Modulating the wayward T cell: New horizons with immune checkpoint inhibitor treatments in autoimmunity, transplant, and cancer.
The T-cell response is regulated by the balance between costimulatory and coinhibitory signals. Immune checkpoints are essential for efficient T-cell activation, but also for maintaining self-tolerance and protecting tissues from damage caused by the immune system, and for providing protective immunity. Modulating immune checkpoints can serve diametric goals, such that blocking a coinhibitory molecule can unleash anti-cancer immunity whereas stimulating the same molecule can reduce an over-reaction in autoimmune disease. The purpose of this review is to examine the regulation of T-cell costimulation and coinhibition, which is central to the processes underpinning autoimmunity, transplant rejection and immune evasion in cancer. We will focus on the immunomodulation agents that regulate these unwanted over- and under-reactions. The use of such agents has led to control of symptoms and slowing of progression in patients with rheumatoid arthritis, reduced rejection rates in transplant patients, and prolonged survival in patients with cancer. The management of immune checkpoint inhibitor treatment in certain challenging patient populations, including patients with pre-existing autoimmune conditions or transplant patients who develop cancer, as well as the management of immune-related adverse events in patients receiving antitumor therapy, is examined. Finally, the future of immune checkpoint inhibitors, including examples of emerging targets that are currently in development, as well as recent insights gained using new molecular techniques, is discussed. T-cell costimulation and coinhibition play vital roles in these diverse therapeutic areas. Targeting immune checkpoints continues to be a powerful avenue for the development of agents suitable for treating autoimmune diseases and cancers and for improving transplant outcomes. Enhanced collaboration between therapy area specialists to share learnings across disciplines will improve our understanding of the opposing effects of treatments for autoimmune disease/transplant rejection versus cancer on immune checkpoints, which has the potential to lead to improved patient outcomes.


[Vesiculectomy with laparoscopic partial prostatectomy in the treatment of primary adenocarcinoma of the seminal vesicle with carcinomatous transformation of the ejaculatory duct].
Primary adenocarcinoma of the seminal vesicle is an extremely rare condition. Some cases have been described in relation to congenital seminal vesicle cysts, which is often also associated with agenesia or ipsilateral renal disgenesia. The rareness of this type of lesions makes it difficult to plan a regulated surgical approach for them, although they are often treated by simple exeresis or exenteration, depending on their stage at the beginning. We present a new surgical technique that consists of radical vesiculectomy associated with laparoscopic partial prostatectomy (total segmentary) of the central area to successfully treat primary seminal vesicle adenocarcinoma in a young man who was diagnosed through an azoospermia study. A study of the scan (MRI) with diffusion and the transrectal biopsy of the mass allowed us to make a thorough preoperative evaluation of the case, confirming the malignity and precociousness of the lesion. The laparoscopic approach allowed us to perform a pelvic lymphadenectomy and transperitoneal exeresis, including the central prostate area and suture of the posterior face of the urethra at the height of the apex of the prostate. The wall of the seminal cyst lesion confirmed infiltrating clear cell adenocarcinoma and non-invasive adenocarcinoma in the prostate segment of the central gland in the light of the ejaculatory conduct with "in situ" growth. Thus, the surgical specimen allowed radical exeresis with negative margins, guaranteeing minimally invasive surgery with preservation of continence and erection. We describe a new integral approach for the radical surgery of localized primary adenocarcinoma of the seminal vesicle. Despite its exceptional nature, the case allowed for a double reflection: a) The study of diffusion with MRI may suggest the diagnosis of malignity in this type of lesions; and b) Radical surgical treatment must include exeresis of the central portion of the prostate gland.


Quantification of training load across two competitive seasons in elite senior and youth male soccer players from an English Premiership club.
This study aimed to compare the daily training load (TL) in first-team and U-18 soccer players from an English Premiership club. 36 first-team (age 23.2 ± 5.9 years, weight 75.2 ± 8.1 kg, height 1.83 ± 0.06 m), and 22 U-18 players (age 17.5 ± 1.1 years, weight 71.1 ± 8.2 kg, height 1.78 ± 0.08 m) participated. GPS metrics were measured during all pitch training sessions throughout the 2020-21 and 2021-22 seasons. Linear mixed-effect model analyses revealed that, irrespective of training day, U-18 players covered greater total and explosive distance than first-team players, and performed a higher number of accelerations and decelerations, whereas first-team players covered greater sprint distance. Irrespective of the team, all examined variables were greater at match-day (MD)-3, while the number of accelerations and decelerations were higher at MD-4. Significant team-by-training day interactions revealed that U-18 players covered greater total and high-intensity distances than first-team players at MD-4, MD-2, and MD-1, whereas first-team players covered greater total and high-intensity distances at MD-3. Sprint distance was greater for first-team players at MD-3 and MD-4, while explosive distance was greater for U-18 players at MD-2. Also, U-18 players performed a higher number of accelerations than first-team players at MD-3 and MD-2, and a higher number of decelerations at MD-4. The present results provide novel information on TL patterns in English Premiership soccer and contribute to understanding how training methods to physically develop players are implemented in different countries and leagues.


IGF-1 deficiency impairs neurovascular coupling in mice: implications for cerebromicrovascular aging.
Aging is associated with marked deficiency in circulating IGF-1, which has been shown to contribute to age-related cognitive decline. Impairment of moment-to-moment adjustment of cerebral blood flow (CBF) via neurovascular coupling is thought to play a critical role in the genesis of age-related cognitive impairment. To establish the link between IGF-1 deficiency and cerebromicrovascular impairment, neurovascular coupling mechanisms were studied in a novel mouse model of IGF-1 deficiency (Igf1(f/f) -TBG-Cre-AAV8) and accelerated vascular aging. We found that IGF-1-deficient mice exhibit neurovascular uncoupling and show a deficit in hippocampal-dependent spatial memory test, mimicking the aging phenotype. IGF-1 deficiency significantly impaired cerebromicrovascular endothelial function decreasing NO mediation of neurovascular coupling. IGF-1 deficiency also impaired glutamate-mediated CBF responses, likely due to dysregulation of astrocytic expression of metabotropic glutamate receptors and impairing mediation of CBF responses by eicosanoid gliotransmitters. Collectively, we demonstrate that IGF-1 deficiency promotes cerebromicrovascular dysfunction and neurovascular uncoupling mimicking the aging phenotype, which are likely to contribute to cognitive impairment.


Comparative study of analgesic effectiveness using three different anesthetic techniques for intravitreal injection of bevacizumab.
To compare the analgesic effectiveness and aesthetic appearance associated with topical, subconjunctival, and peribulbar anesthesia for intravitreal bevacizumab injection. Sixty consecutive patients undergoing their first intravitreal bevacizumab injection were randomized to receive one of three forms of anesthesia: proxymetacaine eye drops, subconjunctival injection of 2% xylocaine, and peribulbar injection of 2% xylocaine. Pain associated with the intravitreal injection and with the entire procedure (including anesthesia administration) was recorded using a Visual Analog Scale 15 minutes after intravitreal injection. Anterior segment evaluation was performed 24 hours after injection to measure the number of clock hours of subconjunctival hemorrhage. Median injection-related pain score was significantly lower in the peribulbar group compared with the topical and subconjunctival groups (P < .05). Median entire procedure pain score was significantly higher in the peribulbar group compared with the topical and subconjunctival groups (P < .05). The median extent of subconjunctival hemorrhage was significantly lower in the topical group compared with the other groups (P < .05). Among the three anesthetic techniques, peribulbar anesthesia was associated with greater effectiveness in controlling injection-related pain but was least effective in controlling entire procedure pain. There was no significant difference in pain scores between the topical and subconjunctival groups, and topical anesthesia was associated with less subconjunctival hemorrhage.


The evolution of human and ape hand proportions.
Human hands are distinguished from apes by possessing longer thumbs relative to fingers. However, this simple ape-human dichotomy fails to provide an adequate framework for testing competing hypotheses of human evolution and for reconstructing the morphology of the last common ancestor (LCA) of humans and chimpanzees. We inspect human and ape hand-length proportions using phylogenetically informed morphometric analyses and test alternative models of evolution along the anthropoid tree of life, including fossils like the plesiomorphic ape Proconsul heseloni and the hominins Ardipithecus ramidus and Australopithecus sediba. Our results reveal high levels of hand disparity among modern hominoids, which are explained by different evolutionary processes: autapomorphic evolution in hylobatids (extreme digital and thumb elongation), convergent adaptation between chimpanzees and orangutans (digital elongation) and comparatively little change in gorillas and hominins. The human (and australopith) high thumb-to-digits ratio required little change since the LCA, and was acquired convergently with other highly dexterous anthropoids.


Studies on the regulation of avidity at the level of the single antibody-forming cell. The effect of antigen dose and time after immunization.
The hemolytic plaque formation of cells producing antibody against heterologous albumins was tested for sensitivity to specific inhibition by free antigen. The inhibition characteristics of plaques in this system were found to be a measure for the avidity of the antibody produced by the plaque-forming cells (PFC:s). High avidity-producing PFC:s were more sensitive to inhibition than low avidity PFC:s. Immunization with a high dose of antigen induced PFC:s that produced antibody with a lower avidity as compared to PFC:s from animals immunized with a low dose. The avidity was increased with time. Determinations of avidity at the serum level were also made, and the results were in agreement with the findings at the cellular level. The present method made it possible to demonstrate differences in avidity of antibody at the level of the single antibody-forming cell. It may also constitute a useful tool for the analysis of the cellular events leading to the production of antibodies with varying affinities during the immune response.


Fractal Dimension Analysis of Widefield Choroidal Vasculature as Predictor of Stage of Macular Degeneration.
To evaluate the fractal dimension (Df ) of the choroidal vasculature using widefield indocyanine green (ICG) angiography and correlate it with the stage of age-related macular degeneration (AMD). Widefield ICG angiography performed on 38 eyes was retrospectively analyzed using the FracLac application within the National Institutes of Health ImageJ software to determine regional fractal dimensions in the macular field and widefield. These values were then associated with a diagnosis of no AMD, non-exudative AMD (subdivided into early/intermediate stage vs. advanced stage), or exudative AMD (subdivided into with or without geographic atrophy). The mean values were compared using Wilcoxon's test. Early/intermediate non-exudative AMD and exudative AMD without geographic atrophy were found to have statistically significantly lower Df values compared to an absence of AMD when examining the macular field. Exudative AMD with geographic atrophy was found to have a statistically significant lower choroidal fractal dimension compared to no AMD when studied in the widefield. Advanced stages of macular degeneration were found to have significantly decreased the fractal dimensions of choroidal vasculature on widefield ICG compared to early/intermediate stages, possibly implying a generalized reduction in complexity and/or vessel caliber of the choroid with advancing stage of AMD. This finding agrees with previous understanding of the development of choriocapillaris atrophy in advanced macular degeneration. These findings suggest that using automated fractal analysis techniques can aid in differentiating stages of macular degeneration and, with further study, may be used to predict advancement of macular degeneration.


