Value of beat-to-beat blood pressure changes, detected by pulse transit time, in the management of the obstructive sleep apnoea/hypopnoea syndrome.
Two important aspects of a respiratory sleep study are a measure of inspiratory effort and an estimate of the number of arousals. These can be derived from an indirect estimate of beat-to-beat blood pressure (BP), pulse transit time (PTT). This study investigated the reproducibility of inspiratory BP falls (reflecting inspiratory effort), and BP arousals derived from PTT, and the contribution they could make to the management of the obstructive sleep apnoea/hypopnoea syndrome (OSAHS). Overnight PTT was recorded at home in 40 patients being investigated for OSAHS, and a second PTT recording was made in the sleep laboratory with full polysomnography. Patients were divided into three groups according to the severity of their sleep disorder, and a third PTT recording was made at home in 13 patients subsequently established on nasal continuous positive airway pressure (CPAP). The reproducibility between the home and laboratory studies was reasonable (r=0.87 for inspiratory BP falls, r=0.81 for BP arousals). Both derivatives showed a clear progression through the three patient groups, which returned to normal on treatment. The differences between the groups were significant (p<0.001 for inspiratory BP falls, p=0.0014 for BP arousals). Receiver operator characteristic curves, used to compare polysomnography variables and PTT variables, confirmed that the PTT variables were as good as apnoea-hypopnoea index (AHI), >4% arterial oxygen saturation dip rate and electroencephalography micro-arousals at dividing patients into two groups, either requiring nasal CPAP or not requiring CPAP. Pulse transit time can provide a noninvasive estimate of inspiratory effort and a measure of arousals that together document disease severity and response to treatment and may be useful in managing obstructive sleep apnoea/hypopnoea syndrome.


Allosteric modulation of glycine receptors.
Inhibitory (or strychnine sensitive) glycine receptors (GlyRs) are anion-selective transmitter-gated ion channels of the cys-loop superfamily, which includes among others also the inhibitory γ-aminobutyric acid receptors (GABA(A) receptors). While GABA mediates fast inhibitory neurotransmission throughout the CNS, the action of glycine as a fast inhibitory neurotransmitter is more restricted. This probably explains why GABA(A) receptors constitute a group of extremely successful drug targets in the treatment of a wide variety of CNS diseases, including anxiety, sleep disorders and epilepsy, while drugs specifically targeting GlyRs are virtually lacking. However, the spatially more restricted distribution of glycinergic inhibition may be advantageous in situations when a more localized enhancement of inhibition is sought. Inhibitory GlyRs are particularly relevant for the control of excitability in the mammalian spinal cord, brain stem and a few selected brain areas, such as the cerebellum and the retina. At these sites, GlyRs regulate important physiological functions, including respiratory rhythms, motor control, muscle tone and sensory as well as pain processing. In the hippocampus, RNA-edited high affinity extrasynaptic GlyRs may contribute to the pathology of temporal lobe epilepsy. Although specific modulators have not yet been identified, GlyRs still possess sites for allosteric modulation by a number of structurally diverse molecules, including alcohols, neurosteroids, cannabinoids, tropeines, general anaesthetics, certain neurotransmitters and cations. This review summarizes the present knowledge about this modulation and the molecular bases of the interactions involved.


Characteristic expression of extracellular matrix in subcutaneous adipose tissue development and adipogenesis; comparison with visceral adipose tissue.
Adipose tissue is a connective tissue specified for energy metabolism and endocrines, but functional differences between subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) have not been fully elucidated. To reveal the physiological role of SAT, we characterized in vivo tissue development and in vitro adipocyte differentiation. In a DNA microarray analysis of SAT and VAT in Wistar rats, functional annotation clusters of extracellular matrix (ECM)-related genes were found in SAT, and major ECM molecules expressed in adipose tissues were profiled. In a histological analysis and quantitative expression analysis, ECM expression patterns could be classified into two types: (i) a histogenesis-correlated type such as type IV and XV collagen, and laminin subunits, (ii) a high-SAT expression type such as type I, III, and V collagen and minor characteristic collagens. Type (i) was related to basal membrane and up-regulated in differentiated 3T3-L1 cells and in histogenesis at depot-specific timings. In contrast, type (ii) was related to fibrous forming and highly expressed in 3T3-L1 preadipocytes. Exceptionally, fibronectin was abundant in developed adipose tissue, although it was highly expressed in 3T3-L1 preadipocytes. The present study showed that adipose tissues site-specifically regulate molecular type and timing of ECM expression, and suggests that these characteristic ECM molecules provide a critical microenvironment, which may affect bioactivity of adipocyte itself and interacts with other tissues. It must be important to consider the depot-specific property for the treatment of obesity-related disorders, dermal dysfunction and for the tissue regeneration.


Atomic force microscopy and Langmuir-Blodgett monolayer technique to assess contact lens deposits and human meibum extracts.
The purpose of this exploratory study was to investigate the differences in meibomian gland secretions, contact lens (CL) lipid extracts, and CL surface topography between participants with and without meibomian gland dysfunction (MGD). Meibum study: Meibum was collected from all participants and studied via Langmuir-Blodgett (LB) deposition with subsequent Atomic Force Microscopy (AFM) visualization and surface roughness analysis. CL Study: Participants with and without MGD wore both etafilcon A and balafilcon A CLs in two different phases. CL lipid deposits were extracted and analyzed using pressure-area isotherms with the LB trough and CL surface topographies and roughness values were visualized using AFM. Meibum study: Non-MGD participant meibum samples showed larger, circular aggregates with lower surface roughness, whereas meibum samples from participants with MGD showed more lipid aggregates, greater size variability and higher surface roughness. CL Study: Worn CLs from participants with MGD had a few large tear film deposits with lower surface roughness, whereas non-MGD participant-worn lenses had many small lens deposits with higher surface roughness. Balafilcon A pore depths were shallower in MGD participant worn lenses when compared to non-MGD participant lenses. Isotherms of CL lipid extracts from MGD and non-MGD participants showed a seamless rise in surface pressure as area decreased; however, extracts from the two different lens materials produced different isotherms. MGD and non-MGD participant-worn CL deposition were found to differ in type, amount, and pattern of lens deposits. Lipids from MGD participants deposited irregularly whereas lipids from non-MGD participants showed more uniformity.


Climate Change Vulnerability Assessment in Mangrove-Dependent Communities of Manoka Island, Littoral Region of Cameroon.
This study was conducted on Manoka Island (Littoral Region of Cameroon) with the aim of analyzing climate change vulnerability and local adaptation strategies based on the local community's perceptions and biophysical evidence. We used household surveys, focus group discussions, field observation, GIS, and remote sensing to collect data on variables of exposure, sensitivity, and adaptive capacity. Historical changes in rainfall and temperature, mangrove cover, and the occurrence of extreme climatic events were used as indicators of exposure. Property losses and income structure were used as indicators of sensitivity, while human, natural, social, financial, and physical assets represented adaptive capacity. 89 households were interviewed in the nine settlements of the island. Results show that Manoka Island is experiencing irregular rainfall patterns (with average annual values deviating from the mean by -1.9 to +1.8 mm) and increasing temperature (with annual values deviating from the mean by -1.2 to +3.12). The dynamics of the coastline between 1975 and 2017 using EPR show average setbacks of more than ±3 m/year, with erosion levels varying depending on the period and location. The number of households perceiving extreme climatic events like seasonal variability, flood, and rain storm was higher. From respondents' perception, housing and health are the sectors most affected by climate change. The reported high dependence of households on fishing for income, their overall low livelihood diversification, and their poor access to climate information reported by 65% of respondents portray their poor adaptive capacity. Local response initiatives are ineffective and include among others constructing buildings on stilts and using car wheels to counter the advancement of seawater inland. The study concludes that households on Manoka Island are vulnerable to the impacts of climate change. Income diversification, mangrove reforestation, the development of sustainable supply chains for wood fuel, and sustainable fish smoking devices are the main pathways for adaptation planning in this area.


30 kW concentrator photovoltaic system using dome-shaped Fresnel lenses.
A 30 kW concentrator photovoltaic power plant was constructed and has started operation with the following new technologies: A new Concentrating PhotoVoltaic (CPV) tracker developed for high wind area like Korea and Japan by Daido Steel. (The power consumption of the tracking motors was only 19.6 W, namely 0.07% of the rated power.) With improved optics that reduce the mismatch losses associated with optical aberrations, an efficiency of 25.8% was achieved under standard testing conditions (STC) even in a large 23.8 m2 array size. A rapid installation sequence was developed. It was designed for long-term power supply to a local sewage center. Peak power corresponds to 10% of the demand. As a result, the system performance ratio was 0.87, and the capacity factor was 11.7%. The energy generation per rated power was 1,020 kWh/kWp. While it is true that CPV systems perform better in dry and high irradiance areas, our 30 kW system installed in a cloudy area like Japan, showed satisfactory performance.


Control of stem cell fate and function by engineering physical microenvironments.
The phenotypic expression and function of stem cells are regulated by their integrated response to variable microenvironmental cues, including growth factors and cytokines, matrix-mediated signals, and cell–cell interactions. Recently, growing evidence suggests that matrix-mediated signals include mechanical stimuli such as strain, shear stress, substrate rigidity and topography, and these stimuli have a more profound impact on stem cell phenotypes than had previously been recognized, e.g. self-renewal and differentiation through the control of gene transcription and signaling pathways. Using a variety of cell culture models enabled by micro and nanoscale technologies, we are beginning to systematically and quantitatively investigate the integrated response of cells to combinations of relevant mechanobiological stimuli. This paper reviews recent advances in engineering physical stimuli for stem cell mechanobiology and discusses how micro- and nanoscale engineered platforms can be used to control stem cell niche environments and regulate stem cell fate and function.


An automated transition state search using classical trajectories initialized at multiple minima.
Very recently, we proposed an automated method for finding transition states of chemical reactions using dynamics simulations; the method has been termed Transition State Search using Chemical Dynamics Simulations (TSSCDS) (E. Martínez-Núñez, J. Comput. Chem., 2015, 36, 222-234). In the present work, an improved automated search procedure is developed, which consists of iteratively running different ensembles of trajectories initialized at different minima. The iterative TSSCDS method is applied to the complex C3H4O system, obtaining a total of 66 different minima and 276 transition states. With the obtained transition states and paths, statistical RRKM calculations and Kinetic Monte Carlo simulations are carried out to study the fragmentation dynamics of propenal, which is the global minimum of the system. The kinetic simulations provide a (three-body dissociation)/(CO elimination) ratio of 1.49 for an excitation energy of 148 kcal mol(-1), which agrees well with the corresponding value obtained in the photolysis of propenal at 193 nm (1.1), suggesting that at least these two channels: three-body dissociation (to give H2 + CO + C2H2) and CO elimination occur on the ground electronic state.


