Retrograde transport of Golgi-localized proteins to the ER.
The ER is uniquely enriched in chaperones and folding enzymes that facilitate folding and unfolding reactions and ensure that only correctly folded and assembled proteins leave this compartment. Here we address the extent to which proteins that leave the ER and localize to distal sites in the secretory pathway are able to return to the ER folding environment during their lifetime. Retrieval of proteins back to the ER was studied using an assay based on the capacity of the ER to retain misfolded proteins. The lumenal domain of the temperature-sensitive viral glycoprotein VSVGtsO45 was fused to Golgi or plasma membrane targeting domains. At the nonpermissive temperature, newly synthesized fusion proteins misfolded and were retained in the ER, indicating the VSVGtsO45 ectodomain was sufficient for their retention within the ER. At the permissive temperature, the fusion proteins were correctly delivered to the Golgi complex or plasma membrane, indicating the lumenal epitope of VSVGtsO45 also did not interfere with proper targeting of these molecules. Strikingly, Golgi-localized fusion proteins, but not VSVGtsO45 itself, were found to redistribute back to the ER upon a shift to the nonpermissive temperature, where they misfolded and were retained. This occurred over a time period of 15 min-2 h depending on the chimera, and did not require new protein synthesis. Significantly, recycling did not appear to be induced by misfolding of the chimeras within the Golgi complex. This suggested these proteins normally cycle between the Golgi and ER, and while passing through the ER at 40 degrees C become misfolded and retained. The attachment of the thermosensitive VSVGtsO45 lumenal domain to proteins promises to be a useful tool for studying the molecular mechanisms and specificity of retrograde traffic to the ER.


Spontaneous pre-stimulus oscillatory activity shapes the way we look: A concurrent imaging and eye-movement study.
Previous behavioural studies have accrued evidence that response time plays a critical role in determining whether selection is influenced by stimulus saliency or target template. In the present work, we investigated to what extent the variations in timing and consequent oculomotor controls are influenced by spontaneous variations in pre-stimulus alpha oscillations. We recorded simultaneously brain activity using magnetoencephalography (MEG) and eye movements while participants performed a visual search task. Our results show that slower saccadic reaction times were predicted by an overall stronger alpha power in the 500 ms time window preceding the stimulus onset, while weaker alpha power was a signature of faster responses. When looking separately at performance for fast and slow responses, we found evidence for two specific sources of alpha activity predicting correct versus incorrect responses. When saccades were quickly elicited, errors were predicted by stronger alpha activity in posterior areas, comprising the angular gyrus in the temporal-parietal junction (TPJ) and possibly the lateral intraparietal area (LIP). Instead, when participants were slower in responding, an increase of alpha power in frontal eye fields (FEF), supplementary eye fields (SEF) and dorsolateral pre-frontal cortex (DLPFC) predicted erroneous saccades. In other words, oculomotor accuracy in fast responses was predicted by alpha power differences in more posterior areas, while the accuracy in slow responses was predicted by alpha power differences in frontal areas, in line with the idea that these areas may be differentially related to stimulus-driven and goal-driven control of selection.


Outcome at 8 years after breast-conserving surgery and radiation therapy for invasive breast cancer: influence of margin status and systemic therapy on local recurrence.
To examine the relationship between pathologic margin status and outcome at 8 years after breast-conserving surgery and radiation therapy. The study population comprised 533 patients with International Union Against Cancer/American Joint Committee on Cancer clinical stage I or II breast cancer who had assessable margins, who received at least 60 Gy to the primary tumor bed, and who had more than 8 years of potential follow-up. Each margin was scored (according to the presence of invasive or in situ disease that touched the inked surgical margin) as one of the following: negative, close, focally positive, or extensively positive. Outcome at 8 years was calculated using crude rates of first site of failure. A polychotomous logistic regression analysis was performed. Median follow-up time was 127 months. At 8 years, patients with close margins and those with negative margins both had a rate of local recurrence (LR) of 7%. Patients with extensively positive margins had an LR rate of 27%, whereas patients with focally positive margins had an intermediate rate of LR of 14%. In the polychotomous logistic regression model, margin status and the use of systemic therapy were the only two variables that had significant effects on the risk ratio of LR to remaining alive and free of disease. Among the 45 patients with focally positive margins who received systemic therapy, the crude LR rate was 7% at 8 years (95% confidence interval, 1% to 20%). Pathologic margin status and the use of adjuvant systemic therapy are the most important factors associated with LR among patients treated with breast-conserving surgery and radiation therapy.


Distribution of echovirus antibody in serum, nasopharynx, rectum, and spinal fluid after natural infection with echovirus type 6.
By employing the techniques of radioimmunodiffusion and autoradiography and by using (32)P-labeled echovirus 6 as the antigen, the distribution of echovirus 6 antibody in serum, nasopharynx, rectum, and spinal fluid was studied in a group of children with naturally acquired echovirus type 6 infection, with or without meningeal involvement. The response in serum was characterized by sequential appearance of gammaM, gammaG, and rarely of gammaA antibody. The secretory response in the nasopharynx and rectum was limited to secretory gammaA immunoglobulin. The response in spinal fluid was limited to gammaG immunoglobulin. gammaG echovirus antibody appeared in the spinal fluid with or without demonstrable replication of virus in the spinal fluid. It is suggested that, after short-term acute infection with enteroviruses, the appearance of gammaG antibody in spinal fluid may largely be a reflection of increased transport of serum antibody, particularly of gammaG immunoglobulin class.


Distribution of genes encoding adhesins and biofilm formation capacity among Uropathogenic Escherichia coli isolates in relation to the antimicrobial resistance.
Escherichia coli is the most predominant pathogen involved in UTIs. Mainly, fimbrial surface appendages are implicated in adherence to urothelium besides non-fimbrial proteins. to determine prevalence of genes encoding fimbrial and non-fimbrial proteins among Uropathogenic Escherichia coli (UPEC). Furthermore, distribution of these genes and biofilm formation capacity were investigated in relation to antimicrobial resistance. Antimicrobial susceptibility of 112 UPEC isolates was performed using disc diffusion method. ESBL production was confirmed by double disc synergy test. Genes encoding fimbrial and non-fimbrial proteins were detected using PCR and biofilm formation was investigated using microtitre plate assay. UPEC isolates exhibited high resistance against doxycyclines (88.39 %), β-lactams (7.14-86.6%), sulphamethoxazole-trimethoprim (53.75%) and fluoro-quinolones (50%). Fifty percent of tested isolates were ESBL producers. PapGII gene was statistically more prevalent among pyelonephritis isolates. SfaS, focG and picU genes were statistically associated with fluoroquinolone (FQs) sensitive isolates and Dr/afaBC gene was statistically associated with ESBL production. Moreover, non-MDR isolates produced sturdier biofilm. PapGII adhesin variant seems to have a critical role in colonization of upper urinary tract. There is a possible link between antimicrobial resistance and virulence being capable of affecting the distribution of some genes besides its negative impact on biofilm formation.


Local excitatory circuits in the intermediate gray layer of the superior colliculus.
We have used photostimulation and whole cell patch-clamp recording techniques to examine local synaptic interactions in slices from the superior colliculus of the tree shrew. Uncaging glutamate 10-75 microm from the somata of neurons in the intermediate gray layer elicited a long-lasting inward current, due to direct activation of glutamate receptors on these neurons, and brief inward currents caused by activation of presynaptic neurons. The synaptic responses occurred as individual currents or as clusters that lasted up to several hundred milliseconds. Excitatory synaptic responses, which reversed at membrane potentials near 0 mV, could be evoked by uncaging glutamate anywhere within 75 microm of an intermediate layer neuron. Our results indicate the presence of extensive local excitatory circuits in the intermediate layer of the superior colliculus and support the hypothesis that such intrinsic circuitry contributes to the development of presaccadic command bursts.


In vivo evaluation of adeno-associated virus gene transfer in airways of mice with acute or chronic respiratory infection.
Patients with cystic fibrosis (CF) often suffer chronic lung infection with concomitant inflammation, a setting that may reduce the efficacy of gene transfer. While gene therapy development for CF often involves viral-based vectors, little is known about gene transfer in the context of an infected airway. In this study, three mouse models were established to evaluate adeno-associated virus (AAV) gene transfer in such an environment. Bordetella bronchiseptica RB50 was used in a chronic, nonlethal respiratory infection in C57BL/6 mice. An inoculum of ∼10(5) CFU allowed B. bronchiseptica RB50 to persist in the upper and lower respiratory tracts for at least 21 days. In this infection model, administration of an AAV vector on day 2 resulted in 2.8-fold reduction of reporter gene expression compared with that observed in uninfected controls. Postponement of AAV administration to day 14 resulted in an even greater (eightfold) reduction of reporter gene expression, when compared with uninfected controls. In another infection model, Pseudomonas aeruginosa PAO1 was used to infect surfactant protein D (SP-D) or surfactant protein A (SP-A) knockout (KO) mice. With an inoculum of ∼10(5) CFU, infection persisted for 2 days in the nasal cavity of either mouse model. Reporter gene expression was approximately ∼2.5-fold lower compared with uninfected mice. In the SP-D KO model, postponement of AAV administration to day 9 postinfection resulted in only a two fold reduction in reporter gene expression, when compared with expression seen in uninfected controls. These results confirm that respiratory infections, both ongoing and recently resolved, decrease the efficacy of AAV-mediated gene transfer.


Optimization of the classical oral cancerization protocol in hamster to study oral cancer therapy.
The hamster carcinogenesis model recapitulates oral oncogenesis. Dimethylbenz[a]anthracene (DMBA) cancerization induces early severe mucositis, affecting animal's welfare and causing tissue loss and pouch shortening. "Short" pouches cannot be everted for local irradiation for boron neutron capture therapy (BNCT). Our aim was to optimize the DMBA classical cancerization protocol to avoid severe mucositis, without affecting tumor development. We evaluated BNCT in animals cancerized with this novel protocol. We studied: Classical cancerization protocol (24 applications) and Classical with two interruptions (completed at the end of the cancerization protocol). BNCT mediated by boronophenylalanine (BPA) was performed in both groups. The twice-interrupted group exhibited a significantly lower percentage of animals with severe mucositis versus the non-interrupted group (17% versus 71%) and a significantly higher incidence of long pouches (100% versus 53%). Tumor development and the histologic characteristics of tumor and precancerous tissue were not affected by the interruptions. For both groups, overall tumor response was more than 80%, with a similar incidence of BNCT-induced severe mucositis. The twice-interrupted protocol reduced severe mucositis during cancerization without affecting tumor development. This favored the animal's welfare and reduced the number of animals to be cancerized for our studies, without affecting BNCT response.