Plakophilin 2: a critical scaffold for PKC alpha that regulates intercellular junction assembly.
Plakophilins (PKPs) are armadillo family members related to the classical cadherin-associated protein p120(ctn). PKPs localize to the cytoplasmic plaque of intercellular junctions and participate in linking the intermediate filament (IF)-binding protein desmoplakin (DP) to desmosomal cadherins. In response to cell-cell contact, PKP2 associates with DP in plaque precursors that form in the cytoplasm and translocate to nascent desmosomes. Here, we provide evidence that PKP2 governs DP assembly dynamics by scaffolding a DP-PKP2-protein kinase C alpha (PKC alpha) complex, which is disrupted by PKP2 knockdown. The behavior of a phosphorylation-deficient DP mutant that associates more tightly with IF is mimicked by PKP2 and PKC alpha knockdown and PKC pharmacological inhibition, all of which impair junction assembly. PKP2 knockdown is accompanied by increased phosphorylation of PKC substrates, raising the possibility that global alterations in PKC signaling may contribute to pathogenesis of congenital defects caused by PKP2 deficiency.


Screening of Visually Impaired Children for Health Problems.
Disability is a significant problem and is accepted globally as a health priority in childhood. Like nonvisually impaired children, visually impaired children also need to use health services during childhood. The purpose of this study was to determine the health problems of visually impaired children. A descriptive design was used. The subjects were 74 children with visual impairment attending primary school (aged 5-14 years), who agreed to participate and whose parents gave permission. Data were collected via physical examination including questionnaires and a physical assessment form. The health screening included physical measurements for height, weight, blood pressure, dental health, hearing, and scoliosis. The mean age of children was 10.43 ± 2.9 years. When the health screening results of children were examined, it was found that 25.7% of the children were overweight or obese, 35.1% of them had dental problems, 27.0% had hearing problems, and 39.2% had scoliosis risk. Systolic and diastolic blood pressures were normal in 91.8% and 93.2% of the children, respectively. These findings showed the important role of school health nurses in performing health screenings directed at visually impaired children who constitute a special group for school health services. Health screening for height, weight, dental health, hearing, and scoliosis is suggested for visually impaired children.


Pancreatico-pleural and bronchial fistulae and associated pseudocysts: case series.
Pancreatico-pleural fistula is rare complication of chronic or acute pancreatitis. Previous studies have reported imaging features and various management options of this condition including conservative/medical management, endoscopic treatments and surgery.This article reviews the myriad of imaging appearances of this condition in multimodality imaging and different strategies for the successful management in a short case series. After obtaining the institutional ethics committee approval, retrospective review of the medical records of five patients of pancreatico-pleural fistulae who were diagnosed and successfully managed in our hospital in 2012 and 2013 was done. Follow up with out patient records of these patients was also included.Findings were compared with the current available literature on this entity. Pancreatico-pleural fistulae presents with massive pleural effusion.A high index of suspicion is essential for accurate diagnosis. Demonstration of the fistulous tracts requires cross sectional imaging with contrast enhanced CT being most commonly used and affords accurate diagnosis. MRI demonstrates the tracts and ductal disruptions with greater detail and are helpful in confirming the CT findings. Endoscopic ultrasound and ERCP also offer potential of diagnosis, although being technically demanding and invasive is reserved for interventions. Management of these conditions should be initially conservative with endoscopic stenting being offered in selected cases with favourable anatomy and not responding to conservative management. Surgery is reserved for cases not responding to conservative and endoscopic management. In conclusion this case series highlights the clinical and imaging spectrum of pancreatico-pleural fistulae and provides insight into the different management strategies that can be adopted for this condition.


The latent class structure of ADHD is stable across informants.
Previous studies have looked at the structure of attention-deficit/hyperactivity disorder (ADHD) using latent class analysis (LCA) of Child Behavior Checklist (CBCL) or Diagnostic and Statistical Manual of Mental Disorders (DSM) symptom structure. These studies have identified distinct classes of children with inattentive, hyperactive, or combined subtypes and have used these classes to refine genetic analyses. The objective of the current report is to determine if the latent class structure of ADHD subtypes is consistent across informant using the Conners' Rating Scales (CRS). LCA was applied to CRS forms from mother, father, and teacher reports of 1837, 1329 and 1048 latency aged Dutch twins, respectively. The optimal solution for boys was a 5-class solution for mothers, a 3-class solution for fathers, and a 4-class solution for teachers. For girls, a 4-class solution for mothers and a 3-class for fathers and teachers was optimal. Children placed into a class by one informant had markedly increased odds ratio of being placed into the same or similar class by the other informants. Results from LCA using Dutch twins with the CRS show stability across informants suggesting that more stable phenotypes may be accessible for genotyping using a multi-informant approach.


Highly Conductive Self-Healing Biocomposites Based on Protein Mediated Self-Assembly of PEDOT:PSS Films.
Composites of conducting polymers offer a broad spectrum of materials for interfacing electronic devices with biological systems. Particularly, material systems based on poly(styrenesulfonate) doped poly(3,4-ethylenedioxythiophene) (PEDOT:PSS) have found applications in many bioelectronic devices as biosensitive transistors, controlled drug delivery media, and strain, temperature, and humidity sensors. The biocompatibility, intercoupled electronic and ionic conductivity, and air stable electrical properties render PEDOT:PSS based material systems indispensable for bioelectronics. However, these materials are commonly used in thin film form since freestanding films of pristine PEDOT:PSS are considered mechanically brittle compared to biological tissues, and unlike biological systems these conductive films cannot restore/heal their physical properties after excessive mechanical deformation. Here we report conductive biocomposites of PEDOT:PSS and tandem repeat proteins with the ability to self-heal once plasticized via water. The tandem repeat proteins acquired from squid ring teeth (SRT) induce structural effects on PEDOT:PSS including improved crystallinity and formation of fibrous network structures. These structural effects lead to electrical conductivity values reaching 120 S/cm for biocomposites with SRT protein concentrations below 20 wt %, which exceeds the conductivity of pristine PEDOT:PSS (∼100 S/cm). More importantly, tandem proteins facilitate consistent self-healing of freestanding biocomposites with SRT protein concentrations beyond 40 wt %. These robust biocomposites with high electrical conductivity and the ability to self-heal can find applications in numerous soft electronic systems spanning from implantable, transient, and epidermal electronics to electronic textiles.


High-resolution metagenomics targets specific functional types in complex microbial communities.
Most microbes in the biosphere remain unculturable. Whole genome shotgun (WGS) sequencing of environmental DNA (metagenomics) can be used to study the genetic and metabolic properties of natural microbial communities. However, in communities of high complexity, metagenomics fails to link specific microbes to specific ecological functions. To overcome this limitation, we developed a method to target microbial subpopulations by labeling DNA through stable isotope probing (SIP), followed by WGS sequencing. Metagenome analysis of microbes from Lake Washington in Seattle that oxidize single-carbon (C1) compounds shows specific sequence enrichments in response to different C1 substrates, revealing the ecological roles of individual phylotypes. We also demonstrate the utility of our approach by extracting a nearly complete genome of a novel methylotroph, Methylotenera mobilis, reconstructing its metabolism and conducting genome-wide analyses. This high-resolution, targeted metagenomics approach may be applicable to a wide variety of ecosystems.


Long Electron-Hole Diffusion Length in High-Quality Lead-Free Double Perovskite Films.
Developing environmentally friendly perovskites has become important in solving the toxicity issue of lead-based perovskite solar cells. Here, the first double perovskite (Cs2 AgBiBr6 ) solar cells using the planar structure are demonstrated. The prepared Cs2 AgBiBr6 films are composed of high-crystal-quality grains with diameters equal to the film thickness, thus minimizing the grain boundary length and the carrier recombination. These high-quality double perovskite films show long electron-hole diffusion lengths greater than 100 nm, enabling the fabrication of planar structure double perovskite solar cells. The resulting solar cells based on planar TiO2 exhibit an average power conversion efficiency over 1%. This work represents an important step forward toward the realization of environmentally friendly solar cells and also has important implications for the applications of double perovskites in other optoelectronic devices.


A qualitative study explaining nurses' perceptions of quality care for older people in long-term care settings in Ireland.
The aim of this research was to explore nurses' perceptions of the attributes of quality care and the factors that facilitate or hinder high-quality nursing care in long-term care. The quality of care for older people living in long-term care has been identified as an issue of concern in many nursing research studies. While many factors have been identified, it is difficult to determine key factors from current research. The study was a qualitative exploration of nurses' perceptions of quality care for older people and the factors that facilitate or hinder quality care. It involved 20 interviews with nurses. Respondents were asked to illustrate their accounts with examples from practice. This phase of the research was guided by the principles of hermeneutic phenomenology and the analysis process by Van Manen. The findings indicated that nurses perceived quality care for older people in Ireland as holistic, individualized and focused on promoting independence and choice. The research revealed, however, that care in many practice areas was not individualized, patient choice and involvement in decision making was limited and some areas engendered dependency. While staffing was identified as a factor which had an impact on the provision of patient choice, other issues, such as the motivation of staff, the role of the ward manager and the dominance of routine were also highlighted. There is a need to review organizational approaches to care, develop patient centred approaches to care and provide educational support for managers. This research focuses on care for older people; it helps practitioners identify key factors in the provision of quality care for older people living in long-term care.