Direct observation of electron confinement in epitaxial graphene nanoislands.
One leading question for the application of graphene in nanoelectronics is how electronic properties depend on the size at the nanoscale. Direct observation of the quantized electronic states is central to conveying the relationship between electronic structures and local geometry. Scanning tunneling spectroscopy was used to measure differential conductance dI/dV patterns of nanometer-size graphene islands on an Ir(111) surface. Energy-resolved dI/dV maps clearly show a spatial modulation, indicating a modulated local density of states due to quantum confinement, which is unaffected by the edge configuration. We establish the energy dispersion relation with the quantized electron wave vector obtained from a Fourier analysis of dI/dV maps. The nanoislands preserve the Dirac Fermion properties with a reduced Fermi velocity.


Expression of the ferrioxamine receptor gene of Erwinia amylovora CFBP 1430 during pathogenesis.
Mutants of Erwinia amylovora CFBP 1430 lacking a functional high-affinity iron transport system mediated by desferrioxamine are impaired in their ability to initiate fire blight symptoms (A. Dellagi, M.-N. Brisset, J.-P. Paulin, and D. Expert. Mol. Plant-Microbe Interact. 11:734-742, 1998). In this study, a chromosomal transcriptional lacZ fusion was used to analyze the expression in planta of the E. amylovora ferrioxamine receptor gene foxR. LacZ activity produced by the strain harboring the fusion was highly induced in iron-restricted conditions and in inoculated apple leaf tissues. Microscopic observation revealed differential expression of this gene in relation to the localization and density of bacterial cells within the diseased tissue. Thus, the ability of bacterial cells to express their iron transport system in accordance with environmental conditions is likely important for disease evolution.


Cellular differentiation in three-dimensional lung cell cultures.
Aspects of human biology that are not sufficiently addressed by current cell culture models are cellular differentiation and three-dimensional (3-D) structural organization. A model that more closely associates the presence and biology of organelles to molecular expressions relevant to these organelles may provide evidence of cellular differentiation and the beginning steps in the construction of a 3-D architecture. The development of a new model--3-D cell cultures--may ultimately provide a better understanding of lung biology and pathobiology. The purpose of this study was to develop both traditional monolayer and 3-D cell cultures of a known and well documented normal lung cell line and then to determine similarities and differences between these cultures in terms of differentiation and molecular marker expression. Electron microscopy identified presence of lipid inclusion, microvilli, extra cellular matrix, and tight junctions in the 3-D cultures; differentiation not seen in ML cultures. The degree of differentiation determined by immunohistochemistry when the cell line was grown as ML or 3-D cultures shows that ultra-structure and marker expressions were more representative of control tissue than when cells were grown in 3-D than as MLs. An immortalized cell line was grown as a traditional monolayer (ML) in culture flasks and as 3-D cultures in rotating walled vessels and incubated under identical conditions. Comparison for presence of differentiation and marker expression between these cultures and control tissue collected from surgical patient specimens was studied. Electron microscopy for identification of ultra structures, and immunohistochemistry (ZO-1, EMA, ICAM-1, villin, tubulin, CK 18, VWF, Collagen IV and human mucin) for phenotypic comparisons between cells in ML and 3-D cell cultures was conducted. The development of 3-D cell cultures will provide for a new and more powerful tool in the study of lung biology and pathobiology.


Imaging and acetylcholinesterase inhibitor response in dementia with Lewy bodies.
Acetylcholinesterase inhibitors are commonly used to treat patients with dementia with Lewy bodies. Hippocampal atrophy on magnetic resonance imaging and amyloid-β load on positron emission tomography are associated with the Alzheimer's disease-related pathology in patients with dementia with Lewy bodies. To date, few studies have investigated imaging markers that predict treatment response in patients with dementia with Lewy bodies. Our objective was to determine whether imaging markers of Alzheimer's disease-related pathology such as hippocampal volume, brain amyloid-β load on (11)C Pittsburgh compound B positron emission tomography predict treatment response to acetylcholinesterase inhibitors in patients with dementia with Lewy bodies. We performed a retrospective analysis on consecutive treatment-naive patients with dementia with Lewy bodies (n = 54) from the Mayo Clinic Alzheimer's Disease Research Centre who subsequently received acetylcholinesterase inhibitors and underwent magnetic resonance imaging with hippocampal volumetry. Baseline and follow-up assessments were obtained with the Mattis Dementia Rating Scale. Subjects were divided into three groups (reliable improvement, stable or reliable decline) using Dementia Rating Scale reliable change indices determined previously. Associations between hippocampal volumes and treatment response were tested with analysis of covariance adjusting for baseline Dementia Rating Scale, age, gender, magnetic resonance field strength and Dementia Rating Scale interval. Seven subjects underwent (11)C Pittsburgh compound B imaging within 12 weeks of magnetic resonance imaging. Global cortical (11)C Pittsburgh compound B retention (scaled to cerebellar retention) was calculated in these patients. Using a conservative psychometric method of assessing treatment response, there were 12 patients with reliable decline, 29 stable cases and 13 patients with reliable improvement. The improvers had significantly larger hippocampi than those that declined (P = 0.02) and the stable (P = 0.04) group. An exploratory analysis demonstrated larger grey matter volumes in the temporal and parietal lobes in improvers compared with those who declined (P < 0.05). The two patients who had a positive (11)C Pittsburgh compound B positron emission tomography scan declined and those who had a negative (11)C Pittsburgh compound B positron emission tomography scan improved or were stable after treatment. Patients with dementia with Lewy bodies who do not have the imaging features of coexistent Alzheimer's disease-related pathology are more likely to cognitively improve with acetylcholinesterase inhibitor treatment.


Relation of triglyceride levels, fasting and nonfasting, to fatal and nonfatal coronary heart disease.
It remains unclear whether hypertriglyceridemia is an independent risk factor for coronary heart disease (CHD), and whether fasting and nonfasting triglyceride (TG) levels are equally predictive. A total of 2809 (of 12 866) men randomized during 1973 through 1975 into the Multiple Risk Factor Intervention Trial with fasting and nonfasting TG levels measured at baseline were followed up for CHD incidence and death. Proportional hazards regression models were used to assess associations of fasting and nonfasting TG levels with CHD. Average fasting and nonfasting TG levels were 187 and 284 mg/dL (2.11 and 3.21 mmol/L), respectively. Prevalence of hypertriglyceridemia (200 mg/dL [2.26 mmol/L] or more) was 31% for fasting and 61% for nonfasting. There were 175 nonfatal or fatal CHD events during 8 years and 328 CHD deaths during 25 years. Compared with TG levels less than 200 mg/dL, risk factor-adjusted hazard ratios for CHD mortality for hypertriglyceridemia were 1.24 (P =.09) for fasting and 1.26 (P =.07) for nonfasting. For nonfatal or fatal CHD, fasting and nonfasting TG levels were similarly predictive with hazard ratios of 1.64 (P =.004) for fasting and 1.46 (P =.03) for nonfasting. These associations for fasting TG levels were assessed to be underestimated by 56% because of regression dilution bias, with attenuation likely greater for nonfasting TG levels. Greater ease of obtaining nonfasting than fasting measurements, greater prevalence of hypertriglyceridemia with nonfasting than fasting values, and similarly increased risk with each indicate that nonfasting TG levels may be more useful than fasting ones for risk stratification.


Cell surface glycoprotein PZR is a major mediator of concanavalin A-induced cell signaling.
PZR is an immunoglobulin superfamily cell surface protein containing a pair of immunoreceptor tyrosine-based inhibitory motifs. As a glycoprotein, PZR displays a strong association with concanavalin A (ConA), a member of the plant lectin family. Treatment of several cell lines with ConA caused tyrosine phosphorylation of a major cellular protein. Immunoblotting and immunoprecipitation studies indicated that this protein corresponded to PZR. Tyrosine phosphorylation of PZR was accompanied by recruitment of SHP-2 and was inhibited by PP1, a selective inhibitor of the Src family tyrosine kinases. Furthermore, c-Src was constitutively associated with PZR and was activated upon treatment of cells with ConA. Moreover, tyrosine phosphorylation of PZR was markedly enhanced in v-Src-transformed NIH-3T3 cells and was predominant in Escherichia coli cells co-expressing c-Src. Expression of an intracellular domain-truncated form of PZR in HT-1080 cells affected cell morphology and had a dominant negative effect on ConA-induced tyrosine phosphorylation of PZR, activation of c-Src, and agglutination of the cells. Together, the data indicate that PZR is a major receptor of ConA and has an important role in cell signaling via c-Src. Considering the various biological activities of ConA, the study of PZR may have major therapeutic implications.


Contact varroacidal efficacy of lithium citrate and its influence on viral loads, immune parameters and oxidative stress of honey bees in a field experiment.
With an almost global distribution, Varroa destuctor is the leading cause of weakening and loss of honey bee colonies. New substances are constantly being tested in order to find those that will exhibit high anti-Varroa efficacy at low doses/concentrations, without unwanted effects on bees. Lithium (Li) salts stood out as candidates based on previous research. The aims of this study were to evaluate Li citrate hydrate (Li-cit) for its contact efficacy against Varroa, but also the effect of Li-cit on honey bees by estimating loads of honey bee viruses, expression levels of immune-related genes and genes for antioxidative enzymes and oxidative stress parameters on two sampling occasions, before the treatment and after the treatment. Our experiment was performed on four groups, each consisting of seven colonies. Two groups were treated with the test compound, one receiving 5 mM and the other 10 mM of Li-cit; the third received oxalic acid treatment (OA group) and served as positive control, and the fourth was negative control (C group), treated with 50% w/v pure sucrose-water syrup. Single trickling treatment was applied in all groups. Both tested concentrations of Li-cit, 5 and 10 mM, expressed high varroacidal efficacy, 96.85% and 96.80%, respectively. Load of Chronic Bee Paralysis Virus significantly decreased (p < 0.01) after the treatment in group treated with 5 mM of Li-cit. In OA group, loads of Acute Bee Paralysis Virus and Deformed Wing Virus significantly (p < 0.05) increased, and in C group, loads of all viruses significantly (p < 0.01 or p < 0.001) increased. Transcript levels of genes for abaecin, apidaecin, defensin and vitellogenin were significantly higher (p < 0.05-p < 0.001), while all oxidative stress parameters were significantly lower (p < 0.05-p < 0.001) after the treatment in both groups treated with Li-cit. All presented results along with easy application indicate benefits of topical Li-cit treatment and complete the mosaic of evidence on the advantages of this salt in the control of Varroa.


