close monitoring. This treatment approach should be considered before a device exchange.


[Demographic profile of the Xavánte Indian population in Sangradouro-Volta Grande, Mato Grosso].
This paper analyzes the demographic profile of the Xavánte population at the Sangradouro-Volta Grande Indigenous Reserve in Mato Grosso, Brazil, from 1993 to 1997. The survey included annual censuses and vital statistics from 7 Xavánte villages. Permanent contact with Brazilian national society, established in the 1940s and 50s, caused a population drop due to epidemics and clashes. In 1995 there were 825 individuals in the community. The crude birth rate (57.7/1,000) and death rate (9.1/1,000) were higher than the national averages. The majority (56%) of the population is under 15 years of age (median: 13 years) and the infant mortality rate is high (87.1 per thousand live births), probably resulting from precarious sanitary conditions in the villages. Other results included the persistence of polygyny; low levels of migration; a dynamic of splits and formation of new villages; traditional housing patterns maintained in the old villages and abandoned in the new ones. The recent demographic recovery in the data from Sangradouro-Volta Grande is similar to that observed in the Pimentel Barbosa community. The study highlights the importance of systematically collecting and analyzing demographic data from indigenous populations.


Structural changes induced in Ca2+-regulated myosin filaments by Ca2+ and ATP.
We have used electron microscopy and proteolytic susceptibility to study the structural basis of myosin-linked regulation in synthetic filaments of scallop striated muscle myosin. Using papain as a probe of the structure of the head-rod junction, we find that this region of myosin is approximately five times more susceptible to proteolytic attack under activating (ATP/high Ca2+) or rigor (no ATP) conditions than under relaxing conditions (ATP/low Ca2+). A similar result was obtained with native myosin filaments in a crude homogenate of scallop muscle. Proteolytic susceptibility under conditions in which ADP or adenosine 5'-(beta, gamma-imidotriphosphate) (AMPPNP) replaced ATP was similar to that in the absence of nucleotide. Synthetic myosin filaments negatively stained under relaxing conditions showed a compact structure, in which the myosin cross-bridges were close to the filament backbone and well ordered, with a clear 14.5-nm axial repeat. Under activating or rigor conditions, the cross-bridges became clumped and disordered and frequently projected further from the filament backbone, as has been found with native filaments; when ADP or AMPPNP replaced ATP, the cross-bridges were also disordered. We conclude (a) that Ca2+ and ATP affect the affinity of the myosin cross-bridges for the filament backbone or for each other; (b) that the changes observed in the myosin filaments reflect a property of the myosin molecules alone, and are unlikely to be an artifact of negative staining; and (c) that the ordered structure occurs only in the relaxed state, requiring both the presence of hydrolyzed ATP on the myosin heads and the absence of Ca2+.


Protecting time for teaching in the ambulatory care setting.
The current drive for efficient clinical teaching threatens the educational mission of academic medical centers. With pressures to increase clinical productivity, protected time and compensation for teaching have become scarce resources for clinical teachers in all settings. Although it may yield new approaches to education, the push for efficiency may ultimately result in insufficient time for teaching and may cause some clinical preceptors to stop teaching completely. Further, it may lead to the illusion that comprehensive teaching truly requires little time. Since the future of American health care depends upon the provision of high-quality clinical education to young physicians, this situation presents a potential national crisis. In this article, the authors discuss the complex nature of teaching, its time requirements, and the special challenges of teaching in outpatient settings. To avoid overemphasizing efficiency to the detriment of education they recommend adhering to two principles: (1) academic medical centers are educational as well as training institutions, and therefore should provide a broad-based education as well as training in clinical skills; and (2) the clinical teaching process is complex and adequate time must be provided for its many phases, including planning, instructing, and reflecting. Finally, the authors make recommendations for ensuring the delivery of high-quality education in ambulatory care settings.


Organogallium- and organozinc-rich palladium and platinum clusters.
The heteroleptic, binuclear compounds [M2(GaCp*)3(PMe3)2] (M = Pd: 1a, Pt: 1b, Cp* = pentamethylcyclopentadienyl) can be obtained if [Pd2(dvds)3] (dvds = 1,3-divinyl-1,1,3,3-tetramethyldisiloxane) or [Pt(COD)2] (COD = cyclooctadiene) were treated with a 3 : 2 ratio of GaCp* and PMe3. A trinuclear complex [Pd3(GaCp*)3(PMe3)3] (2) was isolated from in situ prepared [Pd(PMe3)2Me2] and treatment with GaCp*. The new complexes were used as starting compounds for selective Ga/Zn exchange reactions to afford the dinuclear palladium compound [Pd2(ZnCp*)(ZnMe)3(PMe3)5] (3a) and the mononuclear platinum compound [Pt(ZnCp*)2(ZnMe)2(PMe3)2] (3b). The effects of a final transition metal count in organozinc ligated compounds and stabilization of Lewis acidic fragments like [Pd(PPh3)3], [Ni(C2H4)3] or [Mo(CO)5] were presented and discussed. All compounds have been fully characterized by (1)H, (13)C, and (31)P NMR spectroscopy, mass spectrometry using a liquid injection field desorption ionization (LIFDI) method, elemental analysis, and single crystal X-ray diffraction studies and the coordination polyhedra were analysed by the method of continuous shape measure (CShM).


Suppression of ovarian follicle activation in mice by the transcription factor Foxo3a.
Foxo transcription factors have been implicated in diverse biological processes, including metabolism, cellular stress responses, and aging. Here, we show that Foxo3a-/- female mice exhibit a distinctive ovarian phenotype of global follicular activation leading to oocyte death, early depletion of functional ovarian follicles, and secondary infertility. Foxo3a thus functions at the earliest stages of follicular growth as a suppressor of follicular activation. In addition to providing a molecular entry point for studying the regulation of follicular growth, these results raise the possibility that accelerated follicular initiation plays a role in premature ovarian failure, a common cause of infertility and premature aging in women.


Improving emergency surgical care for patients with right iliac fossa pain at a regional scale: A quality improvement study using the Supported Champions implementation strategy.
Methods to improve clinical systems safety suffer from significant difficulties in implementation and scaling up. We used an upscaling implementation strategy entitled Supported Champions in a quality and safety improvement programme for emergency surgery at regional level, focusing on patients with right iliac fossa pain. A before-after study was conducted across four acute NHS Trusts: A 6 month intervention phase was preceded and followed by 3 months of data collection. An established Human Factors intervention was led at each Trust by a small group of staff selected as Champions. Champions received training in teamwork and systems improvement and were supported by Human Factors experts. The primary improvement aim was to expedite surgery for patients with sepsis, using Royal College of Surgeons emergency surgery guidelines as the measure. Additional outcomes studied included length of inpatient stay and 30-day readmission rates. Breaches of RCS urgency guidelines decreased markedly from 13.7% of operated patients pre-intervention to 3.5% post-intervention (p = 0.000). Mean time from booking to incision decreased in three of the four sites, whilst median length of stay increased in 3 of 4. Overall 30-day readmission rate remained stable (7.84% pre-intervention versus 7.31% post-intervention, p = 0.959). The Supported Champions model allowed all surgical teams to reduce delay for septic patients by more than 50%, using distinct Quality Improvement strategies to address local issues. Improvement was implemented in 4 diverse settings with a quarter of the level of expert input previously used in a single hospital.


Discovery of the novel mTOR inhibitor and its antitumor activities in vitro and in vivo.
The phosphoinositide 3-kinase (PI3-K)/Akt and mTOR signaling pathway plays a critical role in cell survival and proliferation and is often aberrantly activated in many types of cancer. The mTOR kinase protein, one of the key molecules in this pathway, has been shown to be an important target for cancer therapy. In the present study, a ligand docking method was used to screen for novel scaffold mTOR inhibitors. Sixty thousand compounds in the Natural Product Database were screened against the mTOR homologous structure, and 13 commercially available compounds listed in the top-ranked 100 compounds were selected for further examination. Compound [(E)-3-(4-(benzo[d][1,3]dioxol-5-yl)-2-oxobut-3-en-1-yl)- 3-hydroxyindolin-2-one; designated herein as 3HOI-BA-01] was then selected for further study of its antitumor activity. An in vitro study has shown that 3HOI-BA-01 inhibited mTOR kinase activity in a dose-dependent manner by directly binding with mTOR. In a panel of non-small cell lung cancer cells, the compound also attenuated mTOR downstream signaling, including the phosphorylation of p70S6K, S6, and Akt, resulting in G1 cell-cycle arrest and growth inhibition. Results of an in vivo study have shown that intraperitoneal injection of 3HOI-BA-01 in A549 lung tumor-bearing mice effectively suppressed cancer growth without affecting the body weight of the mice. The expression of downstream signaling molecules in the mTOR pathway in tumor tissues was also reduced after 3HOI-BA-01 treatment. Taken together, we identified 3HOI-BA-01 as a novel and effective mTOR inhibitor.


Molecular Imaging and Pharmacokinetic Analysis of Carbon-11 Labeled Antisense Oligonucleotide LY2181308 in Cancer Patients.
Antisense oligonucleotides (ASOs) have potential as anti-cancer agents by specifically modulating genes involved in tumorigenesis. However, little is known about ASO biodistribution and tissue pharmacokinetics (PKs) in humans, including whether sufficient delivery to target tumor tissue may be achieved. In this preliminary study in human subjects, we used combined positron emission and computed tomography (PET-CT) imaging and subsequent modeling analysis of acquired dynamic data, to examine the in vivo biodistribution and PK properties of LY2181308 - a second generation ASO which targets the apoptosis inhibitor protein survivin. Following radiolabeling of LY2181308 with methylated carbon-11 ([(11)C]methylated-LY2181308), micro-doses (<1mg) were administered to three patients with solid tumors enrolled in a phase I trial. Moderate uptake of [(11)C]methylated-LY2181308 was observed in tumors (mean=32.5ng*h /mL, per mg administered intravenously). Highest uptake was seen in kidney and liver and lowest uptake was seen in lung and muscle. One patient underwent repeat analysis on day 15 of multiple dose therapy, during administration of LY2181308 (750mg), when altered tissue PKs and a favorable change in biodistribution was seen. [(11)C]methylated-LY2181308 exposure increased in tumor, lung and muscle, whereas renal and hepatic exposure decreased. This suggests that biological barriers to ASO tumor uptake seen at micro-doses were overcome by therapeutic dosing. In addition, (18)F-labeled fluorodeoxyglucose (FDG) scans carried out in the same patient before and after treatment showed up to 40% decreased tumor metabolism. For the development of anti-cancer ASOs, the results provide evidence of LY2181308 tumor tissue delivery and add valuable in vivo pharmacological information. For the development of novel therapeutic agents in general, the study exemplifies the merits of applying PET imaging methodology early in clinical investigations.