Underwater Single Image Color Restoration Using Haze-Lines and a New Quantitative Dataset.
Underwater images suffer from color distortion and low contrast, because light is attenuated while it propagates through water. Attenuation under water varies with wavelength, unlike terrestrial images where attenuation is assumed to be spectrally uniform. The attenuation depends both on the water body and the 3D structure of the scene, making color restoration difficult. Unlike existing single underwater image enhancement techniques, our method takes into account multiple spectral profiles of different water types. By estimating just two additional global parameters: the attenuation ratios of the blue-red and blue-green color channels, the problem is reduced to single image dehazing, where all color channels have the same attenuation coefficients. Since the water type is unknown, we evaluate different parameters out of an existing library of water types. Each type leads to a different restored image and the best result is automatically chosen based on color distribution. We also contribute a dataset of 57 images taken in different locations. To obtain ground truth, we placed multiple color charts in the scenes and calculated its 3D structure using stereo imaging. This dataset enables a rigorous quantitative evaluation of restoration algorithms on natural images for the first time.


Prospective diary study of nonpainful and painful phantom sensations in a preselected sample of child and adolescent amputees reporting phantom limbs.
To prospectively study factors associated with the occurrence of phantom sensations and pains in a pre-selected sample of child and adolescent amputees reporting phantom limbs. Prospective diary study over 1 month. Fourteen child and adolescent amputees from 10-18 years of age who were missing a limb due to trauma (n = 12) or congenital limb deficiency (n = 2), and who had previously reported having phantom sensations and pain. Diary used to assess the occurrence of non-painful and painful phantom sensations. Items included age, sex, location and cause of amputation, past experience with stump pain and pre-amputation pain, and intensity, quality, duration, and triggers of the sensations and pains. Thirteen amputees reported having 104 incidents of non-painful phantom sensations with an average intensity of 4.17 (SD = 2.14) on a 0-10 rating scale. Fifty-three incidents of phantom pain with an average intensity of 6.43 (SD = 1.76) were recorded by 8 amputees. Both amputees with a congenital limb deficiency reported phantom phenomena. Girls reported more psychosocial triggers than did boys whereas boys were more likely than girls to report that they could not identify a trigger (P = 0.0001). Boys also reported a higher proportion of physical triggers than psychosocial triggers while there were no differences for girls (P = 0.0001). Child and adolescent amputees experience phantom sensations and pains on a regular basis over a 1-month period. Differences in triggers of phantom phenomena between boys and girls may be due to differences in activities, awareness, attribution, and willingness to report psychosocial triggers.


Overexpression of Arabidopsis phytochelatin synthase in tobacco plants enhances Cd(2+) tolerance and accumulation but not translocation to the shoot.
Phytochelatins (PCs) are metal binding peptides involved in heavy metal detoxification. To assess whether enhanced phytochelatin synthesis would increase heavy metal tolerance and accumulation in plants, we overexpressed the Arabidopsis phytochelatin synthase gene (AtPCS1) in the non-accumulator plant Nicotiana tabacum. Wild-type plants and plants harbouring the Agrobacterium rhizogenes rolB oncogene were transformed with a 35S AtPCS1 construct. Root cultures from rolB plants could be easily established and we demonstrated here that they represent a reliable system to study heavy metal tolerance. Cd(2+) tolerance in cultured rolB roots was increased as a result of overexpression of AtPCS1, and further enhanced when reduced glutathione (GSH, the substrate of PCS1) was added to the culture medium. Accordingly, HPLC analysis showed that total PC production in PCS1-overexpressing rolB roots was higher than in rolB roots in the presence of GSH. Overexpression of AtPCS1 in whole seedlings led to a twofold increase in Cd(2+) accumulation in the roots and shoots of both rolB and wild-type seedlings. Similarly, a significant increase in Cd(2+) accumulation linked to a higher production of PCs in both roots and shoots was observed in adult plants. However, the percentage of Cd(2+) translocated to the shoots of seedlings and adult overexpressing plants was unaffected. We conclude that the increase in Cd(2+) tolerance and accumulation of PCS1 overexpressing plants is directly related to the availability of GSH, while overexpression of phytochelatin synthase does not enhance long distance root-to-shoot Cd(2+) transport.


Pharmacokinetics and safety of first-line tuberculosis drugs rifampin, isoniazid, ethambutol, and pyrazinamide during pregnancy and postpartum: results from IMPAACT P1026s.
Physiological changes during pregnancy may alter the pharmacokinetics (PK) of antituberculosis drugs. The International Maternal Pediatric Adolescent AIDS Clinical Trials Network P1026s was a multicenter, phase IV, observational, prospective PK and safety study of antiretroviral and antituberculosis drugs administered as part of clinical care in pregnant persons living with and without HIV. We assessed the effects of pregnancy on rifampin, isoniazid, ethambutol, and pyrazinamide PK in pregnant and postpartum (PP) persons without HIV treated for drug-susceptible tuberculosis disease. Daily antituberculosis treatment was prescribed following World Health Organization-recommended weight-band dosing guidelines. Steady-state 12-hour PK profiles of rifampin, isoniazid, ethambutol, and pyrazinamide were performed during second trimester (2T), third trimester (3T), and 2-8 of weeks PP. PK parameters were characterized using noncompartmental analysis, and comparisons were made using geometric mean ratios (GMRs) with 90% confidence intervals (CI). Twenty-seven participants were included: 11 African, 9 Asian, 3 Hispanic, and 4 mixed descent. PK data were available for 17, 21, and 14 participants in 2T, 3T, and PP, respectively. Rifampin and pyrazinamide AUC0-24 and C max in pregnancy were comparable to PP with the GMR between 0.80 and 1.25. Compared to PP, isoniazid AUC0-24 was 25% lower and C max was 23% lower in 3T. Ethambutol AUC0-24 was 39% lower in 3T but limited by a low PP sample size. In summary, isoniazid and ethambutol concentrations were lower during pregnancy compared to PP concentrations, while rifampin and pyrazinamide concentrations were similar. However, the median AUC0-24 for rifampin, isoniazid, and pyrazinamide met the therapeutic targets. The clinical impact of lower isoniazid and ethambutol exposure during pregnancy needs to be determined.


A methodological and substantive review of the evidence that schools cause pupils to smoke.
The objectives of this review were to examine whether smoking prevalence varies between schools independently of health promotion programmes and pupil composition, to show which school characteristics are responsible for this variation, and to examine the methodological adequacy of such studies. Searches for published studies were performed on medical, educational and social science databases, relevant articles' reference lists, and citation searches. Any study was included that described inter-school variation in smoking prevalence, or related such variation to school characteristics. A model relating pupil smoking to school, neighbourhood, and pupil characteristics unlikely and likely to be influenced by school was used to examine the adequacy of control of confounding by pupil composition. Data from studies were combined qualitatively considering methodological adequacy to examine the relation of smoking prevalence to school characteristics. Theoretical frameworks underpinning the choice of school characteristics and postulated relationships between these characteristics and smoking prevalence were described. There were large variations in smoking prevalence between ostensibly similar schools. Evidence that pupil composition did not cause this was weak, because all studies had methodological problems, including under control of relevant pupil compositional factors and over control of factors likely to represent the mechanism through which schools influence pupils' smoking. There was little evidence that elements of tobacco control policy other than bans and enforcement deterred smoking. Academic practice and school ethos were related to smoking. Academically selective schools did not influence smoking, once pupil composition was controlled. There was one study on neighbourhood influences, which were unrelated to smoking. Studies frequently offered little or no theoretical justification for associating school characteristics with smoking. Some aspects of school influence pupils' smoking, probably independently of pupil composition. However, under-control and over-control of confounding and lack of theoretical underpinning precludes definitive conclusions on how particular school characteristics influence pupils' smoking.


Bilateral germ-cell testicular cancer - long-term experience.
Germ cell testicular cancer (TC) patients have an increased risk of developing a contralateral TC. This paper presents a retrospective analysis of patients with bilateral TC from a database of a single center and aims to highlight the need for an individual approach. In the study period from January 1970 to June 2013, a total of 1,438 patients with primary TC were diagnosed. The group comprised of 1,370 patients with unilateral TC and 68 patients with bilateral TC. Bilateral TC patients were evaluated regarding the age at presentation and the histology of tumor and then compared with the group of patients with unilate-ral TC. The mean age of the patients with unilateral TC was 31.7 years. This group of patients consisted of 408 patients with seminoma and 962 patients with non-seminomatous TC. The group of 68 patients with bilateral TC involved 63 patients with metachronous disease and five patients who developed TCs simultaneously. The mean age at diagnosis of primary tumor in patients with metachronous bilateral TC (25.8 years) was significantly earlier in comparison to patients with unilateral disease. Among 24 patients with seminoma there were 17 patients who subsequently developed a second malignancy of an identical histological type. In the group of 39 patients with initial non-seminomatous TC there were 15 patients with discordant histology of the second tumor. In the metachronous disease group, patients with primary seminoma were significantly older than those with initial non-seminomas. All patients with unilateral TC have an increased risk of developing a contralateral TC, years and even decades after the initial diagnosis. Management of subsequent TC should be individualized for each patient according to the histology and clinical stage of the primary tumor, as well as the therapeutic strategy applied initially. The further management should involve a long-term follow-up and life-long testosterone substitution.


Insights into Crystal Structure and Diffusion of Biphasic Na2Zn2TeO6.
The layered oxide Na2Zn2TeO6 is a fast Na+ ion conductor and a suitable candidate for application as a solid-state electrolyte. We present a detailed study on how synthesis temperature and Na-content affect the crystal structure and thus the Na+ ion conductivity of Na2Zn2TeO6. Furthermore, we report for the first time an O'3-type phase for Na2Zn2TeO6. At a synthesis temperature of 900 °C, we obtain a pure P2-type phase, providing peak performance in Na+ ion conductivity. Synthesis temperatures lower than 900 °C produce a series of mixed P2 and O'3-type phases. The O'3 structure can only be obtained as a pure phase by substituting Li on the Zn-sites to increase the Na-content. Thorough analysis of synchrotron data combined with computational modeling indicates that Li enters the Zn sites and, consequently, the amount of Na in the structure increases to balance the charge according to the formula Na2+xZn2-xLixTeO6 (x = 0.2-0.5). Impedance spectroscopy and computational modeling confirm that reducing the amount of the O'3-type phase enhances the Na+ ion mobility.