Comparison of Physical Activity Between Children With and Without Autism Spectrum Disorder: A Systematic Review and Meta-Analysis.
Lower levels of physical activity (PA) are often observed among children with autism spectrum disorder (ASD) compared to children without ASD; however, some studies have demonstrated few to no PA differences between the two groups. The purpose of this systematic review with meta-analysis was to compare the differences in PA between children (2-18 years) with and without ASD. An exhaustive search of five online databases was completed, and 31 studies met the inclusion criteria. A pooled random-effects Hedges's g model was used to determine differences in PA between children with and without ASD. Children with ASD were found to be significantly less physically active than children without ASD (Δ = -0.62, p < .001). Subgroup analyses revealed significant moderate to large differences in PA by intensity level, age, setting, and measurement methods. Future studies are needed to further explore the underlying mechanisms associated with lower levels of PA among children with ASD.


NF-E2 overexpression delays erythroid maturation and increases erythrocyte production.
The transcription factor Nuclear Factor-Erythroid 2 (NF-E2) is overexpressed in the vast majority of patients with polycythaemia vera (PV). In murine models, NF-E2 overexpression increases proliferation and promotes cellular viability in the absence of erythropoietin (EPO). EPO-independent growth is a hallmark of PV. We therefore hypothesized that NF-E2 overexpression contributes to erythrocytosis, the pathognomonic feature of PV. Consequently, we investigated the effect of NF-E2 overexpression in healthy CD34+ cells. NF-E2 overexpression led to a delay in erythroid maturation, manifested by a belated appearance of glycophorin A-positive erythroid precursors. Maturation delay was similarly observed in primary PV patient erythroid cultures compared to healthy controls. Protracted maturation led to a significant increase in the accumulated number of erythroid cells both in PV cultures and in CD34+ cells overexpressing NF-E2. Similarly, NF-E2 overexpression altered erythroid colony formation, leading to an increase in erythroid burst-forming unit formation. These data indicate that NF-E2 overexpression delays the early phase of erythroid maturation, resulting in an expansion of erythroid progenitors, thereby increasing the number of erythrocytes derived from one CD34+ cell. These data propose a role for NF-E2 in mediating the erythrocytosis of PV.


Optical systems for point-of-care diagnostic instrumentation: analysis of imaging performance and cost.
One of the key elements in point-of-care (POC) diagnostic test instrumentation is the optical system required for signal detection and/or imaging. Many tests which use fluorescence, absorbance, or colorimetric optical signals are under development for management of infectious diseases in resource limited settings, where the overall size and cost of the device is of critical importance. At present, high-performance lenses are expensive to fabricate and difficult to obtain commercially, presenting barriers for developers of in vitro POC tests or microscopic image-based diagnostics. We recently described a compact "hybrid" objective lens incorporating both glass and plastic optical elements, with a numerical aperture of 1.0 and field-of-view of 250 μm. This design concept may potentially enable mass-production of high-performance, low-cost optical systems which can be easily incorporated in the readout path of existing and emerging POC diagnostic assays. In this paper, we evaluate the biological imaging performance of these lens systems in three broad POC diagnostic application areas; (1) bright field microscopy of histopathology slides, (2) cytologic examination of blood smears, and (3) immunofluorescence imaging. We also break down the fabrication costs and draw comparisons with other miniature optical systems. The hybrid lenses provided images with quality comparable to conventional microscopy, enabling examination of neoplastic pathology and infectious parasites including malaria and cryptosporidium. We describe how these components can be produced at below $10 per unit in full-scale production quantities, making these systems well suited for use within POC diagnostic instrumentation.


The search for a unique Raman signature of amyloid-beta plaques in human brain tissue from Alzheimer's disease patients.
Definite Alzheimer's disease (AD) diagnosis is commonly done on ex vivo brain tissue using immuno-histochemical staining to visualize amyloid-beta (Aβ) aggregates, also known as Aβ plaques. Raman spectroscopy has shown its potential for non-invasive and label-free determination of bio-molecular compositions, aiding the post-mortem diagnosis of pathological tissue. Here, we investigated whether conventional Raman spectroscopy could be used for the detection of amyloid beta deposits in fixed, ex vivo human brain tissue, taken from the frontal cortex region. We examined the spectra and spectral maps of three severe AD cases and two healthy control cases and compared their spectral outcome among each other as well as to recent results in the literature obtained with various spectroscopic techniques. After hyperspectral Raman mapping, Aβ plaques were visualized using Thioflavin-S staining on the exact same tissue sections. As a result, we show that tiny diffuse or tangled-like morphological structures, visible under microscopic conditions on unstained tissue and often but erroneously assumed to be deposits of Aβ, are instead usually an aggregation of highly auto-fluorescent lipofuscin granulates without any, or limited, plaque or plaque-like association. The occurrence of these auto-fluorescent particles is equally distributed in both AD and healthy control cases. Therefore, they cannot be used as possible criteria for Alzheimer's disease diagnosis. Furthermore, a unique plaque-specific/Aβ spectrum could not be determined even after possible spectral interferences were carefully removed.


Clinically Relevant Drug-Drug Interactions in Primary Care.
Drug interactions are common in the primary care setting and are usually predictable. Identifying the most important and clinically relevant drug interactions in primary care is essential to patient safety. Strategies for reducing the risk of drug-drug interactions include minimizing the number of drugs prescribed, re-evaluating therapy on a regular basis, considering nonpharmacologic options, monitoring for signs and symptoms of toxicity or effectiveness, adjusting dosages of medications when indicated, and adjusting administration times. Inhibition or induction of cytochrome P450 drug metabolizing isoenzymes is the most common mechanism by which clinically important drug interactions occur. The antimicrobials most likely to affect the international normalized ratio significantly in patients receiving warfarin are trimethoprim/sulfamethoxazole, metronidazole, and fluconazole. An empiric warfarin dosage reduction of 30% to 50% upon initiation of amiodarone therapy is recommended. In patients receiving amiodarone, limit dosages of simvastatin to 20 mg per day and lovastatin to 40 mg per day. Beta blockers should be tapered and discontinued several days before clonidine withdrawal to reduce the risk of rebound hypertension. Spironolactone dosages should be limited to 25 mg daily when coadministered with potassium supplements. Avoid prescribing opioid cough medicines for patients receiving benzodiazepines or other central nervous system depressants, including alcohol. Physicians should consider consultation with a clinical pharmacist when clinical circumstances require the use of drugs with interaction potential.


Requirements of Hsp104p activity and Sis1p binding for propagation of the [RNQ(+)] prion.
The formation and maintenance of prions in the yeast Saccharomyces cerevisiae is highly regulated by the cellular chaperone machinery. The most important player in this regulation is Hsp104p, which is required for the maintenance of all known prions. The requirements for other chaperones, such as members of the Hsp40 or Hsp70 families, vary with each individual prion. [RNQ(+)] cells do not have a phenotype that is amenable to genetic screens to identify cellular factors important in prion propagation. Therefore, we used a chimeric construct that reports the [RNQ(+)] status of cells to perform a screen for mutants that are unable to maintain [RNQ(+)]. We found eight separate mutations in Hsp104p that caused [RNQ(+)] cells to become [rnq(-)]. These mutations also caused the loss of the [PSI(+)] prion. The expression of one of these mutants, Hsp104p-E190K, showed differential loss of the [RNQ(+)] and [PSI(+)] prions in the presence of wild type Hsp104p. Hsp104p-E190K inefficiently propagated [RNQ(+)] and was unable to maintain [PSI(+)]. The mutant was unable to act on other in vivo substrates, as strains carrying it were not thermotolerant. Purified recombinant Hsp104p-E190K showed a reduced level of ATP hydrolysis as compared to wild type protein. This is likely the cause of both prion loss and lack of in vivo function. Furthermore, it suggests that [RNQ(+)] requires less Hsp104p activity to maintain transmissible protein aggregates than Sup35p. Additionally, we show that the L94A mutation in Rnq1p, which reduces its interaction with Sis1p, prevents Rnq1p from maintaining a prion and inducing [PSI(+)].


Transjugular Intrahepatic Portosystemic Shunt Combined with Haemoperfusion in an End-stage Renal Disease Patient with Liver Cirrhosis-related Refractory Portal Hypertensive Variceal Bleeding: A Case Report.
Transjugular intrahepatic portosystemic shunt (TIPS) is recommended as the second-line option for variceal bleeding in liver cirrhosis patients when the bleeding is not well controlled by medical and/or endoscopic therapy. However, due to a high incidence of post-TIPS hepatic encephalopathy, particularly in patients with end-stage renal disease (ESRD), TIPS is rarely used in such cases. We report a case involving the successful and safe use of TIPS combined with haemoperfusion to treat an ESRD patient with liver cirrhosis-related refractory portal hypertensive variceal bleeding. Our case suggests the possibility of inserting TIPS to stop recurrent variceal bleeding in ESRD patients with liver cirrhosis. Haemoperfusion may be used to reduce the risk of post-TIPS hepatic encephalopathy.


Radioimmunotherapy for Brain Metastases: The Potential for Inflammation as a Target of Choice.
Brain metastases (BM) are frequently detected during the follow-up of patients with malignant tumors, particularly in those with advanced disease. Despite a major progress in systemic anti-cancer treatments, the average overall survival of these patients remains limited (6 months from diagnosis). Also, cognitive decline is regularly reported especially in patients treated with whole brain external beam radiotherapy (WBRT), due to the absorbed radiation dose in healthy brain tissue. New targeted therapies, for an earlier and/or more specific treatment of the tumor and its microenvironment, are needed. Radioimmunotherapy (RIT), a combination of a radionuclide to a specific antibody, appears to be a promising tool. Inflammation, which is involved in multiple steps, including the early phase, of BM development is attractive as a relevant target for RIT. This review will focus on the (1) early biomarkers of inflammation in BM pertinent for RIT, (2) state of the art studies on RIT for BM, and (3) the importance of dosimetry to RIT in BM. These two last points will be addressed in comparison to the conventional EBRT treatment, particularly with respect to the balance between tumor control and healthy tissue complications. Finally, because new diagnostic imaging techniques show a potential for the detection of BM at an early stage of the disease, we focus particularly on this therapeutic window.