Goodyschle A, a new butenolide with significant BchE inhibitory activity from Goodyera schlechtendaliana.
Goodyschle A (1), a new butenolide, was isolated from the whole grass of Goodyera schlechtendaliana, an orchidaceous edible medicinal plant. The structure of the new compound was elucidated by 1 D and 2 D NMR experiments in addition to HRESIMS analyses. Compound 1 was evaluated for its bioactivities including cytotoxic activity against human gastric cancer (SGC-7901) and human hepatocellular carcinoma (HepG2) cell lines, inhibitory activity on acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), and DPPH radical scavenging activity. As a result, compound 1 showed potent BChE inhibitory activity (IC50 value = 6.88 ± 1.63 μM), moderate DPPH radical scavenging activity (IC50 value = 16.25 ± 0.21 μM), and slight AChE inhibitory and cytotoxic activities. These findings suggest that compound 1 is worthy for further investigations in terms of its selective BChE inhibitory activity.


Molecular dynamics study of water and Na+ ions in models of the pore region of the nicotinic acetylcholine receptor.
The nicotinic acetylcholine receptor (nAChR) is an integral membrane protein that forms ligand-gated and cation-selective channels. The central pore is lined by a bundle of five approximately parallel M2 helices, one from each subunit. Candidate model structures of the solvated pore region of a homopentameric (alpha7)5 nAChR channel in the open state, and in two possible forms of the closed state, have been studied using molecular dynamics simulations with restraining potentials. It is found that the mobility of the water is substantially lower within the pore than in bulk, and the water molecules become aligned with the M2 helix dipoles. Hydrogen-bonding patterns in the pore, especially around pore-lining charged and hydrophilic residues, and around exposed regions of the helix backbone, have been determined. Initial studies of systems containing both water and sodium ions together within the pore region have also been conducted. A sodium ion has been introduced into the solvated models at various points along the pore axis and its energy profile evaluated. It is found that the ion causes only a local perturbation of the water structure. The results of these calculations have been used to examine the effectiveness of the central ring of leucines as a component of a gate in the closed-channel model.


Cations Regulate Membrane Attachment and Functionality of DNA Nanostructures.
The interplay between nucleic acids and lipids underpins several key processes in molecular biology, synthetic biotechnology, vaccine technology, and nanomedicine. These interactions are often electrostatic in nature, and much of their rich phenomenology remains unexplored in view of the chemical diversity of lipids, the heterogeneity of their phases, and the broad range of relevant solvent conditions. Here we unravel the electrostatic interactions between zwitterionic lipid membranes and DNA nanostructures in the presence of physiologically relevant cations, with the purpose of identifying new routes to program DNA-lipid complexation and membrane-active nanodevices. We demonstrate that this interplay is influenced by both the phase of the lipid membranes and the valency of the ions and observe divalent cation bridging between nucleic acids and gel-phase bilayers. Furthermore, even in the presence of hydrophobic modifications on the DNA, we find that cations are still required to enable DNA adhesion to liquid-phase membranes. We show that the latter mechanism can be exploited to control the degree of attachment of cholesterol-modified DNA nanostructures by modifying their overall hydrophobicity and charge. Besides their biological relevance, the interaction mechanisms we explored hold great practical potential in the design of biomimetic nanodevices, as we show by constructing an ion-regulated DNA-based synthetic enzyme.


National treatment trends for ductal carcinoma in situ of the breast.
To evaluate the national treatment trends for the management of ductal carcinoma in situ as related to the individual characteristics of patients and to the reporting of demographics. National Cancer Data Base review. Patients (N = 39010) who were diagnosed as having ductal carcinoma in situ between 1985 and 1993. Treatment principles, including the use of breast-preserving surgery, axillary lymph node dissection, and radiotherapy, as related to the following variables: age, income level, and ethnicity of the patient; the tumor size, grade, and anatomical subsite; year of diagnosis; geographic location of treatment; and hospital type and caseload. During the 8 years of analysis, the use of breast preservation therapy increased from 31% to 54%. Treatment selection varied to some degree with each of the variables examined. Tumors with favorable sizes and grades were associated with increased rates of breast preservation and lower rates of axillary lymph node dissection and radiotherapy utilization. Overall, only 45% of the patients who were treated with breast preservation received adjuvant radiotherapy. However, during this study, radiotherapy utilization increased from 38% to 54%. Axillary lymph node dissection was performed in 49% of the patients with a 12% reduction in use over time. Breast-preserving surgery now accounts for more than half of all cases of ductal carcinoma in situ followed by the National Cancer Data Base. However, there still remains an inappropriately high rate of axillary lymph node dissection and a low rate of radiotherapy utilization. Clinical trial results and professional education should continue to optimize the management of patients with ductal carcinoma in situ.


Polarisation charges and scattering behaviour of realistically rounded plasmonic nanostructures.
We study the effect of realistically rounding nanorod antennae and gap antennae on their far field and near field properties. The simulations show that both scattering behaviour and polarisation charge distribution depend significantly on rounding. Rounding is also seen to have a major effect on coupling between nanostructures. The results suggest that it is important to incorporate the effect of rounding to be able to design plasmonic nanostructures with desired properties.


Multisensory congruency as a mechanism for attentional control over perceptual selection.
The neural mechanisms underlying attentional selection of competing neural signals for awareness remains an unresolved issue. We studied attentional selection, using perceptually ambiguous stimuli in a novel multisensory paradigm that combined competing auditory and competing visual stimuli. We demonstrate that the ability to select, and attentively hold, one of the competing alternatives in either sensory modality is greatly enhanced when there is a matching cross-modal stimulus. Intriguingly, this multimodal enhancement of attentional selection seems to require a conscious act of attention, as passively experiencing the multisensory stimuli did not enhance control over the stimulus. We also demonstrate that congruent auditory or tactile information, and combined auditory-tactile information, aids attentional control over competing visual stimuli and visa versa. Our data suggest a functional role for recently found neurons that combine voluntarily initiated attentional functions across sensory modalities. We argue that these units provide a mechanism for structuring multisensory inputs that are then used to selectively modulate early (unimodal) cortical processing, boosting the gain of task-relevant features for willful control over perceptual awareness.


Estimating daily methane production in individual cattle with irregular feed intake patterns from short-term methane emission measurements.
Spot measurements of methane emission rate (n = 18 700) by 24 Angus steers fed mixed rations from GrowSafe feeders were made over 3- to 6-min periods by a GreenFeed emission monitoring (GEM) unit. The data were analysed to estimate daily methane production (DMP; g/day) and derived methane yield (MY; g/kg dry matter intake (DMI)). A one-compartment dose model of spot emission rate v. time since the preceding meal was compared with the models of Wood (1967) and Dijkstra et al. (1997) and the average of spot measures. Fitted values for DMP were calculated from the area under the curves. Two methods of relating methane and feed intakes were then studied: the classical calculation of MY as DMP/DMI (kg/day); and a novel method of estimating DMP from time and size of preceding meals using either the data for only the two meals preceding a spot measurement, or all meals for 3 days prior. Two approaches were also used to estimate DMP from spot measurements: fitting of splines on a 'per-animal per-day' basis and an alternate approach of modelling DMP after each feed event by least squares (using Solver), summing (for each animal) the contributions from each feed event by best-fitting a one-compartment model. Time since the preceding meal was of limited value in estimating DMP. Even when the meal sizes and time intervals between a spot measurement and all feeding events in the previous 72 h were assessed, only 16.9% of the variance in spot emission rate measured by GEM was explained by this feeding information. While using the preceding meal alone gave a biased (underestimate) of DMP, allowing for a longer feed history removed this bias. A power analysis taking into account the sources of variation in DMP indicated that to obtain an estimate of DMP with a 95% confidence interval within 5% of the observed 64 days mean of spot measures would require 40 animals measured over 45 days (two spot measurements per day) or 30 animals measured over 55 days. These numbers suggest that spot measurements could be made in association with feed efficiency tests made over 70 days. Spot measurements of enteric emissions can be used to define DMP but the number of animals and samples are larger than are needed when day-long measures are made.


Criterion-related validity of the last 7-day, short form of the International Physical Activity Questionnaire in Swedish adults.
To examine the validity of the short, last 7-day, self-administered form of the International Physical Activity Questionnaire (IPAQ). All subjects wore an accelerometer for seven consecutive days and completed the IPAQ questionnaire on the eighth day. Criterion validity was assessed by linear regression analysis and by modified Bland-Altman analysis. Specificity and sensitivity were calculated for classifying respondents according to the physical activity guidelines of the American College of Sports Medicine/Centers for Disease Control and Prevention. Workplaces in Uppsala, Sweden. One hundred and eighty-five (87 males) participants, aged 20 to 69 years. Total self-reported physical activity (PA) (MET-min day(-1)) was significantly correlated with average intensity of activity (counts min(-1)) from accelerometry (r = 0.34, P < 0.001). Gender, age, education and body mass index did not affect this relationship. Further, subcomponents of self-reported PA (time spent sitting, time in PA, time in moderate and vigorous activity (MVPA)) were significantly correlated with objectively measured PA (P < 0.05). Self-reported time in PA was significantly different from time measured by accelerometry (mean difference: -25.9 min day(-1); 95% limits of agreement: -172 to 120 min day(-1); P < 0.001). IPAQ identified 77% (specificity) of those who met the current PA guidelines of accumulating more than 30 min day(-1) in MVPA as determined by accelerometry, whereas only 45% (sensitivity) of those not meeting the guidelines were classified correctly. Our results indicate that the short, last 7-days version of the IPAQ has acceptable criterion validity for use in Swedish adults. However, the IPAQ instrument significantly overestimated self-reported time spent in PA. The specificity to correctly classify people achieving current PA guidelines was acceptable, whereas the sensitivity was low.


