Strategies for Assessing Acceptable Intakes for Novel N-Nitrosamines Derived from Active Pharmaceutical Ingredients.
The detection of N-nitrosamines, derived from solvents and reagents and, on occasion, the active pharmaceutical ingredient (API) at higher than acceptable levels in drug products, has led regulators to request a detailed review for their presence in all medicinal products. In the absence of rodent carcinogenicity data for novel N-nitrosamines derived from amine-containing APIs, a conservative class limit of 18 ng/day (based on the most carcinogenic N-nitrosamines) or the derivation of acceptable intakes (AIs) using structurally related surrogates with robust rodent carcinogenicity data is recommended. The guidance has implications for the pharmaceutical industry given the vast number of marketed amine-containing drugs. In this perspective, the rate-limiting step in N-nitrosamine carcinogenicity, involving cytochrome P450-mediated α-carbon hydroxylation to yield DNA-reactive diazonium or carbonium ion intermediates, is discussed with reference to the selection of read-across analogs to derive AIs. Risk-mitigation strategies for managing putative N-nitrosamines in the preclinical discovery setting are also presented.


First Report of Plum pox virus Strain M Isolates in Apricot in Sicily, Italy.
Sharka or plum pox disease is one of the most economically important virus diseases of stone fruits. Plum pox virus (PPV), the causal agent, is a member of the genus Potyvirus of the family Potyviridae transmitted by aphids in a non-persistent manner and by grafting. To date, nine PPV strains have been described on the basis of their biological, serological, and molecular properties: M and D are the most widespread and economically important strains, PPV-Rec and PPV-C have been reported mainly in Europe, PPV-EA confined to Egypt, PPV-T to Turkey, PPV-W from Canada, Ukraine, Latvia, and Russia, PPV-CR detected in Russia, and finally a putative PPV strain infecting plum in Albania described as the ancestor of the M. PPV-M is responsible for major epidemics in many Italian regions and despite phytosanitary measures, the infection rate increases each year. The D and Rec isolates are sporadically reported while PPV-C, once signaled in Apulia, has been successfully eradicated. Except for a report from the 1980s, which is no longer traceable, Sicily was considered free from the virus (2). In 2012, two new foci of sharka in a coastal area of Catania in Sicily were first reported by the national plant protection service to the European Commission (DG-SANCO). In spring 2013, plants of different varieties of apricot (Prunus armeniaca) and peach (P. persica) showing typical symptoms of flower color break, yellowing and leaf deformation, chlorotic spots or rings, and malformation on fruits were tested positive to PPV by DAS-ELISA using polyclonal antibodies. In order to characterize two isolates from apricot varieties (Carmen Top and Ninfa), total RNAs, extracted using the RNeasy Plant Mini Kit (Qiagen) from ELISA-positive samples, were analyzed by RT-PCR with primers P1/P2, targeting the 3'-terminal region of the coat protein (CP) gene (5) followed by RFLP analysis after digestion with Rsa1. Subsequently total RNAs were analyzed with the type-specific primers P1/PM and P1/PD (3), P3M/P4b and P3D/P4b amplifying the N-terminal region of the CP gene (1) and, finally, with primers mD5/mD3, mM5/mM3, and mD5/mM3, amplifying the region 3'NIb-5'CP, including the recombination site of Rec isolates (4). Only primer pairs P1/P2, P1/PM, P3M/P4b, and mM5/mM3 produced amplicons of the expected size (243, 198, 466, and 459 bp, respectively). The RFLP assay confirmed both isolates belonging to the M strain. Moreover, no reaction was obtained with primer pair mD5/mM3, excluding isolates belonging to Rec-type. Isolate characterization was completed by direct sequencing in both directions of the of P1/P2 and P3M/P4b amplicons obtained from apricot samples L9-1 (Carmen Top isolate) and 9-335 (Ninfa isolate). The P1/P2 sequences (KJ994235, KJ994237) showed 98% similarity with PPV-M or PPV-Rec isolates. The P3M/P4b sequences (KJ994236, KJ994238) confirmed that Sicilian isolates belong to the PPV-M strain showing 99% similarity with those already present in GenBank, thus ruling out the possibility of an infection with a PPV-Rec isolate. This outbreak of the Marcus strain of PPV in Sicily represents a high risk for the expanding production of stone fruit in southern Italy. An eradication plan was quickly activated by the regional phytosanitary service. References: (1) T. Candresse et al. Phytopathology 101:611, 2011. (2) EPPO. PQR-EPPO database on quarantine pests (available online). http://www.eppo.int , 2014. (3) A. Olmos et al. J. Virol. Methods 68:127, 1997. (4) Z. Subr et al. Acta Virol. 48:173, 2004. (5) T. Wetzel et al. J. Virol. Methods 33:355, 1991.


Immunohistochemical localization of basic fibroblast growth factor in mature and developing retinas of normal and RCS rats.
Basic fibroblast growth factor (bFGF) delays photoreceptor degeneration when injected intraocularly in Royal College of Surgeons (RCS) rats with inherited retinal dystrophy. In the present study, we have determined the localization of endogenous bFGF in retinas of normal and RCS rats during the normal developmental period (postnatal days 0-20) and the period of photoreceptor degeneration in RCS rats (days 20-90). bFGF was localized immunohistochemically by indirect immunoperoxidase using two different polyclonal antibodies and one monoclonal antibody against bFGF. bFGF was present in retinas as early as birth, and remained through adult age. Controls using either PBS, non-immune IgG or antibody preabsorbed with bFGF peptide were devoid of label. In normal rats between the ages of birth and postnatal day (P) 4, bFGF was found in developing ganglion cells, superficial blood vessels, some of the innermost cells in the neuroblastic layer, developing horizontal cells, and retinal pigment epithelial (RPE) cells. Between P0 and P4, the intensity of staining increased significantly in horizontal cells. From P6-P10, some cells in the inner nuclear layer remained positive, but horizontal cell staining became less intense in the central retina. The superficial vessels, ganglion cells and RPE cells also remained positive for bFGF. At P20-25, when the retina was essentially mature, bFGF was found in RPE cells, most cells of the ganglion cell layer, and many cells of the inner nuclear layer, but horizontal cells and blood vessels showed a lower concentration of bFGF than they did at younger ages. At P45 and older, blood vessels no longer showed bFGF immunoreactivity. The staining pattern in RCS rats was indistinguishable from that for normal rats at all ages examined. These results show that bFGF is present in the developing and adult rat retina in some neural cells, in addition to vessels and RPE cells. The transient elevated expression of bFGF immunoreactivity in developing horizontal cells and blood vessels suggests a possible role for this growth factor in retinal development. In addition, if RCS retinas possess any difference in bFGF localization or concentration compared to normal retinas, it must be too small to detect by immunohistochemical means, or at least with the reagents used.


Lens-based fluorescence nanoscopy.
The majority of studies of the living cell rely on capturing images using fluorescence microscopy. Unfortunately, for centuries, diffraction of light was limiting the spatial resolution in the optical microscope: structural and molecular details much finer than about half the wavelength of visible light (~200 nm) could not be visualized, imposing significant limitations on this otherwise so promising method. The surpassing of this resolution limit in far-field microscopy is currently one of the most momentous developments for studying the living cell, as the move from microscopy to super-resolution microscopy or 'nanoscopy' offers opportunities to study problems in biophysical and biomedical research at a new level of detail. This review describes the principles and modalities of present fluorescence nanoscopes, as well as their potential for biophysical and cellular experiments. All the existing nanoscopy variants separate neighboring features by transiently preparing their fluorescent molecules in states of different emission characteristics in order to make the features discernible. Usually these are fluorescent 'on' and 'off' states causing the adjacent molecules to emit sequentially in time. Each of the variants can in principle reach molecular spatial resolution and has its own advantages and disadvantages. Some require specific transitions and states that can be found only in certain fluorophore subfamilies, such as photoswitchable fluorophores, while other variants can be realized with standard fluorescent labels. Similar to conventional far-field microscopy, nanoscopy can be utilized for dynamical, multi-color and three-dimensional imaging of fixed and live cells, tissues or organisms. Lens-based fluorescence nanoscopy is poised for a high impact on future developments in the life sciences, with the potential to help solve long-standing quests in different areas of scientific research.


Five DNA tumor viruses undetectable in human retinoblastomas.
Retinoblastoma (RB) is a childhood eye cancer that arises when a retinal cell lacks a functional RB gene. Recent data indicate the transforming proteins of adenovirus, papillomavirus, and the polyomaviruses BK and JC all can bind to the product of the RB gene. Furthermore, adenovirus 12, JC virus, and simian virus 40 are able to induce RB-like tumors in rodents. In view of these findings, 50 human RBs were tested for the presence of five human DNA tumor viruses: adenovirus 12, BK virus, JC virus, and human papillomaviruses 16 and 18. Using the polymerase chain reaction, no viral sequences were detected in 50 RB DNAs. These data provide no evidence that these viruses have an etiologic role in human RB.


Spicatic acid: A 4-carboxygentisic acid from Gentiana spicata extract with potential hepatoprotective activity.
Due to our interest in bioactive plant derived materials, the hepatoprotective activity of the aqueous alcoholic extract of Gentiana spicata AEGS (Gentianaceae) on carbon tetrachloride treated rats was investigated. CCl4 used at a concentration of 1 mL/kg.b.wt. significantly increased the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). However, pre-treatment with AEGS and its individual components significantly prevented the increase in these enzymes, which are the major indicators of liver injury. Biochemical assays of liver homogenate showed that AEGS and its components restored reduced glutathione (GSH) depletion reduced the level of thiobarbituric acid reactive substances (TBARS). Furthermore, liver histological observation also showed an obvious amelioration in liver cell necrosis, liver lesions, and fatty changes in pre-treated groups. Phytochemical investigation of the extract showed high phenolic content and led to the isolation and identification of the new carboxygentisic acid, 1,4-dicarboxy 2,5-dihydroxybenzene, for which we suggest the name spicatic acid, together with the known flavonoids, quercetin 3-O-[(2,3,4-triacetyl-α-rhamnopyranosyl)1'''→6''] 3-acetyl-β-galactopyranoside and quercetin 3-O-[(2,3,4-triacetyl-α-rhamnopyranosyl)1'''→6'']-4-acetyl-β-galactopyranoside, epicatechin, catechin and their gallolyated derivatives. All structures were elucidated on the basis of conventional analytical methods and confirmed by high resolution ESIMS, 1D- and 2D-NMR data. The new phenolic 4-carboxygentisic acid, spicatic acid is of special interest as it represents the first phenolic acid in nature which bears two carboxyl functions in one aromatic ring.