Protein Nanosheet Mechanics Controls Cell Adhesion and Expansion on Low-Viscosity Liquids.
Adherent cell culture typically requires cell spreading at the surface of solid substrates to sustain the formation of stable focal adhesions and assembly of a contractile cytoskeleton. However, a few reports have demonstrated that cell culture is possible on liquid substrates such as silicone and fluorinated oils, even displaying very low viscosities (0.77 cSt). Such behavior is surprising as low viscosity liquids are thought to relax much too fast (<ms) to enable the stabilization of focal adhesions (with lifetimes on the order of minutes to hours). Here we show that cell spreading and proliferation at the surface of low viscosity liquids are enabled by the self-assembly of mechanically strong protein nanosheets at these interfaces. We propose that this phenomenon results from the denaturation of globular proteins, such as albumin, in combination with the coupling of surfactant molecules to the resulting protein nanosheets. We use interfacial rheology and atomic force microscopy indentation to characterize the mechanical properties of protein nanosheets and associated liquid-liquid interfaces. We identify a direct relationship between interfacial mechanics and the association of surfactant molecules with proteins and polymers assembled at liquid-liquid interfaces. In addition, our data indicate that cells primarily sense in-plane mechanical properties of interfaces, rather than relying on surface tension to sustain spreading, as in the spreading of water striders. These findings demonstrate that bulk and nanoscale mechanical properties may be designed independently, to provide structure and regulate cell phenotype, therefore calling for a paradigm shift for the design of biomaterials in regenerative medicine.


Cone responses in Usher syndrome types 1 and 2 by microvolt electroretinography.
Progressive decline of psychophysical cone-mediated measures has been reported in type 1 (USH1) and type 2 (USH2) Usher syndrome. Conventional cone electroretinogram (ERG) responses in USH demonstrate poor signal-to-noise ratio. We evaluated cone signals in USH1 and USH2 by recording microvolt level cycle-by-cycle (CxC) ERG. Responses of molecularly genotyped USH1 (n = 18) and USH2 (n = 24) subjects (age range, 15-69 years) were compared with those of controls (n = 12). A subset of USH1 (n = 9) and USH2 (n = 9) subjects was examined two to four times over 2 to 8 years. Photopic CxC ERG and conventional 30-Hz flicker ERG were recorded on the same visits. Usher syndrome subjects showed considerable cone flicker ERG amplitude losses and timing phase delays (P < 0.01) compared with controls. USH1 and USH2 had similar rates of progressive logarithmic ERG amplitude decline with disease duration (-0.012 log μV/y). Of interest, ERG phase delays did not progress over time. Two USH1C subjects retained normal response timing despite reduced amplitudes. The CxC ERG method provided reliable responses in all subjects, whereas conventional ERG was undetectable in 7 of 42 subjects. Cycle-by-cycle ERG showed progressive loss of amplitude in both USH1 and USH2 subjects, comparable to that reported with psychophysical measures. Usher subjects showed abnormal ERG response latency, but this changed less than amplitude with time. In USH syndrome, CxC ERG is more sensitive than conventional ERG and warrants consideration as an outcome measure in USH treatment trials.


Evaluation of the Vibe Actigraph in Patients With Chronic Obstructive Pulmonary Disease: A Pilot Study.
To validate the Vibe actigraph in assessing sleep-wake patterns compared to polysomnography (PSG) in patients with COPD. Nine stable COPD patients wore actigraphs while undergoing PSG. The correlation between total sleep time (TST), total sleep period (TSP), sleep onset latency (SOL), wake after sleep onset (WASO), and sleep efficiency was determined for corresponding measurements from the actigraph and PSG. Sensitivity, specificity, and positive and negative predictive values were calculated for the actigraph, considering PSG the gold standard. Levels of agreement between the variables of PSG and the actigraph were estimated using Bland-Altman plots. A strong and statistically significant correlation was noted between PSG and the actigraph in detecting movement during sleep [mean activity score (counts)], TST and TSP (all rs = 0.83; p = 0.005). The median agreement of sleep and wake counts between PSG and the actigraph was 73% and the Cohen's Kappa value was 0.66. The medians of sensitivity and specificity of the actigraph for detecting sleep versus PSG were 84 and 66%, respectively. The median positive and negative predictive values of the actigraph were 74 and 72%, respectively. This study demonstrated that, under controlled laboratory conditions, the Vibe actigraph in its default settings is a promising tool for the detection of sleep-wake parameters in a small number of ambulatory patients with COPD. Clinical Impact: The actigraph used in this pilot study suggests that these devices could provide clinically relevant information in COPD to better understand the relationship between sleep and health in this population.


Transposon-modified antigen-specific T lymphocytes for sustained therapeutic protein delivery in vivo.
A cell therapy platform permitting long-term delivery of peptide hormones in vivo would be a significant advance for patients with hormonal deficiencies. Here we report the utility of antigen-specific T lymphocytes as a regulatable peptide delivery platform for in vivo therapy. piggyBac transposon modification of murine cells with luciferase allows us to visualize T cells after adoptive transfer. Vaccination stimulates long-term T-cell engraftment, persistence, and transgene expression enabling detection of modified cells up to 300 days after adoptive transfer. We demonstrate adoptive transfer of antigen-specific T cells expressing erythropoietin (EPO) elevating the hematocrit in mice for more than 20 weeks. We extend our observations to human T cells demonstrating inducible EPO production from Epstein-Barr virus (EBV) antigen-specific T lymphocytes. Our results reveal antigen-specific T lymphocytes to be an effective delivery platform for therapeutic molecules such as EPO in vivo, with important implications for other diseases that require peptide therapy.


Crystal structure of the processivity clamp loader gamma (gamma) complex of E. coli DNA polymerase III.
The gamma complex, an AAA+ ATPase, is the bacterial homolog of eukaryotic replication factor C (RFC) that loads the sliding clamp (beta, homologous to PCNA) onto DNA. The 2.7/3.0 A crystal structure of gamma complex reveals a pentameric arrangement of subunits, with stoichiometry delta':gamma(3):delta. The C-terminal domains of the subunits form a circular collar that supports an asymmetric arrangement of the N-terminal ATP binding domains of the gamma motor and the structurally related domains of the delta' stator and the delta wrench. The structure suggests a mechanism by which the gamma complex switches between a closed state, in which the beta-interacting element of delta is hidden by delta', and an open form similar to the crystal structure, in which delta is free to bind to beta.


Underreporting of energy intake in repeated 24-hour recalls related to gender, age, weight status, day of interview, educational level, reported food intake, smoking habits and area of living.
The aims of the present study were (1) to evaluate the degree to which underreporting of energy intake by repeated 24-hour recalls was related to gender, age, weight status, day of interview, educational level, smoking habits and area of living, and (2) to compare the dietary characteristics of underreporters with those of others. Cross-sectional study. Ten 24-hour recalls were performed during a one-year period. The Västerbotten intervention programme of cardiovascular disease and diabetes in Northern Sweden. Ninety-four men and 99 women in four age groups: 30, 40, 50 and 60 years. The prevalence of men and women with a food intake level (FIL; reported energy intake divided by estimated basal metabolic rate) below 1.2 was 44% and 47%, respectively. The youngest age group had higher FIL values than the oldest age group for both men (1.5 versus 1.1) and women (1.4 versus 1.1). The prevalence and magnitude of underreporting were directly related to body mass index (BMI; correlation coefficient: -0.47 (men) and -0.55 (women)). Smokers had a lower FIL value (1.1) than non-smokers (1.3). The nutrient density was lower for the group with high FIL values for protein and calcium and higher for fat and sucrose. The upper FIL group often had higher intake frequencies and larger portion sizes than the lower FIL group. Underreporting of energy intake is prevalent when 24-hour recalls are used, but the prevalence differs between sub-groups in the population. BMI was the main predictor of underreporting but also old age and smoking seem to contribute in this aspect. Socially desirable food items were not underreported to the same extent as socially undesirable food items. The intake frequencies and portion sizes partly explained the differences in FIL.


Assessment of global myocardial perfusion reserve using cardiovascular magnetic resonance of coronary sinus flow at 3 Tesla.
Despite increasing clinical use, there is limited data regarding regadenoson in stress perfusion cardiovascular magnetic resonance (CMR). In particular, given its long half-life the optimal stress protocol remains unclear. Although Myocardial Perfusion Reserve (MPR) may provide additive prognostic information, current techniques for its measurement are cumbersome and challenging for routine clinical practice.The aims of this study were: 1) To determine the feasibility of MPR quantification during regadenoson stress CMR by measurement of Coronary Sinus (CS) flow; and 2) to investigate the role of aminophylline reversal during regadenoson stress-CMR. 117 consecutive patients with possible myocardial ischemia were prospectively enrolled. Perfusion imaging was performed at 1 minute and 15 minutes after administration of 0.4 mg regadenoson. A subgroup of 41 patients was given aminophylline (100 mg) after stress images were acquired. CS flow was measured using phase-contrast imaging at baseline (pre CS flow), and immediately after the stress (peak CS flow) and rest (post CS flow) perfusion images. CS flow measurements were obtained in 92% of patients with no adverse events. MPR was significantly underestimated when calculated as peak CS flow/post CS flow as compared to peak CS flow/pre CS flow (2.43±0.20 vs. 3.28±0.32, p=0.03). This difference was abolished when aminophylline was administered (3.35±0.44 vs. 3.30±0.52, p=0.95). Impaired MPR (peak CS flow/pre CS flow<2) was associated with advanced age, diabetes, current smoking and higher Framingham risk score. Regadenoson stress CMR with MPR measurement from CS flow can be successfully performed in most patients. This measurement of MPR appears practical to perform in the clinical setting. Residual hyperemia is still present even 15 minutes after regadenoson administration, at the time of resting-perfusion acquisition, and is completely reversed by aminophylline. Our findings suggest routine aminophylline administration may be required when performing stress CMR with regadenoson.