Papillary tumor of the spinal cord: report of 2 cases.
Intramedullary spinal cord tumors constitute a small fraction of central nervous system tumors in the pediatric population; of these, the majority are ependymomas or astrocytomas. We report 2 pediatric spinal cord tumor cases with unique morphologic and immunohistochemical features. The first patient presented at age 7 with an intramedullary tumor of the thoracic spine. She suffered lumbar, cerebellar, and temporal lobe recurrences despite surgical resection and radiation. The second patient presented at age 17 with an intramedullary tumor of the cervical spine. The tumor recurred locally and in the cerebellum. Magnetic resonance imaging studies demonstrated gadolinium enhancement in each case. Microscopy showed papillary and solid cytoarchitecture with monomorphous epithelioid cells arranged around vascular papillae. Immunohistochemistry in each case revealed diffuse epithelial membrane antigen, cytokeratin, and E-cadherin reactivity. Glial fibrillary acidic protein staining was focal in case 1 and completely negative in case 2. Neural cell adhesion molecule showed patchy membranous reactivity and synaptophysin was negative. Electron microscopy showed ependymal differentiation. The clinical features, including propensity for recurrence and remote subarachnoid spread, and the pathologic features of these tumors are reminiscent of papillary tumor of the pineal region, ependymoma, and choroid plexus papilloma. The cases presented may constitute a new neoplastic entity within the recently described spectrum of central nervous system tumors with ependymal features.


Head-direction cells recorded from the postsubiculum in freely moving rats. I. Description and quantitative analysis.
This paper is a study of the behavioral and spatial firing correlates of neurons in the rat postsubiculum. Recordings were made from postsubicular neurons as rats moved freely throughout a cylindrical chamber, where the major cue for orientation was a white card taped to the inside wall. An automatic video/computer system monitored cell discharge while simultaneously tracking the position of 2 colored light emitting diodes (LEDs) secured to the animal's head. The animal's location was calculated from the position of one of the LEDs and head direction in the horizontal plane calculated from the relative positions of the 2 LEDs. Approximately 26% of the cells were classified as head-direction cells because they discharged as a function of the animal's head direction in the horizontal plane, independent of the animal's behavior, location, or trunk position. For each head-direction cell, vectors drawn in the direction of maximal firing were parallel throughout the recording chamber and did not converge toward a single point. Plots of firing rate versus head direction showed that each firing-rate/head-direction function was adequately described by a triangular function. Each cell's maximum firing rate occurred at only one (the preferred) head direction; firing rates at head directions on either side of the preferred direction decreased linearly with angular deviation from the preferred direction. Results from 24 head-direction cells in 7 animals showed an equal distribution of preferred firing directions over a 360 degrees angle. The peak firing rate of head-direction cells varied from 5 to 115 spikes/sec (mean: 35). The range of head-direction angles over which discharge was elevated (directional firing range) was usually about 90 degrees, with little, if any, discharge at head directions outside this range. Quantitative analysis showed the location of the animal within the cylinder had minimal effect on directional cell firing. For each head-direction cell, the preferred direction, peak firing rate, and directional firing range remained stable for days. These results identify a new cell type that signals the animal's head direction in its environment.


Tenuous structures from disorderly growth processes.
Colloidal aggregation and other random growth processes produce structures that behave differently from ordinary bulk matter. Much of this behavior can be described in terms of the invariance of the aggregates under changes of spatial length scale: they appear to be fractals. There are two types of basic mechanisms for producing fractal aggregates. Those in which aggregation proceeds cluster by cluster can be understood qualitatively in terms of a solvable schematic model. The diffusion-limited aggregation or deposition of individual particles to make a large cluster is not as well understood. It is closely related to several irreversible processes in other areas of physics, such as two-fluid displacement in porous materials and the dielectric breakdown of insulators. More generally, disorderly growth mechanisms provide structures having unique properties, many of which can be understood by using simple statistical principles.


The impact of temperature and pump flow rate during selective cerebral perfusion on regional blood flow in piglets.
Ideal temperature and flow rate for selective cerebral perfusion (SCP) are not known. We examined regional organ perfusion in a piglet SCP model. Three groups underwent SCP at 30 mL/kg/min at different temperatures (15°C, 25°C, and 32°C) and 4 groups remained at 25°C for SCP at different flow rates (10, 30, 50 and 75 mL/kg/min). Fluorescent microspheres were injected at 5 minutes of normothermic cardiopulmonary bypass (CPB), immediately before SCP, SCP 45 minutes, SCP 90 minutes, and 2 hours after CPB. Brain and lower body organs were collected to examine regional blood flow (RBF, mL/min/g). At 2 hours after CPB, RBF of the 32°C group was higher than that of the 15°C group (P < .05) at the caudate nucleus and hippocampus; RBF of the 32°C group was higher than that of the 25°C and 15°C groups (P < .05) at the neocortex. No significant difference in RBF was observed among any of the 25°C groups at different flow rates. Also, there was no significant difference between the RBF to the left and right sides of brain in either the temperature or flow rate groups. RBF did significantly increase with temperature in the liver and quadriceps during SCP (P < .05). At the kidney, RBF at SCP 90 minutes was significantly higher than that at SCP 45 minutes when all temperature groups were combined (P < .05). SCP at 32°C provides higher brain RBF 2 hours after CPB. Increasing SCP flow rate does not increase RBF significantly at 25°C. Higher temperature during SCP results in improved RBF to the liver and quadriceps.


Nkx6 transcription factors and Ptf1a function as antagonistic lineage determinants in multipotent pancreatic progenitors.
The molecular mechanisms that underlie cell lineage diversification of multipotent progenitors in the pancreas are virtually unknown. Here we show that the early fate choice of pancreatic progenitors between the endocrine and acinar cell lineage is restricted by cross-repressive interactions between the transcription factors Nkx6.1/Nkx6.2 (Nkx6) and Ptf1a. Using genetic loss- and gain-of-function approaches, we demonstrate that Nkx6 factors and Ptf1a are required and sufficient to repress the alternative lineage program and to specify progenitors toward an endocrine or acinar fate, respectively. The Nkx6/Ptf1a switch only operates during a critical competence window when progenitors are still multipotent and can be uncoupled from cell differentiation. Thus, cross-antagonism between Nkx6 and Ptf1a in multipotent progenitors governs the equilibrium between endocrine and acinar cell neogenesis required for normal pancreas development.


High levels of serum antibodies to merozoite surface protein 2 of Plasmodium falciparum are associated with reduced risk of clinical malaria in coastal Kenya.
The merozoite surface protein (MSP) 2 is a vaccine candidate antigen of Plasmodium falciparum that is polymorphic in natural populations. In a prospective cohort study in two coastal populations of Kenya using recombinant proteins derived from the two major allelic types of MSP2, high serum levels of IgG to MSP2 were associated with protection from clinical malaria. This protection was independent of that associated with antibodies to another vaccine candidate antigen (AMA1) in these populations. However, low antibody levels to MSP2 appeared to be associated with increased susceptibility to malaria within people who were parasite negative at the time of serum collection. These data suggest that an MSP2 based vaccine should be designed to induce high level antibody responses against the different MSP2 types present globally in P. falciparum populations and that MSP2 could be combined with other P. falciparum antigens to form a multi-component malaria vaccine.


Neonatal and postneonatal mortality in Germany since unification.
After unification, the gap in infant mortality rates between the two parts of Germany widened until 1996 before converging. The reasons for these changes have not, so far, been apparent. To investigate trends in neonatal and postneonatal mortality in the eastern (the new Länder) and western (the old Länder) part of Germany after unification in 1990 and to identify the scope for further improvement. Examination of trends in birth weights, birth weight specific neonatal mortality and cause specific postneonatal mortality in the two parts of Germany from 1990 to 1996 and 1997 by analysing routinely available vital statistics data. In both parts of Germany, neonatal mortality fell considerably, by 33 per cent in the east and 17 per cent in the west, from 4.5 and 3.5 per thousand live births in 1990 to 3.0 and 2.9 in 1997, respectively. This was attributable to an improvement in survival of infants at all birth weights but especially among those with very low birth weights, accounting for an estimated 83 to 85 per cent of the overall improvement. The birth weight distribution showed a slight worsening in the new and the old Länder with an increase in the proportion of those under 1500 g and, in the east, a 24 per cent increase in the proportion of high birth-weight infants of 4000 and more grams. Trends in postneonatal mortality revealed a worsening of about 32 per cent in the east from 1990 to 1991 followed by a decline of over 50 per cent up to 1997, leading to comparable mortality rates of 1.8 per thousand live births in the east and 2.0 in the west. While both parts experienced a decrease of 40 to 48 per cent in deaths from all diseases, the decline in deaths because of accidents and injuries was markedly higher in the new Länder although they are still exceeding the western rate by 3.7 per 100,000 live births in 1997. Since unification, the two parts of Germany underwent a complex process that has led finally to convergence of parameters of infant health that are most likely to have been because of improvements in the quality of perinatal care. To improve infant mortality in Germany, policy measures should focus on preventive rather than curative measures as the proportion of very low birthweight babies is increasing in both parts of Germany.