Adherence to physical activity guidelines in mid-pregnancy does not reduce sedentary time: an observational study.
Physical activity (PA) interventions designed to prevent prenatal complications have focused on increasing moderate PA yielding conflicting results. Minimal attention has focused on the evaluation of sleep, sedentary behavior (SB), light activity or total daily PA during pregnancy. The purpose of this prospective, longitudinal study was to 1) objectively quantify and compare habitual PA and SB during the 2nd and 3rd trimester; and 2) evaluate differences in activity patterns for women meeting prenatal PA guidelines versus those that did not. Forty-six participants wore 2 PA monitors (SenseWear Mini and activPAL) during week 18 and week 35 of pregnancy. We compared differences in sleep duration, postural allocation, daily steps, and PA between the 2nd and 3rd trimester and for women who met and did not meet PA guidelines. During the 2(nd) trimester, 30% of the women's day (24-hours) was total sleep; 52% SB; 13% light; 3% moderate; and 0% vigorous PA. Light (P = 0.05), vigorous (P = 0.02), and moderate-vigorous PA (MET-minutes; P = 0.02), decreased with a trend in increased SB (P = 0.07). Activity of other intensities and sleep duration did not significantly change. Only 39% and 37% of participants slept between 7-9 hours/night at week 18 and 35, respectively. Forty-six percent (n = 21) and 28% (n = 13) of participants met prenatal PA guidelines during the 2(nd) and 3(rd) trimester, respectively. At week 18, no differences in total sleep, SB, or light PA existed for women who met PA guidelines versus those who did not; total PA was significantly greater for women who met guidelines. At week 35, women that met PA guidelines had significantly less SB (P < 0.005) than women who did not. This study demonstrates that pregnant women spend the majority of their day in SB. Significant reductions in total activity across pregnancy may be attributed, in part to shifts in light PA and increased SB. Based on the lifestyle of our sample, regardless of meeting PA guidelines in mid-pregnancy, no significant difference exists in time spent in SB, however meeting PA recommendations in late pregnancy may reduce SB. Future interventions should target reducing SB by increasing light and moderate PA beyond volitional exercise.


Mutagenic and carcinogenic significance and the possible induction of lung cancer by nitro aromatic hydrocarbons in particulate pollutants.
Studies of genotoxicity and carcinogenicity of nitro aromatic hydrocarbons focus on their high mutagenicity for bacteria and mammalian cells. Nitrobenzo[a]pyrenes (NBPs) and related nitroazaarenes also are extraordinarily mutagenic. 3-Nitro-6-azabenzo[a]pyren-N-oxide was found to be a more potent mutagen than 1,8-dinitropyrene. Mutagenicity of NBPs was associated with the position of substitution of the nitro function when nitrogen dioxide (NO2) was substituted at the third position on the benzo[a]pyrene (BP) structure, as in 3,6-dinitrobenzo[a]pyrene but not in 1,6-diNBP. The NBPs were reduced by a rat liver postmitochondrial fraction to nitroso- and subsequently to amino-derivatives. Therefore, tumoral action in rats was induced at significant levels by subcutaneous injection of 3,6-diNBP, but no tumors were observed in rats given 1,6-diNBP. Carcinogenic nitropyrenes were detected in the resected lung of a patient with lung cancer. It is suggested that the presence of nitropyrenes and the resulting tumor were due to exposure to by-products of combustion of heavy oil. The patient was a nonsmoker and farmer who had bred chickens for 40 years. He used heavy oil for heating the chicken house. Similarly, a group of Chinese people at high risk of developing lung cancer was selected to determine the initiator of lung cancer. Lung cancers were obtained from six Chinese female nonsmokers who were living in Fuyuan County, China. Polycyclic aromatic hydrocarbons were detected in resected lung specimens; they were benzo[k]fluoranthene, BP, benzo[g,h,i]perylene, and pyrene. These cases were associated with exposure to soot from combustion of coal usually used for heating and cooking indoors.


Tailoring the emission polarization with metasurface-based emitters designed on a plasmonic ridge waveguide.
Because of low propagation losses and flexible communication paths, inter-chip optical communications based on plasmonic emitters and receivers can overcome the obstacle of the inherent ohmic loss in metallic nanostructures. To increase the communication capacity and integration density in inter-chip optical communications, we propose to tailor the polarization states of the free-radiation fields from the emitters in both the spectral domain and spatial domain by designing the phased and polarized arrangement of subwavelength metallic nanogroove antennas on a two-dimensional plasmonic ridge waveguide. Herein, the utilization of the two-dimensional plasmonic waveguide with tight field confinements considerably decreases the crosstalk to nearby plasmonic devices in plasmonic circuits and chips. In the spectral domain, three different polarization states of the free-radiation fields from the same emitter are experimentally realized at three specific wavelengths. In the spatial domain, the different polarization states as well as the gear polarization states are experimentally demonstrated. Moreover, the separation of the adjacent polarization-tailoring plasmonic emitters is only 5% of that in dielectric emitters because of the ultra-compact size (<0.6λ2) of metasurface-based plasmonic emitters.


A functionally split pathway for lysine synthesis in Corynebacterium glutamicium.
Three different pathways of D,L-diaminopimelate and L-lysine synthesis are known in procaryotes. Determinations of the corresponding enzyme activities in Escherichia coli, Bacillus subtilis, and Bacillus sphaericus verified the fact that in each of these bacteria only one of the possible pathways operates. However, in Corynebacterium glutamicum activities are present which allow in principle the use of the dehydrogenase variant and succinylase variant of lysine synthesis together. Applying gene-directed mutagenesis, various C. glutamicum strains were constructed with interrupted ddh gene. These mutants have an inactive dehydrogenase pathway but are still prototrophic, which is proof that the succinylase pathway of D,L-diaminopimelate synthesis can be utilized. In strains with an increased flow of precursors to D,L-diaminopimelate, however, the inactivation of the dehydrogenase pathway resulted in a reduced formation of lysine, with concomitant accumulation of N-succinyl-diaminopimelate in the cytosol up to a concentration of 25 mM. These data show (i) that both pathways can operate in C. glutamicum for D,L-diaminopimelate and L-lysine synthesis, (ii) that the dehydrogenase pathway is not essential, and (iii) that the dehydrogenase pathway is a prerequisite for handling an increased flow of metabolites to D,L-diaminopimelate.


NO donors inhibit Na,K-ATPase activity by a protein kinase G-dependent mechanism in the nonpigmented ciliary epithelium of the porcine eye.
1. We developed a novel method to isolate nonpigmented epithelial (NPE) cells from porcine eyes in order to examine Na,K-ATPase responses to nitric oxide (NO) donors specifically in the epithelium. 2. Cells were treated with NO donors and other test compounds for 20 min prior to Na,K-ATPase activity measurement. 3. NO donors, sodium nitroprusside (SNP, 1 microM-1 mM), sodium azide (100 nM-1 microM) and S-nitroso-N-acetylpenicillamine (1 microM-1 mM) caused significant concentration-dependent inhibition of Na,K-ATPase activity. Detection of nitrite in the medium of L-arginine and SNP-treated NPE confirmed NO generation. 4. Concentration-dependent inhibition of Na,K-ATPase was also obtained by L-arginine (1-3 mM), a physiological precursor of NO and 8p-CPT-cGMP (1-100 microM), a cell permeable analog of cGMP. The L-arginine effect was abolished when the NO synthesizing enzyme, NO-synthase, was inhibited by L-NAME (100 microM). 5. The inhibitory effect of SNP or sodium azide on Na,K-ATPase activity was suppressed by soluble guanylate cyclase (sGC) inhibitors, ODQ (10 microM) or methylene blue (10 microM). 6. The inhibitory effect of 8p-CPT-cGMP on Na,K-ATPase was abolished by protein kinase G (PKG) inhibitors, H-8 (1 microM) and H-9 (20 microM), but not by the protein kinase A (PKA) inhibitor H-89 (100 nM). H-8 and H-9 partially suppressed the inhibitory effect of SNP on Na,K-ATPase. 7. Taken together the results indicate that Na,K-ATPase inhibition response to NO donors involves activation of sGC, generation of cGMP and activation of PKG. These findings suggest that Na,K-ATPase inhibition in NPE may contribute to the ability of NO donors to reduce aqueous humor secretion.


Impact of Language Experience on Attention to Faces in Infancy: Evidence From Unimodal and Bimodal Bilingual Infants.
Faces capture and maintain infants' attention more than other visual stimuli. The present study addresses the impact of early language experience on attention to faces in infancy. It was hypothesized that infants learning two spoken languages (unimodal bilinguals) and hearing infants of Deaf mothers learning British Sign Language and spoken English (bimodal bilinguals) would show enhanced attention to faces compared to monolinguals. The comparison between unimodal and bimodal bilinguals allowed differentiation of the effects of learning two languages, from the effects of increased visual communication in hearing infants of Deaf mothers. Data are presented for two independent samples of infants: Sample 1 included 49 infants between 7 and 10 months (26 monolinguals and 23 unimodal bilinguals), and Sample 2 included 87 infants between 4 and 8 months (32 monolinguals, 25 unimodal bilinguals, and 30 bimodal bilingual infants with a Deaf mother). Eye-tracking was used to analyze infants' visual scanning of complex arrays including a face and four other stimulus categories. Infants from 4 to 10 months (all groups combined) directed their attention to faces faster than to non-face stimuli (i.e., attention capture), directed more fixations to, and looked longer at faces than non-face stimuli (i.e., attention maintenance). Unimodal bilinguals demonstrated increased attention capture and attention maintenance by faces compared to monolinguals. Contrary to predictions, bimodal bilinguals did not differ from monolinguals in attention capture and maintenance by face stimuli. These results are discussed in relation to the language experience of each group and the close association between face processing and language development in social communication.


