Rho-kinase inactivation prolongs survival of an intermediate SMA mouse model.
Spinal muscular atrophy (SMA) is an inherited disease resulting in the highest mortality of children under the age of two. SMA is caused by mutations or deletions in the survival motor neuron 1 (SMN1) gene, leading to aberrant neuromuscular junction (NMJ) development and the loss of spinal cord alpha-motor neurons. Here, we show that Smn depletion leads to increased activation of RhoA, a major regulator of actin dynamics, in the spinal cord of an intermediate SMA mouse model. Treating these mice with Y-27632, which inhibits ROCK, a direct downstream effector of RhoA, dramatically improves their survival. This lifespan rescue is independent of Smn expression and is accompanied by an improvement in the maturation of the NMJs and an increase in muscle fiber size in the SMA mice. Our study presents evidence linking disruption of actin cytoskeletal dynamics to SMA pathogenesis and, for the first time, identifies RhoA effectors as viable targets for therapeutic intervention in the disease.


Sculpting Dendritic Spines during Initiation and Maintenance of Neuropathic Pain.
Accumulating evidence has established a firm role for synaptic plasticity in the pathogenesis of neuropathic pain. Recent advances have highlighted the importance of dendritic spine remodeling in driving synaptic plasticity within the CNS. Identifying the molecular players underlying neuropathic pain induced structural and functional maladaptation is therefore critical to understanding its pathophysiology. This process of dynamic reorganization happens in unique phases that have diverse pathologic underpinnings in the initiation and maintenance of neuropathic pain. Recent evidence suggests that pharmacological targeting of specific proteins during distinct phases of neuropathic pain development produces enhanced antinociception. These findings outline a potential new paradigm for targeted treatment and the development of novel therapies for neuropathic pain. We present a concise review of the role of dendritic spines in neuropathic pain and outline the potential for modulation of spine dynamics by targeting two proteins, srGAP3 and Rac1, critically involved in the regulation of the actin cytoskeleton.


Risk of complications in patients who are obese following upper limb arthroplasty: A systematic review and meta-analysis.
A systematic review was performed to investigate the impact of obesity on complications following total shoulder arthroplasty (TSA), reverse total shoulder arthroplasty (RTSA) and total elbow arthroplasty (TEA). Electronic databases and grey literature were searched for studies that evaluated the influence of obesity (Body Mass Index[BMI] ≥30kgm2) on upper limb arthroplasty outcomes. Fifteen studies were identified, however only twelve reported predetermined outcomes. Unadjusted data was pooled in statistical meta-analysis where appropriate. Effect sizes were expressed as odds ratios (OR) for categorical data and weighted mean differences for continuous data. Odds of infection increased with increasing BMI, from 2.37 (95%CI [1.653.41]) times in patients who were obese, to greater than five times (OR=5.04; 95%CI [4.705.39]) in patients who were morbidly obese. Furthermore, patients who were obese or morbidly obese had 3.92 (95%CI [3.594.28]) to 5.46 (95%CI [4.916.07]) times greater odds of venous thromboembolism (VTE) compared to their non-obese counterparts, respectively. Conversely, obesity had no influence on the odds of urinary tract infection (OR=0.88; 95%CI [0.481.61], or mortality (OR=1.79; 95%CI [0.794.03]). TSA/RTSA patients who were obese experienced operations 10.00minutes longer (95%CI [6.3113.69]) than patients with a BMI in the normal range, which increased to 12.48min utes (95%CI [8.4016.55]) in patients with a BMI≥35.0. Evidence examining the influence of obesity on blood transfusion was inconclusive, while minimal evidence was available on pneumonia. Surgeons should consider advising patients who are obese of the greater risk of VTE and infection when considering elective upper limb arthroplasty. However, noteworthy limitations surrounded the lack of information regarding prophylaxis regimes and BMI measurement tools used in included studies.