Interfacial Ca2+ environments in nanocrystalline apatites revealed by dynamic nuclear polarization enhanced 43Ca NMR spectroscopy.
The interfaces within bones, teeth and other hybrid biomaterials are of paramount importance but remain particularly difficult to characterize at the molecular level because both sensitive and selective techniques are mandatory. Here, it is demonstrated that unprecedented insights into calcium environments, for example the differentiation of surface and core species of hydroxyapatite nanoparticles, can be obtained using solid-state NMR, when combined with dynamic nuclear polarization. Although calcium represents an ideal NMR target here (and de facto for a large variety of calcium-derived materials), its stable NMR-active isotope, calcium-43, is a highly unreceptive probe. Using the sensitivity gains from dynamic nuclear polarization, not only could calcium-43 NMR spectra be obtained easily, but natural isotopic abundance 2D correlation experiments could be recorded for calcium-43 in short experimental time. This opens perspectives for the detailed study of interfaces in nanostructured materials of the highest biological interest as well as calcium-based nanosystems in general.


Cloning of the gene encoding the Actinobacillus actinomycetemcomitans serotype b OmpA-like outer membrane protein.
The gene encoding an outer membrane protein A (OmpA)-like, heat-modifiable Omp of Actinobacillus actinomycetemcomitans ATCC 43718 (strain Y4, serotype b) was cloned by a PCR cloning procedure. DNA sequence analysis revealed that the gene encodes a protein of 346 amino acid residues with a molecular mass of 36.9 kDa. The protein expressed by the cloned gene reacted with a monoclonal antibody to the previously described 29-kDa Omp (Omp29) of strain Y4. This monoclonal antibody reacted specifically with Omp29 of A. actinomycetemcomitans (serotype b), but not with any Omp of Escherichia coli, including OmpA. This protein exhibited characteristic heat modifiability on sodium dodecyl sulfate-polyacrylamide gels, showing an apparent molecular mass of 29 kDa when unheated and a mass of 34 kDa when heated. The N-terminal amino acid sequence of the protein expressed in E. coli perfectly matched those deduced from the purified Omp29 of strain Y4. The deduced amino acid sequence of the gene coding for Omp29 from serotype b matched completely (except for valine at position 321) that of a recently reported omp34 gene described for A. actinomycetemcomitans serotype c (NCTC 9710). Because of the conserved nature of the gene within these serotypes, we designated the gene described herein from serotype b as omp34.


Mutations affecting GABAergic signaling in seizures and epilepsy.
The causes of epilepsies and epileptic seizures are multifactorial. Genetic predisposition may contribute in certain types of epilepsies and seizures, whether idiopathic or symptomatic of genetic origin. Although these are not very common, they have offered a unique opportunity to investigate the molecular mechanisms underlying epileptogenesis and ictogenesis. Among the implicated gene mutations, a number of GABAA receptor subunit mutations have been recently identified that contribute to several idiopathic epilepsies, febrile seizures, and rarely to certain types of symptomatic epilepsies, like the severe myoclonic epilepsy of infancy. Deletion of GABAA receptor genes has also been linked to Angelman syndrome. Furthermore, mutations of proteins controlling chloride homeostasis, which indirectly defines the functional consequences of GABAA signaling, have been identified. These include the chloride channel 2 (CLCN2) and the potassium chloride cotransporter KCC3. The pathogenic role of CLCN2 mutations has not been clearly demonstrated and may represent either susceptibility genes or, in certain cases, innocuous polymorphisms. KCC3 mutations have been associated with hereditary motor and sensory polyneuropathy with corpus callosum agenesis (Andermann syndrome) that often manifests with epileptic seizures. This review summarizes the recent progress in the genetic linkages of epilepsies and seizures to the above genes and discusses potential pathogenic mechanisms that contribute to the age, sex, and conditional expression of these seizures in carriers of these mutations.


Characterisation of eye-lens DNases: long term persistence of activity in post apoptotic lens fibre cells.
Fibre cells in the ocular lens exhibit a constitutive apoptotic process of nuclear degradation that includes chromatin breakage, generating a ladder pattern of DNA fragments. This process is intrinsic to the normal terminal differentiation program. Despite the loss of nucleus and cytoplasmic organelles, the terminal differentiated fibre cells remain in the lens during the whole life span of the individual. The lens cells thus provide a unique system in which to determine the presence and fate of endonucleases once the chromatin has been cleaved. We report here on the presence of DNase activity in nucleated and anucleated lens cells. Using a nuclease gel assay and double-stranded DNA as substrate, we found active 30 and 60 kDa DNases. The enzymatic activities were Ca(2+), Mg(2+) dependent, and active at neutral pH. The relative amount of these forms changed during development and aging of the lens fibre cells. Both forms were inhibited by Zn(2+), aurintricarboxylic acid, and G-actin. The proteins were also separated by SDS-PAGE, renatured after removing SDS and incubated in the presence of native DNA adsorbed to a membrane. Therefore it was possible to demonstrate, by means of a nick translation reaction, that the enzymes produced single strand cuts. Based on these findings we propose that these chick lens nucleases are probably related to DNase I.


Recreational music-making modulates the human stress response: a preliminary individualized gene expression strategy.
A central component of the complex human biological stress response is the modulation of the neuro-endocrine-immune system with its intricate feedback loops that support homeostatic regulation. Well-documented marked gene expression variability among human and animal subjects coupled with sample collection timing and delayed effects, as well as a host of molecular detection challenges renders the quest for deciphering the human biological stress response challenging from many perspectives. A novel Recreational Music-Making (RMM) program was used in combination with a new strategy for peripheral blood gene expression analysis to assess individualized genomic stress induction signatures. The expression of 45 immune response-related genes was determined using a multiplex preamplification step prior to conventional quantitative Real Time Polymerase Chain Reaction (qRT-PCR) mRNA analysis to characterize the multidimensional biological impact of a 2-phase controlled stress induction/amelioration experimental protocol in 32 randomly assigned individuals. In subjects performing the RMM activity following a 1-hour stress induction protocol, 19 out of 45 markers demonstrated reversal with significant (P = 0.05) Pearson correlations in contrast to 6 out of 45 markers in the resting control group and 0 out of 45 in the ongoing stressor group. The resultant amelioration of stress-induced genomic expression supports the underlying premise that RMM warrants additional consideration as a rational choice within our armamentarium of stress reduction strategies. Modulation of individualized genomic stress induction signatures in peripheral blood presents a new opportunity for elucidating the dynamics of the human stress response.


Protein Structural Memory Influences Ligand Binding Mode(s) and Unbinding Rates.
The binding of small molecules (e.g., natural ligands, metabolites, and drugs) to proteins governs most biochemical pathways and physiological processes. Here, we use molecular dynamics to investigate the unbinding of dimethyl sulfoxide (DMSO) from two distinct states of a small rotamase enzyme, the FK506-binding protein (FKBP). These states correspond to the FKBP protein relaxed with and without DMSO in the active site. Since the time scale of ligand unbinding (2-20 ns) is faster than protein relaxation (100 ns), a novel methodology is introduced to relax the protein without having to introduce an artificial constraint. The simulation results show that the unbinding time is an order of magnitude longer for dissociation from the DMSO-bound state (holo-relaxed). That is, the actual rate of unbinding depends on the state of the protein, with the protein having a long-lived memory. The rate thus depends on the concentration of the ligand as the apo and holo states reflect low and high concentrations of DMSO, respectively. Moreover, there are multiple binding modes in the apo-relaxed state, while a single binding mode dominates the holo-relaxed state in which DMSO acts as hydrogen bond acceptor from the backbone NH of Ile56, as in the crystal structure of the DMSO/FKBP complex. The solvent relaxes very fast (∼1 ns) close to the NH of Ile56 and with the same time scale of the protein far away from the active site. These results have implications for high-throughput docking, which makes use of a rigid structure of the protein target.


Alzheimer disease-specific conformation of hyperphosphorylated paired helical filament-Tau is polyubiquitinated through Lys-48, Lys-11, and Lys-6 ubiquitin conjugation.
One of the key pathological hallmarks of Alzheimer disease (AD) is the accumulation of paired helical filaments (PHFs) of hyperphosphorylated microtubule-associated protein Tau. Tandem mass spectrometry was employed to examine PHF-Tau post-translational modifications, in particular protein phosphorylation and ubiquitination, to shed light on their role in the early stages of Alzheimer disease. PHF-Tau from Alzheimer disease brain was affinity-purified by MC1 monoclonal antibody to isolate a soluble fraction of PHF-Tau in a conformation unique to human AD brain. A large number of phosphorylation sites were identified by employing a data-dependent neutral loss algorithm to trigger MS3 scans of phosphopeptides. It was found that soluble PHF-Tau is ubiquitinated at its microtubule-binding domain at residues Lys-254, Lys-311, and Lys-353, suggesting that ubiquitination of PHF-Tau may be an earlier pathological event than previously thought and that ubiquitination could play a regulatory role in modulating the integrity of microtubules during the course of AD. Tandem mass spectrometry data for ubiquitin itself indicate that PHF-Tau is modified by three polyubiquitin linkages, at Lys-6, Lys-11, and Lys-48. Relative quantitative analysis indicates that Lys-48-linked polyubiquitination is the primary form of polyubiquitination with a minor portion of ubiquitin linked at Lys-6 and Lys-11. Because modification by Lys-48-linked polyubiquitin chains is known to serve as the essential means of targeting proteins for degradation by the ubiquitin-proteasome system, and it has been reported that modification at Lys-6 inhibits ubiquitin-dependent protein degradation, a failure of the ubiquitin-proteasome system could play a role in initiating the formation of degradation-resistant PHF tangles.