Nutritional supplement use among college athletes and their sources of information.
A survey was conducted to examine the source of information and usage of nutritional supplements in 115 male and 88 female varsity athletes at a Division I university. The survey asked each athlete to define supplement, and report supplement use and type, source of information, and reasons for use. Supplement use frequencies were determined, and comparisons were made between gender and sport. Eighty-nine percent of the subjects had or were currently using nutritional supplements. Many athletes did not consider sports drinks and calorie replacement products as supplements. Females were more likely to take calcium and multivitamins, and males had significant intake for ginseng, amino acids, glutamine, hydroxy-methyl-buterate (HMB), weight gainers, whey protein, and Juven. The most frequently used supplements overall were energy drinks (73%), calorie replacement products of all types (61.4%), multivitamin (47.3%), creatine (37.2%), and vitamin C (32.4%). There was also significant supplement use noted per sport. Females were more likely to obtain information from family members regarding supplementation, and males from a store nutritionist, fellow athletes, friends, or a coach. Female athletes were more likely to take supplements for their health or because of an inadequate diet, while men reported taking supplements to improve speed and agility, strength and power, or for weight/muscle gain.


Room Temperature Iron-Catalyzed Transfer Hydrogenation and Regioselective Deuteration of Carbon-Carbon Double Bonds.
An iron catalyst has been developed for the transfer hydrogenation of carbon-carbon multiple bonds. Using a well-defined β-diketiminate iron(II) precatalyst, a sacrificial amine and a borane, even simple, unactivated alkenes such as 1-hexene undergo hydrogenation within 1 h at room temperature. Tuning the reagent stoichiometry allows for semi- and complete hydrogenation of terminal alkynes. It is also possible to hydrogenate aminoalkenes and aminoalkynes without poisoning the catalyst through competitive amine ligation. Furthermore, by exploiting the separate protic and hydridic nature of the reagents, it is possible to regioselectively prepare monoisotopically labeled products. DFT calculations define a mechanism for the transfer hydrogenation of propene with nBuNH2 and HBpin that involves the initial formation of an iron(II)-hydride active species, 1,2-insertion of propene, and rate-limiting protonolysis of the resultant alkyl by the amine N-H bond. This mechanism is fully consistent with the selective deuteration studies, although the calculations also highlight alkene hydroboration and amine-borane dehydrocoupling as competitive processes. This was resolved by reassessing the nature of the active transfer hydrogenation agent: experimentally, a gel is observed in catalysis, and calculations suggest this can be formulated as an oligomeric species comprising H-bonded amine-borane adducts. Gel formation serves to reduce the effective concentrations of free HBpin and nBuNH2 and so disfavors both hydroboration and dehydrocoupling while allowing alkene migratory insertion (and hence transfer hydrogenation) to dominate.


Suppression of lung adenocarcinoma progression by Nkx2-1.
Despite the high prevalence and poor outcome of patients with metastatic lung cancer the mechanisms of tumour progression and metastasis remain largely uncharacterized. Here we modelled human lung adenocarcinoma, which frequently harbours activating point mutations in KRAS and inactivation of the p53 pathway, using conditional alleles in mice. Lentiviral-mediated somatic activation of oncogenic Kras and deletion of p53 in the lung epithelial cells of Kras(LSL-G12D/+);p53(flox/flox) mice initiates lung adenocarcinoma development. Although tumours are initiated synchronously by defined genetic alterations, only a subset becomes malignant, indicating that disease progression requires additional alterations. Identification of the lentiviral integration sites allowed us to distinguish metastatic from non-metastatic tumours and determine the gene expression alterations that distinguish these tumour types. Cross-species analysis identified the NK2-related homeobox transcription factor Nkx2-1 (also called Ttf-1 or Titf1) as a candidate suppressor of malignant progression. In this mouse model, Nkx2-1 negativity is pathognomonic of high-grade poorly differentiated tumours. Gain- and loss-of-function experiments in cells derived from metastatic and non-metastatic tumours demonstrated that Nkx2-1 controls tumour differentiation and limits metastatic potential in vivo. Interrogation of Nkx2-1-regulated genes, analysis of tumours at defined developmental stages, and functional complementation experiments indicate that Nkx2-1 constrains tumours in part by repressing the embryonically restricted chromatin regulator Hmga2. Whereas focal amplification of NKX2-1 in a fraction of human lung adenocarcinomas has focused attention on its oncogenic function, our data specifically link Nkx2-1 downregulation to loss of differentiation, enhanced tumour seeding ability and increased metastatic proclivity. Thus, the oncogenic and suppressive functions of Nkx2-1 in the same tumour type substantiate its role as a dual function lineage factor.


Flexural bending of southern Tibet in a retro foreland setting.
The highest elevation of the Tibetan Plateau, lying 5,700 m above sea level, occurs within the part of the Lhasa block immediately north of the India-Tibet suture zone (Yarlung Zangbo suture zone, YZSZ), being 700 m higher than the maximum elevation of more northern parts of the plateau. Various mechanisms have been proposed to explain this differentially higher topography and the rock uplift that led to it, invoking crustal compression or extension. Here we present the results of structural investigations along the length of the high elevation belt and suture zone, which rather indicate flexural bending of the southern margin of the Lhasa block (Gangdese magmatic belt) and occurrence of an adjacent foreland basin (Kailas Basin), both elements resulting from supra-crustal loading of the Lhasa block by the Zangbo Complex (Indian plate rocks) via the Great Counter Thrust. Hence we interpret the differential elevation of the southern margin of the plateau as due originally to uplift of a forebulge in a retro foreland setting modified by subsequent processes. Identification of this flexural deformation has implications for early evolution of the India-Tibet continental collision zone, implying an initial (Late Oligocene) symmetrical architecture that subsequently transitioned into the present asymmetrical wedge architecture.


Integrins and the activation of latent transforming growth factor beta1 - an intimate relationship.
Integrins are crucial for the ability of cells to sense mechanical perturbations and to transmit intracellular stress to their environment. We here review the more recently discovered role of integrins in activating the pleiotrophic cytokine transforming growth factor beta 1 (TGF-beta1). TGF-beta1 controls tissue homeostasis in embryonic and normal adult tissues and contributes to the development of fibrosis, cancer, autoimmune and vascular diseases when being mis-regulated. In most of these conditions, active TGF-beta1 is generated by dissociation from a large latent protein complex that sequesters latent TGF-beta1 in the extracellular matrix (ECM). Two main models are proposed how integrins contribute to latent TGF-beta1 activation: (1) In a protease-dependent mechanism, integrins alphavbeta8 and alphavbeta3 are suggested to simultaneously bind the latent TGF-beta1 complex and proteinases. This close vicinity at the cell surface improves enzymatic cleavage of the latent complex to release active TGF-beta1. (2) Integrins alphavbeta3, alphavbeta5, alphavbeta6, and alphavbeta8 appear to change the conformation of the latent TGF-beta1 complex by transmitting cell traction forces. This action requires association of the latent complex with a mechanically resistant ECM and is independent from proteolysis. Understanding that different integrins use different mechanisms to activate latent TGF-beta1 opens new possibilities to develop cell-specific therapeutic strategies for TGF-beta1-induced pathologies.


A gastrocnemius heterotopical transplant model with end-to-side neurorraphy.
To present an animal model to assess the effects of end-to-side innervation in the heterotopically transplanted model with reduced chances of neural contamination. The medial portion of the gastrocnemius muscle in wistar male rats was isolated and its pedicle dissected and performed a flap in the abdominal portion. To prevent neural contamination in the abdominal region, the muscle was wrapped with a Goretex(r) sheet. The specimens were divided into 2 groups (G). In G1 was performed an end-to-end suture between tibial nerve of the gastrocnemius and femoral motor nerve and between the saphenous sensory nerve and the motor nerve. In G2 was performed a end-to-side suture between the tibial nerve and the motor femoral and between the tibial nerve and saphenous motor nerve. The specimens were evaluated 60 days later to check the structure of the neurorraphy. Sections were obtained proximal and distal to the coaptation site. The medial gastrocnemius muscle had the advantage of maintaining visible mass after 60 days. No disruption of the coaptation site was found. No major injury to the donor nerve was seen in group 2. The proposed model is simple, reproduciple and prevent the neural contamination in the flap in end-to-side suture.


Thoracolumbar And Lumbopelvic Spinal Alignment During The Deadlift Exercise: A Comparison Between Men And Women.
A neutral spinal alignment is considered important during the execution of the deadlift exercise to decrease the risk of injury. Since male and female powerlifters experience pain in different parts of their backs, it is important to examine whether men and women differ in spinal alignment during the deadlift. The purpose of this study was to quantify the spinal alignment in the upper (thoracolumbar, T11-L2) and lower (lumbopelvic, L2-S2) lumbar spine during the deadlift exercise in male and female lifters. Secondary aims were to compare lumbar spine alignment during the deadlift to standing habitual posture, and determine whether male and female lifters differ in these aspects. Observational, Cross-sectional. Twenty-four (14 men, 10 women) lifters performed three repetitions of the deadlift exercise using 70% of their respective one-repetition maximum. Spinal alignment and spinal range of motion were measured using three inertial measurement units placed on the thoracic, lumbar and sacral spine. Data from three different positions were analyzed; habitual posture in standing, and start and stop positions of the deadlift, i.e. bottom and finish position respectively. During the deadlift, spinal adjustments were evident in all three planes of movement. From standing habitual posture to the start position the lumbar lordosis decreased 13° in the upper and 20° in the lower lumbar spine. From start position to stop position the total range of motion in the sagittal plane was 11° in the upper and 22° in the lower lumbar spine. The decreased lumbar lordosis from standing habitual posture to the start position was significantly greater among men. Men and women adjust their spinal alignment in all three planes of movement when performing a deadlift and men seem to make greater adjustments from their standing habitual posture to start position in the sagittal plane. 3.


Modulation-free optical locking of an external-cavity diode laser to a filter cavity.
Optical locking to a filter cavity is an effective method to eliminate the limitations of an external-cavity diode laser, such as broad spontaneous emission backgrounds and frequency jitters. Stable operation of the optical locking requires simultaneous control of the feedback phase and the diode-laser frequency. Frequency dither is usually used to extract the two error signals, but this causes extra frequency modulation in the output beam. A modulation-free method for deriving the error signals by modulating the laser-cavity coupling strength is demonstrated with a violet diode laser. A modulation-free linewidth upper limit of about 7 kHz for a 1 s measurement is realized by the method.