The importance of soil drying and re-wetting in crop phytohormonal and nutritional responses to deficit irrigation.
Soil drying and re-wetting (DRW) occurs at varying frequencies and intensities during crop production, and is deliberately used in water-saving irrigation techniques that aim to enhance crop water use efficiency. Soil drying not only limits root water uptake which can (but not always) perturb shoot water status, but also alters root synthesis of phytohormones and their transport to shoots to regulate leaf growth and gas exchange. Re-wetting the soil rapidly restores leaf water potential and leaf growth (minutes to hours), but gas exchange recovers more slowly (hours to days), probably mediated by sustained changes in root to shoot phytohormonal signalling. Partial rootzone drying (PRD) deliberately irrigates only part of the rootzone, while the remainder is allowed to dry. Alternating these wet and dry zones (thus re-wetting dry soil) substantially improves crop yields compared with maintaining fixed wet and dry zones or conventional deficit irrigation, and modifies phytohormonal (especially abscisic acid) signalling. Alternate wetting and drying (AWD) of rice can also improve yield compared with paddy culture, and is correlated with altered phytohormonal (including cytokinin) signalling. Both PRD and AWD can improve crop nutrition, and re-wetting dry soil provokes both physical and biological changes which affect soil nutrient availability. Whether this alters crop nutrient uptake depends on competition between plant and microbes for nutrients, with the rate of re-wetting determining microbial dynamics. Nevertheless, studies that examine the effects of soil DRW on both crop nutritional and phytohormonal responses are relatively rare; thus, determining the cause(s) of enhanced crop yields under AWD and PRD remains challenging.


The art of bedside rounds: a multi-center qualitative study of strategies used by experienced bedside teachers.
Medical educators believe bedside rounds are effective for the delivery of patient-centered care, and are necessary in helping trainees acquire competence in clinical care. Although recommendations for bedside rounds have been reported, a recent, systematic assessment of strategies used by current-day bedside teachers was needed to advance knowledge of this teaching method. To identify and understand bedside teachers' 1) preparatory steps, 2) patient selection, and 3) role allocation during the process of bedside rounds. A qualitative inductive thematic analysis using transcripts from audio-recorded, semi-structured telephone interviews. Internal medicine physicians (n = 34) who perform bedside rounds from ten academic US institutions (2010-2011). A purposive sampling strategy was utilized to identify physicians who were active inpatient attending physicians and met specific inclusion criteria for "bedside rounds." A total of 34 interviews were completed, and each was recorded and transcribed verbatim. A thematic analysis extracted key themes pertaining to the three objectives. Most respondents (51 %) were associate or full professors, with an average of 14 years of academic experience. Attending physicians prepared using trainee-specific, patient-specific and disease-specific information, while also mentally preparing for bedside rounds. They sought trainee buy-in and learning objectives, reviewed expectations and methods to ensure patient comfort, and provided early guidance with bedside encounters. Patients were selected if they required immediate care, were new to the service, or had a high educational value, while patients were deferred if unavailable, unwilling, or unable to communicate. The team members' roles during bedside rounds varied, with trainees being given graduated autonomy with increased experience. Bedside teachers' methods for preparation, patient selection, and role allocation during bedside rounds enhance trainees' education within the workplace. Strategies used by experienced bedside teachers can be used for faculty development efforts aimed at promoting this activity.


Pre- and postnatal psychological wellbeing in Africa: a systematic review.
Perinatal mental health disorders are recognised as an important public health issue in low-income countries as well as in developed countries. This paper reviews evidence on the prevalence and risk factors of maternal mental health disorders in African women living in Africa. A systematic review of the literature was conducted. Studies were mainly located through computerised databases, and additionally through hand searching references of identified articles and reviews. Thirty-five studies, with a total of 10,880 participants, were identified that reported prevalence rates of maternal psychological health in eight African countries. Depression was the most commonly assessed disorder with a weighted mean prevalence of 11.3% (95% CI 9.5%-13.1%) during pregnancy and 18.3% (95% CI 17.6%-19.1%) after birth. Only a small number of studies assessed other psychological disorders. Prevalence rates of pre- and postnatal anxiety were 14.8% (95% CI 12.3%-17.4%) and 14.0% (95% CI 12.9%-15.2%), respectively; and one study reported the prevalence of PTSD as 5.9% (95% CI 4.4%-7.4%) following childbirth. Lack of support and marital/family conflict were associated with poorer mental health. Evidence relating sociodemographic and obstetric variables to mental health was inconclusive. Most studies included in this review were cross-sectional and measures of mental health varied considerably. This paper demonstrates that maternal mental health disorders are prevalent in African women, and highlights the importance of maternal mental health care being integrated into future maternal and infant health policies in African countries.