Predictive Factors of Toddlers' Sleep and Parental Stress.
Although most infants consolidate their sleep habits during the first year of life, for many children, sleep is described as disrupted during toddlerhood. Along with individual child variables such as temperamental characteristics, parenting behaviors play a key role in determining children's sleep-wake patterns. The aims of the current study were to evaluate the relationship among toddlers' sleep quality, emotion regulation, bedtime routines, parental bedtime involvement, parental perceived social support and stress, and to integrate a novel combination of the aforementioned dimensions into predictive models of toddlers' sleep quality and parental stress. One hundred and sixty parents with 2-3-year-old children filled out the following self-report questionnaires: the Parent-Child Sleep Interaction Scale; the Emotion Regulation Checklist; the Social Provisions Scale; and an ad-hoc questionnaire to assess parental involvement in everyday and bedtime care for children. Three multiple regression analyses were conducted by regressing maternal and paternal parenting stress and infant's quality sleep onto the independent variables described above. Toddlers' emotion regulation and parental psychosocial functioning were related to parental stress. Toddlers' night awakenings and the time required by toddlers to fall asleep were related to parental distress. The findings evidenced the bidirectional associations among the studied variables, highlighting the protective role of social support in reducing parenting stress and of paternal bedtime involvement in improving toddlers' sleep quality.


Constraints on the formation and structure of Langmuir eigenmodes in the solar wind.
Localized Langmuir waves are commonly observed in space plasmas and are a potential source of radio waves. Using electric field data from STEREO, it is shown that these localized Langmuir waves are eigenmodes of density wells estimated independently. An analytic model is developed for the eigenmode frequencies. The inferred depths and widths of the density wells typically only allow the zeroth-order Langmuir eigenmode to form, explaining the preponderance of single-peaked waveforms. More complicated waveforms are shown to be consistent with single eigenmode solutions of more complicated density profiles. The inferred depth of the density well increases with Langmuir wave intensity, consistent with the ponderomotive force but not wave packet collapse.


Anti-cancer effect of adenovirus p53 on human cervical cancer cell growth in vitro and in vivo.
To evaluate anti-tumor effects of recombinant adenovirus p53, time-course p53, E6 expression, and cell growth inhibition were investigated in vitro and in vivo using cervical cancer cell lines such as CaSki, SiHa, HeLa, HeLaS3, C33A, and HT3. The cell growth inhibition was studied via cell count assay, MTT assay and neutral red assay. After transfecting AdCMVp53 into SiHa cells-xenografted nude mice, the transduction efficiency and anti-tumor effect were investigated for a month. The results showed that adenoviral p53 expression induced significant growth suppression on the cancer cells, in which E6 transcript was strongly repressed, and that the expression of p53 and E6 were remarkably dependent on each cell type. The transduction efficiency was highly maintained in vivo as well as in vitro, and the size of tumor was remarkably decreased in comparison with AdCMVLacZ control. The results suggest that the adenovirus-mediated p53 gene transfection was done very effectively in vitro and in vivo experiment, and the cell growth was suppressed via p53-dependent apoptotic cell death, and that the anti-tumor effect could be related to E6 and p53 expression pattern.


Eye Size Affects Cuteness in Different Facial Expressions and Ages.
Researchers have suggested that infants exhibiting baby schema are considered cute. These similar studies have mainly focused on changes in overall baby schema facial features. However, whether a change in only eye size affects the perception of cuteness across different facial expressions and ages has not been explicitly evaluated until now. In the present study, a paired comparison method and 7-point scale were used to investigate the effects of eye size on perceived cuteness across facial expressions (positive, neutral, and negative) and ages (adults and infants). The results show that stimuli with large eyes were perceived to be cuter than both unmanipulated eyes and small eyes across all facial expressions and age groups. This suggests not only that the effect of baby schema on cuteness is based on changes in a set of features but also that eye size as an individual feature can affect the perception of cuteness.


Fluorescent Molecular Rotor-in-Paraffin Waxes for Thermometry and Biometric Identification.
Novel thermoresponsive sensor systems consisting of a molecular rotor (MR) and paraffin wax (PW) were developed for various thermometric and biometric identification applications. Polydiphenylacetylenes (PDPAs) coupled with long alkyl chains were used as MRs, and PWs of hydrocarbons having 16-20 carbons were utilized as phase-change materials. The PDPAs were successfully dissolved in the molten PWs and did not act as an impurity that prevents phase transition of the PWs. These PDPA-in-PW hybrids had almost the same enthalpies and phase-transition temperatures as the corresponding pure PWs. The hybrids exhibited highly reversible fluorescence (FL) changes at the critical temperatures during phase transition of the PWs. These hybrids were impregnated into common filter paper in the molten state by absorption or were encapsulated into urea resin to enhance their mechanical integrity and cyclic stability during repeated use. The wax papers could be utilized in highly advanced applications including FL image writing/erasing, an array-type thermo-indicator, and fingerprint/palmprint identification. The present findings should facilitate the development of novel fluorescent sensor systems for biometric identification and are potentially applicable for biological and biomedical thermometry.


Cleavage of sterol regulatory element binding proteins (SREBPs) by CPP32 during apoptosis.
Cellular cholesterol homeostasis is controlled by sterol-regulated proteolysis of membrane-bound transcription factors called sterol-regulatory element binding proteins (SREBPs). CPP32, a cysteine protease, was shown previously to cleave SREBP-1 and SREBP-2 in vitro at an aspartic acid between the basic helix-loop-helix leucine zipper domain and the first trans-membrane domain, liberating a transcriptionally active fragment. Here, we show that CPP32 exists in an inactive 32 kDa form in Chinese hamster ovary (CHO) cells. When apoptosis was induced with the protein kinase inhibitor staurosporine, CPP32 was cleaved to subunits of 20 and 10 kDa to form the active protease. Under these conditions membrane-bound SREBP-1 and SREBP-2 were both cleaved, and the transcriptionally active N-terminal fragments were found in nuclear extracts. Similar results were obtained in human U937 cells induced to undergo apoptosis by anti-Fas and etoposide. The apoptosis-induced cleavage of SREBPs was not suppressed by sterols, indicating that apoptosis-induced cleavage and sterol-regulated cleavage are mediated by different proteases. CHO cells expressing a mutant SREBP-2 with an Asp--> Ala mutation at the CPP32 cleavage site showed sterol-regulated cleavage but no apoptosis-induced cleavage. These data are consistent with the emerging concept that CPP32 is a central mediator in apoptosis. They also indicate that SREBPs, like poly (ADP) ribose polymerase, are cleaved by CPP32 during programmed cell death.


An ITS phylogeny of Leccinum and an analysis of the evolution of minisatellite-like sequences within ITS1.
Phylogenetic relationships of the European species of Leccinum (Boletales, Boletaceae) were investigated by maximum parsimony, Bayesian and likelihood analyses of nrITS1-5.8S-ITS2 and 28S sequences. The separate gene trees inferred were largely concordant, and their combined analysis indicates that several traditional sectional and species-level taxonomic schemes are artificial. In Leccinum, the nrITS region ranges in size from 694 to 1480 bp. This extreme length heterogeneity is localized to a part of the ITS1 spacer that contains a minisatellite characterized by the repeated presence of CTATTGAAAAG and CTAATAGAAAG core sequences and mutational derivatives thereof. The number of core sequences present in the minisatellite varied from 12 to 36. Intra-individual sequence variation of the minisatellite was always smaller than between different species, indicating that concerted evolution proceeds rapidly enough to retain phylogenetic signal at the infraspecific level. In contrast, the evolutionary pattern exhibited by the major ITS1 repeat types found was homoplastic when mapped onto the species lineages inferred from the combined 5.8S-ITS2 sequences. The minisatellite therefore appears not to be useful for phylogeny reconstruction at or above the species level.


Origin of notches in CSF: optical or neural?
Grating contrast sensitivity was measured across a range of 1 to 32 cycles per degree (c/deg) in normal observers with a computer-automated method of ascending limits. Monocular contrast sensitivity functions (CSF) were obtained for vertical, oblique and horizontal orientations, with or without full refractive correction. Small amounts of astigmatic error resulted in loss of sensitivity at selective spatial frequencies. Coincident with these CSF "notches" was the presence of monocular diplopia induced, in this study, by the condition of astigmatic error. Experimental manipulation of the selective spatial frequency losses was possible by the introduction of slight cylindrical defocus and by changes in grating orientation. Determination of the angular displacement and orientation of the monocular double images allowed prediction of the spatial frequencies which would show reduced sensitivity due to partial cancellation of contrast. The close fit between the predicted and measured sensitivity loss supports the suggestion that refractive error can affect narrowly-tuned notches. These results indicate that before the presence of a notch in the CSF can be attributed to neural abnormality, an optical cause must be eliminated.


Decreased concentrations of heparinoids are required to inhibit thrombin generation in plasma from newborns and children compared to plasma from adults due to reduced thrombin potential.
Thrombin generation is decreased and delayed in plasma from newborns and children compared to adults. We hypothesized that lower doses of heparinoid anticoagulants are required to give similar thrombin generation in newborn (umbilical cord) and child plasmas compared to that of adults. Thrombin generation was performed in either the absence or presence of unfractionated heparin (UFH), low molecular weight heparin (LMWH) or a covalent antithrombin-heparin complex (ATH). After contact activation and recalcification of each plasma, thrombin activity was measured by periodic sub-sampling into chromogenic substrate. UFH inhibited thrombin generation to a greater extent compared to LMWH in all plasmas. Cord plasma was more sensitive to inhibition and displayed a greater difference in the effectiveness of UFH compared to LMWH than other plasmas. Lower concentrations of UFH and LMWH were required to inhibit thrombin generation in cord and child plasmas compared to adult plasma. In comparison, ATH strongly inhibited thrombin generation in all 3 plasmas. Similar peak thrombin concentrations were observed at lower ATH concentrations (0.1 U/mL) compared to either UFH (0.25 U/mL) or LMWH (0.25 U/mL). As with UFH and LMWH, cord plasma was more sensitive to inhibition by ATH than the other plasmas and lower ATH concentrations inhibited thrombin generation in cord and child plasmas compared to adult plasma. Decreased thrombin generation with heparinoids in cord and child plasmas compared to adult plasma coincided with decreased rates of prothrombin consumption and increased proportion of thrombin-alpha2-macroglobulin inhibitor complexes. In summary, lower doses of UFH, LMWH or ATH result in similar peak thrombin generation in newborn and child plasmas compared to adult plasma. Cord plasma was the most sensitive to inhibition, with ATH being more effective than UFH or LMWH.