31P NMR visibility and characterization of rat liver mitochondrial matrix adenine nucleotides.
Compartmentation and NMR visibility of mitochondrial adenine nucleotides were quantitated in isolated rat liver mitochondria respiring on succinate and glutamate in vitro at 8 and 25 degrees C. Intra- and extramitochondrial nucleotides were discriminated by adding the chelator trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA). T1 values of about 0.2-0.3 s for magnesium-bound matrix nucleotides were determined. Adenine nucleotide T1 values were influenced by the ionic environment; only magnesium-free ATP T1's were affected by temperature. Intra- and extramitochondrial adenine nucleotide ratios were varied in ATP-loaded mitochondria with added ATP and phosphate using the mitochondrial inhibitors oligomycin and carboxyatractyloside, and adenine nucleotides were quantitated by using NMR and enzymatic analysis. There was good agreement between matrix ATP concentrations (magnesium-bound ATP) calculated by using NMR and standard biochemical techniques. Although matrix ADP could be detected by NMR, it was difficult to quantitate accurately by NMR. The data indicate that mitochondrial ATP is NMR-visible in isolated mitochondria in vitro.


Occasional paternal inheritance of the germline-restricted chromosome in songbirds.
Songbirds have one special accessory chromosome, the so-called germline-restricted chromosome (GRC), which is only present in germline cells and absent from all somatic tissues. Earlier work on the zebra finch (Taeniopygia guttata castanotis) showed that the GRC is inherited only through the female line-like the mitochondria-and is eliminated from the sperm during spermatogenesis. Here, we show that the GRC has the potential to be paternally inherited. Confocal microscopy using GRC-specific fluorescent in situ hybridization probes indicated that a considerable fraction of sperm heads (1 to 19%) in zebra finch ejaculates still contained the GRC. In line with these cytogenetic data, sequencing of ejaculates revealed that individual males from two families differed strongly and consistently in the number of GRCs in their ejaculates. Examining a captive-bred male hybrid of the two zebra finch subspecies (T. g. guttata and T. g. castanotis) revealed that the mitochondria originated from a castanotis mother, whereas the GRC came from a guttata father. Moreover, analyzing GRC haplotypes across nine castanotis matrilines, estimated to have diverged for up to 250,000 y, showed surprisingly little variability among GRCs. This suggests that a single GRC haplotype has spread relatively recently across all examined matrilines. A few diagnostic GRC mutations that arose since this inferred spreading suggest that the GRC has continued to jump across matriline boundaries. Our findings raise the possibility that certain GRC haplotypes could selfishly spread through the population via occasional paternal transmission, thereby outcompeting other GRC haplotypes that were limited to strict maternal inheritance, even if this was partly detrimental to organismal fitness.


Single-Copy Knock-In Loci for Defined Gene Expression in Caenorhabditis elegans.
We have generated a single-copy knock-in loci for defined gene expression (SKI LODGE) system to insert any DNA by CRISPR/Cas9 at defined safe harbors in the Caenorhabditis elegans genome. Utilizing a single crRNA guide, which also acts as a Co-CRISPR enrichment marker, any DNA sequence can be introduced as a single copy, regulated by different tissue-specific promoters. The SKI LODGE system provides a fast, economical, and effective approach for generating single-copy ectopic transgenes in C. elegans.


Genes and Pathways Involved in Adult Onset Disorders Featuring Muscle Mitochondrial DNA Instability.
Replication and maintenance of mtDNA entirely relies on a set of proteins encoded by the nuclear genome, which include members of the core replicative machinery, proteins involved in the homeostasis of mitochondrial dNTPs pools or deputed to the control of mitochondrial dynamics and morphology. Mutations in their coding genes have been observed in familial and sporadic forms of pediatric and adult-onset clinical phenotypes featuring mtDNA instability. The list of defects involved in these disorders has recently expanded, including mutations in the exo-/endo-nuclease flap-processing proteins MGME1 and DNA2, supporting the notion that an enzymatic DNA repair system actively takes place in mitochondria. The results obtained in the last few years acknowledge the contribution of next-generation sequencing methods in the identification of new disease loci in small groups of patients and even single probands. Although heterogeneous, these genes can be conveniently classified according to the pathway to which they belong. The definition of the molecular and biochemical features of these pathways might be helpful for fundamental knowledge of these disorders, to accelerate genetic diagnosis of patients and the development of rational therapies. In this review, we discuss the molecular findings disclosed in adult patients with muscle pathology hallmarked by mtDNA instability.


Integrated Biosignal Analysis to Provide Biomarkers for Recognizing Time Perception Difficulties.
Time perception refers to the capability to recognize the passage of time. The cerebellum is located at the back of the brain, underlying the occipital and temporal lobes. Dyschronometria is a cerebellar dysfunction, in which a person cannot precisely estimate the amount of time that has passed. Cardiac indicators such as heart rate (HR) variability have been associated with mental function in healthy individuals. Moreover, time perception has been previously studied concerning cardiac signs. Human time perception is influenced by various factors such as attention and drowsiness. An electroencephalogram (EEG) is a suitable modality for evaluating cortical reactions due to its affordability and usefulness. Because EEG has a high sequential outcome, it offers valuable data to explore variability in psychological situations. An electrocardiogram (ECG) records electrical signals from the heart to examine various heart conditions. The electromyography (EMG) technique detects electrical impulses produced by muscles. EEG, ECG, and EMG are integrated during time perception. This study evaluated the human body's time perception through the neurological, cardiovascular, and muscular systems using a simple neurofeedback exercise after time perception tasks. The three biosignals which are EEG, ECG, and EMG were investigated to use them as biomarkers for recognizing time perception difficulty as the main goal of the study. Five healthy college students with no health issues participated, and their EEG, ECG, and EMG were recorded while relaxing and performing a time wall estimation task and neurofeedback training. Previous research has shown the relationship between EEG frequency bands and the frontal center during time perception. Investigating the connection between ECG, EEG, and EMG under time perception conditions is significant. The results show that ECG (HR), EEG (Delta wave), and EMG (root mean square) are critical features in time perception difficulties. The ability and outcomes of multiple biomarkers might allow for improved diagnosis and monitoring of the progress of any treatment applications such as biofeedback training. Furthermore, those biomarkers could be used as useful for evaluating and treating dyschronometria.


Content-aware image restoration for electron microscopy.
Multiple approaches to use deep neural networks for image restoration have recently been proposed. Training such networks requires well registered pairs of high and low-quality images. While this is easily achievable for many imaging modalities, e.g., fluorescence light microscopy, for others it is not. Here we summarize on a number of recent developments in the fast-paced field of Content-Aware Image Restoration (CARE), in particular, and the associated area of neural network training, more in general. We then give specific examples how electron microscopy data can benefit from these new technologies.


Comparative study of crumpling and folding of thin sheets.
Crumpling and folding of paper are at first sight very different ways of confining thin sheets in a small volume: the former one is random and stochastic whereas the latest one is regular and deterministic. Nevertheless, certain similarities exist. Crumpling is surprisingly inefficient: a typical crumpled paper ball in a waste-bin consists of as much as 80% air. Similarly, if one folds a sheet of paper repeatedly in two, the necessary force becomes so large that it is impossible to fold it more than six or seven times. Here we show that the stiffness that builds up in the two processes is of the same nature, and therefore simple folding models allow us to capture also the main features of crumpling. An original geometrical approach shows that crumpling is hierarchical, just as the repeated folding. For both processes the number of layers increases with the degree of compaction. We find that for both processes the crumpling force increases as a power law with the number of folded layers, and that the dimensionality of the compaction process (crumpling or folding) controls the exponent of the scaling law between the force and the compaction ratio.


Nuclear magnetic resonance analysis of emulsified silicone oil RMN-3 (Oxane HD).
Oxane HD is a mixture of 5700 cs silicone oil and RMN-3 (a partly fluorinated olefin), which has a specific gravity greater than water, thereby enabling endotamponade of inferior retinal breaks. Droplets of emulsified oil were found in the anterior chamber of two patients with complex retinal detachments who had been treated with Oxane HD. Samples removed from these two eyes were analysed in order to investigate the nature of the emulsified substance. Samples were dissolved in CDCl3 and, if necessary, dried using magnesium sulphate. Nuclear magnetic resonance (NMR) was used to compare these samples with the unused commercial product. NMR analysis of the samples removed from the anterior segment of both eyes were mainly silicone oil with only trace amounts of RMN-3. Samples taken from different areas of an unused syringe of Oxane HD demonstrated varying concentrations of the RMN-3 compound within the silicone oil. First, the mixture of RMN-3 and silicone oil is not homogenous either in vitro or in vivo. Second, emulsified oil found in the anterior chamber is composed almost entirely of silicone oil, containing only trace amounts of RMN-3.


Variation Profile of the Orthotospovirus Genome.
Orthotospoviruses are plant-infecting members of the family Tospoviridae (order Bunyavirales), have a broad host range and are vectored by polyphagous thrips in a circulative-propagative manner. Because diverse hosts and vectors impose heterogeneous selection constraints on viral genomes, the evolutionary arms races between hosts and their pathogens might be manifested as selection for rapid changes in key genes. These observations suggest that orthotospoviruses contain key genetic components that rapidly mutate to mediate host adaptation and vector transmission. Using complete genome sequences, we profiled genomic variation in orthotospoviruses. Results show that the three genomic segments contain hypervariable areas at homologous locations across species. Remarkably, the highest nucleotide variation mapped to the intergenic region of RNA segments S and M, which fold into a hairpin. Secondary structure analyses showed that the hairpin is a dynamic structure with multiple functional shapes formed by stems and loops, contains sites under positive selection and covariable sites. Accumulation and tolerance of mutations in the intergenic region is a general feature of orthotospoviruses and might mediate adaptation to host plants and insect vectors.


[Microcirculatory bed of the epithelial layer of human esophageal mucous membrane in postnatal ontogenesis].
The analysis of histological changes in epithelium layer of mucous membrane and its microcirculatory channel of human oesophagus show that development and formation of epithelium and its capillary channel are interwoven processes. Dynamics of their morphological changes strictly correspond to a definite age stage of organ individual development. Changing each other determined four types of capillaries of mucous membrane epithelium layer and determined the degree of its functional development.


