A lysosome targetable versatile fluorescent probe for imaging viscosity and peroxynitrite with different fluorescence signals in living cells.
The lysosome, which acts as the cellular recycling centre, is filled with numerous hydrolases that can degrade most cellular macromolecules. The abnormalities of the lysosome are closely associated with diseases, such as Heřmanský-Pudlák syndrome, Griscelli syndrome and Chédiak-Higashi syndrome. Studies have shown that abnormal viscosity and the accumulation of reactive oxygen species (ROS) in the lysosome will disorder the normal function of the lysosome. In this research, a versatile fluorescent probe Lyso-NA has been developed for the multi-channel imaging of lysosomal viscosity and peroxynitrite (ONOO-). When excited at 550 nm, the Lyso-NA exhibited about a 50-fold increase in fluorescence at 610 nm and also with the increasing viscosity from 1.0 cP to 1410 cP, and about a 3.5-fold increase in fluorescence at 510 nm (excitation at 440 nm) together with the increasing ONOO-. These satisfactory response properties make it possible to use Lyso-NA to monitor changes in both viscosity and ONOO- inside the lysosome. To achieve its practical application, it was further demonstrated that Lyso-NA exhibits low cytotoxicity, and good cell permeability, and could be used to monitor lysosomal viscosity and ONOO- in living cells.


Macrophage killing of Leishmania parasite in vivo is mediated by nitric oxide from L-arginine.
Peritoneal macrophages from CBA mice incubated with rIFN-gamma are effective in killing the protozoal parasite Leishmania major in vitro. This leishmanicidal activity can be completely inhibited by L-NG-monomethyl arginine (L-NMMA), a specific inhibitor of the L-arginine:nitric oxide (NO) pathway. The culture supernatants of macrophage activated by IFN-gamma contain increased levels of NO2-, the production of which is inhibited by L-NMMA, but not by its D-enantiomer. L. major promastigotes are killed when incubated at room temperature in PBS containing NO. These data demonstrate that NO is an effector mechanism in macrophage killing of intracellular protozoa. The importance of NO in vivo is demonstrated by the finding that CBA mice infected with L. major developed exacerbated disease when L-NMMA was injected into the lesions, resulting in 10(4)-fold increases in the number of parasites extractable from the lesions.


Development and adaptability of microvasculature in skeletal muscle.
In skeletal muscle, the size of the capillary bed is adapted to the type of muscle metabolism and can be altered by adaptation to different environments or increased activity. Muscle fibres with high aerobic metabolism have more capillaries, and an increase in aerobic metabolism is usually followed by capillary growth. It is assumed that local hypoxia - created by increased demand for oxygen during growth, cold exposure or increased activity - can stimulate proliferation of capillaries. Capillary density is reduced in parallel with enhanced glycolytic metabolism. The size of the capillary bed can also increase without any apparent change in the oxidative metabolism (e.g. in the early stages of chronic electrical muscle stimulation or as a result of long-term administration of vasodilating drugs), and it is argued that the growth of capillaries in these cases may be due to various mechanical factors connected with increased blood flow.


Allocation of control rights and cooperation efficiency in public-private partnerships: theory and evidence from the Chinese pharmaceutical industry.
This article uses incomplete contract theory to study the allocation of control rights in public-private partnerships (PPPs) between pharmaceutical enterprises and nonprofit organizations; it also investigates how this allocation influences cooperation efficiency. We first develop a mathematic model for the allocation of control rights and its influence on cooperation efficiency, and then derive some basic hypotheses from the model. The results of an empirical test show that the allocation of control rights influences how enterprises invest in PPPs. A proper allocation provides incentives for firms to make fewer self-interested and more public-interested investments. Such an allocation also improves the cooperation efficiency of PPPs.


Comparison of efficacy of intra-articular morphine and steroid in patients with knee osteoarthritis.
Primary therapeutic aim in treatment of osteoarthritis of the knee is to relieve the pain of osteoarthritis. The aim of this study was to compare the efficacy of intra-articular triamcinolone with intra-articular morphine in pain relief due to osteoarthritis of the knee in the elderly population. Patients between 50 and 80 years of age were randomized into three groups. Group M received morphine plus bupivacaine intra-articularly, Group T received triamcinolone plus bupivacaine intra-articularly, and Group C received saline plus bupivacaine intra-articularly. Patients were evaluated before injection and in 2nd, 4th, 6th, and 12th weeks after injection. First-line supplementary analgesic was oral paracetamol 1500 mg/day. If analgesia was insufficient with paracetamol, oral dexketoprofen trometamol 50 mg/day was recommended to patients. After the intra-articular injection, there was statistically significant decrease in visual analog scale (VAS) scores in Groups M and T, when compared to Group C. The decrease of VAS scores seen at the first 2 weeks continued steadily up to the end of 12th week. There was a significant decrease in Groups M and T in the WOMAC scores, when compared to Group C. There was no significant difference in the WOMAC scores between morphine and steroid groups. Significantly less supplementary analgesics was used in the morphine and steroid groups. Intra-articular morphine was as effective as intra-articular triamcinolone for analgesia in patients with osteoarthritis knee. Intra-articular morphine is possibly a better option than intra-articular steroid as it has lesser side effects.


Polarization-induced size control and ablation dynamics of Ge nanostructures formed by a femtosecond laser.
We report a method for controlling the size of a Ge (germanium) nanostructure by changing the angle between the ultrafast laser polarization and the crystal axis of Ge. The nanostructure size dependence on the laser polarization with respect to the Ge crystal axis exhibits a sinusoidal function with a minimum size at (100) axis. Moreover, the measurement of transient reflection reveals the presence of large anisotropies in both its amplitude and its relaxation dynamics with a minimum at (100) crystal axis. This implies that the observed anisotropic dependence of nanostructure size of Ge is followed by a different carrier density as well as its relaxation process, depending on the orientation of the Ge crystal axis only at near and above threshold fluence.


Sequence diversity in the pe_pgrs genes of Mycobacterium tuberculosis is independent of human T cell recognition.
The Mycobacterium tuberculosis genome includes the large family of pe_pgrs genes, whose functions are unknown. Because of precedents in other pathogens in which gene families showing high sequence variation are involved in antigenic variation, a similar role has been proposed for the pe_pgrs genes. However, the impact of immune selection on pe_pgrs genes has not been examined. Here, we sequenced 27 pe_pgrs genes in 94 clinical strains from five phylogenetic lineages of the M. tuberculosis complex (MTBC). We found that pe_pgrs genes were overall more diverse than the remainder of the MTBC genome, but individual members of the family varied widely in their nucleotide diversity and insertion/deletion (indel) content: some were more, and others were much less, diverse than the genome average. Individual pe_pgrs genes also differed in the ratio of nonsynonymous to synonymous mutations, suggesting that different selection pressures act on individual pe_pgrs genes. Using bioinformatic methods, we tested whether sequence diversity in pe_pgrs genes might be selected by human T cell recognition, the major mechanism of adaptive immunity to MTBC. We found that the large majority of predicted human T cell epitopes were confined to the conserved PE domain and experimentally confirmed the antigenicity of this domain in tuberculosis patients. In contrast, despite being genetically diverse, the PGRS domains harbored few predicted T cell epitopes. These results indicate that human T cell recognition is not a significant force driving sequence diversity in pe_pgrs genes, which is consistent with the previously reported conservation of human T cell epitopes in the MTBC. Recognition of Mycobacterium tuberculosis antigens by T lymphocytes is known to be important for immune protection against tuberculosis, but it is unclear whether human T cell recognition drives antigenic variation in M. tuberculosis. We previously discovered that the known human T cell epitopes in the M. tuberculosis complex are highly conserved, but we hypothesized that undiscovered epitopes with naturally occurring sequence variants might exist. To test this hypothesis, we examined the pe_pgrs genes, a large family of genes that has been proposed to function in immune evasion by M. tuberculosis. We found that the pe_pgrs genes exhibit considerable sequence variation, but the regions containing T cell epitopes and the regions of variation are distinct. These findings confirm that the majority of human T cell epitopes of M. tuberculosis are highly conserved and indicate that selection forces other than T cell recognition drive sequence variation in the pe_pgrs genes.


Clinical grade ACE2 as a universal agent to block SARS-CoV-2 variants.
The recent emergence of multiple SARS-CoV-2 variants has caused considerable concern due to reduced vaccine efficacy and escape from neutralizing antibody therapeutics. It is therefore paramount to develop therapeutic strategies that inhibit all known and future SARS-CoV-2 variants. Here we report that all SARS-CoV-2 variants analyzed, including variants of concern (VOC) Alpha, Beta, Gamma, and Delta, exhibit enhanced binding affinity to clinical grade and phase 2 tested recombinant human soluble ACE2 (APN01). Importantly, soluble ACE2 neutralized infection of VeroE6 cells and human lung epithelial cells by multiple VOC strains with markedly enhanced potency when compared to reference SARS-CoV-2 isolates. Effective inhibition of infections with SARS-CoV-2 variants was validated and confirmed in two independent laboratories. These data show that SARS-CoV-2 variants that have emerged around the world, including current VOC and several variants of interest, can be inhibited by soluble ACE2, providing proof of principle of a pan-SARS-CoV-2 therapeutic.


Associations between Fitness Measures and Change of Direction Speeds with and without Occupational Loads in Female Police Officers.
Female police officers may be required to pursue offenders on foot while wearing occupational loads. The aim of this study was to determine relationships between fitness measures and change of direction speed (CODS) in female police officers and the influence of their occupational loads. Retrospective data were provided for 27 female police officers (age = 32.19 ± 5.09 y, height = 162.78 ± 5.01 cm, and mass = 71.31 ± 13.42 kg) and included fitness measures of: lower-body power (standing long jump (SLJ)), upper-body and trunk muscle endurance (push-up (PU) and sit-up (SU)), aerobic power (estimated VO2max), and CODS (Illinois agility test). The CODS test was performed without and with occupational load (10 kg). Paired sample t-tests (between-load conditions) and Pearson's correlations (relationships between measures) were performed with linear regression analysis used to account for the contribution of measures to unloaded and loaded CODS performance. CODS was significantly slower when loaded (unloaded = ~23.17 s, loaded = ~24.14 s, p < 0.001) with a strong, significant relationship between load conditions (r = 0.956, p < 0.001). Moderate to strong, significant relationships were found between all fitness measures ranging from estimated VO2max (r = -0.448) to SU (r = -0.673) in the unloaded condition, with the strength of these relationships increasing in the loaded condition accounting for 61% to 67% of the variance, respectively. While unloaded agility test performance was strongly associated with loaded performance, female police officer CODS was significantly reduced when carrying occupational loads. A variety of fitness measures that influence officer CODS performances become increasingly important when occupational loads are carried.