Functional Electrical Stimulation Leads to Increased Volume of the Aged Thyroarytenoid Muscle.
To reverse sarcopenia and increase the volumes of atrophied laryngeal muscles by functional electrical stimulation (FES) using a minimal invasive surgical procedure in an aged ovine model. Prospective animal study. A stimulation electrode was placed unilaterally near the terminal adduction branch of the recurrent laryngeal nerve (RLN) adjacent to the right cricothyroid joint. The electrode was connected to an implant located subcutaneously at the neck region. Predesigned training patterns were automatically delivered by a bidirectional radio frequency link using a programming device and were repeated automatically by the implant every other day over 11 weeks in the awake animal. Outcome parameters comprised volumetric measurements based on three-dimensional reconstructions of the entire thyroarytenoid muscle (TAM), as well as gene expression analyses. We found significant increases of the volumes of the stimulated TAM of 11% and the TAM diameter at the midmembranous parts of the vocal folds of nearly 40%. Based on gene expression, we did not detect a shift of muscle fiber composition. FES of the terminal branches of the RLN is a secure and effective way to reverse the effects of age-related TAM atrophy and to increase volumes of atrophied muscles. NA Laryngoscope, 128:2852-2857, 2018.


Stepwise increases in FT1 expression regulate seasonal progression of flowering in wheat (Triticum aestivum).
Flowering is regulated by genes that respond to changing daylengths and temperature, which have been well studied using controlled conditions; however, the molecular processes underpinning flowering in nature remain poorly understood. Here, we investigate the genetic pathways that coordinate flowering and inflorescence development of wheat (Triticum aestivum) as daylengths extend naturally in the field, using lines that contain variant alleles for the key photoperiod gene, Photoperiod-1 (Ppd-1). We found flowering involves a stepwise increase in the expression of FLOWERING LOCUS T1 (FT1), which initiates under day-neutral conditions of early spring. The incremental rise in FT1 expression is overridden in plants that contain a photoperiod-insensitive allele of Ppd-1, which hastens the completion of spikelet development and accelerates flowering time. The accelerated inflorescence development of photoperiod-insensitive lines is promoted by advanced seasonal expression of floral meristem identity genes. The completion of spikelet formation is promoted by FLOWERING LOCUS T2, which regulates spikelet number and is activated by Ppd-1. In wheat, flowering under natural photoperiods is regulated by stepwise increases in the expression of FT1, which responds dynamically to extending daylengths to promote early inflorescence development. This research provides a strong foundation to improve yield potential by fine-tuning the photoperiod-dependent control of inflorescence development.


Paramecium Polycomb repressive complex 2 physically interacts with the small RNA-binding PIWI protein to repress transposable elements.
Polycomb repressive complex 2 (PRC2) maintains transcriptionally silent genes in a repressed state via deposition of histone H3K27-trimethyl (me3) marks. PRC2 has also been implicated in silencing transposable elements (TEs), yet how PRC2 is targeted to TEs remains unclear. To address this question, we identified proteins that physically interact with the Paramecium enhancer-of-zeste Ezl1 enzyme, which catalyzes H3K9me3 and H3K27me3 deposition at TEs. We show that the Paramecium PRC2 core complex comprises four subunits, each required in vivo for catalytic activity. We also identify PRC2 cofactors, including the RNA interference (RNAi) effector Ptiwi09, which are necessary to target H3K9me3 and H3K27me3 to TEs. We find that the physical interaction between PRC2 and the RNAi pathway is mediated by a RING finger protein and that small RNA recruitment of PRC2 to TEs is analogous to the small RNA recruitment of H3K9 methylation SU(VAR)3-9 enzymes.


Planococcus versutus sp. nov., isolated from soil.
A taxonomic study was performed on a novel Gram-stain-positive, coccus-shaped, orange-pigmented motile bacterium, designated as strain L10.15T. The organism was isolated from a soil sample collected in Lagoon Island (close to Adelaide Island, western Antarctic Peninsula) using a quorum-quenching enrichment medium. Growth occurred at 4-30 °C, pH 6-11 and at moderately high salinity (0-15 %, w/v, NaCl), with optimal growth at 26 °C, at pH 7-8 and with 6 % (w/v) NaCl. 16S rRNA gene sequence analysis showed that strain L10.15T belonged to the genus Planococcus and was closely related to Planococcus halocryophilus Or1T (99.3 % similarity), Planococcus donghaensis JH1T (99.0 %), Planococcus antarcticus DSM 14505T (98.3 %), Planococcus plakortidis AS/ASP6 (II)T (97.6 %), Planococcus maritimus TF-9T (97.5 %), Planococcus salinarum ISL-6T (97.5 %) and Planococcus kocurii NCIMB 629T (97.5 %). However, the average nucleotide identity-MUMmer analysis showed low genomic relatedness values of 71.1-81.7 % to the type strains of these closely related species of the genus Planococcus. The principal fatty acids were anteiso-C15 : 0, C16 : 1ω7c and anteiso-C17 : 0, and the major menaquinones of strain L10.15T were MK-5 (48 %), MK-6 (6 %) and MK-7 (44 %). Polar lipid analysis revealed the presence of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and aminophospholipid. The DNA G+C content was 39.4 mol%. The phenotypic and genotypic data indicate that strain L10.15T represents a novel species of the genus Planococcus, for which the name Planococcus versutus sp. nov. is proposed. The type strain is L10.15T (=DSM 101994T=KACC 18918T).


Limits to thymidylate synthase and TP53 genes as predictive determinants for fluoropyrimidine sensitivity and further evidence for RNA-based toxicity as a major influence.
The major determinants of 5-flurouracil (5-FU) response would seem, based on accumulated literature, to be thymidylate synthase (TYMS, TS) expression levels, TS gene modifications, and TP53 status. We tested 5-FU sensitivity in yeast and human cancer cell models in which TS or TP53 alleles and expression were varied. Polymorphic TS tandem repeat status, TS expression levels reported, TS intragenic mutations, and TP53 status in outbred and experimental cancer cell lines did not predict 5-FU sensitivity or resistance. Novel observations included a dose-resistant persistence of unbound TS protein in many cancers and, upon 5-FU treatment of the colon cancer cell line, HCT116, evidence of allelic switching favoring transcripts of the mutant TS allele. The reported alleles having an intragenic mutation could not be causally associated with major degrees of 5-FU sensitivity. In yeast, TS protein was altered upon treatment with FdUMP, but 5-FU toxicity seemed to be largely RNA-based, being rescued by uridine rather than by thymidine. Cancer cell lines were also rescued from 5-FU toxicity with uridine rather than thymidine. Additionally, a TS (CDC21) knockout yeast strain, obviating any potential role for TS protein as a target, was hypersensitive to 5-FU. When denatured proteins from cancer cells treated with radiolabeled 5-FU were labeled, species with alternative molecular weights other than TS were visualized, providing further evidence for alternative 5-FU protein targets. These data emphasize that TS and TP53 status do not consistently explain the variance in responses of fluoropyrimidine-treated cancer cells, in part due to RNA-based toxicity.


Comorbidity of overweight and obesity in a nationally representative sample of German adults aged 18-79 years.
Overweight has increased in many countries over the past 20 years and excessive body weight is an established risk factor for adverse health outcomes and chronic diseases. This study aimed to determine comorbidity associated with overweight and obesity in a nationally representative sample of German adults. In the German National Health Interview and Examination Survey 1998 standardized measures of body weight, height and waist circumference (WC) were obtained for 7,124 men and women 18 to 79 years of age. Information on pre-existing health conditions, health-related behaviors, and sociodemographic characteristics was collected using physician-administered computer-assisted interviews and self-administered questionnaires. World Health Organization (WHO) cut-off criteria were applied to define overweight (BMI 25.0-29.9 kg/m2) and obesity (BMI ≥30.0 kg/m2) and abdominal obesity (men: WC ≥102 cm; women: WC ≥88 cm). The crude prevalence of persons with cardiometabolic risk factors, diabetes mellitus, cardiovascular disease (CVD), gall bladder disease, and osteoarthritis showed a significant stepwise increase from the lowest to the highest BMI category in both sexes. In multiple logistic regression models adjusting for age, social status, and smoking, significant associations with overweight and obesity persisted for cardiometabolic risk factors and osteoarthritis. For example, obese persons had a three- to fourfold higher chance of having any cardiometabolic risk factor compared to normal weight persons (odds ratio (OR) = 4.07, 95% CI: 3.16-5.25 for men; OR = 3.40 (2.60-4.46) for women). Only in women, overweight and obesity as well as abdominal obesity, independent of BMI category, were significantly and consistently associated with diabetes (overweight: OR = 1.85 (1.03-3.30); obesity: OR = 2.94 (1.63-5.31); abdominal obesity: OR = 1.44 (1.08-1.92) and gall bladder disease (overweight: OR = 1.65 (1.22-2.25); obesity: OR = 3.06 (2.26-4.14); abdominal obesity: OR = 1.73 (1.25-2.39)). Current estimates of disease burden underline the public health importance and clinical relevance related to overweight and obesity and needs to take into account comorbidity aspects.


Scl regulates the quiescence and the long-term competence of hematopoietic stem cells.
The majority of long-term reconstituting hematopoietic stem cells (LT-HSCs) in the adult is in G(0), whereas a large proportion of progenitors are more cycling. We show here that the SCL/TAL1 transcription factor is highly expressed in LT-HSCs compared with short-term reconstituting HSCs and progenitors and that SCL negatively regulates the G(0)-G(1) transit of LT-HSCs. Furthermore, when SCL protein levels are decreased by gene targeting or by RNA interference, the reconstitution potential of HSCs is impaired in several transplantation assays. First, the mean stem cell activity of HSCs transplanted at approximately 1 competitive repopulating unit was 2-fold decreased when Scl gene dosage was decreased. Second, Scl(+/-) HSCs were at a marked competitive disadvantage with Scl(+/+) cells when transplanted at 4 competitive repopulating units equivalent. Third, reconstitution of the stem cell pool by adult HSCs expressing Scl-directed shRNAs was decreased compared with controls. At the molecular level, we found that SCL occupies the Cdkn1a and Id1 loci in primary hematopoietic cells and that the expression levels of these 2 regulators of HSC cell cycle and long-term functions are sensitive to Scl gene dosage. Together, our observations suggest that SCL impedes G(0)-G(1) transition in HSCs and regulates their long-term competence.


Iron uptake and ferritin synthesis in human erythroblasts.
The erythroblasts from four normal bone marrows were enriched by using an anti-myeloid monoclonal antibody (TG-1), and a polyclonal antibody against mononuclear cells to effect complement mediated lysis of unwanted cells. The erythroblasts were cultured for 2 h in medium containing [59Fe]transferrin and [3H]-leucine, then fractionated on Percoll gradients according to their density. Whole cell iron uptake by early erythroblasts was greater than in the dense late erythroblasts. In all marrows, iron uptake into ferritin was highest in fractions containing the earliest erythroblasts and decreased with increasing erythroblast maturity. In three of the four marrows this paralleled the pattern of ferritin synthesis. It seems likely that apoferritin is synthesized in response to the amount of iron taken into the cell and this iron is incorporated within the protein shell to form ferritin.