Minimally Invasive Surgery for Rectal Cancer: Current Status and Future Perspectives.
Although laparoscopic resection for colon cancer has been proven safe and feasible when compared with open resection, currently no clear evidence is available regarding minimally invasive surgery for rectal cancer. This type of surgery may benefit patients by allowing fast recovery of normal dietary intake and bowel function, reduced postoperative pain, and shorter hospitalization. Therefore, minimally invasive surgeries such as laparoscopic or robot surgery have become the predominant treatment option for colon cancer. Specifically, the proportion of laparoscopic colorectal cancer surgery in Korea increased from 42.6 to 64.7% until 2013. However, laparoscopic surgery for rectal cancer is more difficult and technically demanding. In addition, the procedure requires a prolonged learning curve to achieve equivalent outcomes relative to open surgery. It is very challenging to approach the deep and narrow pelvis using laparoscopic instruments. However, robotic surgery provides better vision with a high definition three-dimensional view, exceptional ergonomics, Endowrist technology, enhanced dexterity of movement, and a lack of physiologic tremor, facilitated by the use of an assistant in the narrow and deep pelvis. Recently, an increasing number of reports have compared the outcomes of laparoscopic and open surgery for colon cancer. Such reports have prompted a discussion of the outcomes of minimally invasive surgery, including robotic surgery, for rectal cancer. The aim of this review is to summarize current data regarding the clinical outcomes, including oncologic outcomes, of minimally invasive surgery for rectal cancer.


CD16+ gammadelta T cells mediate antibody dependent cellular cytotoxicity: potential mechanism in the pathogenesis of multiple sclerosis.
Our overall objective is to understand the role of gammadelta T cells in the pathogenesis of the central nervous system (CNS) autoimmune disease multiple sclerosis (MS). We have demonstrated that gammadelta T cells are directly cytotoxic to CNS cells in vitro. Although the exact mechanism of damage in MS is unknown, recent evidence suggests a role for B cells and antibodies to myelin. We were therefore interested in examining whether gammadelta T cells can injure CNS cells via an indirect mechanism involving antibody dependent cellular cytotoxicity. To study this we developed an in vitro flow cytometric cellular cytotoxicity assay (called "FC(3)A") to quantitate the amount of cytotoxicity. We utilized known target cells (Burkitt's B lymphoma) that express CD20, together with a monoclonal antibody (mAb) to CD20, rituximab, that is being studied as a potential treatment for MS. Target cells are first coated with rituximab followed by co-culture with gammadelta T cells derived from patients with MS. Specific lysis of target cells was determined by quantitation of 7-AAD (which increases only upon nuclear disruption indicating cell death). We determined that this lysis was due to gammadelta T cells that express CD16 (Fc gamma receptor) and were therefore capable of binding the rituximab and mediating cytolysis via ADCC. This specific cell lysis correlated with rituximab concentration, E:T ratio, and the surface expression of CD16 on gammadelta T cells. These findings provide a new perspective with regards to the role of gammadelta T cells in the immunopathogenesis of MS and an insight into one of the potential therapeutic effects of rituximab in the treatment of MS. In addition, this new FC(3)A method we developed could readily be adapted to study other types of immune cells suspected of ADCC-type killing.


Influence of canopy openness, ungulate exclosure, and low-intensity fire for improved oak regeneration in temperate Europe.
Failed oak regeneration is widely reported in temperate forests and has been linked in part to changed disturbance regimes and land-use. We investigated if the North American fire-oak hypothesis could be applicable to temperate European oaks (Quercus robur, Quercus petraea) using a replicated field experiment with contrasting canopy openness, protection against ungulate browsing (fencing/no fencing), and low-intensity surface fire (burn/no burn). Survival, relative height growth (RGRH), browsing damage on naturally regenerated oaks (≤300 cm tall), and changes in competing woody vegetation were monitored over three years. Greater light availability in canopy gaps increased oak RGRH (p = .034) and tended to increase survival (p = .092). There was also a trend that protection from browsing positively affected RGRH (p = .058) and survival (p = .059). Burning reduced survival (p < .001), nonetheless, survival rates were relatively high across treatment combinations at the end of the experiment (54%-92%). Most oaks receiving fire were top-killed and survived by producing new sprouts; therefore, RGRH in burned plots became strongly negative the first year. Thereafter, RGRH was greater in burned plots (p = .002). Burning altered the patterns of ungulate browsing frequency on oaks. Overall, browsing frequency was greater during winter; however, in recently burned plots summer browsing was prominent. Burning did not change relative density of oaks, but it had a clear effect on competing woody vegetation as it reduced the number of individuals (p < .001) and their heights (p < .001). Our results suggest that young, temperate European oaks may respond similarly to fire as their North American congeners. However, disturbance from a single low-intensity fire may not be sufficient to ensure a persistent competitive advantage-multiple fires and canopy thinning to increase light availability may be needed. Further research investigating long-term fire effects on oaks of various ages, species-specific response of competitors and implications for biodiversity conservation is needed.


A demographic snapshot of midwives in Victoria.
Midwifery in Victoria has experienced significant change over the last twenty years. Over more recent years, midwifery workforce issues have become prominent with studies highlighting an ageing midwifery population and a high proportion of part time employment. During a study conducted in 2005 that explored midwives' attitudes to online and computer based learning the demographic data proved to warrant more detailed analysis. This paper presents a discussion on that data. Participants for the study were drawn from the membership of the Australian College of Midwives (ACM) Victorian branch. However, while the sample was small (n=169), the findings presented in the paper add to the existing body of knowledge around the midwifery workforce in Victoria. They reinforce that the midwifery workforce remained an ageing one, predominantly female, and a significant percentage were employed on part time or casual bases. With graduates emerging from recently introduced undergraduate midwifery programs, it is argued that it is timely for larger studies of the midwifery workforce to be conducted. Studies should seek to explore a range of factors such as reasons why midwives are choosing not to work in full time employment.


Age at Onset and Social Cognitive Impairment in Clinically Stabilized Patients with Schizophrenia: An Ecological Cross-Sectional Study.
Objective: Purposes of the present study were to assess the social cognitive impairment in schizophrenia and to detect if some clinical variables (particularly age at onset) are predictive of general/social cognitive deficit in schizophrenia patients. Method: Thirty-five clinically stabilized schizophrenia outpatients were assessed by the Brief Assessment of Cognition in Schizophrenia (BACS) and by Torralva's social cognition battery. Binary logistic models were performed to find an eventual association between continuous clinical variables and cognitive test failures. The total sample was divided in groups according to dichotomous variables (gender, diagnostic subtypes and type of abuse) and the presence of cognitive deficits was compared between groups by χ2 tests. Results: An earlier age at onset was found to be predictive of frontal cognitive impairment (Tower of London p=0.038, OR=0.702). Female gender was more probably associated with mistakes at MET-HV (χ2= 4.80, p=0.05, phi=0.40) and HOTEL tests (χ2= 5.25, p=0.04, phi=0.4) than male one. Cannabis abusers showed more frequently deficits on verbal fluency (χ2= 9.35, p=0.04, phi=0.52) and executive functioning (Tower of London) (χ2= 11.67, p=0.02, phi=0.58) than alcohol/cocaine ones. Conclusion: Female patients with an early age at onset and cannabis abuse seem to have the worst general and social cognitive profile among patients suffering from schizophrenia.


Alcea rosea root extract as a preventive and curative agent in ethylene glycol-induced urolithiasis in rats.
Alcea rosea L. is used in Asian folk medicine as a remedy for a wide range of ailments. The aim of the present study was to investigate the effect of hydroalcoholic extract of Alcea rosea roots on ethylene glycol-induced kidney calculi in rats. Male Wistar rats were randomly divided into control, ethylene glycol (EG), curative and preventive groups. Control group received tap drinking water for 28 days. Ethylene glycol (EG), curative and preventive groups received 1% ethylene glycol for induction of calcium oxalate (CaOx) calculus formation; preventive and curative subjects also received the hydroalcoholic extract of Alcea rosea roots in drinking water at dose of 170 mg/kg, since day 0 or day 14, respectively. Urinary oxalate concentration was measured by spectrophotometer on days 0, 14 and 28. On day 28, the kidneys were removed and examined histopathologically under light microscopy for counting the calcium oxalate deposits in 50 microscopic fields. In both preventive and curative protocols, treatment of rats with hydroalcoholic extract of Alcea rosea roots significantly reduced the number of kidney calcium oxalate deposits compared to ethylene glycol group. Administration of Alcea rosea extract also reduced the elevated urinary oxalate due to ethylene glycol. Alcea rosea showed a beneficial effect in preventing and eliminating calcium oxalate deposition in the rat kidney. This effect is possibly due to diuretic and anti-inflammatory effects or presence of mucilaginous polysaccharides in the plant. It may also be related to lowering of urinary concentration of stone-forming constituents.


Data generation for the Cox proportional hazards model with time-dependent covariates: a method for medical researchers.
The proliferation of longitudinal studies has increased the importance of statistical methods for time-to-event data that can incorporate time-dependent covariates. The Cox proportional hazards model is one such method that is widely used. As more extensions of the Cox model with time-dependent covariates are developed, simulations studies will grow in importance as well. An essential starting point for simulation studies of time-to-event models is the ability to produce simulated survival times from a known data generating process. This paper develops a method for the generation of survival times that follow a Cox proportional hazards model with time-dependent covariates. The method presented relies on a simple transformation of random variables generated according to a truncated piecewise exponential distribution and allows practitioners great flexibility and control over both the number of time-dependent covariates and the number of time periods in the duration of follow-up measurement. Within this framework, an additional argument is suggested that allows researchers to generate time-to-event data in which covariates change at integer-valued steps of the time scale. The purpose of this approach is to produce data for simulation experiments that mimic the types of data structures applied that researchers encounter when using longitudinal biomedical data. Validity is assessed in a set of simulation experiments, and results indicate that the proposed procedure performs well in producing data that conform to the assumptions of the Cox proportional hazards model.


Quantitative solid-state analysis of three solid forms of ranitidine hydrochloride in ternary mixtures using Raman spectroscopy and X-ray powder diffraction.
The aim of the study was to develop a reliable quantification procedure for mixtures of three solid forms of ranitidine hydrochloride using X-ray powder diffraction (XRPD) and Raman spectroscopy combined with multivariate analysis. The effect of mixing methods of the calibration samples on the calibration model quality was also investigated. Thirteen ternary samples of form 1, form 2 and the amorphous form of ranitidine hydrochloride were prepared in triplicate to build a calibration model. The ternary samples were prepared by three mixing methods (a) manual mixing (MM) and ball mill mixing (BM) using two (b) 5 mm (BM5) or (c) 12 mm (BM12) balls for 1 min. The samples were analyzed with XRPD and Raman spectroscopy. Principal component analysis (PCA) was used to study the effect of mixing method, while partial least squares (PLS) regression was used to build the quantification models. PCA score plots showed that, in general, BM12 resulted in the narrowest sample clustering indicating better sample homogeneity. In the quantification models, the number of PLS factors was determined using cross-validation and the models were validated using independent test samples with known concentrations. Multiplicative scattering correction (MSC) without scaling gave the best PLS regression model for XPRD, and standard normal variate (SNV) transformation with centering gave the best model for Raman spectroscopy. Using PLS regression, the root mean square error of prediction (RMSEP) values of the best models were 5.0-6.9% for XRPD and 2.5-4.5% for Raman spectroscopy. XRPD and Raman spectroscopy in combination with PLS regression can be used to quantify the amount of single components in ternary mixtures of ranitidine hydrochloride solid forms. Raman spectroscopy gave better PLS regression models than XRPD, allowing a more accurate quantification.