Purinergic signaling and blood vessels in health and disease.
Purinergic signaling plays important roles in control of vascular tone and remodeling. There is dual control of vascular tone by ATP released as a cotransmitter with noradrenaline from perivascular sympathetic nerves to cause vasoconstriction via P2X1 receptors, whereas ATP released from endothelial cells in response to changes in blood flow (producing shear stress) or hypoxia acts on P2X and P2Y receptors on endothelial cells to produce nitric oxide and endothelium-derived hyperpolarizing factor, which dilates vessels. ATP is also released from sensory-motor nerves during antidromic reflex activity to produce relaxation of some blood vessels. In this review, we stress the differences in neural and endothelial factors in purinergic control of different blood vessels. The long-term (trophic) actions of purine and pyrimidine nucleosides and nucleotides in promoting migration and proliferation of both vascular smooth muscle and endothelial cells via P1 and P2Y receptors during angiogenesis and vessel remodeling during restenosis after angioplasty are described. The pathophysiology of blood vessels and therapeutic potential of purinergic agents in diseases, including hypertension, atherosclerosis, ischemia, thrombosis and stroke, diabetes, and migraine, is discussed.


Unique features of the loblolly pine (Pinus taeda L.) megagenome revealed through sequence annotation.
The largest genus in the conifer family Pinaceae is Pinus, with over 100 species. The size and complexity of their genomes (∼20-40 Gb, 2n = 24) have delayed the arrival of a well-annotated reference sequence. In this study, we present the annotation of the first whole-genome shotgun assembly of loblolly pine (Pinus taeda L.), which comprises 20.1 Gb of sequence. The MAKER-P annotation pipeline combined evidence-based alignments and ab initio predictions to generate 50,172 gene models, of which 15,653 are classified as high confidence. Clustering these gene models with 13 other plant species resulted in 20,646 gene families, of which 1554 are predicted to be unique to conifers. Among the conifer gene families, 159 are composed exclusively of loblolly pine members. The gene models for loblolly pine have the highest median and mean intron lengths of 24 fully sequenced plant genomes. Conifer genomes are full of repetitive DNA, with the most significant contributions from long-terminal-repeat retrotransposons. In depth analysis of the tandem and interspersed repetitive content yielded a combined estimate of 82%.


Is the process of delivery of an individually tailored lifestyle intervention associated with improvements in LDL cholesterol and multiple lifestyle behaviours in people with familial hypercholesterolemia?
More insight in the association between reach, dose and fidelity of intervention components and effects is needed. In the current study, we aimed to evaluate reach, dose and fidelity of an individually tailored lifestyle intervention in people with Familial Hypercholesterolemia (FH) and the association between intervention dose and changes in LDL-Cholesterol (LDL-C), and multiple lifestyle behaviours at 12-months follow-up. Participants (n = 181) randomly allocated to the intervention group received the PRO-FIT intervention consisting of computer-tailored lifestyle advice (PRO-FIT*advice) and counselling (face-to-face and telephone booster calls) using Motivational Interviewing (MI). According to a process evaluation plan, intervention reach, dose delivered and received, and MI fidelity were assessed using the recruitment database, website/counselling logs and the Motivational Interviewing Treatment Integrity (MITI 3.1.1.) code. Regression analyses were conducted to explore differences between participant and non-participant characteristics, and the association between intervention dose and change in LDL-C, and multiple lifestyle behaviours. A 34% (n = 181) representative proportion of the intended intervention group was reached during the recruitment phase; participants did not differ from non-participants (n = 623) on age, gender and LDL-C levels. Of the participants, 95% received a PRO-FIT*advice log on account, of which 49% actually logged on and completed at least one advice module. Nearly all participants received a face-to-face counselling session and on average, 4.2 telephone booster calls were delivered. None of the face-to-face sessions were implemented according to MI guidelines. Overall, weak non-significant positive associations were found between intervention dose and LDL-C and lifestyle behaviours. Implementation of the PRO-FIT intervention in practice appears feasible, particularly PRO-FIT*advice, since it can be relative easily implemented with a high dose delivered. However, only less than half of the intervention group received the complete intervention-package as intended. Strategies to let participants optimally engage in using web-based computer-tailored interventions like PRO-FIT*advice are needed. Further, more emphasis should be put on more extensive MI training and monitoring/supervision.


Developing a methodology for carbon isotope analysis of lacustrine diatoms.
Stable isotope analysis of sedimentary carbon in lakes can help reveal changes in terrestrial and aquatic carbon cycles. A method based on a single, photosynthetic organism, where host effects are minimised, should offer more precision than carbon isotope studies of bulk lake sediments. Here we report the development of a systematic method for use on fossil lacustrine diatom frustules, adapted from previous studies in marine environments. A step-wise cleaning experiment on diatomaceous lake sediments from Lake Challa, near Mount Kilimanjaro, was made to demonstrate the necessary treatment stages to remove external sedimentary carbon. Changes in soluble carbon compounds during these cleaning experiments were measured using gas chromatography/mass spectrometry (GC/MS). The mass spectrometry methods were refined to measure the small percentage of carbon in these samples and details of these methods are presented. Samples of cleaned diatoms containing <1% carbon yielded robust results. Carbon isotope analyses of diatom samples containing different species mixtures were performed and suggested that differences existed, although the effects lay within current experimental error and require further work. Unlike what was found in work on oxygen and silicon isotopes from diatom frustules, mineral contamination had no discernible impact on the diatom carbon isotope ratios from these sediments. The range of values found in the lakes investigated thus far can be interpreted with reference to the supply and nature of carbon from the catchment as well as to the demand generated from lake primary productivity.


Opportunities for genomic prediction for fertility using endocrine and classical fertility traits in dairy cattle.
Endocrine fertility traits, defined from progesterone concentration levels in milk, have been suggested as alternative indicators for fertility in dairy cows because they are less biased by farm management decisions and more directly reflect a cow's reproductive physiology than classical traits derived from insemination and calving data. To determine the potential use of endocrine fertility traits in genomic evaluations, the improvement in accuracy from using endocrine fertility traits concurrent with classical traits in the genomic prediction of fertility was quantified. The impact of recording all traits on all training animals was also investigated. Endocrine and classical fertility records were available on 5,339 lactations from 2,447 Holstein cows in Ireland, the Netherlands, Sweden, and the United Kingdom. The endocrine traits were commencement of luteal activity (C-LA) and proportion of samples with luteal activity (PLA); the classical trait was the interval from calving to first service (CFS). The interval from C-LA to first service (C-LAFS), which is a combination of an endocrine trait and a classical trait, was also investigated. The target (breeding goal) trait for fertility was CFS or C-LAFS, whereas C-LA and PLA served as predictor traits. Genomic EBV (GEBV) for fertility were derived using genomic BLUP in bivariate models with 85,485 SNP. Genomic EBV for the separate fertility traits were also computed, in univariate models. The accuracy of GEBV was evaluated by 5-fold cross-validation. The highest accuracy of GEBV was achieved using bivariate predictions, where both an endocrine fertility trait and the classical fertility trait were used. Accuracy of GEBV for predicting adjusted phenotypes for CFS in the univariate model was 0.04, but when predicting CFS using a bivariate model with C-LA, the accuracy increased to 0.14 when all training animals were phenotyped for C-LA and (or not) for CFS. On phenotyping all training animals for both C-LA and CFS, accuracy for CFS increased to 0.18; however, when validation animals were also phenotyped for C-LA, there was no substantial increase in accuracy. When predicting CFS in bivariate analysis with PLA, accuracy ranged from 0.07 to 0.14. This first study on genomic predictions for fertility using endocrine traits suggests some improvement in the accuracy of prediction over using only the classical traits. Further studies with larger training populations may show greater improvements.


TLR4 Deficiency Protects against Hepatic Fibrosis and Diethylnitrosamine-Induced Pre-Carcinogenic Liver Injury in Fibrotic Liver.
The development of hepatocellular carcinoma (HCC) is a common consequence of advanced liver fibrosis but the interactions between fibrogenesis and carcinogenesis are still poorly understood. Recently it has been shown that HCC promotion depends on Toll-like receptor (TLR) 4. Pre-cancerogenous events can be modelled in mice by the administration of a single dose of diethylnitrosamine (DEN), with HCC formation depending amongst others on interleukin (IL) 6 production. Mice lacking the hepatocanalicular phosphatidylcholine transporter ABCB4 develop liver fibrosis spontaneously, resemble patients with sclerosing cholangitis due to mutations of the orthologous human gene, and represent a valid model to study tumour formation in pre-injured cholestatic liver. The aim of this study was to investigate DEN-induced liver injury in TLR4-deficient mice with biliary fibrosis. ABCB4-deficient mice on the FVB/NJ genetic background were crossed to two distinct genetic backgrounds (TLR4-sufficient C3H/HeN and TLR4-deficient C3H/HeJ) for more than 10 generations. The two congenic knockout and the two corresponding wild-type mouse lines were treated with a single dose of DEN for 48 hours. Phenotypic differences were assessed by measuring hepatic collagen contents, inflammatory markers (ALT, CRP, IL6) as well as hepatic apoptosis (TUNEL) and proliferation (Ki67) rates. Hepatic collagen accumulation is significantly reduced in ABCB4-/-:TLR4-/-double-deficient mice. After DEN challenge, apoptosis, proliferation and inflammatory markers are decreased in TLR4-deficient in comparison to TLR4-sufficient mice. When combining ABCB4 and TLR4 deficiency with DEN treatment, hepatic IL6 expression and proliferation rates are lowest in fibrotic livers from the double-deficient line. Consistent with these effects, selective digestive tract decontamination in ABCB4-/- mice also led to reduced tumor size and number after DEN. This study demonstrates that liver injury upon DEN challenge depends on pre-existing fibrosis and genetic background. The generation of ABCB4-/: TLR4-/- double-deficient mice illustrates that TLR4-deficiency protects against hepatic injury in a preclinical mouse model of chronic liver disease.