Evaluations of training and education interventions for improved infectious disease management in low-income and middle-income countries: a systematic literature review.
To identify most vital input and outcome parameters required for evaluations of training and education interventions aimed at addressing infectious diseases in low-income and middle-income countries. Systematic review. PubMed/Medline, Web of Science and Scopus were searched for eligible studies between January 2000 and November 2021. Health economic and health-outcome studies on infectious diseases covering an education or training intervention in low-income and middle-income countries were included. A total of 59 eligible studies covering training or education interventions for infectious diseases were found; infectious diseases were categorised as acute febrile infections (AFI), non-AFI and other non-acute infections. With regard to input parameters, the costs (direct and indirect) were most often reported. As outcome parameters, five categories were most often reported including final health outcomes, intermediate health outcomes, cost outcomes, prescription outcomes and health economic outcomes. Studies showed a wide range of per category variables included and a general lack of uniformity across studies. Further standardisation is needed on the relevant input and outcome parameters in this field. A more standardised approach would improve generalisability and comparability of results and allow policy-makers to make better informed decisions on the most effective and cost-effective interventions.


Mechanical loading thresholds for lamellar and woven bone formation.
Bone formation was measured in rat tibiae after 12 days of applied loading. Bending forces were applied using a four-point loading apparatus. Sham loads were applied at the same magnitudes as bending forces but the loading pads were arranged so that bending was minimized. Bending and sham loading were applied to the right tibiae of rats and the left tibiae served as contralateral controls. Loading was applied as a sine wave with a frequency of 2 Hz for 18 s (36 cycles) per day. The peak magnitude of applied load was 27, 33, 40, 52, and 64 N. Woven bone was observed on the periosteal surface in all animals subjected to loads of 40 N or greater. Periosteal woven bone formation occurred in both bending and sham loading groups. Woven bone formation on the periosteal surface was either absent or responded at a maximal rate if the stimulus threshold was surpassed. The amount of new woven bone and the woven bone-forming surface were independent of the magnitude of applied strain. Bone formation on the endocortical surface was exclusively lamellar. Lamellar bone formation was stimulated by applied bending of the tibia but not by sham loading. Bending strains above a loading threshold of 40 N or about 1050 mu strain increased both bone-forming surface and the mineral apposition rate and subsequently increased the bone formation rate as much as sixfold. No evidence of increased bone formation was seen for applied strains below 1050 mu strain. Examination of bulk stained sections from animals exposed to the highest applied loads showed no evidence of microcracks.(ABSTRACT TRUNCATED AT 250 WORDS)


Intracameral bevacizumab (Avastin) for neovascular glaucoma: a pilot study in 6 patients.
To describe the use of intracameral bevacizumab (ICB) Avastin in neovascular glaucoma (NVG) as the first maneuver before pan retinal photocoagulation and/or filtering surgery. Between June 2006 and May 2007, 6 consecutive patients with NVG underwent intracameral injection of bevacizumab (1.25 mg/0.05 mL) as the initial treatment of NVG. Pre-ICB and post-ICB anterior segment photography, iris fluoresceingraphy when possible, gonioscopy with peripheral anterior synechiae (PAS), neovascular membrane (NVM) extension grading, as well as intraocular pressure (IOP) changes during treatment were recorded. All patients were followed for at least 7 months. ICB resulted in a marked regression of anterior segment neovascularization with IOP control without filtering surgery in 2 cases. When PAS extended <330 degrees without previous glaucoma, no filtering surgery was needed to control IOP<18 mm Hg. Iris neovascularization extension had no prognostic value in terms of IOP control. After vascular regression following the administration of ICB, filtering surgery with drainage implants or trabeculectomy were performed when needed with no added difficulties owing to the underlying NVG. No macroscopic signs of corneal toxicity were detected, even when ICB injection had to be repeated. In this case, the time elapsed for the neovascular membrane to reappear at the anterior segment was 3 months. ICB resulted in a rapid regression of the iris and angle neovascularization, which permitted to halt the progression of PAS process. This pilot study shows that intracameral injection of bevacizumab may be a helpful adjunct for the surgical treatment of NVG.