Molecular mechanisms of the protective role of wheat germ oil against cyclosporin A-induced hepatotoxicity in rats.
Cyclosporin A (CsA) is one of the most important immunosuppressive agents. However, its clinical use is strongly limited by several side effects including hepatotoxicity which remains a major clinical problem. Involvement of reactive oxygen species (ROS) in CsA-induced hepatotoxicity has been reported. This study investigates the potential protective role of wheat germ oil (WGO) as an antioxidant against CsA-induced hepatotoxicity. Twenty-four male Wistar albino rats (six animals in each group) received castor oil, the vehicle of CsA i.p. (control) or either CsA (25 mg/kg/d i.p.), WGO (900 mg/kg/d by oral gavage), or CsA in combination with WGO daily for 21 d. CsA administration significantly increased serum levels of the liver enzymes alanine aminotransferase (ALT) and aspartate aminotransferase (AST). In addition, an increase in lipid peroxidation, inducible NO-synthase (iNOS), and NF-κB expression were observed in hepatic tissues of CsA-alone-treated rats. Furthermore, significant reduction in the hepatic content of reduced glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) was also observed in CsA-alone-treated animals. Moreover, histopathological changes occurred in CsA-alone-treated rats. Concomitant administration of WGO along with CsA improved all these parameters. Most interestingly, the immunosuppressive effect of CsA was not affected by WGO. The present study suggests that concomitant use of WGO might be useful in reducing liver toxicity induced by CsA via inhibition of ROS, iNOS, and NF-κB expression.


Is human leukocyte antigen-DR and intercellular adhesion molecule-1 expression on human thyrocytes constitutive in papillary thyroid cancer? Comparative studies in human thyroid xenografts in severe combined immunodeficient and nude mice.
We have studied human leukocyte antigen (HLA)-DR and intercellular adhesion molecule (ICAM)-1 expression on thyroid epithelial cells (TEC) from papillary thyroid carcinoma (PTC) tissues xenografted into two different mouse strains [the severe combined immunodeficient (SCID) mouse, which accepts human tissue with lymphocytes; and the nude mouse, which accepts the tissue but destroys all passenger lymphocytes]. Human PTC [PTC/TIL (PTC with tumor infiltrating lymphocytes) and PTC/PTC (PTC without tumor infiltrating lymphocytes)], Graves' disease (GD), and normal thyroid (N) tissues were xenografted sc into 22 SCID and 21 nude mice. Blood samples were taken every 2 weeks for measurement of human IgG and thyroid antibodies. Seven weeks after xenografting, xenografted thyroid tissues were analyzed for thyrocyte HLA-DR and ICAM-1 expression. SCID mice xenografted with PTC/TIL (PTC/TIL-SCID) manifested IgG production for 6 weeks, but nude mice showed diminished and disappearing IgG production from these xenografts. Thyroperoxidase (TPO)-antibody (Ab)(TPO-Ab) was not detectable in PTC/TIL-SCID despite the presence of TPO-Ab in some donors. Thyroglobulin-Ab (Tg-Ab) was detectable in all mice of PTC/TIL-SCID. Thyrocyte HLA-DR expression from PTC-SCID was markedly increased, compared with that from nude mice xenografts or from N xenografts in SCID mice. In addition, thyrocyte HLA-DR expression from PTC-nude was markedly increased, compared with the expression seen in GD-nude and N-nude xenografts. ICAM-1 expression on TEC from PTC xenografts in the SCID mouse was markedly increased, compared with N xenografts. ICAM-1 expression on TEC from PTC did not show any difference between SCID and nude mice. ICAM-1 expression on TEC from PTC xenografts in the nude mice was markedly increased, compared with those from GD and N xenografts. In conclusion, TIL in PTC produce Tg-Ab but do not produce TPO-Ab. HLA-DR expression on TEC from PTC is strongly constitutive, but it is also affected by TIL. TIL might have some role in control of PTC through partial expression of HLA-DR on TEC. ICAM-1 expression on TEC from PTC seems to be entirely constitutive, and it is not affected by the presence of local lymphocytes, in contrast to autoimmune thyroid disease.


Work empowerment and organizational commitment.
As organizations struggle to deliver the same level and quality of services with fewer resources, administrators are challenged with redesigning workplaces to maximize nurses' commitment. This study used Kanter's Structural Theory of Organizational Behavior to examine the relationship between job-related empowerment perceptions of staff nurses and their commitment to the organization. Strategies for creating more empowered work environments are discussed.


[New methods for the diagnosis of cytomegalovirus].
Despite remarkable progress made in the fields of antiviral therapy and virological laboratory diagnosis, infections with the human cytomegalovirus (CMV) still pose considerable problems in certain patient groups. The variety of available test methods - mostly not standardised - with their different test characteristics, sample types etc. is often confusing for the clinician. There is often great uncertainty as to whether a given laboratory result is pathognomonic. In addition, not only is the diagnostic repertoire growing constantly - for instance through genome quantification and measurement of virus-specific T lymphocytes - but also the available antiviral therapeutic options with the need for adequate surveillance. This paper gives an up-to-date overview about the available diagnostic means with their specific advantages and disadvantages. Concrete recommendations regarding a rational and comprehensive, yet economic approach to CMV diagnosis have to be targeted at the individual patient group and specific diagnostic problem; this is beyond the scope of this paper and the reader is referred to the pertinent expert groups' publications. This review article deals with general and fundamental aspects.


Smoking Cessation Strategies Targeting Specific Populations.
Quitting smoking helps smokers maintain their health and extend their lifespan by 10 or more years. Treatment strategies for smoking cessation should be tailored to individual smokers with special needs based on their specific circumstances. It is recommended that pregnant women adopt smoking cessation through counseling and behavioral interventions because the safety of medications has yet to be established. Counseling is the main strategy for smoking cessation in adolescents and nicotine replacement therapy can be used with caution in individuals with serious nicotine dependence. It is important for smokers with psychiatric diseases to quit smoking following accurate assessment of their depression status. Nicotine replacement therapy, varenicline, and bupropion can be used for smoking cessation in smokers with psychiatric disorders. The incidence of cardiovascular disease decreased according to the smoking status and the duration of smoking cessation. In smokers with chronic obstructive pulmonary disease (COPD) who used a combination of counseling and pharmacotherapy the quitting rate was more than twice as high as subjects who used behavioral interventions alone. Varenicline can be used as the most effective anti-smoking drug by most smokers including those with psychiatric disorders, cardiovascular disease, and COPD.


High school incompletion and childhood maltreatment among street-involved young people in Vancouver, Canada.
While the link between educational attainment and future health and wellness is well understood, little investigation has considered the potential impacts of distinct forms of childhood maltreatment on high school completion. In the present study, the relationship between five categories of childhood maltreatment (physical, emotional, and sexual abuse, and physical and emotional neglect) and completion of high school education were examined using the Childhood Trauma Questionnaire (CTQ). From September 2005 to May 2013, data were collected for the At-Risk Youth Study (ARYS), a cohort of street-involved young people who use illicit drugs in Vancouver, Canada. We used logistic regression to examine the relationship between childhood maltreatment and high school completion, while controlling for a range of potential confounding variables. Specifically, five separate models for each category of maltreatment and two combined models were employed to examine the relative associations between, and cumulative impact of, different forms of childhood maltreatment and educational attainment. Among 974 young people, 737 (76%) reported not completing high school. In separate multivariable analyses physical abuse, emotional abuse, physical neglect, and emotional neglect remained positively and independently associated with an incomplete high school education. In a combined multivariable model with all forms of childhood maltreatment considered together, emotional abuse (adjusted odds ratio = 2.08; 95% confidence interval: 1.51-2.86) was the only form of maltreatment that remained significantly associated with an incomplete high school education. The cumulative impact assessment indicated a moderate dose-dependent trend where the greater the number of different forms of childhood maltreatment the greater the risk of not completing a high school education. These findings point to the need for trauma-informed interventions to improve educational attainment among vulnerable young people, as well as evidence-based prevention programmes, such as the Nurse-Family Partnership, aimed at supporting at-risk families before maltreatment occurs.


A largely random AAV integration profile after LPLD gene therapy.
The clinical application of adeno-associated virus vectors (AAVs) is limited because of concerns about AAV integration-mediated tumorigenicity. We performed integration-site analysis after AAV1-LPL(S447X) intramuscular injection in five lipoprotein lipase-deficient subjects, revealing random nuclear integration and hotspots in mitochondria. We conclude that AAV integration is potentially safe and that vector breakage and integration may occur from each position of the vector genome. Future viral integration-site analyses should include the mitochondrial genome.


An embryonic demethylation mechanism involving binding of transcription factors to replicating DNA.
In vertebrates, transcriptionally active promoters are undermethylated. Since the transcription factor Sp1, and more recently NF-kappaB, have been implicated in the demethylation process, we examined the effect of transcription factors on demethylation by injecting in vitro methylated plasmid DNA into Xenopus fertilized eggs. We found that various transactivation domains, including a strong acidic activation domain from the viral protein VP16, can enhance demethylation of a promoter region when fused to a DNA binding domain which recognizes the promoter. Furthermore, demethylation occurs only after the midblastula transition, when the general transcription machinery of the host embryo becomes available. Nevertheless, transcription factor binding need not be followed by actual transcription, since demethylation is not blocked by alpha-amanitin treatment. Finally, replication of the target DNA is a prerequisite for efficient demethylation since only plasmids that carry the bovine papilloma virus sequences which support plasmid replication after the midblastula transition are demethylated. No demethylation is detectable in the oocyte system where DNA is not replicated. These results suggest that, in the Xenopus embryo, promoters for which transcription factors are available are demethylated by a replication-dependent, possibly passive mechanism.