Digestion, Purification, and Enrichment of Protein Samples for Mass Spectrometry.
Proteomic studies rely heavily on the use of liquid chromatography (LC)-mass spectrometry (MS and MS/MS) analyses to provide information about protein composition and function. Profiling the proteome can be the first step to understanding biological pathways, but the challenges scientists face with the complex nature of proteins and proteolysis products can be daunting. Techniques involving fractionation, immunoprecipitation, and phosphopeptide enrichment can simplify complex protein mixtures and enhance the amount of target proteins that are important to the investigator. Emphasis on sample preparation for LC-MS/MS analyses is essential to acquisition of high-quality data for proteomic research. Certain classes of reagents, materials, and contaminants that can be introduced during sample processing may limit the effectiveness of LC-MS/MS analysis. These protocols outline methods for proteolytic digestion of proteins that are compatible with LC-MS/MS, along with procedures that allow for simplification of complex protein matrices.


Clinical safety and tolerance of mitoxantrone.
Mitoxantrone has been administered to more than 4000 patients worldwide. From this data base, a summary of the latest information is presented on the effect of mitoxantrone on bone marrow, on its nonmyelosuppressive acute toxicities, on its local tolerability, and on measurements of organ function. Most of these patients had solid tumors and were treated with an initial dose of mitoxantrone (12 to 14 mg/m2). Predictable leucopenia was dose limiting, but clinically significant suppression of RBC or platelet count was rare. Other hematologic or blood chemistry abnormalities were infrequent. The most common adverse clinical effects were nausea and vomiting, stomatitis, and alopecia, though the majority of these cases were mild. Many patients experienced no adverse reactions to mitoxantrone; there have been no reports of cellulitis, vesication, or tissue necrosis following extravasation. From the global experience with mitoxantrone, it seems that for patients who have not previously received anthracycline therapy, the risk of congestive heart failure is minimal up to a cumulative mitoxantrone dose of 160 mg/m2. In randomized, comparative clinical trials in advanced breast cancer, using mitoxantrone or doxorubicin either as single agents or in combination with other standard drugs, the incidence of moderate or severe acute toxic side effects was much lower for patients treated with mitoxantrone or mitoxantrone-containing combinations. From these data, it is clear that mitoxantrone has an exceptional safety profile, particularly with regard to acute nonmyelosuppressive toxicity and local tolerability. This offers the patient a better quality of life during antineoplastic chemotherapy.


[The effect of neutral polymers on glutamate dehydrogenase activity of mouse liver mitochondria upon administration of endotoxin].
It is shown that neutral polymers administered intraperitoneally to intact animals considerably affect glutamate dehydrogenase activity in the liver cell mitochondria as well as in the supernatant. Of the tested polymers, only polyvinyl methylacetamide and dextran inhibit a decrease in the level of mitochondrial enzyme activity which develops with administration of endotoxin. Polyvinyl pyrrolidone with molecular weight of 100 kDa, polyvinyl methylacetamide, dextran and polyvinyl caprolactam prevent an increase of glutamate dehydrogenase activity in the supernatant in case of endotoxin administration to animals. It is possibly a result of the effect of the mitochondrial structure stabilization by the above polymers. A physiological effect of polyvinyl pyrrolidone revealed as an effect on the activity level of mitochondrial glutamate dehydrogenase and in the supernatant after endotoxin administration to animals, depends on the molecular weight of the polymer.


Colostomy Irrigation: Issues Most Important to Wound, Ostomy and Continence Nurses.
The purpose of this study was to describe issues that WOC nurses find most important related to colostomy irrigation (CI). This is an additional analysis of a study focusing on qualitative responses to a survey querying WOC nurses about CI practices. The target population was members of the Wound, Ostomy and Continence Nurses Society. Of the 985 nurses who responded to the survey, 338 (34.3%) answered the optional open-ended question asking for additional comments. A 1-time online survey was conducted. In addition to demographic, educational information, and forced-choice questions about CI, an open-ended question asked for any additional comments about their experience with irrigation and WOC practice. Content analysis was used to identify common themes identified by WOC nurses. Three hundred thirty-eight out of 985 WOC nurses (34.3%) answered the optional open-ended question asking for additional comments; analysis for this study is based on these responses. WOC nurses who responded to the open-ended question had similar characteristics to those who responded to the entire survey but were significantly more experienced (15.1 vs 11.0 years; P < .001). Multiple themes were identified that were related to forced-choice questions in the survey, such as equipment, lack of teaching time, and increased control, while several new themes emerged, including age-related recommendations, economic/ecologic benefits, cultural implications, and misconceptions of CI. Many WOC nurses feel that CI is a valued modality of ostomy care and should be reestablished, although dissenting opinions were expressed. Additional research to increase the evidence base for this procedure is needed. New opportunities to educate WOC nurses and other healthcare professionals and teach eligible patients irrigation techniques are recommended.


Advanced glycation end products enhance expression of pro-apoptotic genes and stimulate fibroblast apoptosis through cytoplasmic and mitochondrial pathways.
Both aging and diabetes are characterized by the formation of advanced glycation end products (AGEs). Both exhibit other similarities including deficits in wound healing that are associated with higher rates of fibroblast apoptosis. In order to investigate a potential mechanism for enhanced fibroblast apoptosis in diabetes and aged individuals, experiments were carried out to determine whether the predominant advanced glycation end product in skin, N-epsilon-(carboxymethyl) lysine (CML)-collagen, could induce fibroblast apoptosis. In vivo experiments established that CML-collagen but not unmodified collagen induced fibroblast apoptosis and that apoptosis was dependent upon caspase-3, -8, and -9 activity. In vitro experiments demonstrated that CML-collagen but not control collagen induced a time- and dose-dependent increase in fibroblast apoptosis. By use of blocking antibodies, apoptosis was shown to be mediated through receptor for AGE signaling. AGE-induced apoptosis was largely dependent on the effector caspase, caspase-3, which was activated through both cytoplasmic (caspase-8-dependent) and mitochondrial (caspase-9) pathways. CML-collagen had a global effect of enhancing mRNA levels of pro-apoptotic genes that included several classes of molecules including ligands, receptors, adaptor molecules, mitochondrial proteins, and others. However, the pattern of expression was not identical to the pattern of apoptotic genes induced by tumor necrosis factor alpha.


Pathologic splenic rupture in a patient with follicular lymphoma.
A middle aged man presented with abdominal pain and fever, with progressive dyspnea for the past one week. He had generalized lymphadenopathy with hepatosplenomegaly and a left sided pleural effusion on admission. Further evaluation revealed that he had lymphocytosis on peripheral blood. He then developed increasing abdominal pain and fall in hemoglobin which was confirmed on imaging to be due to a splenic rupture and he underwent a splenectomy. The diagnosis on lymph node biopsy and peripheral blood immunophenotyping was grade 1 follicular lymphoma. He has completed his 6 cycles of chemotherapy (R-CVP) and is on maintenance rituximab and doing well. The case highlights the fact that splenic rupture can even be caused by indolent lymphomas.


Nationwide analysis of whole blood hemostatic resuscitation in civilian trauma.
Renewed interest in whole blood (WB) resuscitation in civilians has emerged following its military use. There is a paucity of data on its role in civilians where balanced component therapy (CT) resuscitation is the standard of care. The aim of this study was to assess nationwide outcomes of using WB as an adjunct to CT versus CT alone in resuscitating civilian trauma patients. We analyzed the (2015-2016) Trauma Quality Improvement Program. We included adult (age, ≥18 years) trauma patients presenting with hemorrhagic shock and requiring at least 1 U of packed red blood cells (pRBCs) within 4 hours. Patients were stratified into WB-CT versus CT only. Primary outcomes were 24-hour and in-hospital mortality. Secondary outcomes were hospital length of stay and major complications. Hierarchical logistic regression was performed to account for clustering effect within hospitals and adjusting for patient- and hospital-level potential confounding factors. A total of 8,494 patients were identified, of which 280 received WB-CT (WB, 1 [1-1]; pRBC, 16 [10-23]; FFP, 9 [6-16]; platelets, 3 [2-5]) and 8,214 received CT only (pRBC, 15 [10-24]; FFP, 10 [6-16]; platelets, 2 [1-4]). Mean ± SD age was 34 ± 16 years, 79% were male, Injury Severity Score was 33 (24-43), and 63% had penetrating injuries. Patients who received WB-CT had a lower 24-hour mortality (17% vs. 25%; p = 0.002), in-hospital mortality (29% vs. 40%; p < 0.001), major complications (29% vs. 41%; p < 0.001), and a shorter length of stay (9 [7-12] vs. 15 [10-21]; p = 0.011). On regression analysis, WB was independently associated with reduced 24-hour mortality (odds ratio [OR], 0.78 [0.59-0.89]; p = 0.006), in-hospital mortality (OR, 0.88 [0.81-0.90]; p = 0.011), and major complications (OR, 0.92 [0.87-0.96]; p = 0.013). The use of WB as an adjunct to CT is associated with improved outcomes in resuscitation of severely injured civilian trauma patients. Further studies are required to evaluate the role of adding WB to massive transfusion protocols. Therapeutic, level IV.