Dispersed repetitive DNA has spread to new genomes since polyploid formation in cotton.
Polyploid formation has played a major role in the evolution of many plant and animal genomes; however, surprisingly little is known regarding the subsequent evolution of DNA sequences that become newly united in a common nucleus. Of particular interest is the repetitive DNA fraction, which accounts for most nuclear DNA in higher plants and animals and which can be remarkably different, even in closely related taxa. In one recently formed polyploid, cotton (Gossypium barbadense L.; AD genome), 83 non-cross-hybridizing DNA clones contain dispersed repeats that are estimated to comprise about 24% of the nuclear DNA. Among these, 64 (77%) are largely restricted to diploid taxa containing the larger A genome and collectively account for about half of the difference in DNA content between Old World (A) and New World (D) diploid ancestors of cultivated AD tetraploid cotton. In tetraploid cotton, FISH analysis showed that some A-genome dispersed repeats appear to have spread to D-genome chromosomes. Such spread may also account for the finding that one, and only one, D-genome diploid cotton, Gossypium gossypioides, contains moderate levels of (otherwise) A-genome-specific repeats in addition to normal levels of D-genome repeats. The discovery of A-genome repeats in G. gossypioides adds genome-wide support to a suggestion previously based on evidence from only a single genetic locus that this species may be either the closest living descendant of the New World cotton ancestor, or an adulterated relic of polyploid formation. Spread of dispersed repeats in the early stages of polyploid formation may provide a tag to identify diploid progenitors of a polyploid. Although most repetitive clones do not correspond to known DNA sequences, 4 correspond to known transposons, most contain internal subrepeats, and at least 12 (including 2 of the possible transposons) hybridize to mRNAs expressed at readily discernible levels in cotton seedlings, implicating transposition as one possible mechanism of spread. Integration of molecular, phylogenetic, and cytogenetic analysis of dispersed repetitive DNA may shed new light on evolution of other polyploid genomes, as well as providing valuable landmarks for many aspects of genome analysis.


Impact of the Presence of Select Cardiovascular Risk Factors on Cognitive Changes among Dementia Subtypes.
Studies have shown select associations between cardiovascular risk factors and dementia, but mostly focused on Alzheimer's Disease (AD). We enhance these works by evaluating the relationship between the presence of cardiovascular risk factors and the rate of cognitive decline, measured using the Mini-Mental State Examination (MMSE) and Clinical Dementia Rating Sum of Boxes (CDR-SUM) on four common dementia subtypes (AD, dementia with Lewy bodies (DLB), frontotemporal dementia (FTD), and vascular dementia (VaD), as well as non-demented elderly individuals (normal)). We used generalized linear mixed models with random intercepts to account for correlation at the patient and center levels for each dementia subtype adjusting for time since initial visit, baseline cognitive score, age, and demographic factors. The cardiovascular risk factors evaluated included body mass index, diabetes, years of smoking, atrial fibrillation, hypertension, and hypercholesterolemia. Patients diagnosed with AD (n=1899), DLB (n=65), FTD (n=168), or VaD (n=13); or lacked cognitive impairment (normal) (n=3583) were evaluated using data from the National Alzheimer's Coordinating Centers. Cardiovascular risk factors were associated with select dementia subtypes including AD and FTD. Using MMSE and CDR-SUM, recent or active hypertension and hypercholesterolemia were associated with a slower cognitive decline for AD patients, while higher body mass index and years of smoking were associated with a slower cognitive decline for FTD patients. However, several cardiovascular factors demonstrated associations with more rapid cognitive decline. These results demonstrate disease specific associations and can provide clinicians guidance on predicted cognitive changes at the group level using information about cardiovascular risk factors.


Survival from synchronous bilateral breast cancer: the experience of surgeons participating in the breast audit of the Society of Breast Surgeons of Australia and New Zealand.
Previous studies generally indicate that synchronous bilateral breast cancers (SBBC) have an equivalent or moderately poorer survival compared with unilateral cases. The prognostic characteristics of SBBC would be relevant when planning adjuvant therapies and follow-up medical surveillance. The frequency of SBBC among early breast cancers in clinical settings in Australia and New Zealand were investigated, plus their prognostic significance, using the Breast Cancer Audit Database of the Society of Breast Surgeons of Australia and New Zealand, which covered an estimated 60% of early invasive lesions in those countries. Rate ratios (95% confidence limits) of SBBC were investigated among 35,370 female breast cancer cases by age of woman, histology type, grade, tumour diameter, nodal status, lymphatic/vascular invasion and oestrogen receptor status. Univariate and multivariable disease-specific survival analyses were undertaken. 2.3% of cases were found to be SBBC (i.e., diagnoses occurring within 3 months). The figure increased from 1.4% in women less than 40 years to 4.1% in those aged 80 years or more. Disease-specific survivals did not vary by SBBC status (p=0.206). After adjusting for age, histology type, diameter, grade, nodal status, lymphatic/vascular invasion, and oestrogen receptor status, the relative risk of breast cancer death for SBBC was 1.17 (95% CL: 0.91, 1.51). After adjusting for favourable prognostic factors more common in SBBC cases (i.e., histology type, grade, lymphatic/ vascular invasion, and oestrogen receptor status), the relative risk of breast cancer death for SBBC was 1.42 (95% CL: 1.10, 1.82). After adjusting for unfavourable prognostic factors more common in SBBC cases (i.e., older age and large tumour diameter), the relative risk of breast cancer death for SBBC was 0.98 (95% CL: 0.76, 1.26). Results confirm previous findings of an equivalent or moderately poorer survival for SBBC but indicate that SBBC status is likely to be an important prognostic indicator for some cases.


Dissociation of the neural correlates of implicit and explicit memory.
One presentation of a word to a subject is enough to change the way in which the word is processed subsequently, even when there is no conscious (explicit) memory of the original presentation. This phenomenon is known as implicit memory. The neural correlates of implicit memory have been studied previously, but they have never been compared with the correlates of explicit memory while holding task conditions constant or while using a procedure that ensured that the neural correlates were not 'contaminated' by explicit memory. Here we use scalp-recorded event-related brain potentials to identify neural activity associated with implicit and explicit memory during the performance of a recognition memory task. Relative to new words, recently studied words produced activity in three neuroanatomically and functionally dissociable neural populations. One of these populations was activated whether or not the word was consciously recognized, and its activity therefore represents a neural correlate of implicit memory. Thus, when task and memory contamination effects are eliminated, the neural correlates of explicit and implicit memory differ qualitatively.


Interleukin-6 Induces Vascular Endothelial Growth Factor-C Expression via Src-FAK-STAT3 Signaling in Lymphatic Endothelial Cells.
Elevated serum interleukin-6 (IL-6) levels correlates with tumor grade and poor prognosis in cancer patients. IL-6 has been shown to promote tumor lymphangiogenesis through vascular endothelial growth factor-C (VEGF-C) induction in tumor cells. We recently showed that IL-6 also induced VEGF-C expression in lymphatic endothelial cells (LECs). However, the signaling mechanisms involved in IL-6-induces VEGF-C induction in LECs remain incompletely understood. In this study, we explored the causal role of focal adhesion kinase (FAK) in inducing VEGF-C expression in IL-6-stimulated murine LECs (SV-LECs). FAK signaling blockade by NSC 667249 (a FAK inhibitor) attenuated IL-6-induced VEGF-C expression and VEGF-C promoter-luciferase activities. IL-6's enhancing effects of increasing FAK, ERK1/2, p38MAPK, C/EBPβ, p65 and STAT3 phosphorylation as well as C/EBPβ-, κB- and STAT3-luciferase activities were reduced in the presence of NSC 667249. STAT3 knockdown by STAT3 siRNA abrogated IL-6's actions in elevating VEGF-C mRNA and protein levels. Moreover, Src-FAK signaling blockade reduced IL-6's enhancing effects of increasing STAT3 binding to the VEGF-C promoter region, cell migration and endothelial tube formation of SV-LECs. Together these results suggest that IL-6 increases VEGF-C induction and lymphangiogenesis may involve, at least in part, Src-FAK-STAT3 cascade in LECs.


Microcolinearity in sh2-homologous regions of the maize, rice, and sorghum genomes.
Large regions of genomic colinearity have been demonstrated among grass species by recombinational mapping, but the degree of chromosomal conservation at the sub-centimorgan level has not been extensively investigated. We cloned the rice and sorghum genes homologous to the sh2 locus of maize on bacterial artificial chromosomes (BACs), and observed that a homologue of the maize a1 gene was also present on each of these BACs. In sorghum, we found a direct duplication of a1 homologues separated by about 10 kb. In maize, sh2 and a1 are approximately 140 kb apart and transcribed in the same direction, with sh2 upstream of a1. In rice and sorghum, this arrangement is fully conserved. However, the sh2 and a1 homologues are separated by about 19 kb in both rice and sorghum. We found low-copy-number and repetitive DNAs between the sh2 and a1 homologues of sorghum and rice. The sh2 and a1 homologues cross-hybridized, but the repetitive DNA and most low-copy-number sequences between these genes did not. These results indicate that maize, sorghum, and rice have conserved gene order and composition in the sh2-a1 region, but have acquired extensive qualitative and quantitative differences in the sequences between these genes.


Genetic diversity of Mycobacterium tuberculosis isolates from a tertiary care tuberculosis hospital in South Korea.
Tuberculosis (TB) remains an immense public health problem in the Republic of Korea despite a more than fivefold decrease in the prevalence of the disease over the last 3 decades. The rise in drug-resistant TB has compounded the situation. We analyzed 208 clinical isolates of M. tuberculosis from the National Masan Tuberculosis Hospital by spoligotyping, IS6110 restriction fragment length polymorphism (RFLP), and 24-locus-based mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) typing to assess the diversity and transmission dynamics of the tubercle bacilli in the Republic of Korea. The majority of the isolates (97.1%) belonged to the Beijing genotype. Cluster analysis by MIRU-VNTR yielded a low clustering rate of 22.3%, with most of the clusters comprising isolates with diverse drug resistance patterns. The discriminatory capacity of the typing methods was high for RFLP and MIRU-VNTR (allelic diversity [h] = 0.99) but low for spoligotyping (h = 0.31). Although analysis of 19 MIRU-VNTR loci was needed to achieve maximum discrimination, an informative set of 8 loci (960, 1955, 2163b, 2165, 2996, 3192, 4052, and 4348) (h = 0.98) that was able to differentiate most of the closely related strains was identified. These findings suggest that 24-locus-based MIRU-VNTR typing is a likely suitable alternative to RFLP to differentiate clinical isolates in this setting, which is dominated by M. tuberculosis Beijing strains. Within the study limits, our results also suggest that the problem of drug-resistant TB in the Republic of Korea may be largely due to acquired resistance as opposed to transmission.