An immunoperoxidase assay for the detection of specific IgA antibody in Epstein-Barr virus infections.
A technique using indirect immunoperoxidase antibody was developed for the detection of specific serum IgA antibody to Epstein-Barr virus capsid antigen and early antigen. The IgA technique was compared with an immunofluorescence antibody method. Epstein-Barr virus IgA antibody against viral capsid antigen was detected in all nine patients with Epstein-Barr virus associated undifferentiated nasopharyngeal carcinoma, in 13 (72.2%) of 18 patients with infectious mononucleosis, in 21 (28.3%) of 74 patients with acute lymphoblastic leukaemia, and in six (20%) of 30 patients who had recently had kidney transplants. Epstein-Barr virus IgA antibody against viral capsid antigen was also detected in four (10%) of 40 healthy subjects, but it was not found in any of 20 cord blood samples. Epstein-Barr virus IgA antibody to early antigen was detected in six (66.6%) patients with nasopharyngeal carcinoma and in two (2.7%) patients with acute lymphoblastic leukaemia. The immunoperoxidase assay for Epstein-Barr virus specific IgA was simple, reliable, and rapid and correlated well (r = 0.94) with the immunofluorescence antibody technique.


Cell surface localization of 5'AMP nucleotidase in prestalk cells of Dictyostelium discoideum.
5'AMP nucleotidase activity was localized by electron microscopy in Dictyostelium discoideum during cell differentiation. In addition, the activity was assayed by micro enzymic methods in sections dissected from specific cellular regions of lyophilized individuals. The results of the 2 procedures were in agreement, demonstrating that at the culmination stage of development the activity is strikingly localized in the prestalk cells adjacent to the prespore region. The cytochemically stained reaction product appeared only along the plasma membrane of the cells. As prestalk cells migrate into the stalk sheath and undergo differentiation, the activity is rapidly lost. Examination of stained cells at high magnification revealed the product accumulation to be primarily at the cell surface, suggesting that the enzyme functions extracellularly. Occasionally, cells having the morphological appearance of prestalk cells were found within the prespore region. These cells demonstrated 5'AMP nucleotidase activity at their plasma membrane in sharp contrast with neighbouring prespore cells. The strategic localization of 5'AMP nucleotidase may reflect a mechanism for establishing and maintaining regulatory levels of extracellular 5'AMP and/or adenosine during pattern formation in this model system.


Structure and function of sphingosine-1-phosphate lyase, a key enzyme of sphingolipid metabolism.
Sphingosine-1-phosphate lyase (SPL), a key enzyme of sphingolipid metabolism, catalyzes the irreversible degradation of sphingoid base phosphates. Its main substrate sphingosine-1-phosphate (S1P) acts both extracellularly, by binding G protein-coupled receptors of the lysophospholipid receptor family, and inside the cell, as a second messenger. There, S1P takes part in regulating various cellular processes and its levels are tightly regulated. SPL is a pivotal enzyme regulating S1P intracellular concentrations and a promising drug target for the design of immunosuppressants. We structurally and functionally characterized yeast SPL (Dpl1p) and its first prokaryotic homolog, from Symbiobacterium thermophilum. The Dpl1p structure served as a basis for a very reliable model of Homo sapiens SPL. The above results, together with in vitro and in vivo studies of SPL mutants, reveal which residues are involved in activity and substrate binding and pave the way to studies aimed at controlling the activity of this pivotal enzyme.


Pseudomonas mosselii improves cold tolerance of Asian rice (Oryza sativa L.) in a genotype-dependent manner by increasing proline in japonica and reduced glutathione in indica varieties.
Cold stress is an important factor limiting rice production and distribution. Identifying factors that contribute to cold tolerance in rice is of primary importance. While some plant specific genetic factors involved in cold tolerance have been identified, the role of the rice microbiome remains unexplored. In this study, we evaluated the influence of plant growth promoting bacteria (PGPB) with the ability of phosphate solubilization on rice cold tolerance and survival. To reach this goal, inoculated and uninoculated 2-week-old seedlings were cold stressed and evaluated for survival and other phenotypes such as electrolyte leakage (EL) and necessary elements for cold tolerance. The results of this study showed that of the five bacteria, Pseudomonas mosselii, improved both indica and japonica varietal plants' survival and decreased EL, indicating increased membrane integrity. We observed different possible cold tolerance mechanisms in japonica and indica plants such as increases in proline and reduced glutathione levels, respectively. This bacterium also improved the shoot growth of cold exposed indica plants during the recovery period. This study confirmed the host genotype dependent activity of P. mosselii and indicated that there is an interaction between specific plant genes and bacterial genes that causes different plant responses to cold stress.


Ethosuximide: from bench to bedside.
Ethosuximide, 2-ethyl-2-methylsuccinimide, has been used extensively for "petit mal" seizures and it is a valuable agent in studies of absence epilepsy. In the treatment of epilepsy, ethosuximide has a narrow therapeutic profile. It is the drug of choice in the monotherapy or combination therapy of children with generalized absence (petit mal) epilepsy. Commonly observed side effects of ethosuximide are dose dependent and involve the gastrointestinal tract and central nervous system. Ethosuximide has been associated with a wide variety of idiosyncratic reactions and with hematopoietic adverse effects. Typical absence seizures are generated as a result of complex interactions between the thalamus and the cerebral cortex. This thalamocortical circuitry is under the control of several specific inhibitory and excitatory systems arising from the forebrain and brainstem. Corticothalamic rhythms are believed to be involved in the generation of spike-and-wave discharges that are the characteristic electroencephalographic signs of absence seizures. The spontaneous pacemaker oscillatory activity of thalamocortical circuitry involves low threshold T-type Ca2+ currents in the thalamus, and ethosuximide is presumed to reduce these low threshold T-type Ca2+ currents in thalamic neurons. Ethosuximide also decreases the persistent Na+ and Ca2+ -activated K+ currents in thalamic and layer V cortical pyramidal neurons. In addition, there is evidence that in a genetic absence epilepsy rat model ethosuximide reduces cortical gamma-aminobutyric acid (GABA) levels. Also, elevated glutamate levels in the primary motor cortex of rats with absence epilepsy (but not in normal animals) are reduced by ethosuximide.


Survey investigating homosexual behaviour among adult males used to estimate the prevalence of HIV and AIDS among men who have sex with men in Japan.
This study investigated the prevalence of male homosexual behaviour among adult men and of HIV and AIDS among men who have sex with men (MSM) and non-MSM in Japan. An anonymous self-administered postal questionnaire, and national HIV and AIDS notifications. Same-sex sexual experience was reported by 2.0% of respondents. The prevalence of HIV and AIDS was 0.8818% among MSM and 0.0130% among non-MSM, indicating that HIV and AIDS are 68 times more prevalent among MSM. Our findings underestimate homosexual and HIV prevalence due to several methodological limitations. The high prevalence of HIV and AIDS among MSM in comparison with non-MSM indicates the urgent need to prioritise funding and programs targeting MSM in Japan.


The impact of pre-transplant dialysis on simultaneous pancreas-kidney versus living donor kidney transplant outcomes.
Pre-transplant dialysis is known to affect kidney graft survival. Here, we report the impact of pre-transplant dialysis on patient and graft survival of type 1 diabetic recipients of either a simultaneous pancreas-kidney (SPK) or living donor kidney (LDK) transplant. Using the Organ Procurement Transplant Network/United Network for Organ Sharing (OPTN/UNOS) database, 6822 adult type 1 diabetic recipients transplanted through 2000-2011 were identified. Patients were categorized based on pre-transplant dialysis time (DT): preemptive recipients (P-LDK, n = 498; P-SPK, n = 1529), recipients with <1 year of DT (0-1 year DT; LDK n = 582, SPK n = 1700), and those with 1-2 years DT (1-2 year DT; LDK n = 301, SPK n = 2212). Seven-year patient and kidney survival were examined. Compared with the P-SPK group, both 0-1 year DT and 1-2 year DT SPK recipients had lower 7-year patient survival (89, 84 & 84% respectively; log-rank P-value versus P-SPK = 0.01 & <0.001). For LDK groups, DT > 1 year was associated with inferior patient survival (7-year survival 76% versus 87% for P-LDK, P-value versus P-LDK = 0.009). Comparing P-LDK to all other SPK groups, there was no significant difference in 7-year patient or kidney survival. Preemptive transplantation is associated with the highest patient survival in both LDK and SPK. Compared with the P-LDK group, DT > 1 year is associated with lower patient survival among LDK recipients, but there is no difference in survival with dialysis up to 2 years with SPK. These results highlight the differential impact of DT on LDK and SPK transplantation.


Bleomycin causes alveolar macrophages from cigarette smokers to release hydrogen peroxide.
Alveolar macrophages (AM) were retrieved by bronchoalveolar lavage from 20 patients. Following AM purification by glass adherence, the effect of bleomycin on hydrogen peroxide and lactic dehydrogenase release was assessed by colorimetric methods. All AM from nine nonsmokers had no detectable spontaneous release of hydrogen peroxide. The AM did not release hydrogen peroxide when stimulated with bleomycin (25 mU/mL). AM from all eleven smokers spontaneously released hydrogen peroxide (36 +/- 5.3 nm/10(6) AM, mean +/- SEM). AM from eight of the eleven smokers released more hydrogen peroxide when incubated with bleomycin (smokers 55 +/- 6.8 nm/10(6) AM, p less than 0.05). There was no difference in the amount of lactic dehydrogenase released by AM spontaneously or when incubated with bleomycin at 25 mU/mL, suggesting that this dose of bleomycin was not directly toxic to the AM. Results from this study further support the premise that bleomycin pulmonary toxicity may result from the generation of reactive oxygen metabolites and that cigarette smoking may predispose to bleomycin pulmonary toxicity.


Prevention of Arterial Elastocalcinosis: Differential Roles of the Conserved Glutamic Acid and Serine Residues of Matrix Gla Protein.
Inactivating mutations in matrix Gla protein (MGP) lead to Keutel syndrome, a rare disease hallmarked by ectopic calcification of cartilage and vascular tissues. Although MGP acts as a strong inhibitor of arterial elastic lamina calcification (elastocalcinosis), its mode of action is unknown. Two sets of conserved residues undergoing posttranslational modifications-4 glutamic acid residues, which are γ-carboxylated by gamma-glutamyl carboxylase; and 3 serine residues, which are phosphorylated by yet unknown kinase(s)-are thought to be essential for MGP's function. We pursued a genetic approach to study the roles of MGP's conserved residues. First, a transgenic line (SM22a-GlamutMgp) expressing a mutant form of MGP, in which the conserved glutamic acid residues were mutated to alanine, was generated. The transgene was introduced to Mgp-/- mice to generate a compound mutant, which produced the mutated MGP only in the vascular tissues. We generated a second mouse model (MgpS3mut/S3mut) to mutate MGP's conserved serine residues to alanine. The initiation and progression of vascular calcification in these models were analyzed by alizarin red staining, histology, and micro-computed tomography imaging. On a regular diet, the arterial walls in the Mgp-/-; SM22α-GlamutMgp mice were not calcified. However, on a high phosphorus diet, these mice showed wide-spread arterial calcification. In contrast, MgpS3mut/S3mut mice on a regular diet recapitulated arterial calcification traits of Mgp-/- mice, although with lesser severity. For the first time, we show here that MGP's conserved serine residues are indispensable for its antimineralization function in the arterial tissues. Although the conserved glutamic acid residues are not essential for this function on a regular diet, they are needed to prevent phosphate-induced arterial elastocalcinosis.