Quantitative linkage for autism spectrum disorders symptoms in attention-deficit/hyperactivity disorder: significant locus on chromosome 7q11.
We studied 261 ADHD probands and 354 of their siblings to assess quantitative trait loci associated with autism spectrum disorder symptoms (as measured by the Children's Social Behavior Questionnaire (CSBQ)) using a genome-wide linkage approach, followed by locus-wide association analysis. A genome-wide significant locus for the CSBQ subscale addressing social interaction was found on chromosome 7q11, with suggestive signals supporting this locus on three other CSBQ subscales. We identified two other suggestive loci for the CSBQ total scale and individual subscales on chromosomes 4q35 and 7p12. Fine-mapping the significantly linked locus resulted in interesting candidate genes, although their association was not significant after permutation testing.


Patterning of globus pallidus local field potentials differs between Parkinson's disease and dystonia.
Here we test the hypothesis that there are distinct temporal patterns of synchronized neuronal activity in the pallidum that characterize untreated and treated parkinsonism and dystonia. To this end we recorded local field potentials (LFPs) from the caudal and rostral contact pairs of macroelectrodes implanted into the pallidum of patients for the treatment of Parkinson's disease (12 cases recorded on and off medication, 17 macroelectrodes) and dystonia (10 cases, 19 macroelectrodes). Percentage LFP power in the 11-30 Hz band was decreased and that in the 4-10 Hz band increased across both contact pairs in treated Parkinson's disease compared with untreated Parkinson's disease. Dystonic patients had even less 11-30 Hz power and greater 4-10 Hz power compared with untreated or treated Parkinson's disease patients. The change in the 4-10 Hz band in patients with dystonia was particularly manifest in the more rostral contact pair, presumed to be within or bridging the globus pallidus externus. We conclude that untreated and treated Parkinson's disease and dystonia are characterized by different spatiotemporal patterns of activity in the human pallidum.


Electronic remote blood issue: a combination of remote blood issue with a system for end-to-end electronic control of transfusion to provide a "total solution" for a safe and timely hospital blood transfusion service.
The rapid provision of red cell (RBC) units to patients needing blood urgently is an issue of major importance in transfusion medicine. The development of electronic issue (sometimes termed "electronic crossmatch") has facilitated rapid provision of RBC units by avoidance of the serologic crossmatch in eligible patients. A further development is the issue of blood under electronic control at blood refrigerator remote from the blood bank. This study evaluated a system for electronic remote blood issue (ERBI) developed as an enhancement of a system for end-to-end electronic control of hospital transfusion. Practice was evaluated before and after its introduction in cardiac surgery. Before the implementation of ERBI, the median time to deliver urgently required RBC units to the patient was 24 minutes. After its implementation, RBC units were obtained from the nearby blood refrigerator in a median time of 59 seconds (range, 30 sec to 2 min). The study also found that unused requests were reduced significantly from 42 to 20 percent, the number of RBC units issued reduced by 52 percent, the number of issued units that were transfused increased from 40 to 62 percent, and there was a significant reduction in the workload of both blood bank and clinical staff. This study evaluated a combination of remote blood issue with an end-to-end electronically controlled hospital transfusion process, ERBI. ERBI reduced the time to make blood available for surgical patients and improved the efficiency of hospital transfusion.


Leptin-STAT3-G9a Signaling Promotes Obesity-Mediated Breast Cancer Progression.
Obesity has been linked to breast cancer progression but the underlying mechanisms remain obscure. Here we report how leptin, an obesity-associated adipokine, regulates a transcriptional pathway to silence a genetic program of epithelial homeostasis in breast cancer stem-like cells (CSC) that promotes malignant progression. Using genome-wide ChIP-seq and RNA expression profiling, we defined a role for activated STAT3 and G9a histone methyltransferase in epigenetic silencing of miR-200c, which promotes the formation of breast CSCs defined by elevated cell surface levels of the leptin receptor (OBR(hi)). Inhibiting the STAT3/G9a pathway restored expression of miR-200c, which in turn reversed the CSC phenotype to a more differentiated epithelial phenotype. In a rat model of breast cancer driven by diet-induced obesity, STAT3 blockade suppressed the CSC-like OBR(hi) population and abrogated tumor progression. Together, our results show how targeting STAT3-G9a signaling regulates CSC plasticity during obesity-related breast cancer progression, suggesting a novel therapeutic paradigm to suppress CSC pools and limit breast malignancy.