Reproducibility and repeatability of six high-throughput 16S rDNA sequencing protocols for microbiota profiling.
Culture-independent molecular techniques and advances in next generation sequencing (NGS) technologies make large-scale epidemiological studies on microbiota feasible. A challenge using NGS is to obtain high reproducibility and repeatability, which is mostly attained through robust amplification. We aimed to assess the reproducibility of saliva microbiota by comparing triplicate samples. The microbiota was produced with simplified in-house 16S amplicon assays taking advantage of large number of barcodes. The assays included primers with Truseq (TS-tailed) or Nextera (NX-tailed) adapters and either with dual index or dual index plus a 6-nt internal index. All amplification protocols produced consistent microbial profiles for the same samples. Although, in our study, reproducibility was highest for the TS-tailed method. Five replicates of a single sample, prepared with the TS-tailed 1-step protocol without internal index sequenced on the HiSeq platform provided high alpha-diversity and low standard deviation (mean Shannon and Inverse Simpson diversity was 3.19 ± 0.097 and 13.56 ± 1.634 respectively). Large-scale profiling of microbiota can consistently be produced by all 16S amplicon assays. The TS-tailed-1S dual index protocol is preferred since it provides repeatable profiles on the HiSeq platform and are less labour intensive.


Brief report: Conveying subjective experience in conversation: production of mental state terms and personal narratives in individuals with high functioning autism.
Mental state terms and personal narratives are conversational devices used to communicate subjective experience in conversation. Pre-adolescents with high-functioning autism (HFA, n = 20) were compared with language-matched typically-developing peers (TYP, n = 17) on production of mental state terms (i.e., perception, physiology, desire, emotion, cognition) and personal narratives (sequenced retelling of life events) during short conversations. HFA and TYP participants did not differ in global use of mental state terms, nor did they exhibit reduced production of cognitive terms in particular. Participants with HFA produced significantly fewer personal narratives. They also produced a smaller proportion of their mental state terms during personal narratives. These findings underscore the importance of assessing and developing qualitative aspects of conversation in highly verbal individuals with autism.


Differential effects of the novel kappa opioid receptor antagonist, JDTic, on reinstatement of cocaine-seeking induced by footshock stressors vs cocaine primes and its antidepressant-like effects in rats.
Stress and depression have been linked to relapse of cocaine abuse. Antagonism of the kappa opioid receptor (KOR) has been reported to attenuate some effects of stressors, and antagonism of the KOR has been reported to have antidepressant-like properties. Our objective was to determine whether the potent and selective KOR antagonist, (3R)-7-hydroxy-N-{(1S)-1-{[(3R,4R)-4-(3-hydroxyphenyl)-3,4-dimethyl-1-piperidinyl]methyl}-2-methylpropyl}-1,2,3,4-tetrahydro-3-isoquinoline-carboxamide (JDTic), can reduce the ability of a stressor (intermittent footshock) to reinstate cocaine-seeking behavior and to have antidepressant-like effects in the forced swim test (FST). Male Long-Evans hooded rats were trained to lever-press, reinforced with 0.5 mg/kg i.v. infusion of cocaine, according to fixed ratio 1 reinforcement schedules during daily 2-h experimental sessions. After performance had stabilized, lever pressing was extinguished for 12 consecutive sessions, and doses of 0 (vehicle), 3, 10, and 30 mg/kg JDTic were then administered i.g. to separate groups of 12 rats. Twenty four hours later, the rats were given 15 min of intermittent footshock (0.87 mA, 0.5 s activation time, average inter-activation interval of 40 s) or a 17-mg/kg i.p. administration of cocaine prime followed by a 2-h reinstatement test session. JDTic was also evaluated for its ability to block diuresis induced by the KOR agonist, U50,488H (10 mg/kg, s.c.), during 5-h test sessions beginning 1 h after footshock reinstatement tests to verify its KOR antagonist activity. In the FST, male Sprague-Dawley rats were treated with either nor-binaltorphimine (nor-BNI) or JDTic (both at 0.3, 1, 3, or 10 mg/kg, injected s.c. 23 h before), or desipramine (5.6, 10, or 17 mg/kg, injected i.p. 23, 5, and 1 h before) and placed in a cylinder of water, during which the predominance of immobility, swimming, and climbing were scored during 5-s intervals for 5 min. The 10- and 30-mg/kg doses of JDTic significantly reduced footshock-induced reinstatement of responding previously reinforced by cocaine and significantly attenuated U50,488H-induced diuresis. In contrast, JDTic did not affect cocaine-prime-induced reinstatement. Both nor-BNI and JDTic decreased immobility and increased swimming time in the FST, similar to the antidepressant desipramine. Depression and stress are two states during cocaine abstinence which users identify as precipitating relapse, and JDTic may have properties which attenuate both.


Collective intelligence for translational medicine: Crowdsourcing insights and innovation from an interdisciplinary biomedical research community.
Translational medicine bridges the gap between discoveries in biomedical science and their safe and effective clinical application. Despite the gross opportunity afforded by modern research for unparalleled advances in this field, the process of translation remains protracted. Efforts to expedite science translation have included the facilitation of interdisciplinary collaboration within both academic and clinical environments in order to generate integrated working platforms fuelling the sharing of knowledge, expertise, and tools to align biomedical research with clinical need. However, barriers to scientific translation remain, and further progress is urgently required. Collective intelligence and crowdsourcing applications offer the potential for global online networks, allowing connection and collaboration between a wide variety of fields. This would drive the alignment of biomedical science with biotechnology, clinical need, and patient experience, in order to deliver evidence-based innovation which can revolutionize medical care worldwide. Here we discuss the critical steps towards implementing collective intelligence in translational medicine using the experience of those in other fields of science and public health.


Glomerular lipidosis accompanied by renal tubular oxalosis in wild and laboratory-reared Japanese rock ptarmigans (Lagopus mutus japonicus).
Glomerular lipidosis is a disease characterized by lipid accumulation in mesangial cells but that has not been fully investigated in avian species. We examined four wild and two laboratory-reared Japanese rock ptarmigans (Lagopus mutus japonicus)--an endangered avian species--presenting vacuolar deposits in the glomeruli. All cases had vacuolar deposits in the glomeruli. In the wild cases, fewer than 30% of all glomeruli were affected, compared with more than 90% in the laboratory-reared cases. In the wild cases, most deposits were mild and restricted to the mesangial areas of glomeruli. In the laboratory-reared cases, nearly all of the deposits covered entire glomeruli. Electron microscopy of mild deposits revealed vacuoles in the cytoplasm of mesangial cells. These vacuoles were positive for Sudan III, Sudan black B, oil red O, Nile blue, periodic acid-Schiff, Schultz test, and digitonin stain and were negative for performaric acid-Schiff stains. Based on these results, we diagnosed the glomerular lesion as glomerular lipidosis caused by uptake of low-density lipoprotein in mesangial cells. Except for one wild case, all cases exhibited renal tubular oxalosis. The severity of tubular oxalosis tended to be related to the severity of glomerular lipidosis: In cases of mild glomerular lipidosis, tubular oxalosis was also mild or absent. We therefore diagnosed the primary lesion as glomerular lipidosis accompanied by tubular oxalosis. The four wild cases came from different zones and therefore had no opportunities to interbreed and no common relatives. We believe these data support the hypothesis that glomerular lipidosis is a disease of the general population ofJapanese rock ptarmigans. This is the first report of glomerular lipidosis accompanied by renal tubular oxalosis in an avian species.


Smad4 loss in mice causes spontaneous head and neck cancer with increased genomic instability and inflammation.
Smad4 is a central mediator of TGF-beta signaling, and its expression is downregulated or lost at the malignant stage in several cancer types. In this study, we found that Smad4 was frequently downregulated not only in human head and neck squamous cell carcinoma (HNSCC) malignant lesions, but also in grossly normal adjacent buccal mucosa. To gain insight into the importance of this observation, we generated mice in which Smad4 was deleted in head and neck epithelia (referred to herein as HN-Smad4-/- mice) and found that they developed spontaneous HNSCC. Interestingly, both normal head and neck tissue and HNSCC from HN-Smad4-/- mice exhibited increased genomic instability, which correlated with downregulated expression and function of genes encoding proteins in the Fanconi anemia/Brca (Fanc/Brca) DNA repair pathway linked to HNSCC susceptibility in humans. Consistent with this, further analysis revealed a correlation between downregulation of Smad4 protein and downregulation of the Brca1 and Rad51 proteins in human HNSCC. In addition to the above changes in tumor epithelia, both normal head and neck tissue and HNSCC from HN-Smad4-/- mice exhibited severe inflammation, which was associated with increased expression of TGF-beta1 and activated Smad3. We present what we believe to be the first single gene-knockout model for HNSCC, in which both HNSCC formation and invasion occurred as a result of Smad4 deletion. Our results reveal an intriguing connection between Smad4 and the Fanc/Brca pathway and highlight the impact of epithelial Smad4 loss on inflammation.


Experimental Legionella longbeachae infection in intratracheally inoculated mice.
This study established an experimental model of replicative Legionella longbeachae infection in A/J mice. The animals were infected by intratracheal inoculation of 10(3)-10(9) c.f.u. L. longbeachae serogroup 1 (USA clinical isolates D4968, D4969 and D4973). The inocula of 10(9), 10(8), 10(7) and 10(6) c.f.u. of all tested L. longbeachae serogroup 1 isolates were lethal for A/J mice. Inoculation of 10(5) c.f.u. L. longbeachae caused death in 90 % of the animals within 5 days, whilst inoculation of 10(4) c.f.u. caused sporadic death of mice. All animals that received 10(3) c.f.u. bacteria developed acute lower respiratory disease, but were able to clear Legionella from the lungs within 3 weeks. The kinetics of bacterial growth in the lungs was independent of inoculum size and reached a growth peak about 3 logarithms above the initial inoculum at 72 h after inoculation. The most prominent histological changes in the lungs were observed at 48-72 h after inoculation in the form of a focal, neutrophil-dominant, peribronchiolar infiltration. The inflammatory process did not progress towards the interstitial or alveolar spaces. Immunohistological analyses revealed L. longbeachae serogroup 1 during the early phase of infection near the bronchiolar epithelia and later co-localized with inflammatory cells. BALB/c and C57BL/6 mice strains were also susceptible to infection with all L. longbeachae serogroup 1 strains tested and very similar changes were observed in the lungs of infected animals. These results underline the infection potential of L. longbeachae serogroup 1, which is associated with high morbidity and lethality in mice.