Interlimb differences in control of movement extent.
The current study was designed to examine potential interlimb asymmetries in controlling movement extent. Subjects made repetitive single-joint elbow extension movements while the arm was supported on a horizontal, frictionless, air-jet system. Four targets of 10, 20, 35, and 45 degrees excursions were randomly presented over the course of 150 trials. For both arms, peak tangential hand velocity scaled linearly with movement distance. There was no significant difference between either peak velocities or movement accuracies for the two arms. However, the mechanisms responsible for achieving these velocities and extents were quite distinct for each arm. For the dominant arm, peak tangential finger acceleration varied systematically with movement distance. In contrast, nondominant-arm peak tangential acceleration varied little across targets and, as such, was a poor predictor of movement distance. Instead the velocities of the nondominant arm were determined primarily by variation in the duration of the initial acceleration impulse, which corresponds to the time of peak velocity. These different strategies reflect previously identified mechanisms in controlling movement distance: pulse-height control and pulse-width control. The former is characterized by a variation in peak acceleration and has been associated with preplanning mechanisms. The latter occurs after peak acceleration and has been shown to depend on peripheral sensory feedback. Our findings indicate that the dominant-arm system controls movement extent largely through planning mechanisms that specify pulse-height control, whereas the nondominant system does so largely through feedback mediated pulse-width control.


Xanthone dimers: a compound family which is both common and privileged.
Covering: up to 2014. Xanthone dimers are a widespread, structurally-diverse family of natural products frequently found in plants, fungi and lichens. They feature an intriguing variety of linkages between the component xanthones (benzannulated chromanones). These synthetically elusive secondary metabolites are of great interest due to their broad array of bioactivities, which has led to the xanthones being designated as 'privileged structures'. We seek herein to give an overview of all reliably-described xanthone dimers, their structures, occurrence, and the bioactivities established to date. The possible biosynthetic pathways leading to members of this family are also discussed in light of our current knowledge.


Allele-specific PCR shows that genetic exchange occurs among genetically diverse Nodularia (cyanobacteria) filaments in the Baltic Sea.
Some cyanobacteria have been shown to exchange genetic information under laboratory conditions, but it has not been clear whether such genetic exchange occurs in the natural environment. To address this, a population genetic study was carried out on the filamentous diazotrophic cyanobacterium Nodularia in the Baltic Sea. Nodularia filaments were collected from 20 widely distributed sampling stations in the Baltic Sea during June and July 1998. Allele-specific PCR (AS-PCR) was used to characterize over 2000 filaments at three loci: a non-coding spacer between adjacent copies of the main structural gas vesicle gene gvpA (gvpA-IGS), the phycocyanin intergenic spacer (PC-IGS) and the rDNA internal transcribed spacer (rDNA-ITS). The three loci were all found to be polymorphic in the 1998 population: two alternative alleles were distinguished at the gvpA-IGS and PC-IGS loci, and three at the rDNA-ITS locus. All 12 possible combinations of alleles were found in the filaments studied, but some were much more common than others. The index of association (I:(A)) for all possible pairwise combinations of isolates was found to differ significantly from zero, which implies that there is some linkage disequilibrium between loci. The I:(A) values for 16 out of 20 individual sampling stations also differed significantly from zero: this shows that the observed linkage disequilibrium is not due to pooling data from genetically distinct subpopulations. Monte-Carlo simulations with random subsets of the data confirmed that some combinations showed significantly more linkage disequilibrium than expected by chance alone. It is concluded that genetic exchange occurs in the natural Nodularia population, but the frequency is not high enough for the loci to be in linkage equilibrium. The distribution of the 12 genotypes across the Baltic Sea was found to be non-random, but did not correlate with temperature, salinity or major nutrient concentrations. A significant relationship was found between the gene diversity among filaments at each station and the distance of the station from the centre of the sampling area: possible reasons for this trend are discussed.


Isolation of a Novel Lytic Bacteriophage against a Nosocomial Methicillin-Resistant Staphylococcus aureus Belonging to ST45.
Methicillin-resistant Staphylococcus aureus (MRSA) can cause a wide range of infections from mild to life-threatening conditions. Its enhanced antibiotic resistance often leads to therapeutic failures and therefore alternative eradication methods must be considered. Potential candidates to control MRSA infections are bacteriophages and their lytic enzymes, lysins. In this study, we isolated a bacteriophage against a nosocomial MRSA strain belonging to the ST45 epidemiologic group. The phage belonging to Caudovirales, Siphoviridae, showed a narrow host range and stable lytic activity without the emergence of resistant MRSA clones. Phylogenetic analysis showed that the newly isolated Staphylococcus phage R4 belongs to the Triavirus genus in Siphoviridae family. Genetic analysis of the 45 kb sequence of R4 revealed 69 ORFs. No remnants of mobile genetic elements and traces of truncated genes were observed. We have localized the lysin (N-acetylmuramoyl-L-alanine amidase) gene of the new phage that was amplified, cloned, expressed, and purified. Its activity was verified by zymogram analysis. Our findings could potentially be used to develop specific anti-MRSA bacteriophage- and phage lysin-based therapeutic strategies against major clonal lineages and serotypes.


Immune response to functionalized mesoporous silica nanoparticles for targeted drug delivery.
Multifunctional mesoporous silica nanoparticles (MSN) have attracted substantial attention with regard to their high potential for targeted drug delivery. For future clinical applications it is crucial to address safety concerns and understand the potential immunotoxicity of these nanoparticles. In this study, we assess the biocompatibility and functionality of multifunctional MSN in freshly isolated, primary murine immune cells. We show that the functionalized silica nanoparticles are rapidly and efficiently taken up into the endosomal compartment by specialized antigen-presenting cells such as dendritic cells. The silica nanoparticles showed a favorable toxicity profile and did not affect the viability of primary immune cells from the spleen in relevant concentrations. Cargo-free MSN induced only very low immune responses in primary cells as determined by surface expression of activation markers and release of pro-inflammatory cytokines such as Interleukin-6, -12 and -1β. In contrast, when surface-functionalized MSN with a pH-responsive polymer capping were loaded with an immune-activating drug, the synthetic Toll-like receptor 7 agonist R848, a strong immune response was provoked. We thus demonstrate that MSN represent an efficient drug delivery vehicle to primary immune cells that is both non-toxic and non-inflammagenic, which is a prerequisite for the use of these particles in biomedical applications.


Islamic views on artificial nutrition and hydration in terminally ill patients.
Withholding and withdrawing artificial nutrition and hydration from terminally ill patients poses many ethical challenges. The literature provides little information about the Islamic beliefs, attitudes, and laws related to these challenges. Artificial nutrition and hydration may be futile and reduce quality of life. They can also harm the terminally ill patient because of complications such as aspiration pneumonia, dyspnea, nausea, diarrhea, and hypervolemia. From the perspective of Islam, rules governing the care of terminally ill patients are derived from the principle that injury and harm should be prevented or avoided. The hastening of death by the withdrawal of food and drink is forbidden, but Islamic law permits the withdrawal of futile, death-delaying treatment, including life support. Nutritional support is considered basic care and not medical treatment, and there is an obligation to provide nutrition and hydration for the dying person unless it shortens life, causes more harm than benefit, or is contrary to an advance directive that is consistent with Islamic law. The decision about withholding or withdrawing artificial nutrition and hydration from the terminally ill Muslim patient is made with informed consent, considering the clinical context of minimizing harm to the patient, with input from the patient, family members, health care providers, and religious scholars.


Low-dose electron image reconstruction of negatively stained contractile phage sheath from Bacillus subtilis (PBS-Z).
The structure of the contractile sheath of the defective phage from B. subtilis (PBS-Z) has been investigated by low-dose electron microscopy and image reconstruction. The extended and contracted sheath particles were imaged by means of two negative stains which consisted of uranyl- and phosphotungstate-containing solutions of a pH of 4.2 and 7.0 respectively. Images of identical parts of the same type of specimen were recorded at a total electron dose of 80 C/m2 (5 electrons/A2) and 4 x 10(3) C/m2 (250 electrons/A2). The low-dose reconstructions of the extended and contracted sheath structure in the two stains show good correspondence and made it possible to draw the following structural conclusions. The sheath protein in both types of structure has an elongated shape, and in both structures the long molecular axis lies in a plane perpendicular to the helical sheath axis. The orientation of the protein in the extended and contracted sheath is different; the long axes differ by about 35 degrees in orientation. The reconstructions did not permit conclusions about different conformational states of the protein in both structures. These data, together with the packing parameters of the protein subunits in the contractile sheath [1], form the complete structural analysis of this biological structure by electron microscopy. The radiation damage effects which have been monitored in analyzing image pairs to the full extent may be summarized as follows. (1) Diameters of the sheath structure increase, which indicate flattening. (2) There is no loss in resolution, and layerline altitudes of the Fourier-transformed images do not change. (3) Uranyl stain behaves differently compared to phosphotungstate. In both negative stains the structural noise level increases upon irradiation as follows from the increase in phase residuals of the digital layerline data. In uranyl-stained images also more aperiodic noise appears. (4) The Fourier amplitudes of the principal layerline maxima shift towards lower spatial frequencies; phases of corresponding maxima generally remain constant. This pattern is more pronounced in the extended sheath data; there is no rationale describing these positional shifts. Moreover, in the case of contracted sheath the amplitudes of Fourier components also change more in absolute value. Therefore the damage effects also seem to depend on the type of structure embedded in the stain. (5) In the reconstructed images these radiation effects create artificial stain-excluded volumes of a type and at a radius which depend on the stain and structure.