Spondyloarthropathy in progressive ankylosis mice: ultrastructural features of the intervertebral disk.
Progressive ankylosis mice (ank/ank) spontaneously develop a joint disorder characterized by calcium hydroxyapatite deposition and bony ankylosis of the joints of the axial and the appendicular skeleton. Mice in this study were used to determine the extent of mineralization of the intervertebral disks and to determine their suitability as a model for ankylosing spondyloarthropathies. Intervertebral disks taken from affected mice from 4 to 18 weeks of age were studied by electron-microscopic, electron energy-dispersive X-ray microanalysis and selected-area electron diffraction techniques. Necrotic foci seen in light microscope studies were found to be massive accumulations of calcium hydroxyapatite crystals deposited within the extracellular matrix. Chondrocytes of older animals demonstrated what appeared to be postmortem changes and contained numerous large vacuoles.


Rhabdoneuroglial heterotopias of the pontine leptomeninges in trisomy 13.
An unusual heterotopia of striated skeletal muscle and glial tissue occurred in the pontine meninges of a stillborn male showing feature of trisomy 13 and an Arnold-Chiari malformation. In two other reports of this type of heterotopia, the location and association with trisomy 13 has been the same. An error in tissue differentiation at the pontine flexure is suggested to account for this relationship.


A case of structure determination using pseudosymmetry.
Here, a case is presented of an unusual structure determination which was facilitated by the use of pseudosymmetry. Group A streptococcus uses cysteine protease Mac-1 (also known as IdeS) to evade the host immune system. Native Mac-1 was crystallized in the orthorhombic space group P2(1)2(1)2. Surprisingly, crystals of the inactive C94A mutant of Mac-1 displayed monoclinic symmetry with space group P2(1), despite the use of native orthorhombic Mac-1 microcrystals for seeding. Attempts to solve the structure of the C94A mutant by MAD phasing in the monoclinic space group did not produce an interpretable map. The native Patterson map of the C94A mutant showed two strong peaks along the (1 0 1) diagonal, indicating possible translational pseudosymmetry in space group P2(1). Interestingly, one-third of the monoclinic reflections obeyed pseudo-orthorhombic P2(1)2(1)2 symmetry similar to that of the wild-type crystals and could be indexed and processed in this space group. The pseudo-orthorhombic and monoclinic unit cells were related by the following vector operations: a(m) = b(o) - c(o), b(m) = a(o) and c(m) = -2c(o) - b(o). The pseudo-orthorhombic subset of data produced good SAD phases, leading to structure determination with one monomer in the asymmetric unit. Subsequently, the structure of the Mac-1 mutant in the monoclinic form was determined by molecular replacement, which showed six molecules forming three translationally related dimers aligned along the (1 0 1) diagonal. Knowing the geometric relationship between the pseudo-orthorhombic and the monoclinic unit cells, all six molecules can be generated in the monoclinic unit cell directly without the use of molecular replacement. The current case provides a successful example of the use of pseudosymmetry as a powerful phase-averaging method for structure determination by anomalous diffraction techniques. In particular, a structure can be solved in a higher pseudosymmetry subcell in which an NCS operator becomes a crystallographic operator. The geometrical relationships between the subcell and parental cell can be used to generate a complete molecular representation of the parental asymmetric unit for refinement.


Mass cytometric analysis unveils a disease-specific immune cell network in the bone marrow in acquired aplastic anemia.
Idiopathic acquired aplastic anemia (AA) is considered an immune-mediated syndrome of bone marrow failure since approximately 70% of patients respond to immunosuppressive therapy (IST) consisting of a course of anti-thymocyte globulin (ATG) followed by long-term use of ciclosporin. However, the immune response that underlies the pathogenesis of AA remains poorly understood. In this study, we applied high-dimensional mass cytometry on bone marrow aspirates of AA patients pre-ATG, AA patients post-ATG and healthy donors to decipher which immune cells may be implicated in the pathogenesis of AA. We show that the bone marrow of AA patients features an immune cell composition distinct from healthy donors, with significant differences in the myeloid, B-cell, CD4+ and CD8+ T-cells lineages. Specifically, we discovered that AA pre-ATG is characterized by a disease-specific immune cell network with high frequencies of CD16+ myeloid cells, CCR6++ B-cells, Th17-like CCR6+ memory CD4+ T-cells, CD45RA+CCR7+CD38+ CD8+ T-cells and KLRG1+ terminally differentiated effector memory (EMRA) CD8+ T-cells, compatible with a state of chronic inflammation. Successful treatment with IST strongly reduced the levels of CD16+ myeloid cells and showed a trend toward normalization of the frequencies of CCR6++ B-cells, CCR6+ memory CD4+ T-cells and KLRG1+EMRA CD8+ T-cells. Altogether, our study provides a unique overview of the immune landscape in bone marrow in AA at a single-cell level and proposes CCR6 as a potential new therapeutic target in AA.


Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy.
Ex vivo human skin has been used extensively for cosmeceutical and drug delivery studies, transplantable skin allografts, or skin flaps. However, it has a half-life of a few days due to ischemic necrosis. Traditional methods of assessing viability can be time-consuming and provide limited metabolic information. Using multiphoton tomography and fluorescence lifetime imaging (MPT-FLIM) we assess ischemic necrosis of ex vivo skin by NAD(P)H autofluorescence intensity and fluorescence lifetime. Ex vivo skin is stored in the presence and absence of nutrient media (Dulbecco Modified Eagle Medium) at -20, 4, and 37 degrees C and room temperature over a 7-day time course to establish different rates of metabolic deterioration. At higher temperatures we observe a decrease in NAD(P)H autofluorescence, higher image noise, and a significant increase in the average fluorescence lifetime (tau(m)) from approximately 1000 to 2000 ps. Additionally, significant distortions in NAD(P)H fluorescence lifetime histograms correspond to the reduction in autofluorescence. Skin kept at 4 degrees C, with or without media, showed the least change. Our findings suggest that MPT-FLIM enables useful noninvasive optical biopsies to monitor the metabolic state and deterioration of human skin for research and clinical purposes.


Severity of alcohol dependence in patients with alcoholic liver disease.
To determine the severity of dependence on alcohol in patients with alcoholic liver disease the severity of alcohol dependence questionnaire was administered to 193 patients with various types of alcoholic liver disease established histologically, in whom a detailed history of lifetime alcohol consumption was also obtained. Only 34 patients (18%) were classified as being severely dependent compared with 56% of patients without overt liver disease who were attending a neighbouring alcohol treatment unit. There was a significant correlation between the severity of dependence and mean daily alcohol consumption (r = 0.45 and 0.39 for men and women, respectively) but not duration of drinking. Dependence scores tended to be lower in patients with cirrhosis than in those with precirrhotic liver disease, but this difference reached significance only in women. These findings confirm that patients who develop chronic alcoholic liver disease are usually only mildly dependent on alcohol and support the hypothesis that patients who escape florid symptoms of alcohol dependence are at greater risk of developing liver damage because they are able to sustain a continual consumption of alcohol over many years.


Diagnostic utility and limitations of glutamine synthetase and serum amyloid-associated protein immunohistochemistry in the distinction of focal nodular hyperplasia and inflammatory hepatocellular adenoma.
Inflammatory hepatocellular adenoma can show overlapping histological features with focal nodular hyperplasia, including inflammation, fibrous stroma, and ductular reaction. Expression of serum amyloid-associated protein in inflammatory hepatocellular adenoma and map-like pattern of glutamine synthetase in focal nodular hyperplasia can be helpful in this distinction, but the pitfalls and limitations of these markers have not been established. Morphology and immunohistochemistry were analyzed in 54 inflammatory hepatocellular adenomas, 40 focal nodular hyperplasia, and 3 indeterminate lesions. Morphological analysis demonstrated that nodularity, fibrous stroma, dystrophic blood vessels, and ductular reaction were more common in focal nodular hyperplasia, while telangiectasia, hemorrhage, and steatosis were more common in inflammatory hepatocellular adenoma, but there was frequent overlap of morphological features. The majority of inflammatory hepatocellular adenomas demonstrated perivascular and/or patchy glutamine synthetase staining (73.6%), while the remaining cases had diffuse (7.5%), negative (3.8%), or patchy pattern of staining (15%) that showed subtle differences from the classic map-like staining pattern and was designated as pseudo map-like staining. Positive staining for serum amyloid-associated protein was seen in the majority of inflammatory hepatocellular adenomas (92.6%) and in the minority of focal nodular hyperplasia (17.5%). The glutamine synthetase staining pattern was map-like in 90% of focal nodular hyperplasia cases, with the remaining 10% of cases showing pseudo map-like staining. Three cases were labeled as indeterminate and showed focal nodular hyperplasia-like morphology but lacked map-like glutamine synthetase staining pattern; these cases demonstrated a patchy pseudo map-like glutamine synthetase pattern along with the expression of serum amyloid-associated protein. Our results highlight the diagnostic errors that can be caused by variant patterns of staining with glutamine synthetase and serum amyloid-associated protein in inflammatory hepatocellular adenoma and focal nodular hyperplasia.


The 52 degrees-55 degrees segment of the Bacillus subtilis chromosome: a region devoted to purine uptake and metabolism, and containing the genes cotA, gabP and guaA and the pur gene cluster within a 34960 bp nucleotide sequence.
Within the framework of the international project for sequencing the entire Bacillus subtilis genome, we have determined the complete sequence of the segment flanking the purE-D gene cluster (55 degrees) as far as cotA (52 degrees). This segment (34960 bp) contains, as well as 12 genes already identified as part of the pur operon, 17 putative ORFs and one partial one. Two of them (gabP and guaA) are known B. subtilis genes. The gene product of cotA (formerly pig) shows significant similarity to oxidoreductases (phenoxazine synthase and bilirubin oxidase). The putative products of ORFs yeaB (Czd protein), yeaC (MoxR), yebA (CNG-channel and cGMP-channel proteins from eukaryotes), yebB (hypothetical 32.9 kDa protein of Escherichia coli), yecA (amino acid permease) and yecB (adenine deaminase) were similar to proteins in data banks.


Investigating four 'myths' surrounding dysphagia in patients with metastatic esophageal cancer. A multi-institutional study from the North Central Cancer Treatment Group.
Eighty-five to 95% of esophageal cancer patients suffer dysphagia. Yet, few studies have focused on this symptom, and four 'myths' persist: (i) dysphagia cannot be measured; (ii) chemotherapy cannot palliate it; (iii) dysphagia predicts a poor prognosis; (iv) dysphagia is associated with a frustratingly insatiable appetite. Forty-four patients with metastatic esophageal cancer participated in this quality of life/translational component of a previously reported clinical trial. All were monitored for chemotherapy efficacy and toxicity and completed questionnaires on dysphagia and appetite at baseline and every 6 weeks. The appetite hormones, leptin and neuropeptide y, were also assessed. Forty-five per cent of patients could easily swallow solid foods; all others had varying dysphagia, thus enabling exploration of these four 'myths.' First, a single-item visual analog scale (Swallowing Scale), demonstrated excellent agreement with a previously validated questionnaire (81% at baseline), thus reminding us that dysphagia is measurable. Second, chemotherapy was associated with a trend towards improved dysphagia (P = 0.059). Third, dysphagia did not predict tumor response or survival. Fourth, dysphagia was not associated with appetite, leptin or neuropeptide y. This study helps to dispel these four 'myths' and underscores the need for further quality of life research on dysphagia.