Tetrahydrocannabinol and Cannabidiol Use in an Outpatient Palliative Medicine Population.
Palliative medicine physicians are challenged by lack of guidance regarding effectiveness and dosing of cannabis products in the setting of their emerging popularity. The aim of this study was to describe early patterns of tetrahydrocannabinol (THC) and cannabidiol (CBD) use in Florida following passage of the state's first medical marijuana law. We describe here the perceived benefits, side effects, and beliefs expressed by patients in a single outpatient academic palliative medicine practice. A cross-sectional survey was performed of a sequential convenience sample of patients who presented to an outpatient academic palliative medicine clinic over a 3-month period. In all, 24% (14/58) of respondents reported THC use, with half using THC on a daily basis. Patients reported improvements in pain, appetite, and nausea. In all, 71% (10/14) began using THC after the diagnosis of their chronic illness, and the most common form of usage was vaping. In all, 24% (14/58) of patients reported CBD use. Patients reported improvements in pain, and the most common form of usage was topical application. None of the patients had used CBD prior to the onset of their chronic illness. In all, 21% (3/14) of THC users and 21% (3/14) of CBD users thought that their substance was helping to cure their illness. Individual reported side effects in both groups were minimal. Approximately a quarter of outpatient palliative care patients use THC or CBD, often on a daily basis. Palliative care providers should be aware of the frequency, diverse usage, and beliefs behind cannabis product use in this patient population.


Investigating 'place effects' on mental health: implications for population-based studies in psychiatry.
Background. Interest in features of our local environments that may promote better mental health and wellbeing continues to rise among decision makers. Our purpose was to highlight a selection of these challenges and some promising avenues for enhancing the quality of evidence. Method. An analysis of approximately 267, 000 people was used to test the local relative deprivation hypothesis, wherein the shortfall of a person's socioeconomic circumstances from their neighbours is said to impact negatively upon mental health. This case was used to anchor further discussion of challenges to identifying and interpreting genuine 'place effects' from spurious correlations. Results. A Median Odds Ratio of 1.29 computed via multilevel logistic regression showed that the odds of experiencing psychological distress (as measured by the Kessler score) varied by geographical area. Approximately 67% of this was attributed to a cross-classified measure of household income and neighbourhood deprivation. Compared to people on high incomes living in affluent neighbourhoods, the odds ratio of psychological distress for people on low incomes in affluent areas was 4.73 (95% confidence interval (95% CI) 4.39, 5.09), whereas that for people on low incomes in deprived areas was significantly higher at 5.83 (95% CI 5.41, 6.28). Conclusions. While no evidence was found to support local relative deprivation hypothesis, the pattern suggests that more affluent areas may contain features that are conducive to better mental health. Selection of bespoke geographical boundaries, use of directed acyclic graphs and more evaluations of natural experiments are likely to be important in taking the field of enquiry onwards.


Lennox-Gastaut syndrome: impact on the caregivers and families of patients.
Lennox-Gastaut syndrome (LGS) has a major impact on the health-related quality of life (HRQL) of the affected children as well as their caregivers. The primary caregiver in the family is generally the mother, with support from the father and siblings. The burden of care and the effects of the disease on the child necessitate adjustments in virtually all aspects of the lives of their family. These adjustments inevitably affect the physical, emotional, social, and financial health of the whole family. Numerous sources of support for families can help to ease the burden of care. Improvements in the treatment of LGS, in addition to helping the child with LGS, would likely help improve the HRQL of the family members. This pilot parent survey was designed to explore the impact of epilepsy on caregiver HRQL. Parents of children with epilepsy who had contacted the Epilepsy Information Service at the Wake Forest University School of Medicine, Winston-Salem, NC, USA, were sent questionnaires comprising open- and closed-ended questions. A total of 200 surveys were distributed, with a return rate of 48%. The results revealed that 74% of the parents believed that having a child with epilepsy brought them and their partner closer together. However, when the parents were asked to explain the manner in which epilepsy affected their families, answers included continuous stress, major financial distress, and lack of time to spend with other children. Information and resources for the families of children with LGS could help improve the HRQL of both the patients and their relatives.


First-admission psychiatric ward milieu: treatment process and outcome.
A two-year, prospective study of 123 first-admission patients examined the impact of different inpatient treatment decision-making styles on patient outcome on a therapeutic community ward and a medically oriented psychiatric ward. Ward philosophy, programmatic and administrative structure, and ward atmosphere defined the ward milieus. Data on length of stay and the use of various therapeutic modalities provided comparisons of treatment process. Standardized ratings of psychopathologic condition and levels of function, readmissions, and illness attitudes were used to assess two-year outcomes. The major finding was the absence of outcome differences despite longer lengths of stay on the therapeutic community ward. Hence, differences in ward decision-making style may not influence outcome, but may affect the process of care among diagnostically heterogeneous, first-admission patients treated in a university setting.


Dogma derailed: the many influences of RNA on the genome.
Epigenetic control of gene expression is a critical component of transcriptional regulation. Remarkably, the deposition of epigenetic modifications is often guided by noncoding RNAs. Although noncoding RNAs have been most often implicated in posttranscriptional gene silencing, these molecules are now emerging as critical regulators of gene expression and genomic stability at the transcriptional level. Here, we review recent efforts to understand the mechanisms by which RNA controls the expression or content of DNA. We discuss the role of both small RNAs and long noncoding RNAs in directing chromatin changes through histone modifications and DNA methylation. Furthermore, we highlight the function of RNA in mediating DNA cleavage during genome rearrangements and pathogen defense. In understanding the mechanisms of RNA control over DNA, the power of RNA may one day be harnessed to impact gene expression in a therapeutic setting.


Effect of dysfunctional vascular endothelium on myocardial performance in isolated papillary muscles.
Vascular endothelium has been shown to modify the contractile characteristics of vascular smooth muscle, and endocardial endothelium has been shown to modify the contractile characteristics of adjacent myocardium. In this study, whether vascular endothelium also modifies the contractile characteristics of adjacent myocardium and whether these effects are additive to those of endocardial endothelium were investigated. Rabbit hearts (n = 54) were excised and mounted in a Langendorff preparation. Vascular reactivity was verified by acetylcholine infusion. One group of these hearts had Triton X-100 injected as a bolus into the coronaries to render the vascular endothelium dysfunctional. The other portion served as control hearts. Triton X-100 bolus injection resulted in little or no pathological changes on morphological examination; however, the vasodilatory response to acetylcholine in these hearts was abolished, suggesting vascular endothelial dysfunction. Vascular smooth muscle reactivity was verified in Triton X-100-injected hearts by nitroprusside infusion. In the control Langendorff-perfused hearts, there was little evidence of vascular endothelial dysfunction, with the coronary perfusion rate increasing from 8.9 +/- 0.4 to 11.0 +/- 0.3 ml/g per minute (p < 0.01) in response to acetylcholine. All hearts were then removed, and right ventricular papillary muscles were excised for myocardial mechanical studies. Control Langendorff-perfused hearts had myocardial mechanical characteristics similar to those of muscles from 18 other control hearts without Langendorff perfusion, indicating that the Langendorff perfusion itself had little effect on myocardial mechanics. The muscles from the Triton X-100-injected Langendorff hearts had marked changes: a shortening of twitch duration (363 +/- 16 versus 449 +/- 9 msec, p < 0.01) and decreases in total tension (2.2 +/- 0.2 versus 2.9 +/- 0.2 g/mm2, p < 0.01), dT/dt (9 +/- 1 versus 12 +/- 1 g/mm2 per second, p < 0.05), and maximum velocity of unloaded muscle shortening (Vmax) (0.89 +/- 0.06 versus 1.14 +/- 0.07 length at which maximum developed tension occurred [Lmax]/sec, p < 0.05). Endocardial endothelial removal of the papillary muscles in the two control groups (with and without Langendorff perfusion) by Triton X-100 caused the same changes in twitch characteristics as occurred in muscles from the Langendorff-perfused hearts injected with Triton X-100 but with intact endocardial endothelium, suggesting that vascular endothelial dysfunction had similar effects on contractile characteristics as endocardial endothelial removal.(ABSTRACT TRUNCATED AT 400 WORDS)


Design of a small personal air monitor and its application in aircraft.
A small air sampling system using standard air filter sampling technology has been used to monitor the air in aircraft. The device is a small ABS constructed cylinder 5 cm in diameter and 9 cm tall and can be operated by non technical individuals at an instant notice. It is completely self contained with a 4 AAA cell power supply, DC motor, a centrifugal fan, and accommodates standard 37 mm filters and backup pads. The monitor is totally enclosed and pre assembled in the laboratory. A 45 degrees twist of the cap switches on the motor and simultaneously opens up the intake ports and exhaust ports allowing air to pass through the filter. A reverse 45 degrees twist of the cap switches off the motor and closes all intake and exhaust ports, completely enclosing the filter. The whole monitor is returned to the laboratory by standard mail for analysis and reassembly for future use. The sampler has been tested for electromagnetic interference and has been approved for use in aircraft during all phases of flight. A set of samples taken by a BAe-146-300 crew member during two flights in the same aircraft and analyzed by GC-MS, indicated exposure to tricresyl phosphate (TCP) levels ranging from 31 to 83 nanograms/m(3) (detection limit <4.5 nanograms/m(3)). The latter elevated level was associated with the use of the auxiliary power unit (APU) in the aircraft. It was concluded that the air sampler was capable of monitoring air concentrations of TCP isomers in aircraft above 4.5 nanogram/m(3).