Salivary cortisol response to infant distress in pregnant women with depressive symptoms.
The Hypothalamic-Pituitary-Adrenal (HPA) axis has been proposed as a potential underlying biological mechanism linking prenatal depression with adverse offspring outcomes. However, it is unknown whether the reactivity of this system to stress is altered in pregnant women experiencing depression. The objective of this study was to investigate whether salivary cortisol response to a distressed infant film is enhanced in pregnant women with symptoms of depression compared with non-depressed controls. Salivary cortisol and subjective mood responses to the film were measured in 53 primiparous women, between 11 and 18 weeks gestation. Both groups showed similar increases in state anxiety in response to the film, but there was a significantly increased cortisol response in women experiencing symptoms of depression. Depression during pregnancy is associated with increased reactivity of the HPA axis. This is consistent with altered HPA axis functioning being a key mechanism by which prenatal mood disturbance can impact upon fetal development.


Ethanol-induced alterations in fatty acid-related lipids in serum and tissues in mice.
Chronic alcohol consumption is a major factor for several human diseases, and alcoholism is associated with a host of societal problems. One of the major alcohol-induced metabolic changes is the increased NADH levels, which reduces glucose synthesis and increases fatty acid (FA) synthesis. Probably more important is the induction of FA synthesizing enzymes under the control of sterol regulatory element binding proteins (SREBP), plus increased malonyl-CoA, which blocks FA entry to the mitochondria for oxidation. The changes in FA-related lipids, particularly lysophospholipids and ceramides (Cers), in different tissues in ethanol-fed mice have not been reported. We systematically determined the levels of FA-related lipids, including FAs, phosphatidylcholines, phosphatidylethanolamines, lysophosphatidic acid, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylinositol, sphingomyelins, and ceramides (Cers), in the serum and different tissues by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS). The study was performed in C57BL/6J mice fed with Lieber-DeCarli diet, in which ethanol was added to account for 27.5% of total calories. The serum and tissues were collected from these mice at the time of killing, and the results were compared to pair-fed controls. The important observation was that ethanol-induced tissue-specific changes, which were related to different FA chains. Several 22:6 FA, 18:0 FA, 18:0 to 18:3 FA-containing lipids were significantly increased in the serum, liver, and skeletal muscle, respectively. In the kidney, all 22:6 FA-containing lipids detected were increased. In addition, alterations in other lipids in tissues, except adipose tissue, were also observed. We found tissue-specific alterations in the levels of FA-related lipids after ethanol administration. The implications of these findings pertinent to human physiology/pathology warrant further investigation. More studies are needed to explore the mechanisms on the different effects of ethanol on certain lipids in different tissues.


The human lipid phosphate phosphatase-3 decreases the growth, survival, and tumorigenesis of ovarian cancer cells: validation of the lysophosphatidic acid signaling cascade as a target for therapy in ovarian cancer.
Lysophosphatidic acid (LPA) is present at elevated concentrations in the ascites and plasma of ovarian cancer patients. Ovarian cancer cells produce and release LPA both constitutively and after stimulation. LPA can induce proliferation, survival, invasiveness, and resistance to chemotherapy of ovarian cancer cells. This suggests that LPA may be critically important for the development or progression of ovarian cancer and is thus a potential target for therapy. In this study, we demonstrate that introduction of the integral membrane protein, human lipid phosphate phosphohydrolase-3 (hLPP-3) enzyme, which hydrolyzes phosphatidic acid, LPA, sphingosine, and ceramide phosphate in vitro with selectivity for LPA, into SKOV3 and OVCAR-3 ovarian cancer cells decreases colony-forming activity, increases apoptosis, and decreases tumor growth in vitro and in vivo. Strikingly, coculture of hLPP-3-expressing cells with nontransfected parental cells decreased the colony-forming activity of the parental cells, compatible with hLPP-3 decreasing levels of an extracellular mediator, likely LPA. Compatible with this contention, the expression of hLPP-3 was associated with increased rates of extracellular LPA hydrolysis. The effects of hLPP-3 on colony-forming activity were substantially reversed by the LPP-resistant LPA analogue, O-methylphosphothionate. The ability of O-methylphosphothionate to ameliorate the effects of hLPP-3, combined with the inability of an enzymatically inactive hLPP-3 to alter cellular function, suggests that the major effect of hLPP-3 was to increase the hydrolysis of extracellular LPA. Thus genetic or pharmacological manipulation of LPA metabolism, receptor activation, or downstream signaling is an attractive approach for therapy of ovarian cancer.


Early complement genes are associated with visual system degeneration in multiple sclerosis.
Multiple sclerosis is a heterogeneous disease with an unpredictable course and a wide range of severity; some individuals rapidly progress to a disabled state whereas others experience only mild symptoms. Though genetic studies have identified variants that are associated with an increased risk of developing multiple sclerosis, no variants have been consistently associated with multiple sclerosis severity. In part, the lack of findings is related to inherent limitations of clinical rating scales; these scales are insensitive to early degenerative changes that underlie disease progression. Optical coherence tomography imaging of the retina and low-contrast letter acuity correlate with and predict clinical and imaging-based outcomes in multiple sclerosis. Therefore, they may serve as sensitive phenotypes to discover genetic predictors of disease course. We conducted a set of genome-wide association studies of longitudinal structural and functional visual pathway phenotypes in multiple sclerosis. First, we assessed genetic predictors of ganglion cell/inner plexiform layer atrophy in a discovery cohort of 374 patients with multiple sclerosis using mixed-effects models adjusting for age, sex, disease duration, optic neuritis and genetic ancestry and using a combination of single-variant and network-based analyses. For candidate variants identified in discovery, we conducted a similar set of analyses of ganglion cell/inner plexiform layer thinning in a replication cohort (n = 376). Second, we assessed genetic predictors of sustained loss of 5-letters in low-contrast letter acuity in discovery (n = 582) using multivariable-adjusted Cox proportional hazards models. We then evaluated candidate variants/pathways in a replication cohort. (n = 253). Results of both studies revealed novel subnetworks highly enriched for connected genes in early complement activation linked to measures of disease severity. Within these networks, C3 was the gene most strongly associated with ganglion cell/inner plexiform layer atrophy (P = 0.004) and C1QA and CR1 were top results in analysis of sustained low-contrast letter acuity loss. Namely, variant rs158772, linked to C1QA, and rs61822967, linked to CR1, were associated with 71% and 40% increases in risk of sustained LCLA loss, respectively, in meta-analysis pooling discovery and replication cohorts (rs158772: hazard ratio: 1.71; 95% confidence interval 1.30-2.25; P = 1.3 × 10-4; rs61822967: hazard ratio: 1.40; 95% confidence interval: 1.16-1.68; P = 4.1 × 10-4). In conclusion, early complement pathway gene variants were consistently associated with structural and functional measures of multiple sclerosis severity. These results from unbiased analyses are strongly supported by several prior reports that mechanistically implicated early complement factors in neurodegeneration.


Contact With Young Children Increases the Risk of Respiratory Infection in Older Adults in Europe-the RESCEU Study.
Knowledge about how older adults get a respiratory infection is crucial for planning preventive strategies. We aimed to determine how contact with young children living outside of the household affects the risk of acute respiratory tract infections (ARTI) in community-dwelling older adults. This study is part of the European RESCEU older adult study. Weekly surveillance was performed to detect ARTI throughout 2 winter seasons (2017-2018, 2018-2019). Child exposure, defined as having regular contact with children under 5 living outside of the subject's household, was assessed at baseline. The average attributable fraction was calculated to determine the fraction of ARTI explained by exposure to these children. We prospectively established that 597/1006 (59%) participants experienced at least 1 ARTI. Child exposure increased the risk of all-cause ARTI (adjusted odds ratio [aOR], 1.58; 95% confidence interval [CI], 1.21 -2.08; P = .001). This risk was highest in those with the most frequent contact (aOR, 1.80; 95% CI, 1.23-2.63; P = .003). The average attributable fraction of child exposure explaining ARTI was 10% (95% CI, 5%-15%). One of 10 ARTI in community-dwelling older adults is attributable to exposure to preschool children living outside of the household. NCT03621930.


Adult living-donor liver transplantation for a recipient with a high preoperative 1,3-beta-D-glucan level and positive test result for Aspergillus antigen.
The patient was a 45-year-old man with underlying alcoholic liver cirrhosis. Two years prior, he was repeatedly hospitalized for liver failure symptoms and requested a living-donor liver transplantation (LDLT) because of end-stage cirrhosis. A pretransplantation blood test revealed a high 1,3-beta-d-glucan (BDG) value of 102.0 pg/mL (reference value <20.0 pg/mL) and a high blood Aspergillus antigen (AsAg) value of 1.6 cutoff index (COI; reference value <0.5 COI). Contrast-enhanced thoracoabdominal-pelvic computed tomography (CT) and cranial magnetic resonance imaging revealed no fungal infection. However, latent fungal infection could not be ruled out, hence preoperative antifungal agent treatment was administered. BDG and AsAg levels showed a decreasing trend after treatment initiation. However, normalization did not occur; the BDG and AsAg levels were 25.8 pg/mL and 1.0 COI, respectively. Although the possibility of latent fungal infection was judged low, we prophylactically administered antifungal agents after LDLT. The BDG level consistently increased at 35-39 pg/mL until postoperative day 5 but subsequently normalized. The AsAg level was higher than the limit of detection at 5.0 COI on postoperative day 3 but normalized to 0.2 COI on postoperative day 5 and did not subsequently increase. The postoperative course was uneventful despite bacterial pneumonia and the patient was discharged on postoperative day 35. A histopathologic examination (Grocott methenamine silver staining) and a fungal polymerase chain reaction assay were performed for the resected liver, but the results of both were negative. At 9 postoperative months, the patient was making ambulatory follow-up visits. Currently, the BDG and AsAg values remain normal and clinical progress is favorable. We found no reports of LDLT for a recipient with a high preoperative BDG level and positive test result for AsAg. Thus, we report on such a case with a discussion of the literature on the causes of high preoperative BDG and AsAg values.