Incomplete resupination during floral development leads to pollination failure in a slipper orchid.
In many families, plants undergo floral resupination by twisting through approximately 180° during floral development so that the flower is effectively positioned upside down. In most orchids, resupination results in the median petal (i.e., the labellum) becoming lowermost, which plays a crucial role in pollination by serving as a landing platform or as a trapping device, or both. Incomplete resupination is predicted to lead to reduced pollination, although tests of this assumption are still lacking. We investigated the effect of resupination using Phragmipedium vittatum, a rare lady's slipper orchid whose specialized labellum forms a trapping device. First, we surveyed the natural occurrence of incomplete resupination. Then we manipulated flowers into non- (≈0°), half- (≈90°), and fully resupinate (≈180°) positions to test the effect of orientation on pollen smear removal and deposition by pollinators (female hoverflies). We found that ca. 10% of flowers in the natural population were not fully resupinate, being either non- (upward, 0-60°) or half-resupinate (sideward, 60-120°). The change in orientation prevented the effectiveness of pollination by hoverflies since no pollen smear removal or deposition were found in flowers from non- and half-resupinate treatments. Although these flowers still attracted hoverflies, they were not trapped effectively. As this orchid is incapable of autonomous self-pollination, flowers that do not resupinate fail to set fruits. These results highlight the importance of correct floral orientation provided by resupination to ensure pollination in orchids and other resupinate flowers.


Characterization of two conformational forms of the major DNA-binding protein encoded by herpes simplex virus 1.
We have resolved two electrophoretic species of the major DNA-binding protein, infected cell polypeptide 8 (ICP8), encoded by herpes simplex virus 1. In pulse-chase experiments, we observed the conversion of the ICP8a form, the slower migrating species, to the faster migrating form, ICP8b. Thus, the two species appear to be related as precursor-product. The conversion was not due to proteolytic cleavage, because higher concentrations of reducing agents in the sample buffer shifted the faster moving form to the slower moving species. Also, the two forms have identical peptide patterns as analyzed by partial proteolysis in sodium dodecyl sulfate. Thus, the faster moving species appears to be a conformational isomer containing intramolecular disulfide bonds. The functional significance of the two forms of the protein is discussed.


Application of thymic factor "Thymalin" in complex treatment of endometrial cancer patients.
Indications have been received of cell immunity in 111 primary endometrial cancer patients in the process of complex treatment: hormonotherapy+surgery. These indices have revealed correlation with pathogenetic variants of disease. Suppression has also been registered of T-lymphocytes under the influence of complex treatment. 35 patients received thymic factor "Thymalin" as a component of complex treatment. Its positive effects on cell immunity and some clinico-morphological parameters have been registered.


Ethics issues identified by applicants and ethics experts in Horizon 2020 grant proposals.
Background: We assessed the ethics review of proposals selected for funding under the Marie Skłodowska-Curie Actions (MSCA) and the European Research Council (ERC) in Horizon 2020, EU's framework programme for research and innovation, 2014-2020. Methods: We analysed anonymized datasets for 3,054 MSCA individual fellowships (IF), 417 MSCA Innovative Training Networks (ITN), and 1,465 ERC main-listed proposals with ethics conditional clearance, over four years (2016 to 2019). The datasets included the information on ethics issues identified by applicants in their proposal and ethics issues and requirements identified by ethics experts during the ethics review. Results: 42% of proposals received ethical clearance. For proposals with conditional ethics clearance (n=3546), most of the identified ethics issues by both applicants and ethics experts were in the ethics categories related to humans; protection of personal data; environment, health and safety; and non-EU countries. Ethics experts identified twice as many ethics issues compared to applicants across funding schemes, years, and from high- and low-research performing countries. ERC grants had the highest number of ethics requirements per proposal (median (Md)=8, interquartile range (IQR=4-14), compared to ITN (Md=6, IQR=3-13) and IF grants (Md=3, IQR=2-6). The majority of requirements had to be fulfilled after grant agreement: 99.4% for IF, 99.5% for ITN, and 26.0% for ERC. For 9% of the proposals, the requirements included the appointment of an independent ethics advisor and 1% of the proposals had to appoint an ethics advisory board. Conclusions: Many applicants for highly competitive H2020 funding schemes lack awareness of ethics issues raised by their proposed research. There is a need for better training of researchers at all career stages about ethics issues in research, more support to researchers from research organizations to follow the funding agencies requirements, as well as further development and harmonization of the ethics appraisal process during grant assessment.


Otitis Media Diagnosis for Developing Countries Using Tympanic Membrane Image-Analysis.
Otitis media is one of the most common childhood diseases worldwide, but because of lack of doctors and health personnel in developing countries it is often misdiagnosed or not diagnosed at all. This may lead to serious, and life-threatening complications. There is, thus a need for an automated computer based image-analyzing system that could assist in making accurate otitis media diagnoses anywhere. A method for automated diagnosis of otitis media is proposed. The method uses image-processing techniques to classify otitis media. The system is trained using high quality pre-assessed images of tympanic membranes, captured by digital video-otoscopes, and classifies undiagnosed images into five otitis media categories based on predefined signs. Several verification tests analyzed the classification capability of the method. An accuracy of 80.6% was achieved for images taken with commercial video-otoscopes, while an accuracy of 78.7% was achieved for images captured on-site with a low cost custom-made video-otoscope. The high accuracy of the proposed otitis media classification system compares well with the classification accuracy of general practitioners and pediatricians (~64% to 80%) using traditional otoscopes, and therefore holds promise for the future in making automated diagnosis of otitis media in medically underserved populations.


Block of inward rectification by intracellular H+ in immature oocytes of the starfish Mediaster aequalis.
Intracellular pH was recorded in immature starfish oocytes using pH-sensitive microelectrodes, and inwardly rectifying potassium currents were measured under voltage clamp. When the intracellular pH was lowered using acetate-buffered artificial sea water from the normal value of 7.09 to 5.9, inward rectification was completely blocked. The relationship between inward rectification and internal pH between 7.09 and 5.9 could be fit by a titration curve for the binding of three H ions to a site with a pK of 6.26 to block the channel. The H+ block showed no voltage dependence, and the activation kinetics of the inwardly rectifying currents were not affected by the changes in internal pH.


Hop Phytochemicals and Their Potential Role in Metabolic Syndrome Prevention and Therapy.
Historically, hop cones (Humulus lupulus) have been used since ancient times as a remedy for many ailments and, as a source of polyphenols and bitter acids, is very effective in the treatment of metabolic syndrome (MS). Hop flavonoids, particularly xanthohumol (XN), are substances with hypoglycemic, antihyperlipidemic, and antiobesity activities. Iso-α-acids (IAA) and matured hop bitter acids (MHBA) improve health by influencing lipid metabolism, glucose tolerance, and body weight. The modulatory effect of IAA and MHBA on lipid metabolism may also be responsible for a loss in body weight. These results suggest promising applications for IAA, MHBA, and XN in humans, particularly in the prevention of diet-induced obesity and diabetes.


Heterochromatin dynamics during the differentiation process revealed by the DNA methylation reporter mouse, MethylRO.
In mammals, DNA is methylated at CpG sites, which play pivotal roles in gene silencing and chromatin organization. Furthermore, DNA methylation undergoes dynamic changes during development, differentiation, and in pathological processes. The conventional methods represent snapshots; therefore, the dynamics of this marker within living organisms remains unclear. To track this dynamics, we made a knockin mouse that expresses a red fluorescent protein (RFP)-fused methyl-CpG-binding domain (MBD) protein from the ROSA26 locus ubiquitously; we named it MethylRO (methylation probe in ROSA26 locus). Using this mouse, we performed RFP-mediated methylated DNA immunoprecipitation sequencing (MeDIP-seq), whole-body section analysis, and live-cell imaging. We discovered that mobility and pattern of heterochromatin as well as DNA methylation signal intensity inside the nuclei can be markers for cellular differentiation status. Thus, the MethylRO mouse represents a powerful bioresource and technique for DNA methylation dynamics studies in developmental biology, stem cell biology, as well as in disease states.


[The cervico-cranial syndrome in the practice of the otorhinolaryngologist].
The craniocervical syndrome is an entity whose symptoms: vertigo, cephalea, tinnitus, facial pain, otalgia, dysphagia, pain of the carotid artery are thought to be caused by cervical factors. In the majority of cases the cranio-cervical syndrome is caused by a spondylarthrogenic segmental dysfunction whose pathophysiology is explained. In the pathogenesis lesions of the joints of the skull which may be responsible for pain and dysfunction in the segmental areas are of great importance. The neurology of the joints of the skull, as well as the pathological mechanisms of spondylarthrogenic disturbances, responsible for the different kinds of dysfunction of the equilibrium and for cephalea are discussed. The pathophysiological basis of manual diagnosis is explained; also the radiological findings of the upper cervical vertebral column are principally discussed. A short review of therapeutic recommendations is given.


A truly early starter model of antisocial behavior revisited.
This paper revisits a developmental model of the origins of early conduct problems. Several of the model's primary tenets have now been validated in two samples of at-risk children followed prospectively from infancy to school-age. In both cohorts, child, family, and sociodemographic factors all play a significant role in the development of early conduct problems. In particular, the quality of the caregiving environment during the child's second year differentiates clinical impairment according to both parent and teacher report 6 years later. We conclude by making recommendations for future studies. Research that utilizes a developmental framework, incorporates more sophisticated measurement of infant negative emotionality, and addresses the influences of neighborhood and culture, is suggested.