Supporting the diffusion of healthy public policy in Canada: the Prevention Policies Directory.
Healthy public policy plays an essential role in a comprehensive public health approach to preventing cancer and chronic disease. Public policies spread through the 'policy diffusion' process, enabling governments to learn from another's enacted policy solutions. The Prevention Policies Directory (the Directory), an online database of municipal, provincial/territorial, and federal cancer and chronic disease prevention policies from across Canada, was developed to facilitate the diffusion of healthy public policies and support the work of prevention researchers, practitioners, and policy specialists. This information technology solution was implemented, through a participatory engagement approach, as a communication channel or policy knowledge transfer tool. It also addressed the intrinsic shortcomings of environmental scanning for policy surveillance and monitoring. A combination of quantitative web metrics and qualitative anecdotal evidence have illustrated that the Directory is becoming an important tool for healthy public policy surveillance and policy diffusion in Canada.


The Cappadocia mesothelioma epidemic: its influence in Turkey and abroad.
The epidemic of mesothelioma in Cappadocia, Turkey, is unprecedented in medical history. In three Cappadocian villages, Karain, Tuzkoy and "old" Sarihidir, about 50% of all deaths (including neonatal deaths and traffic fatalities) have been caused by mesothelioma. No other epidemic in medical history has caused such a high incidence of death. This is even more unusual when considering that (I) epidemics are caused by infectious agents, not cancer, and (II) mesothelioma is a rare cancer. World-wide mesothelioma incidence varies between 1/106 in areas with no asbestos industry to about 10-30/106 in areas with asbestos industry. This article reviews how the mesothelioma epidemic was discovered in Cappadocia by Dr. Baris (my mentor), how we initially linked the epidemic to erionite exposure, and later (with Dr. Carbone) to the interaction between genetic predisposition and environmental exposure. Our team's work had an important positive impact on the lives of those living in Cappadocia and also in many genetically predisposed families living around the world. I will discuss how the work that started in three remote Cappadocian villages led to the award of a NCI P01 grant to support our studies. Our studies proved that genetics modulates mineral fiber carcinogenesis and led to the discovery that carriers of germline BAP1 mutations have a very high risk of developing mesothelioma and other malignancies. A new, very active field of research developed following our discoveries to elucidate the mechanism by which BAP1 modulates mineral fiber carcinogenesis as well as to identify additional genes that when mutated increase the risk of mesothelioma and other environmentally related cancers. I am the only surviving member of this research team who saw all the phases of this research and I believe it is important to provide an accurate report, which hopefully will inspire others.


Iron-induced Lipid Peroxidation in Human Skin-derived Cell Lines: Protection by a Red Orange Extract.
Although photodamage and photoprotection have already been extensively studied in cultured cells, few data have been reported in the literature regarding the in vitro behaviour of skin cells toward a chemical stress, such as iron-induced lipid peroxidation. We investigated the susceptibilities of two human skin-derived cell lines (NCTC 2544 keratinocytes and HFFF2 fibroblasts) to lipid peroxidation induced by FeSO4/histidine, FeSO4/ascorbate and Fe2(SO4)3/ADP. NCTC 2544 cells were more susceptible than HFFF2 cells to lipid peroxidation (assessed by measuring the content of malondialdehyde [MDA]) with iron/ascorbate and iron/ADP as pro-oxidants whereas, with iron/histidine, the same level of MDA production was achieved (about 10nmol/mg protein) in the two cell populations. On the basis of these results, one experimental model ( (iron/histidine) was selected to assess the protective effect of a mixture of two classical antioxidants, Trolox C™ (50μM) and Vitamin C (1mM), added to the cell cultures according to various protocols. The maximal decrease of MDA production in both cell lines was obtained when the antioxidant mixture and the pro-oxidant were added simultaneously to the cultures. By using the same experimental design, NCTC 2544 and HFFF2 cells were exposed to a standardised extract of red oranges (ROE; 0.025-0.5mg/ml), the main active principles of which (anthocyanins 3.1%, hydroxycinnamic acid 2.07%, and flavanone glycosides 8.1%) possess antioxidant activity. Cells treated with ROE, that were still over 90% viable, as evaluated by means of neutral red uptake and tetrazolium salt reduction tests, showed a significant and dose-dependent inhibition of MDA production. This study provides new information about the behaviour of cultured skin cells exposed to chemically induced oxidative stress, and provides further support to the possibility of using skin-derived human cell lines in the evaluation of the effectiveness of antioxidant ingredients for new drugs and/or cosmetics.