Mitonuclear interactions modulate nutritional preference.
In nature, organisms are faced with constant nutritional options which fuel key life-history traits. Studies have shown that species can actively make nutritional decisions based on internal and external cues. Metabolism itself is underpinned by complex genomic interactions involving components from both nuclear and mitochondrial genomes. Products from these two genomes must coordinate how nutrients are extracted, used and recycled. Given the complicated nature of metabolism, it is not well understood how nutritional choices are affected by mitonuclear interactions. This is under the rationale that changes in genomic interactions will affect metabolic flux and change physiological requirements. To this end we used a large Drosophila mitonuclear genetic panel, comprising nine isogenic nuclear genomes coupled to nine mitochondrial haplotypes, giving a total of 81 different mitonuclear genotypes. We use a capillary-based feeding assay to screen this panel for dietary preference between carbohydrate and protein. We find significant mitonuclear interactions modulating nutritional choices, with these epistatic interactions also being dependent on sex. Our findings support the notion that complex genomic interactions can place a constraint on metabolic flux. This work gives us deeper insights into how key metabolic interactions can have broad implications on behaviour.


[The synaptic changes in the structures of the hyperstriatal section of the chicken brain during imprinting, visual deprivation and intermittent photic stimulation].
Morphometric analysis of synapses was conducted in medial areas of the ventral and accessory hyperstriatum in the right and left hemispheres of chickens brain after: 1) rhythmic optical stimulation beginning from the 18-th day of incubation, 2) presentation of flashing stimuli on the 10-12-th hours of life, 3) keeping under constant illumination in the first day or 4) in darkness during 24 hours after hatching. On electronograms of ultrathin slices parameters were recorded of synapses numerical density, mean length and width of postsynaptic condensations, number of synaptic vesicles per active zone in the presynapse and mean length of the whole axodendrite contact. The results of the studies have shown that early visual experience leads to significant changes of synaptic complex in the studied brain regions. Plasticity of synapses is discussed during stimulation and morphofunctional interrelations between hyperstriatal areas which are connected with different aspects of processing and storage of the visual information.


Oxytocin and oxytocin receptors in cancer cells and proliferation.
The hypothalamic nonapeptide oxytocin plays a crucial role in many reproductive and behavioural functions. However, in recent years, an additional new role for oxytocin has been identified in neoplastic pathology. In tumours, oxytocin acts as a growth regulator, through the activation of a specific G-coupled transmembrane receptor, the oxytocin receptor. In vitro, oxytocin inhibits proliferation of neoplastic cells of either epithelial (mammary and endometrial), nervous or bone origin, all expressing oxytocin receptor. Furthermore, an oxytocin growth-inhibiting effect was also tested and confirmed in vivo in mouse and rat mammary carcinomas. In neoplastic cells derived from two additional oxytocin target tissues, trophoblast and endothelium, oxytocin was found to promote cell proliferation, an effect opposite to that previously described in all other neoplastic oxytocin-responsive cells. The signal transduction pathways coupled to the biological effects of oxytocin are different in oxytocin growth-inhibited or growth-stimulated cells, and may depend on the membrane localization of the oxytocin receptor itself. The inhibitory effect of oxytocin is apparently mediated by activation of the cAMP-protein kinase A pathway, a nonconventional oxytocin signalling pathway, whereas the mitogenic effect is coupled to the increase of intracellular [Ca(2+)] and tyrosine phosphorylation, 'classical' oxytocin transducers. Moreover, the oxytocin receptor localization in lipid rafts enriched in caveolin-1 turns the inhibition of cell growth into a proliferative response, eliciting different epidermal growth factor receptor/mitogen-activated protein kinase activation patterns. This unexpected role of oxytocin (and oxytocin analogues) in regulating cell proliferation, as well as the widespread expression of oxytocin receptors in neoplastic tissues of different origin, opens up new perspectives on the biological role of the oxytocin-oxytocin receptor system in cancer.


Ice nucleation forced by transient electric fields.
Icing affects many technical systems, like aircraft or high-voltage power transmission and distribution in cold regions. Ice accretion is often initiated by ice nucleation in sessile supercooled water droplets and is influenced by several influencing factors, of which the impact of electric fields on ice nucleation is still not completely understood. Especially the influence of transient electric fields is rarely or not at all investigated, even though it is of great interest, e.g., for high-voltage transmission lines or for the food industry. In the present study the impact of transient electric fields on ice nucleation in supercooled sessile water droplets is experimentally investigated under well-defined conditions. A set of droplets is cooled down to a certain temperature and is subsequently exposed to electric fields generated from standard lightning or standard switching impulse voltages, which are commonly used for testing of high-voltage equipment. The nucleation behavior of individual droplets is captured using a high-speed camera and the effect of the transient electric field on ice nucleation is analyzed by considering both the singular and the stochastic nature of nucleation. While the singular nature of nucleation is referred to during analysis of the relative number of droplets remaining liquid long times after the impulse voltage, its stochastic nature is accounted for in the analysis of the temporal evolution of the relative number of frozen droplets. It is shown that low electric field strengths (E[over ̂]≤6.52kV/cm) only have a negligible impact on ice nucleation, independent of the supercooling. In contrast, high electric field strengths (E[over ̂]≥9.78kV/cm) promote significantly ice nucleation. It is also shown that depending on the supercooling, the freezing delay of the different droplets in the ensemble may vary over several magnitudes for the same conditions. It is demonstrated that the electric field appears to indirectly affect the nucleation rate by generating droplet oscillations, finally promoting ice nucleation. The experiments clearly demonstrate the possibility to actively force ice nucleation by applying transient electric fields. These results improve the understanding of ice accretion on high-voltage insulators and may also lend insight into freezing processes in food industry. We expect that these results will be a valuable contribution in formulating and/or validating new nucleation models.


Effects of magnetic resonance imaging in patients with implanted deep brain stimulation systems.
The aim of this study was to study the effects of MR imaging on the electrical settings of deep brain stimulation (DBS) systems and their clinical consequences. The authors studied the effects of 1.5-T MR imaging on the electrical settings of implanted DBS systems, including 1 or more monopolar or quadripolar leads, extension leads, and single- or dual-channel implantable pulse generators (IPGs). The IPG was switched off during the procedure and the voltage was set to 0. The impedances were checked before and after MR imaging. Five hundred seventy patients were treated with DBS for movement disorders and underwent brain MR imaging after lead implantation and before IPG implantation. None of the patients experienced any adverse events. Thirty-one of these patients underwent 61 additional MR imaging sessions after the entire DBS system had been implanted. The authors report neither local cutaneous nor neurological disorders during or after the MR imaging session. No change in the IPG settings occurred when the magnet reed switch function remained disabled during the procedure. This study demonstrates that 1.5-T MR imaging can be performed safely with continuous monitoring in patients with a DBS system. The ability to disable the magnet reed switch function of the IPG prevents any change in the electrical settings and thus any side effects. The increasing number of DBS indications and the widespread use of MR imaging indicates the need for defining safety guidelines for the use of MR imaging in patients with implanted neurostimulators.


The molecular biology of head and neck cancer.
Head and neck squamous cell carcinomas (HNSCCs) are caused by tobacco and alcohol consumption and by infection with high-risk types of human papillomavirus (HPV). Tumours often develop within preneoplastic fields of genetically altered cells. The persistence of these fields after treatment presents a major challenge, because it might lead to local recurrences and second primary tumours that are responsible for a large proportion of deaths. Aberrant signalling pathways have been identified in HNSCCs and inhibition of epidermal growth factor receptor (EGFR) has proved a successful therapeutic strategy. In this Review, we discuss the recent literature on tumour heterogeneity, field cancerization, molecular pathogenesis and the underlying causative cancer genes that can be exploited for novel and personalized treatments of patients with HNSCC.


Sun-induced leaf fluorescence retrieval in the O2-B atmospheric absorption band.
Sun-induced leaf fluorescence was inferred by using high resolution (0.5 cm(-1)) radiance measurements and simulated spectra of the solar irradiance at the ground level, in the region of the O(2)-B absorption band. The minimization of a cost function was performed in the Fourier transform domain in order to make an accurate fit of the Instrumental Line- Shape that convoluted the simulated spectrum. Second- order polynomials were used to fit the leaf fluorescence and reflectance in the 100-cm(-1)-wide spectral window. The scale and the instrumental conversion factor were also fitted in order to obtain an accuracy that could not be attained by using the radiance measurements alone.