Anaemia in diabetes: Is there a rationale to TREAT?
Anaemia is a common finding in patients with diabetes, particularly in those with overt nephropathy or renal impairment. In tertiary clinics, at least one outpatient in five with diabetes has anaemia, for whom it constitutes a significant additional burden. Anaemia is associated with an increased risk of diabetic complications including nephropathy, retinopathy and macrovascular disease. Anaemia may also be significant in determining the outcome of heart failure and hypoxia-induced organ damage in diabetes. While several factors contribute to the increased prevalence of anaemia in diabetes, the failure of the kidney to increase erythropoietin in response to falling haemoglobin appears to be the dominant factor. Although there is a clear rationale for correcting anaemia in people with diabetes, it remains to be established whether this will lead to improved outcomes. Moreover, the balance of risks, costs, and benefits remains to be established in patients with diabetes. The Trial to Reduce Cardiovascular Events with Aranesp (darbepoetin alpha) Therapy (TREAT) is a randomised controlled trial designed to determine the impact of anaemia correction on mortality and non-fatal cardiovascular events in patients with type 2 diabetes and stage 3-4 nephropathy. It is anticipated that TREAT will help to define the optimal approach to the management of anaemia in diabetes.


Context-dependent encoding in the auditory brainstem subserves enhanced speech-in-noise perception in musicians.
Musical training strengthens speech perception in the presence of background noise. Given that the ability to make use of speech sound regularities, such as pitch, underlies perceptual acuity in challenging listening environments, we asked whether musicians' enhanced speech-in-noise perception is facilitated by increased neural sensitivity to acoustic regularities. To this aim we examined subcortical encoding of the same speech syllable presented in predictable and variable conditions and speech-in-noise perception in 31 musicians and nonmusicians. We anticipated that musicians would demonstrate greater neural enhancement of speech presented in the predictable compared to the variable condition than nonmusicians. Accordingly, musicians demonstrated more robust neural encoding of the fundamental frequency (i.e., pitch) of speech presented in the predictable relative to the variable condition than nonmusicians. The degree of neural enhancement observed to predictable speech correlated with subjects' musical practice histories as well as with their speech-in-noise perceptual abilities. Taken together, our findings suggest that subcortical sensitivity to speech regularities is shaped by musical training and may contribute to musicians' enhanced speech-in-noise perception.


Strain Relief during Ice Growth on a Hexagonal Template.
Heterogeneous ice nucleation at solid surfaces impacts many areas of science, from environmental processes, such as precipitation, to microbiological systems and food processing, but the microscopic mechanisms underpinning nucleation remain unclear. Discussion of ice growth has often focused around the role of the surface in templating the structure of water, forcing the first layer to adopt the registry of the underlying substrate rather than that of ice. To grow a thick ice film, water in the first few ice layers must accommodate this strain, but understanding how this occurs requires detailed molecular-scale information that is lacking. Here we combine scanning tunneling microscopy, low-energy electron diffraction, and work-function measurements with electronic structure calculations to investigate the initial stages of ice growth on a Pt alloy surface, having a lattice spacing 6% larger than ice. Although the first layer of water forms a strictly commensurate hexagonal network, this behavior does not extend to the second layer. Instead, water forms a 2D structure containing extended defect rows made from face-sharing pentamer and octamer rings. The defect rows allow the majority of second-layer water to remain commensurate with the solid surface while compensating lateral strain by increasing the water density close to that of an ice surface. The observation of octamer-pentamer rows in ice films formed on several surfaces suggests that the octamer-pentamer defect motif acts as a flexible strain relief mechanism in thin ice films, providing a mechanism that is not available during the growth of strained films in other materials, such as semiconductors.


A method for labeling proteins with tags at the native genomic loci in budding yeast.
Fluorescent proteins and epitope tags are often used as protein fusion tags to study target proteins. One prevailing technique in the budding yeast Saccharomyces cerevisiae is to fuse these tags to a target gene at the precise chromosomal location via homologous recombination. However, several limitations hamper the application of this technique, such as the selectable markers not being reusable, tagging of only the C-terminal being possible, and a "scar" sequence being left in the genome. Here, we describe a strategy to solve these problems by tagging target genes based on a pop-in/pop-out and counter-selection system. Three fluorescent protein tag (mCherry, sfGFP, and mKikGR) and two epitope tag (HA and 3×FLAG) constructs were developed and utilized to tag HHT1, UBC13 or RAD5 at the chromosomal locus as proof-of-concept.


Evaluation and strategy for use of MIRU-VNTRplus, a multifunctional database for online analysis of genotyping data and phylogenetic identification of Mycobacterium tuberculosis complex isolates.
Because of its portable data, discriminatory power, and recently proposed standardization, mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) typing has become a major method for the epidemiological tracking of Mycobacterium tuberculosis complex (MTBC) clones. However, no public MIRU-VNTR database based on well-characterized reference strains has been available hitherto for easy strain identification. Therefore, a collection of 186 reference strains representing the primary MTBC lineages was used to build a database, which is freely accessible at http://www.MIRU-VNTRplus.org. The geographical origin and the drug susceptibility profile of each strain were stored together with comprehensive genetic lineage information, including the 24-locus MIRU-VNTR profile, the spoligotyping pattern, the single-nucleotide- and large-sequence-polymorphism profiles, and the IS6110 restriction fragment length polymorphism fingerprint. Thanks to flexible import functions, a single or multiple user strains can be analyzed, e.g., for lineage identification with or without the use of reference strains, by best-match or tree-based analyses with single or combined marker data sets. The results can easily be exported. In the present study, we evaluated the database consistency and various analysis parameters both by testing the reference collection against itself and by using an external population-based data set comprising 629 different strains. Under the optimal conditions found, lineage predictions based on typing by 24-locus MIRU-VNTR analysis optionally combined with spoligotyping were verified in >99% of the cases. On the basis of this evaluation, a user strategy was defined, which consisted of best-match analysis followed, if necessary, by tree-based analysis. The MIRU-VNTRplus database is a powerful tool for high-resolution clonal identification and has little equivalent in terms of functionalities among the bacterial genotyping databases available so far.


Inverse Scattering of the Zakharov-Shabat System Solves the Weak Noise Theory of the Kardar-Parisi-Zhang Equation.
We solve the large deviations of the Kardar-Parisi-Zhang (KPZ) equation in one dimension at short time by introducing an approach which combines field theoretical, probabilistic, and integrable techniques. We expand the program of the weak noise theory, which maps the large deviations onto a nonlinear hydrodynamic problem, and unveil its complete solvability through a connection to the integrability of the Zakharov-Shabat system. Exact solutions, depending on the initial condition of the KPZ equation, are obtained using the inverse scattering method and a Fredholm determinant framework recently developed. These results, explicit in the case of the droplet geometry, open the path to obtain the complete large deviations for general initial conditions.


Comparison of health-related quality of life (HRQoL) between ileal conduit diversion and orthotopic neobladder based on validated questionnaires: a systematic review and meta-analysis.
The question of whether orthotopic neobladder (ONB) reconstruction is superior to ileal conduit diversion (ICD) with respect to health-related quality of life (HRQoL) remains controversial. The goal of this study is to perform a meta-analysis to compare post-ICD and post-ONB HRQoL in patients with bladder cancer. A systematic search of Medline, Embase, the Cochrane Central Register of Controlled Trials, and the annual congress abstracts of the European Association of Urology (EAU), the American Urological Association (AUA) and the Société Internationale d'Urologie (SIU) up to June 2017 was conducted to identify all relevant clinical trials using validated questionnaires to assess HRQoL. A systematic review and meta-analysis were then performed. A total of 2507 patients from 26 eligible studies were included. Meta-analyses showed significant differences favouring ONB patients in global health status (WMD + 9.13, p = 0.004), physical functioning (WMD + 11.57, p = 0.0001), role functioning (WMD + 9.64, p = 0.002), and social functioning (WMD + 6.81, p = 0.03) based on the EORTC-QLQ-C30 questionnaire and in the total score of FACT questionnaire (WMD + 6.80, p = 0.001). However, ONB patients were more likely to have postoperative urinary symptoms than ICD patients (WMD - 22.19, p = 0.0001). ONB patients are more likely to have a better global health status than ICD patients. Regardless of the type of urinary diversion (UD) surgery, a gradual improvement in HRQoL over preoperative status tended to stabilise after 12 months postoperatively.


A red-flag-based approach to risk management of EHR-related safety concerns.
Although electronic health records (EHRs) have a significant potential to improve patient safety, EHR-related safety concerns have begun to emerge. Based on 369 responses to a survey sent to the memberships of the American Society for Healthcare Risk Management and the American Health Lawyers Association and supplemented by our previous work in EHR-related patient safety, we identified the following common EHR-related safety concerns: (1) incorrect patient identification; (2) extended EHR unavailability (either planned or unplanned); (3) failure to heed a computer-generated warning or alert; (4) system-to-system interface errors; (5) failure to identify, find, or use the most recent patient data; (6) misunderstandings about time; (7) incorrect item selected from a list of items; and (8) open or incomplete orders. In this article, we present a "red-flag"-based approach that can be used by risk managers to identify potential EHR safety concerns in their institutions. An organization that routinely conducts EHR-related surveillance activities, such as the ones proposed here, can significantly reduce risks associated with EHR implementation and use.


Systematic Analysis of Metabolic Pathway Distributions of Bacterial Energy Reserves.
Previous bioinformatics studies have linked gain or loss of energy reserves with host-pathogen interactions and bacterial virulence based on a comparatively small number of bacterial genomes or proteomes. Thus, understanding the theoretical distribution patterns of energy reserves across bacterial species could provide a shortcut route to look into bacterial lifestyle and physiology. So far, five major energy reserves have been identified in bacteria due to their capacity to support bacterial persistence under nutrient deprivation conditions. These include polyphosphate (polyP), glycogen, wax ester (WE), triacylglycerol (TAG), and polyhydroxyalkanoates (PHAs). Although the enzymes related with metabolism of energy reserves are well understood, there is a lack of systematic investigations into the distribution of bacterial energy reserves from an evolutionary point of view. In this study, we sourced 8282 manually reviewed bacterial reference proteomes and combined a set of hidden Markov sequence models (HMMs) to search homologs of key enzymes related with the metabolism of energy reserves. Our results revealed that specific pathways like trehalose-related glycogen metabolism and enzymes such as wax ester synthase/acyl-CoA:diacylglycerol acyltransferase (WS/DGAT) are mainly restricted within specific types of bacterial groups, which provides evolutionary insights into the understanding of their origins and functions. In addition, the study also confirms that loss of energy reserves like polyP metabolism absence in Mollicutes is correlated with bacterial genome reduction. Through this analysis, a clearer picture about the metabolism of energy reserves in bacteria is presented, which could serve as a guide for further theoretical and experimental analyses of bacterial energy metabolism.