Effects of spatial and nonspatial cognitive activity on postural stability.
Is postural stability controlled automatically, or is it affected by concurrent cognitive activity? Are the effects influenced by the nature of the cognitive activity required, and do they increase in old age? To address these questions, 70 participants aged 20-79 years were asked to stand as still as possible on a force platform (postural control task) while performing (a) no cognitive task, (b) a spatial memory task, and (c) a nonspatial memory task. The memory tasks were also performed while seated as a comparison condition. Both spatial and nonspatial memory recall declined with increasing age but were unaffected by position (standing vs. seated). Postural stability declined with age; moreover, there was support for an earlier finding that age decline was greatest when performing the spatial memory task. Each recording period was split into two phases which, for the spatial and nonspatial memory tasks, corresponded to encoding and maintaining the stimuli. In comparison with no task, participants were more stable when encoding stimuli (particularly in the spatial task), but they were less stable when maintaining stimuli (particularly in the nonspatial task). The results suggest that postural stability can be affected by cognitive activity in complex ways, depending on the age of participants, the type of cognitive task (spatial vs. nonspatial), and the cognitive processing required (encoding vs. maintenance).


Alendronate-eluting polyglucose-lignol composite (POGLICO): a new biomaterial for fracture fixating implants.
Due to the known drawbacks of metal implants, new biomaterials for internal fracture fixation are attracting increasing interest, among them poly(lactic-co-glucolic) acids (PLGAs) and the recently developed silk-tenoin derived materials (STDMs). In accordance with the new philosophy of bio-derived biomaterials (BIODERIBIOs), I describe a novel innovative technology for use in fracture fixation. Screws (2 mm dia.) were manufactured from cylindrical bars of polyglucose-lignol composite (POGLICO) in the form of birch toothpicks from the hospital canteen, dip-coated with alendronate (1 mg/mL, n=6) or saline (n=6), and inserted in the proximal tibias of rats for 4 weeks. Fixation was evaluated by mechanical pullout testing. POGLICO nails were inserted in the contralateral tibia for microCT and histology. All POGLICO implants remained fixed in the bone (p<0.001) with a mean pullout force of 37 (SD 5.5) N. MicroCT showed that the control nails were surrounded by a thin layer of new bone, while all bisphosphonate-treated implants were surrounded by a thick layer of cancellous bone. Bisphosphonates more than doubled the bone density around the nails (p=0.004). POGLICO is biocompatible, remains in situ, and appears to provide a higher resistance to pullout forces than bulk silk protein. The material is light, strong, and bio-derived. BIODERIBIO-POGLICO can be sterilized by autoclaving, and has a porous surface that can serve for slow release of drugs applied by simple dip-coating, as demonstrated by the effect of the alendronate treatment. As the raw material for the screws is readily available from the toothpick industry, I believe that the possibilities for commercial development of the material for fracture fixation are promising.


Therapeutic effects of zerumbone in an alkali-burned corneal wound healing model.
Cornea is an avascular transparent tissue. Ocular trauma caused by a corneal alkali burn induces corneal neovascularization (CNV), inflammation, and fibrosis, leading to vision loss. The purpose of this study was to examine the effects of Zerumbone (ZER) on corneal wound healing caused by alkali burns in mice. CNV was induced by alkali-burn injury in BALB/C female mice. Topical ZER (three times per day, 3μl each time, at concentrations of 5, 15, and 30μM) was applied to treat alkali-burned mouse corneas for 14 consecutive days. Histopathologically, ZER treatment suppressed alkali burn-induced CNV and decreased corneal epithelial defects induced by alkali burns. Corneal tissue treated with ZER showed reduced mRNA levels of pro-angiogenic genes, including vascular endothelial growth factor, matrix metalloproteinase-2 and 9, and pro-fibrotic factors such as alpha smooth muscle actin and transforming growth factor-1 and 2. Immunohistochemical analysis demonstrated that the infiltration of F4/80 and/or CCR2 positive cells was significantly decreased in ZER-treated corneas. ZER markedly inhibited the mRNA and protein levels of monocyte chemoattractant protein-1 (MCP-1) in human corneal fibroblasts and murine peritoneal macrophages. Immunoblot analysis revealed that ZER decreased the activation of signal transducer and activator of transcription 3 (STAT3), with consequent reduction of MCP-1 production by these cells. In conclusion, topical administration of ZER accelerated corneal wound healing by inhibition of STAT3 and MCP-1 production.


Interpretation and estimates of mean residence time with statistical moment theory.
The definitions of mean residence time of drug molecules in the body (MRT) from the literature are reviewed. A formal definition of MRT, based on excretion of drug molecules and amount of drug, a parameter which is independent of constancy of both clearance and volume of distribution, is introduced and compared to other existing definitions of MRTs, that is, MRT for the two-compartment model, MRT for the stochastic model, and MRT based on amount in the body. The inherent assumptions of the methods for determining the various MRTs are discussed. Published information on coefficients and constants (Collier) which describe a biexponential drug decay in the systemic circulation were utilized to illustrate the similarities and differences in MRTs. MRTs were calculated from theoretical relationships as well as estimated from simulated data based on equivalent conditions. The MRTs based on area under the moment curve/area under the curve (AUMC/AUC) for the two-compartment model and AUC/C(0) for the stochastic model assume a constancy in clearance and in volume of distribution, respectively.


Peripheral pulmonary artery stenosis in a four-month-old West Highland White Terrier.
A 4-month-old West Highland White Terrier was evaluated for dyspnea. Severe cardiac silhouette enlargement was present radiographically. Severe right ventricular hypertrophy and dilation with a dilated pulmonary trunk and a stenosis of the right pulmonary artery were diagnosed via echocardiography. Additional areas of peripheral pulmonary artery stenosis were diagnosed with nonselective computed tomography (CT) angiography and selective fluoroscopic angiography. Balloon dilation therapy was unsuccessful and the imaging findings were confirmed at necropsy.


Ethical values in caring encounters on a geriatric ward from the next of kin's perspective: an interview study.
The aim of this study was to identify and describe the governing ethical values that next of kin experience in interaction with nurses who care for elderly patients at a geriatric clinic. Interviews with 14 next of kin were conducted and data were analysed by constant comparative analysis. Four categories were identified: receiving, showing respect, facilitating participation and showing professionalism. These categories formed the basis of the core category: 'Being amenable', a concept identified in the next of kin's description of the ethical values that they and the elderly patients perceive in the caring encounter. Being amenable means that the nurses are guided by ethical values; taking into account the elderly patient and the next of kin. Nurses' focusing on elderly patients' well-being as a final criterion affects the next of kin and their experience of this fundamental condition for high-quality care seems to be fulfilled.


The role of granulocytes in the activation of intravascular coagulation and the precipitation of soluble fibrin by endotoxin.
This study examines the role of neutrophils (PMN) in the pathogenesis of endotoxin-induced microclot formation. It is intended to clarify whether granulocytes are involved in endotoxin-induced activation of intravascular coagulation (generation of soluble fibrin) and/or in endotoxin-induced precipitation of soluble fibrin. Precipitation of soluble fibrin was achieved by injection of endotoxin into ancrod-infused rabbits with circulating soluble fibrin (first model). Activation of intravascular coagulation was elicited by two intravenous injections of endotoxin into rabbits (second model). Seventy-two and ninety-six hours after injection of nitrogen mustard, leukopenic rabbits had PMN counts between 0 and 50 cells per mul. Neutropenia did not prevent the occurrence of glomerular microclots after infusion of ancrod and injection of endotoxin (first model). Neutropenia influenced neither the decrease in mean fibrinogen concentrations nor the drop in mean platelet counts after ancrod and endotoxin administration. In contrast to the first model, neutropenia prevented the occurrence of glomerular microclots and of circulating soluble fibrin after two injections of endotoxin (second model). It did not, however, protect rabbits from the decrease in mean platelet counts after endotoxin administration. These data indicate that granulocytes are involved in endotoxin-induced activation of intravascular coagulation and the production of soluble fibrin but are not essential to endotoxin-induced precipitation of soluble fibrin.


Detection of movement in bed using unobtrusive load cell sensors.
Quality of sleep is an important attribute of an individual's health state and its assessment is therefore a useful diagnostic feature. Changes in the patterns of motor activities during sleep can be a disease marker, or can reflect various abnormal physiological and neurological conditions. Presently, there are no convenient, unobtrusive ways to assess quality of sleep outside of a clinic. This paper describes a system for unobtrusive detection of movement in bed that uses load cells installed at the corners of a bed. The system focuses on identifying when a movement occurs based on the forces sensed by the load cells. The movement detection approach estimates the energy in each load cell signal over short segments to capture the variations caused by movement. The accuracy of the detector is evaluated using data collected in the laboratory. The detector is capable of detecting voluntary movements in bed while the subjects were awake, with an average equal error rate of 3.22% (+/-0.54). Its performance is invariant with respect to the individual's characteristics, e.g., weight, as well as those of the bed. The simplicity of the resulting algorithms and their relative insensitivity to the weight and height of the monitored individual make the approach practical and easily deployable in residential and clinical settings.


Ten ways remote sensing can contribute to conservation.
In an effort to increase conservation effectiveness through the use of Earth observation technologies, a group of remote sensing scientists affiliated with government and academic institutions and conservation organizations identified 10 questions in conservation for which the potential to be answered would be greatly increased by use of remotely sensed data and analyses of those data. Our goals were to increase conservation practitioners' use of remote sensing to support their work, increase collaboration between the conservation science and remote sensing communities, identify and develop new and innovative uses of remote sensing for advancing conservation science, provide guidance to space agencies on how future satellite missions can support conservation science, and generate support from the public and private sector in the use of remote sensing data to address the 10 conservation questions. We identified a broad initial list of questions on the basis of an email chain-referral survey. We then used a workshop-based iterative and collaborative approach to whittle the list down to these final questions (which represent 10 major themes in conservation): How can global Earth observation data be used to model species distributions and abundances? How can remote sensing improve the understanding of animal movements? How can remotely sensed ecosystem variables be used to understand, monitor, and predict ecosystem response and resilience to multiple stressors? How can remote sensing be used to monitor the effects of climate on ecosystems? How can near real-time ecosystem monitoring catalyze threat reduction, governance and regulation compliance, and resource management decisions? How can remote sensing inform configuration of protected area networks at spatial extents relevant to populations of target species and ecosystem services? How can remote sensing-derived products be used to value and monitor changes in ecosystem services? How can remote sensing be used to monitor and evaluate the effectiveness of conservation efforts? How does the expansion and intensification of agriculture and aquaculture alter ecosystems and the services they provide? How can remote sensing be used to determine the degree to which ecosystems are being disturbed or degraded and the effects of these changes on species and ecosystem functions?