P,O-Phosphinophenolate zinc(II) species: synthesis, structure and use in the ring-opening polymerization (ROP) of lactide, ε-caprolactone and trimethylene carbonate.
The P,O-type phosphinophenol proligands (1·H, 2-PPh2-4-Me-6-Me-C6H2OH; 2·H, 2-PPh2-4-Me-6-(t)Bu-C6H2OH) readily react with one equiv. of ZnEt2 to afford in high yields the corresponding Zn(II)-ethyl dimers of the type [(κ(2)-P,O)Zn-Et]2 (3 and 4) with two μ-OPh bridging oxygens connecting the two Zn(II) centers, as determined by X-ray diffraction (XRD) studies in the case of 3. Based on diffusion-ordered NMR spectroscopy (DOSY), both species 3 and 4 retain their dimeric structures in solution. The alcoholysis reaction of Zn(II) alkyls 3 and 4 with BnOH led to the high yield formation of the corresponding Zn(II) benzyloxide species [(κ(2)-P,O)Zn-OBn]2 (5 and 6), isolated in a pure form as colorless solids. The centrosymmetric and dimeric nature of Zn(II) alkoxides 5 and 6 in solution was deduced from DOSY NMR experiments and multinuclear NMR data. Though the heteroleptic species 5 is stable in solution, its analogue 6 is instable in CH2Cl2 solution at room temperature to slowly decompose to the corresponding homoleptic species 8via the transient formation of (κ(2)-P,O)2Zn2(μ-OBn)(μ–κ(1):κ(1)-P,O) (6′). Crystallization of compound 6 led to crystals of 6′, as established by XRD analysis. The reaction of ZnEt2 with two equiv. of 1·H and 2·H allowed access to the corresponding homoleptic species of the type [Zn(P,O)2] (7 and 8). All gathered data are consistent with compound 7 being a dinuclear species in the solid state and in solution. Data for species 8, which bears a sterically demanding P,O-ligand, are consistent with a mononuclear species in solution. The Zn(II) alkoxide species 5 and the [Zn(P,O)2]-type compounds 7 and 8 were evaluated as initiators of the ring-opening polymerization (ROP) of lactide (LA), ε-caprolactone (ε-CL) and trimethylene carbonate (TMC). Species 5 is a well-behaved ROP initiator for the homo-, co- and ter-polymerization of all three monomers with the production of narrow disperse materials under living and immortal conditions. Though species 7 and 8 are ROP inactive on their own, they readily polymerize LA in the presence of a nucleophile such as BnOH to produce narrow disperse PLA, presumably via an activated-monomer ROP mechanism.


Effectiveness of non-surgical interventions for rotator cuff calcific tendinopathy: A systematic review.
To evaluate the effectiveness of non-surgical interventions for rotator cuff calcific tendinopathy. Medline, EMBASE, CINAHL, Cochrane Register of Clinical Trials, PEDro and SPORTDiscus from inception to March 2018, and accompanying reference lists. Peer-reviewed randomized clinical trials of non-surgical interventions for adults with rotator cuff calcific tendinopathy were included. The same 2 reviewers independently evaluated eligibility, extracted data and evaluated risk of bias of the included randomized clinical trials. A system to resolve any disagreements was established a priori. Short-term, medium-term and long-term outcomes for pain, shoulder function and calcific morphology related to rotator cuff calcific tendinopathy were extracted. Due to diversity in outcome measures a meta-analyses was not conducted. Of the 2,085 articles identified, 18 met the inclusion criteria, all of which had high risk of bias. Five non-surgical interventions were identified (extracorporeal shockwave therapy, ultrasound-guided percutaneous intervention, pulsed ultrasound, acetic acid iontophoresis, and transcutaneous electrical nerve stimulation). There was moderate evidence for high-energy extracorporeal shockwave therapy over low-energy extracorporeal shockwave therapy for pain and function between 3 and 6 months, and over placebo for function in the first 6 months. There was moderate evidence for ultrasound-guided percutaneous intervention over medium/high-energy extracorporeal shockwave therapy for reduced pain and calcific morphology over a 1-year period. Methodological concerns preclude definitive recommendations.


A randomised controlled trial of cognitive-behaviour therapy for behavioural insomnia of childhood in school-aged children.
Chronic sleep problems can lead to the development of Behavioural Insomnia of Childhood - a sleep disorder involving problematic sleep-onset associations (i.e., parental presence), and resulting in impairments for children and family members. The aim of the present paper was to perform a controlled evaluation of cognitive-behaviour therapy (CBT) for Behavioural Insomnia. 42 children (M = 9.3 ± 1.9 yrs, range 7-13 yrs, 18f, 24 m) were randomised to CBT (N = 21) or waitlist control (N = 21). CBT consisted of 6 sessions, and combined behavioural sleep medicine techniques (e.g., sleep restriction) with anxiety treatment techniques (e.g., cognitive restructuring). Compared to waitlist controls, children receiving CBT showed significant improvements in sleep latency, wake after sleep onset, and sleep efficiency (all p ≤ .003), but not total sleep time (p > .05). CBT was also associated with a reduction in problematic sleep associations (p ≤ .001), child-reported total and separation anxiety (both p ≤ .01), with all gains being maintained 6 months post-treatment. This is the first controlled study to demonstrate that multi-component CBT can be effective for the sleep, insomnia, and anxiety symptoms of Behavioural Insomnia of Childhood in school-aged children. Future research is needed to ascertain active treatment components.


Cord blood perfluoroalkyl substances in mothers exposed to the World Trade Center disaster during pregnancy.
Perfluoroalkyl substances (PFAS) may have been released during the collapse of the World Trade Center (WTC) on 9/11. Evidence suggests PFAS can cross the placental barrier in humans and cause harm to the developing fetus; however, no studies have measured PFAS in mothers exposed to the WTC disaster during pregnancy. We measured PFAS in maternal plasma (n = 48) or cord blood (n = 231) from pregnant women in the Columbia University WTC birth cohort, enrolled between December 13, 2001 and June 26, 2002 at one of three hospitals located near the WTC site. In order to maximize sample size, we used a linear regression to transform the 48 maternal plasma samples to cord blood equivalents in our study; cord blood and transformed maternal plasma-to-cord blood samples were then analyzed together. We evaluated the association between WTC exposure and PFAS concentrations using three exposure variables: 1) living/working within two miles of WTC; 2) living within two miles of WTC regardless of work location; and 3) working but not living within two miles of WTC. Exposure was compared with those not living/working within two miles of WTC (reference group). Living/working within two miles of WTC was associated with 13% higher perfluorooctanoic acid (PFOA) concentrations compared with the reference group [GMR (95% CI): 1.13 (1.01, 1.27)]. The association was stronger when comparing only those who lived within two miles of WTC to the reference group [GMR (95% CI): 1.17 (1.03, 1.33)], regardless of work location. Our results provide evidence that exposure to the WTC disaster during pregnancy resulted in increases in PFAS concentrations, specifically PFOA. This work identifies a potentially vulnerable and overlooked population, children exposed to the WTC disaster in utero, and highlights the importance of future longitudinal studies in this cohort to investigate later life effects resulting from these early life exposures.


Expansion of CD4+ cytotoxic T lymphocytes with specific gene expression patterns may contribute to suppression of tumor immunity in oral squamous cell carcinoma: single-cell analysis and in vitro experiments.
Cancer immunotherapy targeting CD8+ T cells has made remarkable progress, even for oral squamous cell carcinoma (OSCC), a heterogeneous epithelial tumor without a substantial increase in the overall survival rate over the past decade. However, the therapeutic effects remain limited due to therapy resistance. Thus, a more comprehensive understanding of the roles of CD4+ T cells and B cells is crucial for more robust development of cancer immunotherapy. In this study, we examined immune responses and effector functions of CD4+ T cells, CD8+ T cells and B cells infiltrating in OSCC lesions using single-cell RNA sequencing analysis, T cell receptor (TCR) and B cell receptor (BCR) repertoire sequencing analysis, and multi-color immunofluorescence staining. Finally, two Kaplan-Meier curves and several Cox proportional hazards models were constructed for the survival analysis. We observed expansion of CD4+ cytotoxic T lymphocytes (CTLs) expressing granzymes, which are reported to induce cell apoptosis, with a unique gene expression patterns. CD4+ CTLs also expressed CXCL13, which is a B cell chemoattractant. Cell-cell communication analysis and multi-color immunofluorescence staining demonstrated potential interactions between CD4+ CTLs and B cells, particularly IgD- CD27- double negative (DN) B cells. Expansion of CD4+ CTLs, DN B cells, and their contacts has been reported in T and B cell-activated diseases, including IgG4-related disease and COVID-19. Notably, we observed upregulation of several inhibitory receptor genes including CTLA-4 in CD4+ CTLs, which possibly dampened T and B cell activity. We next demonstrated comprehensive delineation of the potential for CD8+ T cell differentiation towards dysfunctional states. Furthermore, prognostic analysis revealed unfavorable outcomes of patients with a high proportion of CD4+ CTLs in OSCC lesions. Our study provides a dynamic landscape of lymphocytes and demonstrates a systemic investigation of CD4+ CTL effects infiltrating into OSCC lesions, which may share some pathogenesis reported in severe T and B cell-activated diseases such as autoimmune and infectious diseases.


Mechanism of phosphorylation of protein kinase B/Akt by a constitutively active 3-phosphoinositide-dependent protein kinase-1.
Phosphorylation of Thr(308) in the activation loop and Ser(473) at the carboxyl terminus is essential for protein kinase B (PKB/Akt) activation. However, the biochemical mechanism of the phosphorylation remains to be characterized. Here we show that expression of a constitutively active mutant of mouse 3-phosphoinositide-dependent protein kinase-1 (PDK1(A280V)) in Chinese hamster ovary cells overexpressing the insulin receptor was sufficient to induce PKB phosphorylation at Thr(308) to approximately the same extent as insulin stimulation. Phosphorylation of PKB by PDK1(A280V) was not affected by treatment of cells with inhibitors of phosphatidylinositol 3-kinase or by deletion of the pleckstrin homology (PH) domain of PKB. C(2)-ceramide, a cell-permeable, indirect inhibitor of PKB phosphorylation, did not inhibit PDK1(A280V)-catalyzed PKB phosphorylation in cells and had no effect on PDK1 activity in vitro. On the other hand, co-expression of full-length protein kinase C-related kinase-1 (PRK1/PKN) or 2 (PRK2) inhibited PDK1(A280V)-mediated PKB phosphorylation. Replacing alanine at position 280 with valine or deletion of the PH domain enhanced PDK1 autophosphorylation in vitro. However, deletion of the PH domain of PDK1(A280V) significantly reduced PDK1(A280V)-mediated phosphorylation of PKB in cells. In resting cells, PDK1(A280V) localized in the cytosol and at the plasma membrane. However, PDK1(A280V) lacking the PH domain localized predominantly in the cytosol. Taken together, our findings suggest that the wild-type PDK1 may not be constitutively active in cells. In addition, activation of PDK1 is sufficient to phosphorylate PKB at Thr(308) in the cytosol. Furthermore, the PH domain of PDK1 may play both positive and negative roles in regulating the in vivo function of the enzyme. Finally, unlike the carboxyl-terminal fragment of PRK2, which has been shown to bind PDK1 and allow the enzyme to phosphorylate PKB at both Thr(308) and Ser(473), full-length PRK2 and its related kinase PRK1/PKN may both play negative roles in PKB-mediated downstream biological events.