[Serotonin metabolism in the rat brain during water deprivation and hydration].
Water deprivation (WD) decreased the serotonin (5-HT) level and significantly increased the 5-hydroxyindoleacetic acid (5-HIAA) in the rat midbrain and hypothalamus, the catabolic 5-HIAA/5-HT ratio increasing three-fold. Hydration (H) produced a moderate increase in the 5-HT and 5-HIAA levels in the hypothalamus with no changes in the 5-HIAA/5-HT ratio. Hydration exerted no significant effect upon the 5-HT level and metabolism in the midbrain. A two-fold increase of corticosterone concentration in water deprivation and its decrease in hydration were shown to occur in peripheral blood plasma.


FAR-RED INSENSITIVE219 modulates CONSTITUTIVE PHOTOMORPHOGENIC1 activity via physical interaction to regulate hypocotyl elongation in Arabidopsis.
FAR-RED INSENSITIVE219 (FIN219) in Arabidopsis (Arabidopsis thaliana) is involved in phytochrome A-mediated far-red (FR) light signaling. Previous genetic studies revealed that FIN219 acts as an extragenic suppressor of CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1). However, the molecular mechanism underlying the suppression of COP1 remains unknown. Here, we used a transgenic approach to study the regulation of COP1 by FIN219. Transgenic seedlings containing ectopic expression of the FIN219 amino (N)-terminal domain in wild-type Columbia (named NCox for the expression of the N-terminal coiled-coil domain and NTox for the N-terminal 300-amino acid region) exhibited a dominant-negative long-hypocotyl phenotype under FR light, reflected as reduced photomorphogenic responses and altered levels of COP1 and ELONGATED HYPOCOTYL5 (HY5). Yeast two-hybrid, pull-down, and bimolecular fluorescence complementation assays revealed that FIN219 could interact with the WD-40 domain of COP1 and with its N-terminal coiled-coil domain through its carboxyl-terminal domain. Further in vivo coimmunoprecipitation study confirms that FIN219 interacts with COP1 under continuous FR light. Studies of the double mutant fin219-2/cop1-6 indicated that HY5 stability requires FIN219 under darkness and FR light. Moreover, FIN219 levels positively regulated by phytochrome A can modulate the subcellular location of COP1 and are differentially regulated by various fluence rates of FR light. We conclude that the dominant-negative long-hypocotyl phenotype conferred by NCox and NTox in a wild-type background was caused by the misregulation of COP1 binding with the carboxyl terminus of FIN219. Our data provide a critical mechanism controlling the key repressor COP1 in response to FR light.


Concurrent impact of de novo mutations on cranial and cortical development in nonsyndromic craniosynostosis.
Nonsyndromic craniosynostosis (nsCS), characterized by premature cranial suture fusion, is considered a primary skull disorder in which impact on neurodevelopment, if present, results from the mechanical hindrance of brain growth. Despite surgical repair of the cranial defect, neurocognitive deficits persist in nearly half of affected children. Therefore, the authors performed a functional genomics analysis of nsCS to determine when, where, and in what cell types nsCS-associated genes converge during development. The authors integrated whole-exome sequencing data from 291 nsCS proband-parent trios with 29,803 single-cell transcriptomes of the prenatal and postnatal neurocranial complex to inform when, where, and in what cell types nsCS-mutated genes might exert their pathophysiological effects. The authors found that nsCS-mutated genes converged in cranial osteoprogenitors and pial fibroblasts and their transcriptional networks that regulate both skull ossification and cerebral neurogenesis. Nonsyndromic CS-mutated genes also converged in inhibitory neurons and gene coexpression modules that overlapped with autism and other developmental disorders. Ligand-receptor cell-cell communication analysis uncovered crosstalk between suture osteoblasts and neurons via the nsCS-associated BMP, FGF, and noncanonical WNT signaling pathways. These data implicate a concurrent impact of nsCS-associated de novo mutations on cranial morphogenesis and cortical development via cell- and non-cell-autonomous mechanisms in a developmental nexus of fetal osteoblasts, pial fibroblasts, and neurons. These results suggest that neurodevelopmental outcomes in nsCS patients may be driven more by mutational status than surgical technique.


[Kocher and goitre].
Emile Theodor Kocher (1841-1917) studied and spent his entire career as surgeon (since 1866) and professor (since 1872) in his native Berne, apart from a 'grand tour' of surgical institutions in Europe. The discipline of surgery rapidly expanded, not least through the introduction of anaesthesia and antisepsis. Kocher's expertise ranged from ankle fractures to hypophysectomy; he wrote an authoritative textbook on surgical technique. He became most famous through his treatment of goitre, an endemic condition in the mountainous parts of Switzerland. Kocher developed a meticulous technique for thyroidectomy, without major haemorrhage or damage to the recurrent nerve. The 'Kocher clamp' was developed for haemostasis. On discovering that patients might develop cretinism or 'cachexia' many years after total thyroidectomy, he took care to leave part of the gland intact. As the relation with myxoedema became clear, he experimented with thyroid transplantations and attempted to find biochemical function tests. In 1909 he was awarded the Nobel prize for Physiology or Medicine. Five years later thyroxin was isolated.


A systematic review of acupuncture for chemotherapy-induced peripheral neuropathy.
In cancer patients, chemotherapy-induced peripheral neuropathy (cipn) is a common complication, characterized by pain, loss of sensation, and numbness. Medical treatment for peripheral neuropathies has been shown to be ineffective for cipn. Acupuncture has been shown to be safe and effective in treating cancer-related symptoms and other peripheral neuropathies. For the present review, we aimed to evaluate the efficacy of acupuncture for the treatment of cipn. Comprehensive searches for relevant studies were conducted in Ovid embase, the Web of Science, Ovid medline, the Cochrane Central Register of Controlled Trials (central), cinahl (ebsco Information Services, Ipswich, MA, U.S.A.), and the ClinicalTrials.gov Web site. References from previous systematic reviews were also searched. Additional trials were found in the reference lists of relevant papers and in searches of Google Scholar and acupuncture-specific Web sites. Included studies were randomized controlled trials (rcts) of any type of acupuncture used to treat patients with cipn. Three clinical trials (203 participants) were included. Two studies found acupuncture to be effective in alleviating cipn pain and improving quality of life. One study found no benefit in improving neuropathic pain, symptoms, or quality of life. Study quality was variable and included a moderate overall risk of bias. The evidence is insufficient to recommend acupuncture for the treatment or prevention of cipn. Further research is needed to evaluate the effects of acupuncture in the treatment of cipn. Given that acupuncture is considered safe and might provide relief for patients, it can be considered at the clinician's discretion.


A quantitative method for measurement of HL-60 cell apoptosis based on diffraction imaging flow cytometry technique.
A quantitative method for measurement of apoptosis in HL-60 cells based on polarization diffraction imaging flow cytometry technique is presented in this paper. Through comparative study with existing methods and the analysis of diffraction images by a gray level co-occurrence matrix algorithm (GLCM), we found 4 GLCM parameters of contrast (CON), cluster shade (CLS), correlation (COR) and dissimilarity (DIS) exhibit high sensitivities as the apoptotic rates. It was further demonstrated that the CLS parameter correlates significantly (R(2) = 0.899) with the degree of nuclear fragmentation and other three parameters showed a very good correlations (R(2) ranges from 0.69 to 0.90). These results demonstrated that the new method has the capability for rapid and accurate extraction of morphological features to quantify cellular apoptosis without the need for cell staining.


Neonatal outcomes according to different degrees of maternal morbidity: cross-sectional evidence from the Perinatal Information System (SIP) of the CLAP network.
The burden of maternal morbidity in neonatal outcomes can vary with the adequacy of healthcare provision and tool implementation to improve monitoring. Such information is lacking in Latin American countries, where the decrease in severe maternal morbidity and maternal death remains challenging. To determine neonatal outcomes according to maternal characteristics, including different degrees of maternal morbidity in Latin American health facilities. This is a secondary cross-sectional analysis of the Perinatal Information System (SIP) database from eight health facilities in five Latin American and Caribbean countries. Participants were all women delivering from August 2018 to June 2021, excluding cases of abortion, multiple pregnancies and missing information on perinatal outcomes. As primary and secondary outcome measures, neonatal near miss and neonatal death were measured according to maternal/pregnancy characteristics and degrees of maternal morbidity. Estimated adjusted prevalence ratios (PRadj) with their respective 95% CIs were reported. In total 85,863 live births were included, with 1,250 neonatal near miss (NNM) cases and 695 identified neonatal deaths. NNM and neonatal mortality ratios were 14.6 and 8.1 per 1,000 live births, respectively. Conditions independently associated with a NNM or neonatal death were the need for neonatal resuscitation (PRadj 16.73, 95% CI [13.29-21.05]), being single (PRadj 1.45, 95% CI [1.32-1.59]), maternal near miss or death (PRadj 1.64, 95% CI [1.14-2.37]), preeclampsia (PRadj 3.02, 95% CI [1.70-5.35]), eclampsia/HELPP (PRadj 1.50, 95% CI [1.16-1.94]), maternal age (years) (PRadj 1.01, 95% CI [<1.01-1.02]), major congenital anomalies (PRadj 3.21, 95% CI [1.43-7.23]), diabetes (PRadj 1.49, 95% CI [1.11-1.98]) and cardiac disease (PRadj 1.65, 95% CI [1.14-2.37]). Maternal morbidity leads to worse neonatal outcomes, especially in women suffering maternal near miss or death. Based on SIP/PAHO database all these indicators may be helpful for routine situation monitoring in Latin America with the purpose of policy changes and improvement of maternal and neonatal health.


Experimental use of a gas sensor-based instrument for differentiation of Escherichia coli O157:H7 from non-O157:H7 Escherichia coli field isolates.
The rapid and economical detection of human pathogens in animal and food production systems would enhance food safety efforts. An instrument based on gas sensors coupled with an artificial neural network (ANN) was developed for the detection of and differentiation between laboratory isolates of Escherichia coli O157:H7 and non-O157:H7 E. coli. The purpose of this study was to use field isolates of E. coli to further evaluate the sensor system. This gas sensor-based, computer-controlled detection system was used to monitor gas emissions from 12 isolates of E. coli O157:H7 and 8 non-O157:H7 E. coli isolates. A standard concentration of each isolate was grown in 10 ml of nutrient broth at 37 degrees C for 16 h, and gas sampling was carried out every 5 min. Readings were continuously plotted to generate gas signatures. A back-propagation ANN algorithm was used to interpret the gas patterns. By analysis of the response of the ANN, the sensitivity and specificity of the instrument were calculated. Detectable differences between the gas signatures of the E. coli O157:H7 isolates and the non-O157:H7 isolates were observed. The instruments degree of sensitivity was high for E. coli O157:H7 isolates, but a lower degree of accuracy was observed for non-O157:H7 isolates because of increased strain variation. The sensitivity of the detection system was improved by the normalization of the data generated from the gas sensors. Because of its ability to detect differences in gas patterns, this instrument has a broad range of potential food safety applications.