Race and colorectal cancer disparities: health-care utilization vs different cancer susceptibilities.
It is unclear whether the disproportionately higher incidence and mortality from colorectal cancer among blacks compared with whites reflect differences in health-care utilization or colorectal cancer susceptibility. A total of 60, 572 non-Hispanic white and black participants in the ongoing Prostate, Lung, Colorectal, and Ovarian (PLCO) Cancer Screening Trial underwent trial-sponsored screening flexible sigmoidoscopy (FSG) without biopsy at baseline in 10 geographically dispersed centers from November 1993 to July 2001. Subjects with polyps or mass lesions detected by FSG were referred to their physicians for diagnostic workup, the cost of which was not covered by PLCO. The records of follow-up evaluations were collected and reviewed. We used log binomial modeling with adjustment for age, education, sex, body mass index, smoking, family history of colorectal cancer, colon examination within previous 3 years, personal history of polyps, and screening center to examine whether utilization of diagnostic colonoscopy and yield of neoplasia differed by race. Among 57 561 whites and 3011 blacks who underwent FSG, 13,743 (23.9%) and 767 (25.5%) had abnormal examinations, respectively. A total of 9944 (72.4%) whites and 480 (62.6%) blacks had diagnostic colonoscopy within 1 year following the abnormal FSG screening. When compared with whites, blacks were less likely to undergo diagnostic evaluation (adjusted risk ratio = 0.88, 95% confidence interval = 0.83 to 0.93). Overall, among subjects with diagnostic colonoscopy (n = 10 424), there was no statistically significant difference by race in the prevalence of adenoma, advanced adenoma, advanced pathology in small adenomas (high-grade dysplasia or villous histology in adenomas <10 mm), or colorectal cancer. We observed a lower follow-up for screen-detected abnormalities among blacks when compared with whites but little difference in the yield of colorectal neoplasia. Health-care utilization may be playing more of a role in colorectal cancer racial disparity than biology.


Dissecting the molecular network of virus-plant interactions: the complex roles of host factors.
A successful infection by a plant virus results from the complex molecular interplay between the host plant and the invading virus. Thus, dissecting the molecular network of virus-host interactions advances the understanding of the viral infection process and may assist in the development of novel antiviral strategies. In the past decade, molecular identification and functional characterization of host factors in the virus life cycle, particularly single-stranded, positive-sense RNA viruses, have been a research focus in plant virology. As a result, a number of host factors have been identified. These host factors are implicated in all the major steps of the infection process. Some host factors are diverted for the viral genome translation, some are recruited to improvise the viral replicase complexes for genome multiplication, and others are components of transport complexes for cell-to-cell spread via plasmodesmata and systemic movement through the phloem. This review summarizes current knowledge about host factors and discusses future research directions.


Can out-of-hours operating in gynaecology be reduced?
Against a background of concerns about future hospital medical staffing and the safety of unsupervised operations at night, this study examines which gynaecological operations now undertaken at night may be safely postponed until the following day. In the first phase of the study, the operations taking place at night in four hospitals were identified. In the second phase a nominal group technique was used to seek the extent of agreement among professionals about the appropriateness of postponing surgery to the following day in certain circumstances. There were considerable variations between hospitals in the volume of work and the grade of staff involved. The panel concluded that most operations now performed at night should not be postponed. This study suggests that there is limited scope for postponing gynaecological operations currently undertaken at night.


Predator diversity and abundance provide little support for the enemies hypothesis in forests of high tree diversity.
Predatory arthropods can exert strong top-down control on ecosystem functions. However, despite extensive theory and experimental manipulations of predator diversity, our knowledge about relationships between plant and predator diversity--and thus information on the relevance of experimental findings--for species-rich, natural ecosystems is limited. We studied activity abundance and species richness of epigeic spiders in a highly diverse forest ecosystem in subtropical China across 27 forest stands which formed a gradient in tree diversity of 25-69 species per plot. The enemies hypothesis predicts higher predator abundance and diversity, and concomitantly more effective top-down control of food webs, with increasing plant diversity. However, in our study, activity abundance and observed species richness of spiders decreased with increasing tree species richness. There was only a weak, non-significant relationship with tree richness when spider richness was rarefied, i.e. corrected for different total abundances of spiders. Only foraging guild richness (i.e. the diversity of hunting modes) of spiders was positively related to tree species richness. Plant species richness in the herb layer had no significant effects on spiders. Our results thus provide little support for the enemies hypothesis--derived from studies in less diverse ecosystems--of a positive relationship between predator and plant diversity. Our findings for an important group of generalist predators question whether stronger top-down control of food webs can be expected in the more plant diverse stands of our forest ecosystem. Biotic interactions could play important roles in mediating the observed relationships between spider and plant diversity, but further testing is required for a more detailed mechanistic understanding. Our findings have implications for evaluating the way in which theoretical predictions and experimental findings of functional predator effects apply to species-rich forest ecosystems, in which trophic interactions are often considered to be of crucial importance for the maintenance of high plant diversity.


Cytochrome P450 2E1 is a cell surface autoantigen in halothane hepatitis.
Recent studies have shown that cytochrome P450 2E1 (CYP2E1) is a major catalyst of formation of trifluoroacetylated proteins, which have been implicated as target antigens in the mechanism of halothane hepatitis. In the present investigation, trifluoroacetylated CYP2E1 was detected immunochemically in livers of rats treated with halothane. Furthermore, high levels of autoantibodies that recognized purified rat CYP2E1 but not purified rat CYP3A were detected by enzyme-linked immunosorbent assay in 14 of 20 (70%) sera from patients with halothane hepatitis. Only very low levels of such antibodies were detected in sera from healthy controls, from patients anesthetized with halothane without developing hepatitis, or from patients with other liver diseases. The intracellular distribution of CF3CO-adducts was studied in highly differentiated FGC4 rat hepatoma cell cultures. High levels of adducts were found after 22-hr culture in the presence of halothane, and their generation was dependent on the expression of CYP2E1. Adducts were predominantly located in the endoplasmic reticulum but also, to a minor extent, on the cell surface, as detected by immunofluorescence. A very similar distribution was found for CYP2E1 in FGC4 cells, and immunoprecipitation experiments performed in cultures of FGC4-related Fao hepatoma cells suggest that surface immunoreactivity originates from a small fraction of intact CYP2E1 apoprotein. Human CYP2E1, expressed in V79 cells after cDNA transfection, was also detected to a minor extent in the plasma membrane, whereas no immunofluorescence was evident in parental V79 cells. It is suggested that immune responses to cell surface CYP2E1 could be involved in the pathogenesis of halothane hepatitis.


A caspase-6-cleaved fragment of Glial Fibrillary Acidic Protein as a potential serological biomarker of CNS injury after cardiac arrest.
Blood levels of Glial Fibrillary Acidic protein (GFAP) reflect processes associated with different types of CNS injury. Evidence suggests that GFAP is cleaved by caspases during CNS injury, hence positioning GFAP fragments as potential biomarkers of injury-associated processes. We set out to develop an assay detecting the neo-epitope generated by caspase-6 cleavage of GFAP (GFAP-C6), and to assess the ability of GFAP-C6 to reflect pathological processes in patients suffering a cardiac arrest and subsequent global cerebral ischemia. Anti-GFAP-C6 antibodies recognized their specific target sequence, and dilution and spike recoveries in serum were within limits of ±20% reflecting high precision and accuracy of measurements. Intra- and inter-assay CVs were below limits of 10% and 15%, respectively. Serological levels of GFAP-C6 were significantly elevated 72 hours after CA (Mean±SD) (20.39±10.59 ng/mL) compared to time of admission (17.79±10.77 ng/mL, p<0.0001), 24 hours (17.40±7.99 ng/mL, p<0.0001) and 48 hours (17.87±8.56 ng/mL, p<0.0001) after CA, but were not related to neurological outcome at day 180. GFAP-C6 levels at admission, 24, 48, and 72 hours after cardiac arrest correlated with two proteolytic fragments of tau, tau-A (r = 0.30, r = 0.40, r = 0.50, r = 0.53, p < 0.0001) and tau-C (r = 54, r = 0.48, r = 0.55, r = 0.54, p < 0.0001), respectively. GFAP-C6 levels did not correlate with other markers of CNS damage; total tau, NSE and S100B. In conclusion, we developed the first assay detecting a caspase-6 cleaved fragment of GFAP in blood. Increased levels at 72 hours after cardiac arrest as well as moderate correlations between GFAP-C6 and two other blood biomarkers of neurodegeneration suggest the ability of GFAP-C6 to reflect pathological processes of the injured brain. Investigations into the potential of GFAP-C6 in other types of CNS injury are warranted.


Vaccinia virus hemagglutinin. A novel member of the immunoglobulin superfamily.
Striking similarities between vaccinia virus hemagglutinin (VVHA) and proteins belonging to the Ig superfamily clearly indicate that VVHA, a 315-amino acid glycoprotein expressed on the surface of the infected cells, is a novel viral protein that can be added to the expanding list of the Ig superfamily. Its deduced amino acid sequence contains one Ig-like domain at the NH2 terminus, followed by two tandem repeating units and a hydrophobic region, suggestive of membrane spanning. The results offer an opportunity for the further study of the probable evolutionary and possible functional relationship between VVHA and other members of the Ig superfamily. Our observation, together with a recent finding that human CMV possibly encodes a protein similar to the MHC class I antigens (13), provides evidence supporting the fact that the viral capture of cellular Ig-related genes is more common than expected in vaccinia and other viruses, and that usage of an Ig-like domain as recognition signals might be extended from higher animals to animal viruses.


[Infantile epileptic encephalopathies: what matters is genetics].
Epileptic encephalopathies in infancy are defined as conditions where the sustained epileptic activity itself may contribute to the severe neurological and cognitive impairment. These epileptic encephalopathies include Ohtahara syndrome, early myoclonic epileptic encephalopathy, West syndrome, Dravet syndrome, and malignant migrating epilepsy in infancy. These syndromes result from identifiable primary causes, such as structural, neurodegenerative, metabolic, or genetic defects. To present and discuss current knowledge regarding genetic findings in epileptic encephalopathies in infancy, phenotype-genotype correlations in different forms of paediatric epileptic encephalopathies, and the impact of these new findings in clinical practice. Patients with unclear etiologies after performing a brain magnetic resonance imaging should be considered for a further workup, which should include an evaluation for genetic defects. Nowadays, more than 50 genes have been associated with epileptic encephalopathies in infancy. Targeted next-generation sequencing panels show a high diagnostic yield in patients with epileptic encephalopathies. Genetic knowledge about epileptic encephalopathies in infancy has revolutionized the diagnostic approach to these disorders, and an increasing number of gene mutations have been related to their pathogenesis. A more detailed classification of epileptic encephalopathies genotypes will improve the accuracy of genotype-phenotype correlation and genetic counseling. All these developments could yield therapeutic applications such as gene therapy or antiepileptic drugs 'tailored' to the specific genetic markers or targets.