Connexins in Cardiovascular and Neurovascular Health and Disease: Pharmacological Implications.
Connexins are ubiquitous channel forming proteins that assemble as plasma membrane hemichannels and as intercellular gap junction channels that directly connect cells. In the heart, gap junction channels electrically connect myocytes and specialized conductive tissues to coordinate the atrial and ventricular contraction/relaxation cycles and pump function. In blood vessels, these channels facilitate long-distance endothelial cell communication, synchronize smooth muscle cell contraction, and support endothelial-smooth muscle cell communication. In the central nervous system they form cellular syncytia and coordinate neural function. Gap junction channels are normally open and hemichannels are normally closed, but pathologic conditions may restrict gap junction communication and promote hemichannel opening, thereby disturbing a delicate cellular communication balance. Until recently, most connexin-targeting agents exhibited little specificity and several off-target effects. Recent work with peptide-based approaches has demonstrated improved specificity and opened avenues for a more rational approach toward independently modulating the function of gap junctions and hemichannels. We here review the role of connexins and their channels in cardiovascular and neurovascular health and disease, focusing on crucial regulatory aspects and identification of potential targets to modify their function. We conclude that peptide-based investigations have raised several new opportunities for interfering with connexins and their channels that may soon allow preservation of gap junction communication, inhibition of hemichannel opening, and mitigation of inflammatory signaling.


Small molecule binders recognize DNA microstructural variations via an induced fit mechanism.
Regulation of gene-expression by specific targeting of protein-nucleic acid interactions has been a long-standing goal in medicinal chemistry. Transcription factors are considered "undruggable" because they lack binding sites well suited for binding small-molecules. In order to overcome this obstacle, we are interested in designing small molecules that bind to the corresponding promoter sequences and either prevent or modulate transcription factor association via an allosteric mechanism. To achieve this, we must design small molecules that are both sequence-specific and able to target G/C base pair sites. A thorough understanding of the relationship between binding affinity and the structural aspects of the small molecule-DNA complex would greatly aid in rational design of such compounds. Here we present a comprehensive analysis of sequence-specific DNA association of a synthetic minor groove binder using long timescale molecular dynamics. We show how binding selectivity arises from a combination of structural factors. Our results provide a framework for the rational design and optimization of synthetic small molecules in order to improve site-specific targeting of DNA for therapeutic uses in the design of selective DNA binders targeting transcription regulation.


An effective intervention to reduce intravaginal practices among HIV-1 uninfected Kenyan women.
Intravaginal practices (IVP) are common among African women and are associated with HIV acquisition. A behavioral intervention to reduce IVP is a potential new HIV risk-reduction strategy. Fifty-eight HIV-1-uninfected Kenyan women reporting IVP and 42 women who denied IVP were followed for 3 months. Women using IVP attended a skill-building, theory-based group intervention occurring weekly for 3 weeks to encourage IVP cessation. Vaginal swabs at each visit were used to detect yeast, to detect bacterial vaginosis, and to characterize the vaginal microbiota. Intravaginal insertion of soapy water (59%) and lemon juice (45%) was most common among 58 IVP women. The group-counseling intervention led to a decrease in IVP from 95% (54/58) at baseline to 0% (0/39) at month 3 (p=0.001). After 3 months of cessation, there was a reduction in yeast on vaginal wet preparation (22% to 7%, p=0.011). Women in the IVP group were more likely to have a Lactobacillus iners-dominated vaginal microbiota at baseline compared to controls [odds ratio (OR), 6.4, p=0.006] without significant change in the microbiota after IVP cessation. The group counseling intervention was effective in reducing IVP for 3 months. Reducing IVP may be important in itself, as well as to support effective use of vaginal microbicides, to prevent HIV acquisition.


Schiff bases formed from retinal and phosphatidylethanolamine, phosphatidylserine, ethanolamine or serine.
1. Conditions were established for the reaction of retinal with phosphatidylethanolamine, phosphatidylserine, ethanolamine and serine in chloroform, ethanol or ethanol-water solutions to form retinylidene compounds, or Schiff bases. 2. The Schiff bases were reduced to retinyl compounds with sodium borohydride. 3. Absorption maxima and molar extinction coefficients were determined for the various retinylidene and retinyl compounds and for the corresponding coloured products formed by their reaction with antimony trichloride.


Epidemiology of osteoarthritis.
Osteoarthritis (OA) is the most common joint disorder in the United States. Symptomatic knee OA occurs in 10% men and 13% in women aged 60 years or older. The number of people affected with symptomatic OA is likely to increase due to the aging of the population and the obesity epidemic. OA has a multifactorial etiology, and can be considered the product of an interplay between systemic and local factors. Old age, female gender, overweight and obesity, knee injury, repetitive use of joints, bone density, muscle weakness, and joint laxity all play roles in the development of joint OA, particularly in the weight-bearing joints. Modifying these factors may reduce the risk of OA and prevent subsequent pain and disability.


Percutaneous tracheostomy, a systematic review.
Percutaneous dilatational tracheostomy (PDT) is a common procedure in intensive care units and the identification of the best technique is very important. We performed a systematic review and meta-analysis of randomized studies comparing different PDT techniques in critically ill adult patients to investigate if one technique is superior to the others with regard to major and minor intraprocedural complications. BioMedCentral and other database of clinical trials were searched for pertinent studies. Inclusion criterion was random allocation to at least two PDT techniques. Exclusion criteria were duplicate publications, nonadult studies, and absence of outcome data. Population, clinical setting, and complications were extracted. Data from 1130 patients in 13 randomized trials were analyzed. Multiple dilators, single-step dilatation, guide wire dilating forceps, rotational dilation, retrograde tracheostomy, and balloon dilation techniques were always performed in the intensive care unit. The different techniques and devices appeared largely equivalent, with the exception of retrograde tracheostomy, which was associated with more severe complications and more frequent need of conversion to other techniques when compared with guide wire dilating forceps and single-step dilatation techniques. Single-step dilatation technique was associated with fewer failures than rotational dilation, and fewer mild complications in comparison with balloon dilation and guide wire dilating forceps (all P < 0.05). Among the six analyzed techniques, single-step dilatation technique appeared the most reliable in terms of safety and success rate. However, the number of available randomized trials was insufficient to confidently assess the best PDT technique.


Development and validation of a multiplex RT-PCR method for the simultaneous detection of five grapevine viroids.
Grapevine yellow speckle viroid 1 (GYSVd-1), Grapevine yellow speckle viroid 2 (GYSVd-2), Australian grapevine viroid (AGVd), Hop stunt viroid (HSVd) and Citrus exocortis viroid (CEVd) are the five viroids known to infect naturally grapevines. We developed a multiplex RT-PCR (mRT-PCR) method for the simultaneous detection of these five viroids and the amplification of the cDNA fragment of a host-derived mRNA (actin mRNA) as an internal positive control. Specific primers for each targeted viroid were designed by taking into account the sequence variability within and between the viroid species and tested in silico. The method was validated by testing 57 grapevine samples from Iran and showed reliability and high sensitivity. The RT-PCR-negative samples were further assayed by Northern-blot hybridization. For this, a method was developed for the simultaneous detection of three different grapevine viroids on a single hybridization membrane. In this survey, HSVd, GYSVd-1, AGVd, and GYSVd-2 were detected in 100, 95, 93, and 65% of the samples tested, respectively, confirming the wide distribution of these viroids in Iran. CEVd was not detected in any of the samples collected. Based on these results, HSVd is proposed as a positive internal control for mRT-PCR in the areas where this viroid is widespread, so as to reduce the time and costs of DNase treatment, which is required when a host-derived internal control is used. The mRT-PCR method has the potential to be used routinely for large-scale surveys and certification programs.


Early outcome after paclitaxel-eluting stents in patients with acute and subacute myocardial infarction. A clinical study.
Paclitaxel-eluting stents (PES) have been proven to prevent in-stent restenosis in patients submitted to elective percutaneous coronary intervention. No data are so far available about the safety and efficacy of PES in acute and subacute myocardial infarction. The aim of the present investigation was to evaluate the occurrence of in-hospital adverse events in patients with acute and subacute myocardial infarction submitted to PES implantation. From June 1 to July 31, 2003, we implanted 53 PES in 43 consecutive patients with acute (34 patients) and subacute (9 patients) myocardial infarction. In 65.1% of the patients the culprit lesion was located in the left anterior descending artery. Direct stenting was performed in 27.9% and glycoprotein IIb/IIIa inhibitors were used in 74.4%. Before the procedure a TIMI flow 0-1 was present in 46.5% while post-procedural TIMI flow 3 was achieved in all patients. A pre-procedural TIMI thrombus grade 2 to 5 was present in 67.8%. No death, reinfarction, early post-infarction angina or any other episode referable to in-stent thrombosis were observed during hospitalization. No patient was submitted to target lesion revascularization. At short-term follow-up with a mean duration of 118 +/- 75 days from discharge no cardiac or noncardiac death, reinfarction or any other major adverse coronary events were reported in the study population and no target lesion revascularization was performed. In this study, PES implant in patients with acute and subacute myocardial infarction was safe, with an early outcome comparable to conventional standard stents and no adverse events related to acute or subacute thrombosis.


Incidence and Predictors of Hypertension Among HIV Patients Receiving ART at Public Health Facilities, Northwest Ethiopia: A One-Year Multicenter Prospective Follow-Up Study.
The introduction of highly active retroviral therapy has dramatically reduced mortality and improved survival among HIV patients. However, there is a possible risk of comorbid complications such as hypertension. Little evidence is available regarding the incidence of hypertension among HIV patients receiving anti-retroviral therapy in Ethiopia. To assess the incidence and predictors of hypertension among HIV positive patients receiving ART at Public Health Facilities, Northwest Ethiopia. A one-year prospective follow-up study was conducted among a cohort of 302 new adult individuals initiating on a standard anti-retroviral therapy regimen with a median (IQR) age of 35 years (IQR=30-41). A pretested data extraction checklist was used to extract baseline patient records. The collected data were entered into Epi-Data version 3.1 and exported to STATA version 14 for analysis. The incidence rate was calculated, and a Kaplan-Meier survival curve was used to estimate the survival probabilities of developing hypertension. Cox proportional hazards model was fitted to identify the predictors of hypertension. About 40 (13.25) new hypertensive cases were observed during the follow-up period, and the remaining 262 (86.75%) were censored. The overall incidence rate of hypertension was 16.35 per 1000 person-month with 2447 patient-month observations. Male sex (AHR = 2.45, 95% CI: 1.02, 6.14), old age (AHR = 2.83, 95% CI: 1.08, 7.45), high BMI (AHR = 6.54, 95% CI: 2.03, 21.13), diabetic comorbidity (AHR = 2.36, 95% CI: 1.07, 5.22), and patients who were on Zidovudine (AZT)-based ART regimen (AHR =3.47, 95% CI: 1.10, 10.94) were significant predictors for the development of hypertension. The findings of this study revealed that incident hypertension is a common problem among HIV patients receiving ART. Routine monitoring of blood pressure and screening and treating high blood pressure should be an integral part of follow-up for HIV patients in ART clinics.