Effect of a Computerized Reminder on Splinting of Pediatric Upper Limb Fractures in the Emergency Department.
The primary objective was to assess if a computerized pop-up reminder increased splint application before X-ray in pediatric emergency department patients with deformed upper limb fractures. The secondary objective was to evaluate whether this same reminder improved pharmaceutical analgesia provision in this population. This was a prospective study of 518 pediatric emergency department patients with upper limb fractures after the implementation of a computerized pop-up reminder to give analgesia and apply a splint. The records of those with fractures requiring manipulation were identified and reviewed for data on demographics, clinical characteristics, splint application, and analgesic use. These data were compared with that in a previous retrospective study of 1407 patients in the same population before implementation of the computerized reminder. There were 86 patients with upper limb fractures requiring manipulation identified during the study period. After the computerized intervention, splint application rates improved from 22% to 49% (P = <0.001). There was no significant change to analgesia provision in the first hour after presentation. A computerized reminder at the point of X-ray ordering significantly improved splint application rates for children with deformed upper limb fractures, though it did not affect pharmaceutical analgesia provision. The computerized pop-up is cheap to implement, easy to use, and potentially transferrable to other institutions and for other uses.


Chiral coordination polymers based on d10 metals and 2-aminonicotinate with blue fluorescent/green phosphorescent anisotropic emissions.
Herein, we report the detailed structural characterization and photoluminescence (PL) properties of two new isostructural coordination polymers (CPs) based on 2-aminonicotinate (2ani) ligand and zinc(ii) or cadmium(ii) metal ions, namely, [M(μ-2ani)2]n [MII = Zn(1) and Cd(2)]. These compounds are characterized as chiral 2D-layered structures, resulting from the bridging of the 2ani ligands in the MN2O4 coordination shell. Also, strong supramolecular interactions are found to sustain the overall crystal structure, which endows these materials with high stability. The single crystals of 1 exhibit waveguiding behaviour along some preferred orientations. The examination of the polycrystalline samples of both compounds under UV light also shows that the samples exhibit remarkably strong PL emissions, which consist of blue fluorescence and green phosphorescence involving long-lasting phosphorescence (LLP) at low temperature. Fitting of the decay curves confirms the occurrence of short (in the order of nanoseconds) and very large lifetimes (τ ≈ 320 and 560 ms for 1 and 2, respectively) of the wavelengths responsible for their blue and green emissions, respectively, which have been studied using DFT calculations.


Leaf Form Evolution in Viburnum Parallels Variation within Individual Plants.
Few studies have critically evaluated how morphological variation within individual organisms corresponds to variation within and among species. Subindividual variation in plants facilitates such studies because their indeterminate modular growth generates multiple serially homologous structures along growing axes. Focusing on leaf form, we evaluate how subindividual trait variation relates to leaf evolution across Viburnum, a clade of woody angiosperms. In Viburnum we infer multiple independent origins of wide/lobed leaves with toothed margins from ancestors with elliptical, smooth-margined leaves. We document leaf variation along the branches of individual plants of 28 species and among populations across the wide range of Viburnum dentatum. We conclude that when novel leaf forms evolved in Viburnum, they were intercalated at the beginning of the seasonal leaf sequence, which then generated a repeated spectrum of leaf forms along each branch (seasonal heteroblasty). We hypothesize that the existence of such a spectrum then facilitated additional evolutionary shifts, including reversions to more ancestral forms. We argue that the recurrent production of alternative phenotypes provides opportunities to canalize the production of particular forms and that this phenomenon has played an important role in generating macroscale patterns.


On the function of BMP-4 in patterning the marginal zone of the Xenopus embryo.
Bone morphogenetic protein 4 (BMP-4) is expressed in the ventral marginal zone of the gastrulating embryo. At late gastrula stage this gene is expressed in the ventral-most part of the slit blastopore and in tissues that derive from it. At tailbud stages BMP-4 is expressed in the spinal cord roof plate, neural crest, eye and auditory vesicle. The interactions of BMP-4 with dorsal genes such as goosecoid (gsc) and Xnot-2 were studied in vivo. In embryos ventralized by UV irradiation and suramin treatment, BMP-4 zygotic transcripts accumulate prematurely and the entire marginal zone expresses this gene. The patterning effect of BMP-4 on ventro-posterior development can be revealed by a sensitive assay involving the injection of BMP-4 mRNA in the ventral marginal zone of embryos partially dorsalized with LiCl, which leads to the complete rescue of trunk and tail structures. The experiments presented here argue that BMP-4 may act in vivo as a ventral signal for the proper patterning of the marginal zone, actively interacting with dorsal genes such as gsc and Xnot-2. A model is proposed in which the timing of expression of various marginal zone-specific genes plays a central role in patterning the mesoderm.


Production of colony-stimulating activity by human natural killer cells: analysis of the conditions that influence the release and detection of colony-stimulating activity.
Highly purified natural killer (NK) cell suspensions were tested for their capacity to release colony-stimulating activity (CSA) in vitro. NK cell suspensions comprised primarily CD16+ cells and were devoid of CD3+ T cells, CD15+ monocytes, and of B cells. CSA was detected in the NK cell supernatants and sustained the growth of myeloid colonies from both normal peripheral blood and bone marrow. CSA could be in part inhibited by pretreating NK cell culture supernatants with a specific goat anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) antiserum. The inhibition, however, was never complete, a finding that suggests that additional factors were responsible for CSA. Incubation of NK cells with K562 cells (an NK-sensitive target) or with normal bone marrow cells resulted in the appearance of a strong colony-inhibiting activity (CIA) in the culture supernatants. Such CIA was demonstrable in an experimental system where bone marrow or peripheral blood progenitors were induced to form myeloid colonies in the presence of conditioned medium by CSA-producing giant cell tumor (GCT) cells. Stimulation of NK cells with NK-insensitive targets failed to induce CIA production. Neutralizing antitumor necrosis factor (TNF) monoclonal antibodies (MoAbs) were found capable of inhibiting CIA present in the supernatants of NK cells stimulated with K562 cells. Following treatment with anti-TNF antibodies, CSA was again detectable in the same supernatants. This finding indicates that induction of TNF production did not concomitantly switch off CSA production by NK cells. Pretreatment of NK cells with recombinant interleukin-2 (rIL-2) or gamma interferon (r gamma IFN) did not change the amount of CSA released. However, treatment with rIL-2 caused the appearance of a factor in the NK cell supernatants capable of sustaining the formation of colonies of a larger size.


Electronic gaming machines and gambling disorder: A cross-cultural comparison between treatment-seeking subjects from Brazil and the United States.
The objective of this paper is to perform a cross-cultural comparison of gambling disorder (GD) due to electronic gaming machines (EGM), a form of gambling that may have a high addictive potential. Our goal is to investigate two treatment-seeking samples of adults collected in Brazil and the United States, countries with different socio-cultural backgrounds. This comparison may lead to a better understanding of cultural influences on GD. The total studied sample involved 733 treatment-seeking subjects: 353 men and 380 women (average age=45.80, standard deviation ±10.9). The Brazilian sample had 517 individuals and the American sample 216. Subjects were recruited by analogous strategies. We found that the Brazilian sample was younger, predominantly male, less likely to be Caucasian, more likely to be partnered, tended to have a faster progression from recreational gambling to GD, and were more likely to endorse chasing losses. This study demonstrated that there are significant differences between treatment-seeking samples of adults presenting GD due to EGM in Brazil and in the United States. These findings suggest that cultural aspects may have a relevant role in GD due to EGM.


Toward Successful and Sustainable Statewide Screening for Social Determinants of Health: Testing the Interest of Hospitals.
The impact of social and behavioral factors on health outcomes are well defined in the field of public health. Additionally, characteristics such as race, ethnicity, and language have been proven to affect an individual's capacity to address health care needs. While these nonclinical components affect care, variations in screening methodology between organizations make it difficult to analyze data broadly. Standardized approaches can mitigate the impact of these factors but may be difficult to incorporate into an established workflow. The Connecticut Hospital Association identified social determinants of health (SDOH) as a factor affecting patient outcomes during a statewide collaborative on asthma. The goal of this quality improvement project was to explore change in workflow as a barrier to screening for SDOH in hospitals. Four hospitals participated in the pilot using a standardized screening tool to assess 662 patients; 62% (n = 410) were White, 11% (n = 76) were Black, 5% (n = 31) were classified as other, and 22% (n = 145) were in unknown race categories. Of those reporting needs, 438 (66%) had food-, housing-, or transportation-related needs. Qualitative interviews with staff from pilot hospitals were conducted. There were 3 main themes: the screening tool was easy to use; patients could be reluctant to reveal SDOH information; and lack of a standardized referral process made patient screening difficult to sustain or justify. The volume and magnitude of SDOH needs identified, along with the sense of helplessness expressed in qualitative interviews, reinforced the decision to implement a technology platform for screening, closed-loop referral, and outcome measurement.


The circular RNA circCPE regulates myoblast development by sponging miR-138.
Skeletal muscle development, a long-term and complex process, is controlled by a set of the myogenic genes. Circular RNAs (circRNAs), a class of noncoding RNA, have been shown to regulate various biological processes. Recent studies indicate circRNAs may be involved in myogenesis, but the role and regulatory mechanism of circRNAs in myogenesis is largely unknown. In the present study, circCPE was firstly found to promote the bovine myoblast proliferation and inhibit cell apoptosis and differentiation by influencing the expression of FOXC1 in a miR138-mediated manner. And in vivo experiments revealed that overexpression of circCPE attenuates skeletal muscle regeneration. We identified a novel circular RNA circCPE by analyzing circRNAs sequencing data of bovine muscle tissue. Sequencing verification, RNase R treatment and Actinomycin D treatment confirmed the circular nature of circCPE in bovine muscle. Functional assays showed that overexpression of circCPE could inhibit bovine myoblast apoptosis and differentiation, as well as facilitate cell proliferation. Moreover, in vivo experiments revealed that overexpression of circCPE attenuates skeletal muscle regeneration. In consideration of circRNA action as miRNAs sponge, we found that circCPE harbors miR-138 binding sites and absorbed miR-138. Mechanistically, the rescue experiments showed that the overexpression of circCPE can counteract the inhibitory effect of miR-138 on the cell proliferation and the accelerated effects on the differentiation and apoptosis. Subsequently, we found that circCPE sequester the inhibitory effect of miR-138 on FOXC1 so as to involve in myogenesis. Collectively, we constructed a novel circCPE/miR-138/FOXC1 regulatory network in bovine myogenesis, which further provide stronger evidence that circRNA involved in muscle development acting as miRNA sponge.