Keratinolytic abilities of Micrococcus luteus from poultry waste.
Keratinolytic microorganisms have become the subject of scientific interest due to their ability to biosynthesize specific keratinases and their prospective application in keratinic waste management. Among several bacterial classes, actinobacteria remain one of the most important sources of keratin-degrading strains, however members of the Micrococcaceae family are rarely scrutinized in regard to their applicatory keratinolytic potential. The tested Micrococcus sp. B1pz isolate from poultry feather waste was identified as M. luteus. The strain, grown in the medium with 1-2% chicken feathers and a yeast extract supplement, produced keratinases of 32 KU and lower level of proteases, 6 PU. It was capable to effectively decompose feathers or "soft" keratin of stratum corneum, in contrast to other "hard" hair-type keratins. The produced keratinolytic enzymes were mainly a combination of alkaline serine or thiol proteases, active at the optimum pH 9.4, 55 °C. Four main protease fractions of 62, 185, 139 and 229 kDa were identified in the crude culture fluid. The research on the auxiliary role of reducing factors revealed that reducing sulfur compounds could be applied in keratinolysis enhancement during enzymatic digestion of keratin, rather than in culture conditions. The presented M. luteus isolate exhibits a significant keratinolytic potential, which determines its feasible applicatory capacity towards biodegradation of poultry by-products or formulation of keratin-based feed components.


Genomics and metabolomics as markers for the interaction of diet and health: lessons from lipids.
Foods are not purified compounds acting on single molecular targets, but complex mixtures of molecules that modulate many biochemical pathways simultaneously. Diet affects the probability of developing various diseases. Nevertheless, specific recommendations for individual diets are not simple. Recommending nutrient intakes above and beyond those needed to provide adequacy requires scientific knowledge and regulatory scrutiny to ensure the efficacy and safety even of essential nutrients. Designing a diet to improve metabolic health is a bold and ambitious goal. It is possible to design foods that will alter metabolism, but what change will make everyone who is otherwise healthy even healthier? Changing one aspect of metabolism to lower the risk of one disease does not improve overall health if it comes at the expense of disrupting another aspect of metabolism that increases the risk of another disease. This issue has: 1) frustrated nutritional recommendations that could provide benefits to the health of large subsets of the population, 2) caused the recall of drugs with many beneficial effects and 3) caused harm by implying that single nutrients/foods could be healthy for everyone. An individualized system for metabolic assessment would establish the efficacy and safety of nutrients such as amino acids or fatty acids when these are designed to be consumed at levels providing improved metabolic health. The need to document the lack of an adverse effect of a food or drug on physiology necessitates a global, i.e. metabolomic approach.


Structural and functional characterization of Rv2966c protein reveals an RsmD-like methyltransferase from Mycobacterium tuberculosis and the role of its N-terminal domain in target recognition.
Nine of ten methylated nucleotides of Escherichia coli 16 S rRNA are conserved in Mycobacterium tuberculosis. All the 10 different methyltransferases are known in E. coli, whereas only TlyA and GidB have been identified in mycobacteria. Here we have identified Rv2966c of M. tuberculosis as an ortholog of RsmD protein of E. coli. We have shown that rv2966c can complement rsmD-deleted E. coli cells. Recombinant Rv2966c can use 30 S ribosomes purified from rsmD-deleted E. coli as substrate and methylate G966 of 16 S rRNA in vitro. Structure determination of the protein shows the protein to be a two-domain structure with a short hairpin domain at the N terminus and a C-terminal domain with the S-adenosylmethionine-MT-fold. We show that the N-terminal hairpin is a minimalist functional domain that helps Rv2966c in target recognition. Deletion of the N-terminal domain prevents binding to nucleic acid substrates, and the truncated protein fails to carry out the m(2)G966 methylation on 16 S rRNA. The N-terminal domain also binds DNA efficiently, a property that may be utilized under specific conditions of cellular growth.


Rapid assessment of partograph utilisation in selected maternity units in Kenya.
Prolonged labour causes maternal and perinatal morbidity and mortality. Its sequela include obstructed labour, uterine rupture, maternal exhaustion, postpartum haemorrhage, puerperal sepsis, obstetric fistula, stillbirths, birth asphyxia and neonatal sepsis. These complications can be reduced by using the partograph to assess the progress of labour. The Ministry of Health, Kenya has adopted this tool for labour management in the country and the standardised partograph is recommended for use in all delivery units. To determine the utilisation of the partograph in the management of labour in selected health facilities in Kenya. A descriptive cross sectional study. Nine health facilities -ranging from a tertiary hospital to health centre, including public private and faith based facilities in four provinces in Kenya. All facilities apart from Pumwani Maternity Hospital and one health centre were using the partograph. The correct use was low, the knowledge on the use of the tool was average and there was minimal formal training being provided. Staff shortage was listed as the most common cause of not using the tool. Contractions were recorded 30-80%, foetal heart rate 53-90% and cervical dilatation 70-97%. Documentation of state of the liquor, moulding and descent as well as maternal parameters such as pulse, and blood pressure and urinalysis were minimally recorded. Supplies for monitoring labour such as fetoscopes and blood pressure machines were in short supply and sometimes not functional. Overall, the poor usage was contributed to staff shortages, lack of knowledge especially on interpretation of findings, negative attitudes, conflict between providers as to their roles in filling the partograph, and senior staff themselves not acting as role models with regards to the use, advocacy and implementation of the partograph. The partograph was available in most units. However, accurate recording of parameters to monitor the foetus, the mother and progress of labour as recommended was mostly not done. Shortage of staff, lack of knowledge, lack of team work, lack of supplies and negative attitude among healthcare providers were some of the obstacles noted to hamper partograph use.


Interventions to reduce risks associated with vehicle incompatibility.
Occupants of smaller, lighter passenger cars are more likely to be killed or injured in collisions with larger, heavier sport utility vehicles and light trucks than in collisions with other cars. Interventions are needed to reduce this vehicle "incompatibility" and its consequences. The authors conducted a systematic literature review to identify evaluations of interventions to reduce incompatibility. They reviewed engineering, biomedical, and other technical literature. To be included, a study must have 1) evaluated an intervention to reduce vehicle incompatibility, or its consequences, in a crash; 2) reported new research; and 3) been published in English from 1990 to 2010. Seventeen studies met the inclusion criteria. Interventions were designed to reduce the aggressivity of larger vehicles or improve the crashworthiness of smaller vehicles. Effective interventions included 1) modified bumper heights, 2) improved side strength of smaller vehicles, 3) side-impact air bags, 4) changes to vehicle stiffness, and 5) modifications of other front-end structures. Some of the interventions shown to be effective are now in wide use. However, others have yet to be required by regulators or voluntarily agreed to by manufacturers. If larger, heavier vehicles remain on the nation's roads, countermeasures will be needed to reduce risks for occupants of other vehicles.


Are changes in synaptic function that underlie hyperexcitability responsible for seizure activity?
The synaptic and intrinsic mechanisms responsible for epileptic seizures and briefer interictal epileptic discharges have been characterized in some detail. This chapter will outline some aspects of this work in the context of focal epilepsies, particularly in the temporal lobe, and will identify some of the major questions that remain. Early work, mainly using the actions of convulsant treatments on brain slices in vitro, revealed synaptic circuitry that could recruit populations of neurons into synchronous epileptic discharges. Subsequent investigations into cellular mechanisms of chronic experimental and clinical foci, again often in vitro, have revealed complex changes in synaptic properties, synaptic connectivity, intrinsic neuronal properties and selective losses of neurons: unraveling their roles in generating seizures, interictal discharges and interictal dysfunctions/comorbidities remains a significant challenge. In vivo recordings have revealed aspects of the pathophysiology of epileptic foci that have practical implications, for instance high-frequency oscillations, and potentially high-frequency hypersynchronous neuronal firing, which have been useful in localizing the epileptogenic zone for surgical resection.


Gender differences in mortality and quality of life after septic shock: A post-hoc analysis of the ARISE study.
To assess the impact of gender and pre-menopausal state on short- and long-term outcomes in patients with septic shock. Cohort study of the Australasian Resuscitation in Sepsis Evaluation (ARISE) trial, an international randomized controlled trial comparing early goal-directed therapy (EGDT) to usual care in patients with early septic shock, conducted between October 2008 and April 2014. The primary exposure in this analysis was legal gender and the secondary exposure was pre-menopausal state defined by chronological age (≤ 50 years). 641 (40.3%) of all 1591 ARISE trial participants in the intention-to-treat population were females and overall, 337 (21.2%) (146 females) patients were 50 years of age or younger. After risk-adjustment, we could not identify any survival benefit for female patients at day 90 in the younger (≤50 years) (adjusted Odds Ratio (aOR): 0.91 (0.46-1.89), p = .85) nor in the older (>50 years) age-group (aOR: 1.10 (0.81-1.49), p = .56). Similarly, there was no gender-difference in ICU, hospital, 1-year mortality nor quality of life measures. This post-hoc analysis of a large multi-center trial in early septic shock has shown no short- or long-term survival effect for women overall as well as in the pre-menopausal age-group.


Genetic identification of intestinal microsporidia species in immunocompromised patients in Tunisia.
Stool samples from 86 immunocompromised patients (51 human immunodeficiency virus (HIV)-infected patients and 35 patients with haematologic malignancies) were systematically screened for intestinal microspordiosis by microscopic examination and polymerase chain reaction (PCR) using universal primer V1/PMP2. Nine samples (10.5%) showed amplification with the predictive size of fragment (6 from HIV-infected patients and 3 from patients with myeloma). Only 5 out of them (all HIV-infected patients) were revealed positive by microscopy. By means of amplicons fragment size, species-specific primers (V1/EB450, V1/IS500) and sequencing, 3 microsporidia species were for the first time identified in Tunisia: Enterocytozoon bieneusi (3 isolates), Encephelitozoon intestinalis (2 isolates), and Encephalitozoon hellem (1 isolate). Systematic use of such sensitive and discriminative molecular tools will contribute to determining the true prevalence of microsporidiosis in Tunisia and to better management of infected immunocompromised subjects.