The content of our cooperation, not the color of our skin: an alliance detection system regulates categorization by coalition and race, but not sex.
Humans in all societies form and participate in cooperative alliances. To successfully navigate an alliance-laced world, the human mind needs to detect new coalitions and alliances as they emerge, and predict which of many potential alliance categories are currently organizing an interaction. We propose that evolution has equipped the mind with cognitive machinery that is specialized for performing these functions: an alliance detection system. In this view, racial categories do not exist because skin color is perceptually salient; they are constructed and regulated by the alliance system in environments where race predicts social alliances and divisions. Early tests using adversarial alliances showed that the mind spontaneously detects which individuals are cooperating against a common enemy, implicitly assigning people to rival alliance categories based on patterns of cooperation and competition. But is social antagonism necessary to trigger the categorization of people by alliance--that is, do we cognitively link A and B into an alliance category only because they are jointly in conflict with C and D? We report new studies demonstrating that peaceful cooperation can trigger the detection of new coalitional alliances and make race fade in relevance. Alliances did not need to be marked by team colors or other perceptually salient cues. When race did not predict the ongoing alliance structure, behavioral cues about cooperative activities up-regulated categorization by coalition and down-regulated categorization by race, sometimes eliminating it. Alliance cues that sensitively regulated categorization by coalition and race had no effect on categorization by sex, eliminating many alternative explanations for the results. The results support the hypothesis that categorizing people by their race is a reversible product of a cognitive system specialized for detecting alliance categories and regulating their use. Common enemies are not necessary to erase important social boundaries; peaceful cooperation can have the same effect.


Urinary and serum biomarkers for the diagnosis of acute kidney injury: an in-depth review of the literature.
Acute kidney injury (AKI) remains associated with high morbidity and mortality, despite progress in medical care. Although the RIFLE (Risk, Injury, Failure, Loss, End-Stage Kidney Disease) and AKIN (Acute Kidney Injury Network) criteria, based on serum creatinine and urine output, were a step forward in diagnosing AKI, a reliable tool to differentiate between true parenchymal and pre-renal azotaemia in clinical practice is still lacking. In the last decade, many papers on the use of new urinary and serum biomarkers for the diagnosis and prognostication of AKI have been published. Thus, the question arises which biomarker is a reliable differential diagnostic tool under which circumstances. We searched Medline from inception to April 2012 using medical subject heading and text words for AKI and biomarkers [neutrophil gelatinase-associated lipocalin (NGAL), kidney injury molecule-1 (KIM-1), Cystatin C, interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-18 (IL-18), N-acetyl-glucosaminidase (NAG), glutathione transferases (GST) and liver fatty acid binding protein (LFABP)] to identify relevant papers in five different settings (paediatrics, cardiac surgery, emergency department, critically ill and contrast-induced nephropathy). We included 87 relevant papers, reporting on 74 studies. Depending upon the setting, 7-27 different definitions of AKI were used. Reported diagnostic performance of the different biomarkers was variable from poor to excellent, and no consistent generalizable conclusions can be drawn on their diagnostic value. Early diagnosing of AKI in clinical conditions by using new serum and urinary biomarkers remains cumbersome, especially in those settings where timing and aetiology of AKI are not well defined. Putting too much emphasis on markers that have not convincingly proven reliability might lead to incorrect interpretation of clinical trials. Further research in this field is warranted before biomarkers can be introduced in clinical practice.


Implementing a standardized testing program: preparing students for the NCLEX-RN.
The undergraduate nursing faculty of a large Midwestern university initiated a program of standardized computerized testing for two purposes: to provide students experience with standardized computerized testing prior to taking the National Council Licensure Exam for Registered Nurses (NCLEX-RN) and to increase the students' NCLEX-RN passing rate. This article chronicles the process of implementing a comprehensive testing program developed by the Assessment Technologies Institute (Overland Park, KS). Although the first class of students to have taken the entire testing package has just graduated, midprogram results have demonstrated potential as key indicators for identifying at-risk students. The trends in scores on standardized computerized tests, grades in prerequisite science courses, and grades in medical-surgical courses are used to identify students who are at risk for failure in the program and on the NCLEX-RN. Faculty advisors meet with these students to develop individual plans of study and to provide additional resources. The testing process is going on smoothly, and faculty members are learning to use the extensive information on students' test scores to further assist them in passing the NCLEX-RN.


Interim report of the Medical Research Council/Royal College of Obstetricians and Gynaecologists multicentre randomized trial of cervical cerclage. MRC/RCOG Working Party on Cervical Cerclage.
Overall 905 pregnant women whose obstetricians were 'uncertain' whether to recommend cervical cerclage, chiefly because of a history of early delivery or cervical surgery, were randomly allocated to cerclage or no surgery; 92% were treated as allocated. The overall preterm delivery rate was 30%. The results for those allocated cerclage were marginally statistically significant, more favourable in terms of fewer deliveries before 33 weeks [59 (13%) compared with 82 (18%), P = 0.03] and correspondingly for birthweight under 1500 g [48 (11%) compared with 73 (16%), P = 0.01] and for miscarriage, stillbirth or neonatal death [37 (8%) compared with 54 (12%), P = 0.06]. There were similar numbers of deliveries between 33 and 36 weeks [65 (14%) compared with 64 (14%)]. These results suggest that the operation had an important beneficial effect in one in 20 to 25 cases in the trial. But because the observed differences are not strongly statistically significant and because no such benefit has been seen in other randomized trials, there remains uncertainty about how much (if any) of this apparent benefit is real. So, the trial still remains open for randomization of more women whose obstetricians are uncertain about the advisability of cerclage.


Bio-inspired protonic memristor devices based on metal complexes with proton-coupled electron transfer.
A new type of memristor inspired by bio-membranes is presented, based on the proton movement resulting from proton-coupled electron transfer (PCET) processes in dinuclear Ru complexes, whereby a two-terminal device based on said Ru complexes and a proton-conducting polymer was constructed as a proof-of-concept. Two ITO electrodes were modified separately with dinuclear Ru complexes that bear tetraphosphonic acid linkers at both ends and a 2,6,2',6'-tetrakis(benzimidazol-2-yl)-4,4'-bipyridine (RuNH-OH) or 1,3,1',3'-tetrakis(benzimidazol-2-yl)-5,5'-biphenyl (RuCH-OH) bridging ligand, and both ITO electrodes exhibit PCET processes with different Ru(ii/iii) redox potentials and pKa values. Poly(4-vinylpyridine) (P4VP; pKa = 4-5), a proton-conducting polymer, was sandwiched between the two modified ITO electrodes to construct a two-terminal device of the type ITO|(RuNH-OH)3|P4VP|(RuCH-OH)3|ITO. Initially, the oxidation state of the metal centers in RuNH-OH and RuCH-OH is Ru(ii) and Ru(iii), respectively. Upon applying a bias voltage between the two ITO electrodes, the high and low current states switch at approximately ±1.10 V due to Ru(ii/iii) redox reactions. At the RuNH-OH|P4VP and RuCH-OH|P4VP interfaces, a proton is released from Ru(ii)NH-OH and subsequently captured by Ru(iii)CH-OH through the hydrogen-bonding interaction with the P4VP polymer, which is driven by the changes in the pKa values of the Ru complexes from 4.1-8.8 [Ru(ii)NH-OH] to <3.8 [Ru(iii)NH-OH] and from <8.4 [Ru(ii)CH-OH] to 5.2-9.8 [Ru(iii)CH-OH] under these conditions. The redox reactions on the modified Ru films create a large proton gradient between the two electrodes, enhancing the proton conductivity through the P4VP layer (pKa = 4-5). When the applied bias potential was inverted, the pKa gradient returned to the original state and the current decreased. Such a proton-conductivity enhancement is relevant to the transport of protons by proton gradients in bio-membranes. Therefore, the present protonic coordination-network films containing metal complexes that exhibit PCET should open new avenues for the design of a new type of memristor devices mimicking the function of synapses.


Reconstruction of canine diffuse large B-cell lymphoma gene regulatory network: detection of functional modules and hub genes.
Lymphoma is one of the most common malignancies in dogs. Canine lymphoma is similar to human non-Hodgkin's lymphoma (NHL) with shared clinical presentation and histopathological features. This study reports the construction of a comprehensive gene regulatory network (GRN) for canine diffuse large B-cell lymphoma (DLBCL), the most common type of canine lymphoma, and performs analysis for detection of major functional modules and hub genes (the most important genes in a GRN). The canine DLBCL GRN was reconstructed from gene expression data (NCBI GEO dataset: GSE30881) using the STRING and MiMI interaction databases. Reconstructed GRNs were then assessed, using various bioinformatics programmes, in order to analyze network topology and identify major pathways and hub genes. The resultant network from both interaction databases had a logically scale-free pattern. Gene ontology (GO) analysis revealed cell activation, cell cycle phase, immune effector process, immune system development, immune system process, integrin-mediated signalling pathway, intracellular protein kinase cascade, intracellular signal transduction, leucocyte activation and differentiation, lymphocyte activation and differentiation as major GO terms in the biological processes of the networks. Moreover, bioinformatics analysis showed E2F1, E2F4, PTEN, CDKN1A, PCNA, DKC1, MNAT1, NDUFB4, ATP5J, PRKDC, BRCA1, MYCN, RFC4 and POLA1 as the most important hub genes. The phosphatidyl inositol signalling system, P53 signalling pathway, Rac CycD pathway, G1/S checkpoint, chemokine signalling pathway and telomere maintenance were the main signalling pathways in which the protein products of the hub genes are involved.