In vitro fermentation of carbohydrate by breast fed and formula fed infants.
Unabsorbed carbohydrates are fermented by colonic bacteria to short chain fatty acids (SCFA) which are rapidly absorbed, salvaging energy and reducing stool output. There are marked differences between the faecal flora and SCFA of breast fed (BF) and formula fed (FF) infants which may be related to the higher incidence of diarrhoea in FF infants. Part of this effect may be caused by a difference in the ability of the microflora to ferment carbohydrate. To test the hypothesis that BF and FF have different fermentation capacities for simple and complex carbohydrates, in vitro cultures of faeces from healthy infants (2-10 weeks; 11 BF, 11 FF) containing glucose, lactose, raftilose (a fructo-oligosaccharide), or soybean polysaccharide were incubated anaerobically. Results were compared with those of adult faecal cultures using the same carbohydrates. Cultures of faeces from BF and FF infants produced comparable amounts of total SCFA in all cultures. These cultures produced less SCFA than those from adult faeces and produced very little SCFA from complex carbohydrate. BF cultures produced more acetic acid than FF in all cultures, whereas FF cultures produced more propionate with sugars and more butyrate with raftilose. Both groups of infants produced less butyrate than adults in all cultures. Thus it is unlikely that a lower ability to ferment carbohydrate is a major cause of increased risk of diarrhoea in FF fed infants but individual SCFA production may be important.


Net ecosystem productivity, net primary productivity and ecosystem carbon sequestration in a Pinus radiata plantation subject to soil water deficit.
Tree carbon (C) uptake (net primary productivity excluding fine root turnover, NPP') in a New Zealand Pinus radiata D. Don plantation (42 degrees 52' S, 172 degrees 45' E) growing in a region subject to summer soil water deficit was investigated jointly with canopy assimilation (A(c)) and ecosystem-atmosphere C exchange rate (net ecosystem productivity, NEP). Net primary productivity was derived from biweekly stem diameter growth measurements using allometric relations, established after selective tree harvesting, and a litterfall model. Estimates of A(c) and NEP were used to drive a biochemically based and environmentally constrained model validated by seasonal eddy covariance measurements. Over three years with variable rainfall, NPP' varied between 8.8 and 10.6 Mg C ha(-1) year(-1), whereas A(c) and NEP were 16.9 to 18.4 Mg C ha(-1) year(-1) and 5.0-7.2 Mg C ha(-1) year(-1), respectively. At the end of the growing season, C was mostly allocated to wood, with nearly half (47%) to stems and 27% to coarse roots. On an annual basis, the ratio of NEP to stand stem volume growth rate was 0.24 +/- 0.02 Mg C m(-3). The conservative nature of this ratio suggests that annual NEP can be estimated from forest yield tables. On a biweekly basis, NPP' repeatedly lagged A(c), suggesting the occurrence of intermediate C storage. Seasonal NPP'/A(c) thus varied between nearly zero and one. On an annual basis, however, NPP'/A(c) was 0.54 +/- 0.03, indicating a conservative allocation of C to autotrophic respiration. In the water-limited environment, variation in C sequestration rate was largely accounted for by a parameter integrative for changes in soil water content. The combination of mensurational data with canopy and ecosystem C fluxes yielded an estimate of heterotrophic respiration (NPP' - NEP) approximately 30% of NPP' and approximately 50% of NEP. The estimation of fine-root turnover rate is discussed.


Functional Remodeling Associated With Language Recovery After Repetitive Transcranial Magnetic Stimulation in Chronic Aphasic Stroke.
Repetitive transcranial magnetic stimulation (rTMS) has shown promising efficacy in improving the language functions in poststroke aphasia. However, randomized controlled trials were lacking to investigate the rTMS-related neuroimaging changes underlying the therapeutic effects on language improvement in chronic aphasia. In this study, we aimed to evaluate the effects of low-frequency rTMS (LF-rTMS) on chronic poststroke aphasia. We hypothesized that the deactivation of the right pars triangularis could restore the balance of interhemispheric inhibition and, hence, facilitated the functional remodeling of language networks in both the hemispheres. Furthermore, the rTMS-induced functional reorganization should underpin the language recovery after rTMS. A total of 33 patients (22 males; age: 58.70 ± 13.77 years) with chronic stroke in the left hemisphere and nonfluent aphasia were recruited in this randomized double-blinded study. The ratio of randomization between the rTMS and sham groups is 17:16. All the patients received real 1-Hz rTMS or sham stimulation (placebo coil delivered < 5% of magnetic output with similar audible click-on discharge) at the right posterior pars triangularis for 10 consecutive weekdays (stroke onset to the first stimulation: 10.97 ± 10.35 months). Functional connectivity of language networks measured by resting-state fMRI was calculated and correlated to the scores of the Concise Chinese Aphasia Test by using the stepwise regression analysis. After LF-rTMS intervention, significant improvement in language functions in terms of comprehension and expression abilities was observed compared with the sham group. The rTMS group showed a significant decrease of coupling strength between right pars triangularis and pars opercularis with a strengthened connection between right pars orbitalis and angular gyrus. Furthermore, the LF-rTMS significantly enhanced the coupling strength associated with left Wernicke area. Results of regression analysis showed that the identified functional remodeling involving both the hemispheres could support and predict the language recovery after LF-rTMS treatment. We reported the therapeutic effects of LF-rTMS and corresponding functional remodeling in chronic poststroke aphasia. Our results provided neuroimage evidence reflecting the rebalance of interhemispheric inhibition induced by LF-rTMS, which could facilitate future research in the refinement of rTMS protocol to optimize the neuromodulation efficacy and benefit the clinical management of patients with stroke.


Calcium channels in planar lipid bilayers: insights into mechanisms of ion permeation and gating.
Electrophysiological recordings were used to analyze single calcium channels in planar lipid bilayers after membranes from bovine cardiac sarcolemmal vesicles had been incorporated into the bilayer. In these cell-free conditions, channels in the bilayer showed unitary barium or calcium conductances, gating kinetics, and pharmacological responses that were similar to dihydropyridine-sensitive calcium channels in intact cells. The open channel current varied in a nonlinear manner with voltage under asymmetric (that is, physiological) ionic conditions. However, with identical solutions on both sides of the bilayer, the current-voltage relation was linear. In matched experiments, calcium channels from skeletal muscle T-tubules differed significantly from cardiac calcium channels in their conductance properties and gating kinetics.


A sacro-caudal spinal cord choroid plexus papilloma in a shar-pei dog.
A seven-year-old shar-pei dog was referred because of severe lumbosacral pain and faecal incontinence of 20 days' duration. Neurological examination was characterised by plegic tail, absence of perineal reflex, dilated anus, perineum and tail analgesia, and severe lumbosacral pain. The neurological clinical signs were suggestive of a selective lesion involving sacral and caudal spinal cord segments and/or related nerve roots. A magnetic resonance imaging of lumbosacral spine was performed and was suggestive of an intradural lesion. Primary or secondary neoplasia was considered as the most probable differential diagnosis. The dog was euthanased upon the owner's request. Histopathological examination confirmed the presence of an intradural-extramedullary neoplastic tissue enveloping intradural tract of spinal nerve roots. On the basis of histological and immunohistochemical findings, a diagnosis of well-differentiated choroid plexus papilloma was made. To the authors's knowledge, this is the first case of primary or metastatic spinal choroid plexus papilloma in dogs.


Non-invasive visualization of tumor infiltrating lymphocytes in patients with metastatic melanoma undergoing immune checkpoint inhibitor therapy: a pilot study.
Early in the course of immunotherapy there is frequently a transient enlargement of tumor masses (pseudo-progression) due to tumor infiltration by TILs. Current clinical imaging modalities are not able to distinguished pseudo-progression from true tumor progression. Thus, patients often remain on treatment 4-8 weeks longer to confirm disease progression. Nuclear medicine offers the possibility to image immune cells and potentially discriminate pseudo-progression and progression. We conducted a pilot study in patients with metastatic melanoma receiving ipilimumab (IPI) or pembrolizumab (PEMBRO) to assess safety and feasibility of SPECT/CT imaging with 99mTc- interleukin-2 (99mTc-HYNIC-IL2) to detect TILs and distinguish between true progression from pseudo- progression. Scans were performed prior to and after 12w treatment. After labelling,99mTc-HYNIC-IL2 was purified and diluted in 10 mL of 5% glucose with 0.1% human serum albumin. Of the 5 patients (2 treated with IPI and 3 with PEMBRO) enrolled, two failed to complete the second scan as they discontinued IPI due grade 3 colitis (1 patient) or patient refusal after developing multiple toxicities attributed to IPI (1 patient). Following the first scan, one patient reported to have a grade 1 pruritus with grade 1 pain. No other toxicities attributed to the radiopharmaceutical infusion were reported. Metastatic lesions could be visualized by 99mTc-IL2 imaging and there was positive correlation between size and 99mTc-HYNIC-IL2 uptake, both before and after 12 weeks of therapy. The results of this pilot study demonstrate the safety and feasibility of 99mTc-IL2 imaging and has led to a number of hypotheses to be tested in future studies.


Sound practice: Exploring the benefits of establishing a music group on an acute mental health inpatient unit.
There is evidence supporting the use of formal music therapy in the treatment of mental health consumers. Despite this, it appears to be an intervention which has not been routinely offered to consumers in Australian acute mental health inpatient units, possibly due to the lack of trained music therapists (or inadequate funding to employ them), as well as the challenges posed by the acuity of presentations and the short duration of admissions. Less formal therapeutic music activities may benefit consumers within these settings. This article describes how a music group activity facilitated by clinical staff with no music therapy qualifications was established. The first phase of this evaluation is then described using a descriptive qualitative method. We undertook a series of consumer and staff focus groups to explore the impact of a music group activity on an acute mental health inpatient unit. Five themes emerged from the transcripts of the focus groups' discussions, effects on mood, relationships and engagement, social connectedness and inclusion, the ward atmosphere and noise/agitation. Positive effects were shown across these areas, suggesting that the music group activity we established was beneficial for consumers and staff, and enhanced the ward atmosphere.