N-methyl-norsalsolinol, a putative dopaminergic neurotoxin, passes through the blood-brain barrier in vivo.
In earlier studies the dihydroxylated tetrahydroisoquinoline derivatives salsolinol and 2(N)-methyl-norsalsolinol (NMNorsal), a 2(N)-analogue of salsolinol, were identified as putative endogenous neurotoxins in patients with Parkinson's disease. Since a prominent blood-brain barrier (BBB) was described to exist for salsolinol, in the present study microdialysis experiments were performed to investigate the penetration of NMNorsal through the BBB into the caudate nucleus of the rat brain. After i.p. administration of NMNorsal (20 mg/kg), it could be detected in the dialysate of the caudate nucleus with a mean maximum after 40 min. There was no alteration in extracellular dopamine or 3,4-dihydroxyphenylacetic acid levels. Addition of the monoamine oxidase inhibitor pargyline (10 microM) to the perfusate did not modify NMNorsal levels in the caudate nucleus. To corroborate the microdialysis results, homogenates of the contralateral caudate nucleus were prepared and NMNorsal could also be detected. These findings indicate that NMNorsal is indeed able to pass through the blood-brain barrier of the rat brain.


in silico screening and molecular dynamics simulations study to identify novel potent inhibitors against Mycobacterium tuberculosis DnaG primase.
Tuberculosis remains a major global health threat killing millions of people. Due to existing multiple drug resistance (MDR) and prolonged treatment it becomes necessary to explore novel drug targets in Mycobacterium tuberculosis (Mtb). DnaG primase, having a significant role in primer synthesis during initiation of DNA replication, has emerged as a promising drug target. The three dimensional (3D) model of its catalytic domain (Toprim) was constructed. Further, in silico screening of the three diverse chemical compound libraries against the modeled domain was carried out. Four top screened compounds were identified and evaluated by ADMET analysis. The stability of these compounds in complex with the Toprim domain was validated through 50 ns molecular dynamics simulations. Lys 101, Glu 137 and Asp 188 in the active site predominantly formed the hydrogen bonds with the top screened compounds. Hence, the drug-like compounds identified can be taken up for the further experimental investigation as anti-tubercular agents.


Balloon aortic valvuloplasty: the Texas Heart Institute experience.
Between October 1986 and January 1989, 73 percutaneous catheter balloon aortic valvuloplasty procedures were performed in 68 adult patients (32 men and 36 women; mean age, 77 +/- 9 years) with severe symptomatic aortic stenosis. Following the procedures, significant improvements were documented in aortic valve area, mean transvalvular pressure gradient, peak-to-peak pressure gradient, left ventricular systolic pressure, radionuclide ejection fraction, and left ventricular end-systolic volume index. There were no procedure-related deaths, but 2 patients (3%) required emergency surgery for acute aortic regurgitation. During hospitalization, 4 patients had persistent symptoms (3 died; 1 subsequently underwent repeat valvuloplasty and later, valve replacement). Short-term clinical improvement was noted in 59 of 65 patients (91%). During the follow-up period (mean, 11.6 +/- 8.4 months), 22 patients died (including the 3 who died during hospitalization). Sixteen underwent aortic valve replacement (including the 2 who underwent emergency aortic valve replacement); all 16 are alive. A total of 6 patients (1 with an initial balloon aortic valvuloplasty at an outside institution) underwent repeat valvuloplasty; of those, 4 subsequently underwent aortic valve replacement, and 2 died. Of the remaining 30 patients, 27 continue to experience relief of symptoms, and 3 have clinical symptoms that have not improved or have worsened since the valvuloplasty procedure. Multivariate predictors of clinical outcome (p<0.05) included post-valvuloplasty aortic valve area, pre- and post-valvuloplasty ejection fraction, absence of coronary artery disease, and absolute change in valve area. Overall actuarial and event-free survivals were 83% and 49%, respectively, at 1 year. Although clinical improvement is frequently noted after balloon aortic valvuloplasty, the procedure is associated with a high recurrence of symptoms and restenosis. Balloon aortic valvuloplasty is at best a palliative procedure; when feasible, surgical valve replacement is the more definitive therapy.


Residual Symptoms After Treatment for Depression in Patients With Coronary Heart Disease.
Depression is associated with an increased risk of mortality in patients with coronary heart disease (CHD). The risk may be reduced in patients who remit with adequate treatment, but few patients achieve complete remission. The purpose of this study was to identify the symptoms that persist despite aggressive treatment for depression in patients with CHD. One hundred twenty-five patients with stable CHD who met the DSM-IV criteria for a moderate-to-severe major depressive episode completed treatment with cognitive behavior therapy, either alone or combined with an antidepressant, for up to 16 weeks. Depression symptoms were assessed at baseline and after 16 weeks of treatment. The M (SD) Beck Depression Inventory scores were 30.0 (8.6) at baseline and 8.3 (7.5) at 16 weeks. Seventy seven (61%) of the participants who completed treatment met remission criteria (Hamilton Rating Scale for Depression ≤7) at 16 weeks. Loss of energy and fatigue were the most common posttreatment symptoms both in remitters (n = 44, 57%; n = 34, 44.2%) and nonremitters (n = 42, 87.5%; n = 35, 72.9%). These symptoms were not predicted by baseline depression severity, anxiety, demographic, or medical variables including inflammatory markers or cardiac functioning or by medical events during depression treatment. Fatigue and loss of energy often persist in patients with CHD even after otherwise successful treatment for major depression. These residual symptoms may increase the risks of relapse and mortality. Development of effective interventions for these persistent symptoms is a priority for future research.


Rubidium uptake and accumulation in peripheral myelinated internodal axons and Schwann cells.
To study mechanisms of K+ transport in peripheral nerve, uptake of rubidium (Rb+), a K+ tracer, was characterized in rat tibial nerve myelinated axons and glia. Isolated nerve segments were perfused with zero-K+ Ringer's solutions containing Rb+ (1-20 mM) and x-ray microanalysis was used to measure water content and concentrations of Rb, Na, K, and Cl in internodal axoplasm, mitochondria, and Schwann cell cytoplasm and myelin. Both axons and Schwann cells were capable of removing extracellular Rb+ (Rb+(o)) and exchanging it for internal K+. Uptake into axoplasm, Schwann cytoplasm, and myelin was a saturable process over the 1-10 mM Rb+(o) concentration range, although corresponding axoplasmic uptake rates were higher than respective glial velocities. Mitochondrial accumulation was a linear function of axoplasmic Rb+ concentrations, which suggests involvement of a nonenzymatic process. At 20 mM Rb+(o), a differential stimulatory response was observed; i.e., axoplasmic Rb+ uptake velocities increased more than fivefold relative to the 10 mM rate, and glial cytoplasmic uptake rose almost threefold. Finally, Rb+(o) uptake rate into axons and glia was completely inhibited by ouabain (2-4 mM) exposure or incubation at 4 degrees C. These results suggest that Rb+ uptake into peripheral nerve internodal axons and Schwann cells is mediated by Na+,K+-ATPase activity and implicate the presence of axonal- and glial-specific Na+ pump isozymes.


The adoption of children from public care: a prospective study of outcome in adolescence.
To discover the outcomes for children placed late for adoption (between 5 and 11 years old) from public care and to establish which factors predict poorer outcome. Data were collected prospectively (1993-2003) from a representative sample of domestic U.K. adoptive placements (N = 108) at the start of placement, at 1 year, and 6 years later. Most of the children entered care because of abuse and neglect. Outcome was assessed by the disruption rate, by a twofold classification of the character of continuing placements, and by an assessment of psychological well-being. At the adolescent follow-up, 23% of placements had disrupted, 49% were continuing positively, and 28% were continuing but with substantial ongoing difficulties. Four factors contributed independently to a higher risk of disruption: older age at placement (odds ratio = 1.07), having been singled out from siblings and rejected (5.87), time in care (1.04), and a high level of behavioral problems (1.25). Two factors predicted differences in continuing placements. Late adoption can be successful in that half the children made good progress, but the extent of disruptions and difficulties in continuing placements gives rise to concern. Knowledge of predictors will help in devising planning pre- and postplacement support services.


[Regulatory properties of phosphorylase from chicken skeletal muscle].
The main kinetic parameters for purified phosphorylase kinase from chicken skeletal muscle were determined at pH 8.2: Vm = 18 micromol/min/mg; apparent Km values for ATP and phosphorylase b from rabbit muscle were 0.20 and 0.02 mM, respectively. The activity ratio at pH 6.8/8.2 was 0.1-0.4 for different preparations of phosphorylase kinase. Similar to the rabbit enzyme, chicken phosphorylase kinase had an absolute requirement for Ca2+ as demonstrated by complete inhibition in the presence of EGTA. Half-maximal activation occurred at [Ca2+] = 0.4 microM at pH 7.0. In the presence of Ca2+, the chicken enzyme from white and red muscles was activated 2-4-fold by saturating concentrations of calmodulin and troponin C. The C0.5 value for calmodulin and troponin C at pH 6.8 was 2 and 100 nM, respectively. Similar to rabbit phosphorylase kinase, the chicken enzyme was stimulated about 3-6-fold by glycogen at pH 6.8 and 8.2 with half-maximal stimulation occurring at about 0.15% glycogen. Protamine caused 60% inhibition of chicken phosphorylase kinase at 0.8 mg/ml. ADP (3 mM) at 0.05 mM ATP caused 85% inhibition with Ki = 0.2 mM. Unlike rabbit phosphorylase kinase, no phosphorylation of the chicken enzyme occurred in the presence of the catalytic subunit of cAMP-dependent protein kinase. Incubation with trypsin caused 2-fold activation of the chicken enzyme.