Breastfeeding Rates and Programs in Europe: A Survey of 11 National Breastfeeding Committees and Representatives.
Among the world's regions, the WHO European Region has the lowest rates of exclusive breastfeeding at the age of 6 months with approximately 25%. Low rates and early cessation of breastfeeding have important adverse health consequences for women, infants, and young children. Protecting, promoting, and supporting breastfeeding are a public health priority. National breastfeeding data and monitoring systems among selected European countries and the WHO European Region are compared. Mechanisms for the support, protection, and promotion of breastfeeding are reviewed and successes and challenges in implementation of national programs are presented. National representatives of national breastfeeding committees and initiatives in 11 European countries, including Belgium, Croatia, Denmark, Germany, Ireland, Italy, The Netherlands, Norway, Spain, Sweden, and Switzerland, participated in a standardized survey. Results are evaluated and compared in a narrative review. Variation exists in Europe on breastfeeding rates; methodology for data collection; and mechanisms for support, protection, and promotion of breastfeeding. Directly after birth, between 56% and 98% of infants in all countries were reported to receive any human milk, and at 6 months 38% to 71% and 13% to 39% of infants to be breastfed or exclusively breastfed, respectively. National plans addressing breastfeeding promotion, protection, and support exist in 6 of the 11 countries. National governments should commit to evidence-based breastfeeding monitoring and promotion activities, including financial and political support, to improve breastfeeding rates in the Europe. Renewed efforts for collaboration between countries in Europe, including a sustainable platform for information exchange, are needed.


Spectral receptive field properties of visually active neurons in the caudate nucleus.
Recent studies stress the importance of the caudate nucleus in visual information processing. Although the processing of moving visual signals depends upon the capability of a system to integrate spatial and temporal information, no study has investigated the spectral receptive field organization of the caudate nucleus neurons yet. Therefore, we tested caudate neurons of the feline brain by extracellular single-cell recording applying drifting sinewave gratings of various spatial and temporal frequencies, and reconstructed their spectral receptive fields by plotting their responsiveness as a function of different combinations of spatial and temporal frequencies. The majority of the caudate cells (74%) exhibited peak tuning, which means that their spatio-temporal frequency response profile had a characteristic region of increased activity with a single maximum in the spatio-temporal frequency domain. In one-quarter of the recorded caudate neurons ridge tuning was found, where the region of increased activity, forming an elongated ridge of maximal sensitivity parallel or angled to the spatial or the temporal frequency axis, indicating temporal (16%), spatial (5%) or speed (5%) tuning, respectively. The velocity preference of the ridge tuned caudate nucleus neurons is significantly lower than that of the peak tuned neurons. The peak tuned neuron could encode high velocities, while the ridge tuned neurons were responsible for the detection of moderate and lower velocities. Based upon our results, we suggest that the wide variety of spatio-temporal frequency response profiles might represent different functional neuronal groups within the caudate nucleus that subserve different behaviors to meet various environmental requirements.


Left Main Coronary Artery Revascularization in Patients with Impaired Renal Function; Percutaneous Coronary Intervention vs. Coronary Artery Bypass Grafting.
The evidence about the optimal revascularization strategy in patients with left-main coronary artery (LMCA) disease and impaired renal function is limited. Thus, we aimed to compare the outcomes of LMCA disease revascularization (PCI vs. CABG) in patients with and without impaired renal function. This retrospective cohort study included 2138 patients recruited from 14 centers between 2015 to 2019. We compared patients with impaired renal function who had PCI (n= 316) to those who had CABG (n= 121) and compared patients with normal renal function who had PCI (n= 906) to those who had CABG (n= 795). The study outcomes were in-hospital and follow-up major adverse cardiovascular and cerebrovascular events (MACCE). Multivariable logistic regression analysis showed that the risk of in-hospital MACCE was significantly higher in CABG compared to PCI in patients with impaired renal function [OR: 8.13 (95% CI: 4.19- 15.76), P<0.001] and normal renal function [OR: 2.59 (95% CI: 1.79- 3.73); P<0.001]. There were no differences in follow-up MACCE between CABG and PCI in patients with impaired renal function [HR: 1.14 (95% CI: 0.71- 1.81), P= 0.585] and normal renal function [HR: 1.12 (0.90- 1.39), P= 0.312]. PCI could have an advantage over CABG in revascularization of LMCA disease in patients with impaired renal function regarding in-hospital MACCE. The follow-up MACCE was comparable between PCI and CABG in patients with impaired and normal renal function.


Rhodium-catalyzed reductive cyclization of 1,6-diynes and 1,6-enynes mediated by hydrogen: catalytic C-C bond formation via capture of hydrogenation intermediates.
Catalytic hydrogenation of carbon-, nitrogen- and oxygen-tethered 1,6-diynes 1a-9a and 1,6-enynes 10a-18a using cationic Rh(I) precatalysts at ambient temperature and pressure enables reductive carbocyclization to afford 1,2-dialkylidene cyclopentanes 1b-9b and monoalkylidene cyclopentanes 10b-18b, respectively, in good to excellent yields and as single alkene stereoisomers. Notably, the 1,3-diene and alkene containing cyclization products 1b-9b and 10b-18b are not subject to over-reduction under the conditions of catalytic hydrogenation in which they are formed. Reductive cyclization 1,6-diyne 1a and 1,6-enyne 10a performed under an atmosphere of D(2) provides the carbocyclization products deuterio-1b and deuterio-10b, respectively, which incorporate two deuterium atoms. The collective data are consistent with a catalytic mechanism involving heterolytic activation of elemental hydrogen (H(2) + Rh(+)X(-) --> Rh-H + HX) followed by Rh(I)-mediated oxidative cyclization of the 1,6-diyne or 1,6-enyne substrates to afford (hydrido)Rh(III)-based metallocyclopentadiene and metallocyclopentene intermediates, respectively. These transformations represent the first examples of metal-catalyzed reductive carbocyclization mediated by hydrogen.


Distal Biceps Tendon Anatomic Repair.
Distal biceps injuries, which usually occur in active middle-aged men, can result in chronic pain and loss of supination and flexion strength3,4. Surgical repair of a ruptured distal biceps tendon can reliably decrease pain and improve strength compared with nonoperative management3,4. However, even following successful healing and rehabilitation of a surgically repaired biceps tendon, full supination strength is rarely restored5-7. The expected outcome following distal biceps repair using a traditional anterior approach is a measurable loss of rotational strength, especially from neutral to supinated positions5,7. This deficit can lead to difficulty with occupational and recreational activities5,8. The center of an uninjured biceps tendon inserts into the radial tuberosity 6.7 mm anterior to its apex9,10. This posterior location forces the biceps tendon to wrap around the radial protuberance during pronation, thus utilizing the protuberance as a mechanical cam during forceful forearm supination10,11. The distal biceps tendon comprises a medial short head and lateral long head; the 2 heads are continuations of the proximal muscles2,20,21. The short head inserts distal to the long head on their radial attachment site2,20,21. Performing a distal biceps repair via an anterior approach typically places the center of the reattachment site 12.9 mm anterior to its apex or approximately 6 mm anterior to an uninjured control tendon9. This shifts the repair site from its anatomic location (posterior to the radial protuberance) to a new nonanatomic location (on top of the protuberance). This anterior reattachment location decreases the cam effect of the radial protuberance, resulting in an average supination loss of 10% in neutral rotation and 33% in 60° of supination7,10. A posterior approach to the radial tuberosity using 2 separate intramedullary buttons for the short and long heads reliably positions the distal biceps insertion at its anatomic footprint, which is posterior to the radial protuberance9,10,11. This technique has been named the distal biceps tendon anatomic repair. Not only does it restore the normal supination cam effect of the radial protuberance, but it also provides superior initial fixation strength, with load to failure strength similar to the native tendon1. The distal biceps anatomic repair can be divided into the following 9 key steps: Step 1: Preoperative planning; Step 2: Positioning; Step 3: Identifying and retrieving the tendon; Step 4: Preparing the 2 heads of the tendon; Step 5: Posterior exposure of tendon footprint; Step 6: Drilling the short and long-head drill holes; Step 7: Passage of the tendon; Step 8: Unicortical button fixation; Step 9: Alternative fixation: cortical trough; and Step 10: Postoperative management.


Diverse syntrophic partnerships from deep-sea methane vents revealed by direct cell capture and metagenomics.
Microorganisms play a fundamental role in the cycling of nutrients and energy on our planet. A common strategy for many microorganisms mediating biogeochemical cycles in anoxic environments is syntrophy, frequently necessitating close spatial proximity between microbial partners. We are only now beginning to fully appreciate the diversity and pervasiveness of microbial partnerships in nature, the majority of which cannot be replicated in the laboratory. One notable example of such cooperation is the interspecies association between anaerobic methane oxidizing archaea (ANME) and sulfate-reducing bacteria. These consortia are globally distributed in the environment and provide a significant sink for methane by substantially reducing the export of this potent greenhouse gas into the atmosphere. The interdependence of these currently uncultured microbes renders them difficult to study, and our knowledge of their physiological capabilities in nature is limited. Here, we have developed a method to capture select microorganisms directly from the environment, using combined fluorescence in situ hybridization and immunomagnetic cell capture. We used this method to purify syntrophic anaerobic methane oxidizing ANME-2c archaea and physically associated microorganisms directly from deep-sea marine sediment. Metagenomics, PCR, and microscopy of these purified consortia revealed unexpected diversity of associated bacteria, including Betaproteobacteria and a second sulfate-reducing Deltaproteobacterial partner. The detection of nitrogenase genes within the metagenome and subsequent demonstration of (15)N(2) incorporation in the biomass of these methane-oxidizing consortia suggest a possible role in new nitrogen inputs by these syntrophic assemblages.