Intraepithelial neutrophils in pediatric severe asthma are associated with better lung function.
Neutrophils and IL-17A have been linked mechanistically in models of allergic airways disease and have been associated with asthma severity. However, their role in pediatric asthma is unknown. We sought to investigate the role of neutrophils and the IL-17A pathway in mediating pediatric severe therapy-resistant asthma (STRA). Children with STRA (n = 51; age, 12.6 years; range, 6-16.3 years) and controls without asthma (n = 15; age, 4.75 years; range, 1.6-16 years) underwent clinically indicated fiberoptic bronchoscopy, bronchoalveolar lavage (BAL), endobronchial brushings, and biopsy. Neutrophils, IL-17A, and IL-17RA-expressing cells and levels of IL-17A and IL-22 were quantified in BAL and biopsies and related to clinical features. Primary bronchial epithelial cells were stimulated with IL-17A and/or IL-22, with and without budesonide. Children with STRA had increased intraepithelial neutrophils, which positively correlated with FEV1 %predicted (r = 0.43; P = .008). Neutrophilhigh patients also had better symptom control, despite lower dose maintenance inhaled steroids. Submucosal neutrophils were not increased in children with STRA. Submucosal and epithelial IL-17A-positive cells and BAL IL-17A and IL-22 levels were similar in children with STRA and controls. However, there were significantly more IL-17RA-positive cells in the submucosa and epithelium in children with STRA compared with controls (P = .001). Stimulation of primary bronchial epithelial cells with IL-17A enhanced mRNA expression of IL-17RA and increased release of IL-8, even in the presence of budesonide. A proportion of children with STRA exhibit increased intraepithelial airway neutrophilia that correlated with better lung function. STRA was also characterized by increased airway IL-17RA expression. These data suggest a potential beneficial rather than adverse role for neutrophils in pediatric severe asthma pathophysiology.


Protein disorder and the evolution of molecular recognition: theory, predictions and observations.
Observations going back more than 20 years show that regions in proteins with disordered backbones can play roles in their binding to other molecules; typically, the disordered regions become ordered upon complex formation. Thought-experiments with Schulz Diagrams, which are defined herein, suggest that disorder-to-order transitions are required for natural selection to operate separately on affinity and specificity. Separation of affinity and specificity may be essential for fine-tuning the molecular interaction networks that comprise the living state. For low affinity, high specificity interactions, our analysis suggests that natural selection would parse the amino acids conferring flexibility in the unbound state from those conferring specificity in the bound state. For high affinity, low specificity or for high affinity, multiple specificity interactions, our analysis suggests that the disorder-to-order transitions enable alternative packing interactions between side chains to accommodate the different binding targets. Disorder-to-order transitions upon binding also have significant kinetic implications as well, by having complex effects on both on- and off-rates. Current data are insufficient to decide on these proposals, but sequence and structure analysis on two examples support further investigations of the role of disorder-to-order transitions upon binding.


Quercus infectoria gall extracts reduce quorum sensing-controlled virulence factors production and biofilm formation in Pseudomonas aeruginosa recovered from burn wounds.
Quercus gall extracts' ability to kill pathogens in vitro and even removal of chronic drug-resistant infections has been reported by several studies. The current investigation is focused on the action of extracts of Quercus infectoria gall in their sub-inhibitory concentrations on the corresponding bacterial behaviours instead of killing them. The effect of gall extracts on the quorum sensing (QS) associated virulence of multiple drug resistant Pseudomonas aeruginosa recovered from burns wounds was studied. The influence of different extracts on the production of bacterial virulence and biofilm, and expression of the genes encoding quorum sensing and exotoxin A were investigated. Quorum sensing is a crucial regulator of virulence and biofilm development in Pseudomonas aeruginosa and other medical related microbes. Experiments to characterise and quantify Q. infectoria gall extracts impact on the quorum sensing networks of P.aeruginosa revealed that the expression of las, rhl, and exotoxin A (ETA) genes levels including the associated virulence were reduced by the extracts at their subinhibitory concentrations. The obtained results indicated that extracts of Q. infectoria galls fight infections either by their inhibitory constituents, which vigorously eradicate cells or by disruption of the pathogens quorum sensing system through weakening the virulence and bacterial coordination.


Healing of saccular aneurysms following platinum coil embolization: lack of improved efficacy with vitamin C supplementation.
To test the hypothesis that systemic administration of vitamin C, through its action of stimulating collagen synthesis and crosslinking, would decrease the recurrence and improve the occlusion of experimental aneurysms treated with platinum coils. Experimental aneurysms were created in female rabbits and were embolized with platinum coils (>30% packing density). The animals were divided into three groups: group 1 (n=6) rabbits served as controls, group 2 (n=5) rabbits were fed with a vitamin C supplemented feed and group 3 (n=8) rabbits were medicated with vitamin C pills. Digital subtraction angiography was used to evaluate stability after embolization. Subjects were euthanized at 12 weeks after coil implantation, and serum vitamin C levels were then measured. Histological samples were examined with a grading system (range 0-12) based on the neck and dome features. Masson Trichrome staining was used to evaluate collagen deposition. Parametric data were analyzed with one way analysis of variance and non-parametric data were examined using a Kruskal-Wallis test. There were no significant differences between groups in mean aneurysm size. Mean serum vitamin C concentration was significantly higher in group 3 and group 2 compared with group 1, while vitamin C levels between group 2 and group 3 were statistically comparable. Coil compaction was noted in one of six subjects in group 1 and in three of eight subjects in group 3. All of the remaining aneurysms in the test and control groups showed stable occlusion. There were no significant differences in histological scores or collagen deposition among groups. Vitamin C supplementation following platinum coil embolization does not demonstrate improvement of long term occlusion rates of aneurysms.


Severe tricuspid valve endocarditis related to tunneled catheters in chronic hemodialysis patients: when should the catheter be removed?
Hemodialysis (HD) patients are predisposed to infective endocarditis (IE) mainly due to repeated manipulation of the vascular access. However, catheter seeding and IE may also result from a distant infection site. A diabetic patient who was maintained on regular HD through a permanent catheter in the right internal jugular vein presented with septicemia of urinary origin due to Pseudomonas aeruginosa. He was treated with injectable antibiotics for 15 days. Few days after the end of the treatment period he presented with inflammation of the catheter exit site. Blood cultures revealed Pseudomonas aeruginosa and echocardiography showed a large vegetation on the tricuspid valve, confirming the diagnosis of IE. The patient improved after catheter removal and treatment with ceftazidime, vancomycin and amikacin. Another patient who was maintained on chronic HD through a tunneled catheter in the right internal jugular vein presented with a limited infection in the sub-cutaneous tunnel of the catheter that improved after treatment with injectable antibiotics. Two months later, he presented with severe sepsis without signs of local infection of the catheter. Chest radiography showed right sided infiltrates. Injectable antibiotic therapy was given without improvement. Blood cultures were negative but echocardiography showed a large tricuspid valve vegetation. Antibiotic regimen was modified to include vancomycin, amikacin and ceftazidime and the catheter was removed. Unfortunately, the patient died two days after diagnosing IE. catheter seeding and IE may be a consequence of sepsis originating away from the HD catheter site. Tunneled catheters may need to be preemptively removed in such situations to prevent serious IE.


Gestational weight gain counselling practices among different antenatal health care providers: a qualitative grounded theory study.
Inappropriate gestational weight gain in pregnancy may negatively impact health outcomes for mothers and babies. While optimal gestational weight gain is often not acheived, effective counselling by antenatal health care providers is recommended. It is not known if gestational weight gain counselling practices differ by type of antenatal health care provider, namely, family physicians, midwives and obstetricians, and what barriers impede the delivery of such counselling. The objective of this study was to understand the counselling of family physicians, midwives and obstetricians in Ontario and what factors act as barriers and enablers to the provision of counselling about GWG. Semi-structured interviews were conducted with seven family physicians, six midwives and five obstetricians in Ontario, Canada, where pregnancy care is universally covered. Convenience and purposive sampling techniques were employed. A grounded theory approach was used for data analysis. Codes, categories and themes were generated using NVIVO software. Providers reported that they offered gestational weight gain counselling to all patients early in pregnancy. Counselling topics included gestational weight gain targets, nutrition & exercise, gestational diabetes prevention, while dispelling misconceptions about gestational weight gain. Most do not routinely address the adverse outcomes linked to gestational weight gain, or daily caloric intake goals for pregnancy. The health care providers all faced similar barriers to counselling including patient attitudes, social and cultural issues, and accessibility of resources. Patient enthusiasm and access to a dietician motivated health care providers to provide more in-depth gestational weight gain counselling. Reported gestational weight gain counselling practices were similar between midwives, obstetricians and family physicians. Antenatal knowledge translation tools for patients and health care providers are needed, and would seem to be suitable for use across all three types of health care provider specialties.


The nature of torque "overshoot" in Cybex isokinetic dynamometry.
Transient peaks or "spikes" frequently appear in the initial segments of torque curves recorded with the Cybex isokinetic dynamometer. The purpose of this investigation was to determine whether these spikes represent artifact or transient initial surges of true muscular force output. Cinematographic analysis using both inert weights and a human subject as the source of torque input to the Cybex revealed that the dynamometer's input lever initially exceeded the pre-set angular velocity by an amount ranging from 11% (inert weight; pre-set Cybex velocity = 180 degrees/s) to 200% (human hip abduction; pre-set velocity = 30 degrees/s). The majority of this "overspeeding" occurred in the latter part of the free acceleration period, prior to the engagement of the dynamometer's resistance mechanism. The remainder occurred in the initial part of the elastic loading phase, just after resistance had engaged. A sharp deceleration of the overspeeding lever and the affixed weight or limb then followed in response to the dynamometer's continuing build-up of resistance. Simultaneous with this deceleration, a prominent torque spike was recorded that superseded the correct (mechanical equilibrium) torque value. Within our error of measurement, the deceleration observed in the film quantitatively accounted for all of the "overshoot" torque, i.e., that amount of the spike that exceeded the correct value. It was concluded that such prominent, initial torque spikes represent inertial forces and should not be confused with true muscular tension development.


[Chemical neuro-anatomy of the thalamus: an example of the co-expression of 2 neuropeptides, cholecystokinin and intestinal vasoactive peptide].
Recent studies using hybridization histochemistry have demonstrated the presence of cholecystokinin (CCK) gene expression in thalamocortical and thalamo-striatal neurons. To further understand the chemical anatomy of the thalamus, we used the same method to study the coexpression of CCK and vasoactive intestinal peptide (VIP) genes in the same neurons. Most of the neurons expressing the VIP gene in the rat dorsal thalamus (found especially in the ventrolateral nucleus) also expressed the CCK gene, as demonstrated by autoradiography on emulsion-coated adjacent sections studied with probes recognizing either mRNA. Some further neurons, located in the reticular nucleus, had VIP mRNA at lower levels of expression but did not appear to contain CCK mRNA.