Genetic analyses suggest no immigration of adult females and their offspring into the Sonso community of chimpanzees in the Budongo Forest Reserve, Uganda.
Chimpanzees are frequently used to illustrate the relationship between sex differences in dispersal and sex differences in cooperation in primates and other group-living mammals. Male chimpanzees are highly philopatric, typically remaining in their natal communities for their entire lives to cooperate with related males in competition against less related males from other groups, whereas females typically disperse once at adolescence and cooperate with each other less frequently. However, there have been a few reports of dependent male offspring joining groups when their mothers transferred between communities as adults. Although such events are difficult to document, determining how often they actually occur is important for elucidating the links between philopatry, kinship, and cooperation in both chimpanzees and group-living animals more generally. Here we use genetic analyses to investigate a previous report of a large-scale transfer of many females and their offspring into the Sonso community of chimpanzees in the Budongo Forest Reserve, Uganda. Using autosomal microsatellite genotypes, we assigned a Sonso father to ten of the fourteen putative immigrants, and found that the four putative immigrants for whom we could not assign a Sonso father (perhaps due to incomplete sampling of all Sonso candidate fathers) nevertheless had Y-chromosome microsatellite haplotypes that were common in Sonso males but absent in males from four other chimpanzee communities at Budongo. These results suggest that these putative immigrant females and their offspring were probably actually long-term residents of Sonso whose identifications were delayed by their peripheral or unhabituated status. These results are consistent with other genetic and behavioral evidence showing that male between-community gene flow is exceedingly rare in east African chimpanzees.


Integrative modelling reveals mechanisms linking productivity and plant species richness.
How ecosystem productivity and species richness are interrelated is one of the most debated subjects in the history of ecology. Decades of intensive study have yet to discern the actual mechanisms behind observed global patterns. Here, by integrating the predictions from multiple theories into a single model and using data from 1,126 grassland plots spanning five continents, we detect the clear signals of numerous underlying mechanisms linking productivity and richness. We find that an integrative model has substantially higher explanatory power than traditional bivariate analyses. In addition, the specific results unveil several surprising findings that conflict with classical models. These include the isolation of a strong and consistent enhancement of productivity by richness, an effect in striking contrast with superficial data patterns. Also revealed is a consistent importance of competition across the full range of productivity values, in direct conflict with some (but not all) proposed models. The promotion of local richness by macroecological gradients in climatic favourability, generally seen as a competing hypothesis, is also found to be important in our analysis. The results demonstrate that an integrative modelling approach leads to a major advance in our ability to discern the underlying processes operating in ecological systems.


The antiviral sirtuin 3 bridges protein acetylation to mitochondrial integrity and metabolism during human cytomegalovirus infection.
Regulation of mitochondrial structure and function is a central component of infection with viruses, including human cytomegalovirus (HCMV), as a virus means to modulate cellular metabolism and immune responses. Here, we link the activity of the mitochondrial deacetylase SIRT3 and global mitochondrial acetylation status to host antiviral responses via regulation of both mitochondrial structural integrity and metabolism during HCMV infection. We establish that SIRT3 deacetylase activity is necessary for suppressing virus production, and that SIRT3 maintains mitochondrial pH and membrane potential during infection. By defining the temporal dynamics of SIRT3-substrate interactions during infection, and overlaying acetylome and proteome information, we find altered SIRT3 associations with the mitochondrial fusion factor OPA1 and acetyl-CoA acyltransferase 2 (ACAA2), concomitant with changes in their acetylation levels. Using mutagenesis, microscopy, and virology assays, we determine OPA1 regulates mitochondrial morphology of infected cells and inhibits HCMV production. OPA1 acetylation status modulates these functions, and we establish K834 as a site regulated by SIRT3. Control of SIRT3 protein levels or enzymatic activity is sufficient for regulating mitochondrial filamentous structure. Lastly, we establish a virus restriction function for ACAA2, an enzyme involved in fatty acid beta-oxidation. Altogether, we highlight SIRT3 activity as a regulatory hub for mitochondrial acetylation and morphology during HCMV infection and point to global acetylation as a reflection of mitochondrial health.


Neuropsychologic changes from before transplantation to 1 year in patients receiving myeloablative allogeneic hematopoietic cell transplant.
Research indicates that myeloablative hematopoietic cell transplantation (HCT) impairs neurocognitive function. However, prospective studies on long-term effects are lacking. This longitudinal study examined neurocognitive changes over the first year in 142 adult recipients of allogeneic HC transplants who received neuropsychologic testing before transplantation and again after 80 days and 1 year. Age-, sex-, and education-adjusted population-based standardized scores were used for normative comparisons. Performance on all tests declined from before transplantation to 80 days (P < .05) and improved by 1 year (P < .05), returning to pretransplantation levels on all tests except for grip strength and motor dexterity. Although verbal fluency and memory recovered by 1 year, both were below norms at all 3 testing times (P < .01). Logistic regressions indicated that patients without chemotherapy, other than hydroxyurea, previous to HCT and patients not receiving chronic graft-versus-host disease (GVHD) medication at 1 year had lower risk of impaired function (P < .05). In conclusion, HCT was associated with significant generalized decline in neurocognitive performance at 80 days, with subsequent recovery to pretransplantation levels by 1 year for most survivors, except on motor tasks. Results indicate that long-term cognitive decrements, as distinct from motor disabilities, infrequently derive directly from HCT.


Immune landscape of human placental villi using single-cell analysis.
Maintenance of a healthy pregnancy is reliant on a successful balance between the fetal and maternal immune systems. Although the maternal mechanisms responsible have been well studied, those used by the fetal immune system remain poorly understood. Using suspension mass cytometry and various imaging modalities, we report a complex immune system within the mid-gestation (17-23 weeks) human placental villi (PV). Consistent with recent reports in other fetal organs, T cells with memory phenotypes, although rare in abundance, were detected within the PV tissue and vasculature. Moreover, we determined that T cells isolated from PV samples may be more proliferative after T cell receptor stimulation than adult T cells at baseline. Collectively, we identified multiple subtypes of fetal immune cells within the PV and specifically highlight the enhanced proliferative capacity of fetal PV T cells.


Ultraspecific probes for high throughput HLA typing.
The variations within an individual's HLA (Human Leukocyte Antigen) genes have been linked to many immunological events, e.g. susceptibility to disease, response to vaccines, and the success of blood, tissue, and organ transplants. Although the microarray format has the potential to achieve high-resolution typing, this has yet to be attained due to inefficiencies of current probe design strategies. We present a novel three-step approach for the design of high-throughput microarray assays for HLA typing. This approach first selects sequences containing the SNPs present in all alleles of the locus of interest and next calculates the number of base changes necessary to convert a candidate probe sequences to the closest subsequence within the set of sequences that are likely to be present in the sample including the remainder of the human genome in order to identify those candidate probes which are "ultraspecific" for the allele of interest. Due to the high specificity of these sequences, it is possible that preliminary steps such as PCR amplification are no longer necessary. Lastly, the minimum number of these ultraspecific probes is selected such that the highest resolution typing can be achieved for the minimal cost of production. As an example, an array was designed and in silico results were obtained for typing of the HLA-B locus. The assay presented here provides a higher resolution than has previously been developed and includes more alleles than previously considered. Based upon the in silico and preliminary experimental results, we believe that the proposed approach can be readily applied to any highly polymorphic gene system.


Extreme mortality and reproductive failure of common murres resulting from the northeast Pacific marine heatwave of 2014-2016.
About 62,000 dead or dying common murres (Uria aalge), the trophically dominant fish-eating seabird of the North Pacific, washed ashore between summer 2015 and spring 2016 on beaches from California to Alaska. Most birds were severely emaciated and, so far, no evidence for anything other than starvation was found to explain this mass mortality. Three-quarters of murres were found in the Gulf of Alaska and the remainder along the West Coast. Studies show that only a fraction of birds that die at sea typically wash ashore, and we estimate that total mortality approached 1 million birds. About two-thirds of murres killed were adults, a substantial blow to breeding populations. Additionally, 22 complete reproductive failures were observed at multiple colonies region-wide during (2015) and after (2016-2017) the mass mortality event. Die-offs and breeding failures occur sporadically in murres, but the magnitude, duration and spatial extent of this die-off, associated with multi-colony and multi-year reproductive failures, is unprecedented and astonishing. These events co-occurred with the most powerful marine heatwave on record that persisted through 2014-2016 and created an enormous volume of ocean water (the "Blob") from California to Alaska with temperatures that exceeded average by 2-3 standard deviations. Other studies indicate that this prolonged heatwave reduced phytoplankton biomass and restructured zooplankton communities in favor of lower-calorie species, while it simultaneously increased metabolically driven food demands of ectothermic forage fish. In response, forage fish quality and quantity diminished. Similarly, large ectothermic groundfish were thought to have increased their demand for forage fish, resulting in greater top-predator demands for diminished forage fish resources. We hypothesize that these bottom-up and top-down forces created an "ectothermic vise" on forage species leading to their system-wide scarcity and resulting in mass mortality of murres and many other fish, bird and mammal species in the region during 2014-2017.


Rho GTPases regulate distinct aspects of dendritic arbor growth in Xenopus central neurons in vivo.
The development and structural plasticity of dendritic arbors are governed by several factors, including synaptic activity, neurotrophins and other growth-regulating molecules. The signal transduction pathways leading to dendritic structural changes are unknown, but likely include cytoskeleton regulatory components. To test whether GTPases regulate dendritic arbor development, we collected time-lapse images of single optic tectal neurons in albino Xenopus tadpoles expressing dominant negative or constitutively active forms of Rac, Cdc42 or RhoA. Analysis of images collected at two-hour intervals over eight hours indicated that enhanced Rac activity selectively increased branch additions and retractions, as did Cdc42 to a lesser extent. Activation of endogenous RhoA decreased branch extension without affecting branch additions and retractions, whereas dominant-negative RhoA increased branch extension. Finally, we provide data suggesting that RhoA mediates the promotion of normal dendritic arbor development by NMDA receptor activation.


Mechanism of hyperventilation in acute cerebrovascular accidents.
Lumbar cerebrospinal fluid and arterial blood acid-base state were assessed in 19 patients within 24 hours of an acute cerebrovascular accident. Those with haemorrhage into the C.S.F. showed a lower C.S.F. pH and higher C.S.F. lactate than those without haemorrhage but the Pco(2), was similar in the two groups, suggesting that this greater C.S.F. acidity was not responsible for a greater degree of hyperventilation. In those without haemorrhage an inverse relation was found between C.S.F. pH and arterial Pco(2), suggesting that a non-chemical ventilatory drive-for example, due to central neurological damage-was responsible for the acid-base changes observed.