Semiclassical scattering of an electric dipole source inside a spherical particle.
Semiclassical scattering phenomena appearing in the far-zone scattered intensity of a point source of electromagnetic radiation inside a spherical particle are examined in the context of both ray theory and wave theory, and the evolution of the phenomena is studied as a function of source position. A number of semiclassical effects that do not occur for plane-wave scattering by the sphere appear prominently for scattering by an interior source. These include a series of scattering resonances and a new family of rainbows in regions of otherwise total internal reflection. Diffractive effects accompanying the semiclassical phenomena are also examined.


Next Generation Sequencing Analysis Reveals Segmental Patterns of microRNA Expression in Mouse Epididymal Epithelial Cells.
The functional maturation of mammalian spermatozoa is accomplished as the cells descend through the highly specialized microenvironment of the epididymis. This dynamic environment is, in turn, created by the combined secretory and absorptive activity of the surrounding epithelium and displays an extraordinary level of regionalization. Although the regulatory network responsible for spatial coordination of epididymal function remains unclear, recent evidence has highlighted a novel role for the RNA interference pathway. Indeed, as noncanonical regulators of gene expression, small noncoding RNAs have emerged as key elements of the circuitry involved in regulating epididymal function and hence sperm maturation. Herein we have employed next generation sequencing technology to profile the genome-wide miRNA signatures of mouse epididymal cells and characterize segmental patterns of expression. An impressive profile of some 370 miRNAs were detected in the mouse epididymis, with a subset of these specifically identified within the epithelial cells that line the tubule (218). A majority of the latter miRNAs (75%) were detected at equivalent levels along the entire length of the mouse epididymis. We did however identify a small cohort of miRNAs that displayed highly regionalized patterns of expression, including miR-204-5p and miR-196b-5p, which were down- and up-regulated by approximately 39- and 45-fold between the caput/caudal regions, respectively. In addition we identified 79 miRNAs (representing ~ 21% of all miRNAs) as displaying conserved expression within all regions of the mouse, rat and human epididymal tissue. These included 8/14 members of let-7 family of miRNAs that have been widely implicated in the control of androgen signaling and the repression of cell proliferation and oncogenic pathways. Overall these data provide novel insights into the sophistication of the miRNA network that regulates the function of the male reproductive tract.


Revisiting the Cape Cod bacteria injection experiment using a stochastic modeling approach.
Bromide and resting-cell bacteria tracer tests conducted in a sandy aquifer at the U.S. Geological Survey Cape Cod site in 1987 were reinterpreted using a three-dimensional stochastic approach. Bacteria transport was coupled to colloid filtration theory through functional dependence of local-scale colloid transport parameters upon hydraulic conductivity and seepage velocity in a stochastic advection-dispersion/attachment-detachment model. Geostatistical information on the hydraulic conductivity (K) field that was unavailable at the time of the original test was utilized as input. Using geostatistical parameters, a groundwater flow and particle-tracking model of conservative solute transport was calibrated to the bromide-tracer breakthrough data. An optimization routine was employed over 100 realizations to adjust the mean and variance of the natural-logarithm of hydraulic conductivity (InK) field to achieve best fit of a simulated, average bromide breakthrough curve. A stochastic particle-tracking model for the bacteria was run without adjustments to the local-scale colloid transport parameters. Good predictions of mean bacteria breakthrough were achieved using several approaches for modeling components of the system. Simulations incorporating the recent Tufenkji and Elimelech (Environ. Sci. Technol. 2004, 38, 529-536) correlation equation for estimating single collector efficiency were compared to those using the older Rajagopalan and Tien (AIChEJ. 1976, 22, 523-533) model. Both appeared to work equally well at predicting mean bacteria breakthrough using a constant mean bacteria diameter for this set of field conditions. Simulations using a distribution of bacterial cell diameters available from original field notes yielded a slight improvement in the model and data agreement compared to simulations using an average bacterial diameter. The stochastic approach based on estimates of local-scale parameters for the bacteria-transport process reasonably captured the mean bacteria transport behavior and calculated an envelope of uncertainty that bracketed the observations in most simulation cases.


2D materials for renewable energy storage devices: Outlook and challenges.
Scientists are looking for cost-effective, clean and durable alternative energy devices. Superior charge storage devices can easily meet the demands of our daily needs. In this respect, a material with suitable dimensions for charge storage devices has been considered to be very important. Improved performance of charge storage devices has been derived from whole-body participation and the best are from 2D materials, which provide a viable and acceptable solution.


Bar-coded pyrosequencing reveals shared bacterial community properties along the temperature gradients of two alkaline hot springs in Yellowstone National Park.
An understanding of how communities are organized is a fundamental goal of ecology but one which has historically been elusive for microbial systems. We used a bar-coded pyrosequencing approach targeting the V3 region of the bacterial small-subunit rRNA gene to address the factors that structure communities along the thermal gradients of two alkaline hot springs in the Lower Geyser Basin of Yellowstone National Park. The filtered data set included a total of nearly 34,000 sequences from 39 environmental samples. Each was assigned to one of 391 operational taxonomic units (OTUs) identified by their unique V3 sequence signatures. Although the two hot springs differed in their OTU compositions, community resemblance and diversity changed with strikingly similar dynamics along the two outflow channels. Two lines of evidence suggest that these community properties are controlled primarily by environmental temperature. First, community resemblance decayed exponentially with increasing differences in temperature between samples but was only weakly correlated with physical distance. Second, diversity decreased with increasing temperature at the same rate along both gradients but was uncorrelated with other measured environmental variables. This study also provides novel insights into the nature of the ecological interactions among important taxa in these communities. A strong negative association was observed between cyanobacteria and the Chloroflexi, which together accounted for approximately 70% of the sequences sampled. This pattern contradicts the longstanding hypothesis that coadapted lineages of these bacteria maintain tightly cooccurring distributions along these gradients as a result of a producer-consumer relationship. We propose that they instead compete for some limiting resource(s).


Sleep after Heavy Alcohol Consumption and Physical Activity Levels during Alcohol Hangover.
Alcohol consumption can negatively affect sleep quality. The current study examined the impact of an evening of alcohol consumption on sleep, and next day activity levels and alcohol hangover. n = 25 healthy social drinkers participated in a naturalistic study, consisting of an alcohol and alcohol-free test day. On both days, a GENEactiv watch recorded sleep and wake, and corresponding activity levels. In addition, subjective assessments of sleep duration and quality were made, and hangover severity, and the amount of consumed alcoholic beverages were assessed. Alcohol consumption was also assessed in real-time during the drinking session, using smartphone technology. The results confirmed, by using both objective and subjective assessments, that consuming a large amount of alcohol has a negative impact on sleep, including a significant reduction in objective sleep efficiency and significantly lower self-reported sleep quality. Activity levels during the hangover day were significantly reduced compared to the alcohol-free control day. Of note, next-morning retrospective alcohol consumption assessments underestimated real-time beverage recordings. In conclusion, heavy alcohol consumption impairs sleep quality, which is associated with increased next day hangover severity and reduced activity levels. The outcome of this study underlines that, in addition to retrospectively reported data, real-time objective assessments are needed to fully understand the effects of heavy drinking.


Nutritional significance and health benefits of omega-3, -6 and -9 fatty acids in animals.
The balance between omega-3 (ω-3), omega-6 (ω-6) and omega-9 (ω-9) fatty acids (FAs) is very important because these types of oils constitute essential components for the formation of the cell membrane, also they are precursors for a large number of substances in the body. One of the most important strategies for improving the increment of polyunsaturated FAs in poultry and animal meat is the dietary administration of these FAs. Additionally, the different sources of ω-3 or 6 in the diet improve the performance, public health and physiological aspects including anti-oxidative properties and immunity. ω-3 FAs have anti-inflammatory characteristics due to their ability to reduce cytokines liberation. High-level of ω-6 FAs is always associated with an increased incidence of dangerous disorders like depression and heart disease. These FAs showed a tremendous series of beneficial impacts like improved cholesterol levels and a decreased occurrence of coronary heart diseases. This article includes some information on the use of ω-3, ω-6 and ω-9 FAs in animal and human diets. These oils are vital for the physiological and health aspects, and the information mentioned here will improve our understanding of the functions and roles of these FAs in the body.


Coexistence of eating disorders and autoimmune diseases: Record linkage cohort study, UK.
Recent research indicates that eating disorders (ED) are associated with type 1 diabetes and Crohn's disease. The aim of this study was to determine whether, in a hospitalized population, a range of autoimmune diseases (AIDs) occurred more often than expected in people with anorexia nervosa (AN) or bulimia nervosa (BN), and whether AIDs elevated the risk of ED. Retrospective, record-linkage cohort study using national administrative statistical data on hospital care and mortality in England, 1999-2011. In people admitted when aged 10-44, cohorts of 8,700 females and 651 males with AN, and 4,783 females and 330 males with BN were constructed, along with a control cohort with the same age range. Results were expressed as risk ratios comparing each ED cohort with the control cohort. The overall rate ratio for an AID after admission for AN was 2.04 (95% confidence interval 1.81-2.28) in females, and 1.14 (0.37-2.67) in males; and, for BN, 1.83 (1.56-2.14) in females, and 4.41 (2.11-8.10) in males. Rate ratios for AN after admission for an AID were 3.34 (2.94-3.79) in females, 3.76 (2.06-6.53) in males; and those for BN were 2.57 (2.22-2.97) in females, and 3.10 (1.50-5.90) in males. There were significant associations between ED and several specific individual AIDs. Strong associations between ED and specific AIDs exist, although it is not possible from this study to determine if these are causal. Clinicians should be aware of the co-occurrence of these conditions. © 2016 Wiley Periodicals, Inc.(Int J Eat Disord 2016; 49:663-672).


Surface-sustained permanent gratings in nematic liquid crystals doped with carbon nanotubes.
Response properties of photoinduced gratings were studied in planar cells of liquid crystal doped with multiwalled carbon nanotubes. The grating formation time deduced from beam-coupling measurements was obtained to be ~30 ms. Absorption spectroscopy implies that the permanent gratings are associated with periodically distributed carbonaceous material adsorbed on the inner surfaces of the cell windows under prolonged illumination to the 514.5-nm beams, giving rise to the persistent lightinduced modulation of the easy axis.


Less increase of copeptin and MR-proADM due to intervention with selenium and coenzyme Q10 combined: Results from a 4-year prospective randomized double-blind placebo-controlled trial among elderly Swedish citizens.
Intervention with selenium and coenzyme Q10 have recently been found to reduce mortality and increase cardiac function. The mechanisms behind these effects are unclear. As selenium and coenzyme Q10 is involved in the anti-oxidative defence, the present study aimed to evaluate effects of selenium and coenzyme Q10 on copeptin and adrenomedullin as oxidative stress biomarkers. Therefore 437 elderly individuals were included and given intervention for 4 years. Clinical examination and blood samples were undertaken at start and after 18 and 48 months. Evaluations of copeptin and MR-proADM changes were performed using repeated measures of variance. Cardiovascular mortality was evaluated using a 10-year-period of follow-up, and presented in Kaplan-Meier plots. A significant increase in copeptin level could be seen in the placebo group during the intervention period (from 9.4 pmol/L to 15.3 pmol/L), compared to the active treatment group. The difference between the groups was confirmed in the repeated measurement of variance analyses (P = 0.031) with less copeptin increase in the active treatment group. Furthermore, active treatment appeared to protect against cardiovascular death both in those with high and with low copeptin levels at inclusion. Less increase of MR-proADM could also be seen during the intervention in the active treatment group compared to controls (P = 0.026). Both in those having an MR-proADM level above or below median level, significantly less cardiovascular mortality could be seen in the active treatment group (P = 0.0001, and P = 0.04 respectively). In conclusion supplementation with selenium and coenzyme Q10 during four years resulted in less concentration of both copeptin and MR-proADM. A cardioprotective effect of the supplementation was registered, irrespective of the initial levels of these biomarkers, and this protection was recognized also after 10 years of observation.


Functional attributes of epilithic diatoms for palaeoenvironmental interpretations in South-West Greenland lakes.
Benthic diatoms are commonly used for palaeoenvironmental reconstruction in Arctic regions, but interpretation of their ecology remains challenging. We studied epilithic diatom assemblages from the shallow margins of 19 lakes from three areas (coast-inland-ice sheet margin) along a climate gradient in Kangerlussuaq, West Greenland during two periods; shortly after ice-off (spring) and in the middle of the growth season (summer). We aimed to understand the distribution of Arctic epilithic diatoms in relation to water chemistry gradients during the two seasons, to investigate their incorporation into lake sediments and to assess their applicability as palaeoenvironmental indicators. Diatoms were correlated with nutrients in the spring and alkalinity/major ions in the summer, when nutrients were depleted; approximately half of the variance explained was independent of spatial factors. When categorised by functional attributes, diatom seasonal succession differed among regions with the most obvious changes in inland lakes where summer temperatures are warmer, organic nutrient processing is prevalent and silicate is limiting. These conditions led to small, motile and adnate diatoms being abundant in inland lakes during the summer (Nitzschia spp., Encyonopsis microcephala), as these functional attributes are suited to living within complex mats of non-siliceous microbial biofilms. Seasonal succession in silica-rich lakes at the coast was less pronounced and assemblages included Tabellaria flocculosa (indicating more acidic conditions) and Hannaea arcus (indicating input from inflowing rivers). The nitrogen-fixing diatom Epithemia sorex increased from the coast to the ice sheet, negatively correlating with a gradient of reactive nitrogen. The presence of this diatom in Holocene sediment records alongside cyanobacterial carotenoids during arid periods of low nitrogen delivery, suggests that it is a useful indicator of nitrogen limitation. Nitzschia species appear to be associated with high concentrations of organic carbon and heterotrophy, but their poor representation in West Greenland lake sediments due to taphonomic processes limits their palaeoenvironmental application in this region. Proportions of epilithic taxa in lake sediment records of coastal lakes increased during some wetter periods of the Holocene, suggesting that snowpack-derived nutrient delivery may offer diatom taxa living at lake margins a competitive advantage over planktonic diatoms during the "moating" ice melt period. Thus, further research investigating linkages between epilithic diatoms, snowpack and nutrient delivery in seasonally frozen lakes is recommended as these taxa live on the 'front-line' during the spring and may be especially sensitive to changes in snowmelt conditions.


Seven regions of the genome show evidence of linkage to type 1 diabetes in a consensus analysis of 767 multiplex families.
Type 1 diabetes (T1D) is a genetically complex disorder of glucose homeostasis that results from the autoimmune destruction of the insulin-secreting cells of the pancreas. Two previous whole-genome scans for linkage to T1D in 187 and 356 families containing affected sib pairs (ASPs) yielded apparently conflicting results, despite partial overlap in the families analyzed. However, each of these studies individually lacked power to detect loci with locus-specific disease prevalence/sib-risk ratios (lambda(s)) <1.4. In the present study, a third genome scan was performed using a new collection of 225 multiplex families with T1D, and the data from all three of these genome scans were merged and analyzed jointly. The combined sample of 831 ASPs, all with both parents genotyped, provided 90% power to detect linkage for loci with lambda(s) = 1.3 at P=7.4x10(-4). Three chromosome regions were identified that showed significant evidence of linkage (P<2.2x10(-5); LOD scores >4), 6p21 (IDDM1), 11p15 (IDDM2), 16q22-q24, and four more that showed suggestive evidence (P<7.4x10(-4), LOD scores > or =2.2), 10p11 (IDDM10), 2q31 (IDDM7, IDDM12, and IDDM13), 6q21 (IDDM15), and 1q42. Exploratory analyses, taking into account the presence of specific high-risk HLA genotypes or affected sibs' ages at disease onset, provided evidence of linkage at several additional sites, including the putative IDDM8 locus on chromosome 6q27. Our results indicate that much of the difficulty in mapping T1D susceptibility genes results from inadequate sample sizes, and the results point to the value of future international collaborations to assemble and analyze much larger data sets for linkage in complex diseases.


EPS8 Facilitates Uncoating of Influenza A Virus.
All viruses balance interactions between cellular machinery co-opted to support replication and host factors deployed to halt the infection. We use gene correlation analysis to perform an unbiased screen for host factors involved in influenza A virus (FLUAV) infection. Our screen identifies the cellular factor epidermal growth factor receptor pathway substrate 8 (EPS8) as the highest confidence pro-viral candidate. Knockout and overexpression of EPS8 confirm its importance in enhancing FLUAV infection and titers. Loss of EPS8 does not affect virion attachment, uptake, or fusion. Rather, our data show that EPS8 specifically functions during virion uncoating. EPS8 physically associates with incoming virion components, and subsequent nuclear import of released ribonucleoprotein complexes is significantly delayed in the absence of EPS8. Our study identifies EPS8 as a host factor important for uncoating, a crucial step of FLUAV infection during which the interface between the virus and host is still being discovered.


Evaluation of the Effect of Frequency-Shifting Technique Used in Hearing Aids for Speech Discrimination.
To evaluate the effect of frequency-shifting techniques recently used in hearing aids for speech discrimination scores and hearing thresholds. Thirty individuals (16 men and 14 women) with sensorineural hearing loss who used normal fitting monaural hearing aids and frequency-shifting feature for at least 2 months and whose 4000-8000 Hz hearing threshold was over 70 dB participated. The average age was 69.73 ± 10 (range: 65-80 years). We detected the types and degree of each participant's hearing loss. Measurements with and without hearing aids were made in a free field. For speech discrimination scores, 6 listings consisting of 25 monosyllables have been used. It has been avoided to learn words thanks to presentation of words in different listings to subjects. Pure-tone averages of the participants were measured using a supra-aural headphone, normal fitting hearing aid, and frequency-shifting feature. The results were 55.93 ± 6.89, 40.47 ± 5.48, and 36.73 ± 5.72 dB, respectively (p˂0.05). Speech discrimination scores for hearing aids worn on the right ear were measured as 67.73 ± 12.42%, 77.33 ± 10.33%, and 82.13 ± 10.46% with supra-aural headphones, normal fitting hearing aids, and frequency-shifting feature, respectively. Scores for hearing aids worn on the left ear were 68 ± 7.56%, 76.80 ± 6.96%, and 82.13 ± 6.67% with supra-aural headphone, normal fitting hearing aid, and frequency-shifting feature, respectively (p˂0.05). Elderly individuals using hearing aids had low speech discrimination scores. The frequency-shifting feature recently used in hearing aids significantly increased the scores, making a significant contribution to the solution of speech reception and communication problems in cases of high-frequency hearing loss caused by presbyacusis in elderly individuals..


The novel camptothecin derivative, CPT211, induces cell cycle arrest and apoptosis in models of human breast cancer.
Breast cancer is the second leading cause of cancer deaths in women worldwide and represents a highly aggressive nature with limited therapeutic options; thus, investigating novel therapeutic agents for breast cancer is much needed. In this study, we investigated the anticancer effects of a novel camptothecin derivative, CPT211, against human breast cancer. We used hormone receptor-positive MCF-7, triple-negative (TNBC) MDA-MB-231, and HER2-positive BT-474 human breast cancer cells to examine cytotoxicity of CPT211. We measured cell viability with dose dependence of CPT211 treatments by an MTT assay and investigated the potential underlying mechanism through flow cytometric and Western blot methods. Furthermore, we evaluated the efficacy of the treatment combination of CPT211 and doxorubicin in a mouse model bearing MDA-MB-231 xenografts. CPT211 treatment led to dose-dependent decreases in cell viability of both MCF-7 and MDA-MB-231 cells, but not BT-474 cells. Analysis of the underlying molecular mechanism revealed that CPT211 activated p53-mediated apoptosis, by triggering intrinsic and extrinsic apoptotic pathways in MCF-7 cells. Additionally, CPT211 induced apoptosis and cell cycle arrest of MDA-MB-231 cells by activating Fas/FADD/caspase-8 signaling, suggesting that CPT211-mediated MDA-MB-231 cell apoptosis may occur through an extrinsic apoptosis pathway. CPT211 treatment with doxorubicin in mice bearing MDA-MB-231 xenografts was shown to enhance caspase-8 and caspase-7 activation, resulting in significant inhibition of tumor growth. These results indicate that Fas/FADD/caspase-8 activation plays an important role in CPT211-mediated tumor growth suppression in TNBC, and the novel camptothecin derivative, CPT211, can be exploited for specific targeted therapies and potentially improve approaches to combination treatments for human breast cancer.