A Restricted Repertoire of De Novo Mutations in ITPR1 Cause Gillespie Syndrome with Evidence for Dominant-Negative Effect.
Gillespie syndrome (GS) is characterized by bilateral iris hypoplasia, congenital hypotonia, non-progressive ataxia, and progressive cerebellar atrophy. Trio-based exome sequencing identified de novo mutations in ITPR1 in three unrelated individuals with GS recruited to the Deciphering Developmental Disorders study. Whole-exome or targeted sequence analysis identified plausible disease-causing ITPR1 mutations in 10/10 additional GS-affected individuals. These ultra-rare protein-altering variants affected only three residues in ITPR1: Glu2094 missense (one de novo, one co-segregating), Gly2539 missense (five de novo, one inheritance uncertain), and Lys2596 in-frame deletion (four de novo). No clinical or radiological differences were evident between individuals with different mutations. ITPR1 encodes an inositol 1,4,5-triphosphate-responsive calcium channel. The homo-tetrameric structure has been solved by cryoelectron microscopy. Using estimations of the degree of structural change induced by known recessive- and dominant-negative mutations in other disease-associated multimeric channels, we developed a generalizable computational approach to indicate the likely mutational mechanism. This analysis supports a dominant-negative mechanism for GS variants in ITPR1. In GS-derived lymphoblastoid cell lines (LCLs), the proportion of ITPR1-positive cells using immunofluorescence was significantly higher in mutant than control LCLs, consistent with an abnormality of nuclear calcium signaling feedback control. Super-resolution imaging supports the existence of an ITPR1-lined nucleoplasmic reticulum. Mice with Itpr1 heterozygous null mutations showed no major iris defects. Purkinje cells of the cerebellum appear to be the most sensitive to impaired ITPR1 function in humans. Iris hypoplasia is likely to result from either complete loss of ITPR1 activity or structure-specific disruption of multimeric interactions.


Human cortical excitability increases with time awake.
Prolonged wakefulness is associated not only with obvious changes in the way we feel and perform but also with well-known clinical effects, such as increased susceptibility to seizures, to hallucinations, and relief of depressive symptoms. These clinical effects suggest that prolonged wakefulness may be associated with significant changes in the state of cortical circuits. While recent animal experiments have reported a progressive increase of cortical excitability with time awake, no conclusive evidence could be gathered in humans. In this study, we combine transcranial magnetic stimulation (TMS) and electroencephalography (EEG) to monitor cortical excitability in healthy individuals as a function of time awake. We observed that the excitability of the human frontal cortex, measured as the immediate (0-20 ms) EEG reaction to TMS, progressively increases with time awake, from morning to evening and after one night of total sleep deprivation, and that it decreases after recovery sleep. By continuously monitoring vigilance, we also found that this modulation in cortical responsiveness is tonic and not attributable to transient fluctuations of the level of arousal. The present results provide noninvasive electrophysiological evidence that wakefulness is associated with a steady increase in the excitability of human cortical circuits that is rebalanced during sleep.


Validation of the Abbreviated Socio-Political Control Scale for Youth (SPCS-Y) Among Urban Girls of Color.
This study tested and validated the factor structure of the abbreviated Sociopolitical Control Scale for Youth (SPCS-Y) among a sample of urban girls of color. Participants include (N = 830) urban girls of color from a northeastern United States community. Confirmatory Factor analyses (CFA) were conducted using AMOS Structural Equation Modeling Software. Cluster groups were created using Latent Class Cluster Analysis (LCA) and tested using Multivariate analysis of covariance (MANCOVA) with conceptually related variables. CFA results supported the two-factor structure of the abbreviated SPCS-Y among the sample. Following the creation of cluster groups, MANCOVA analyses revealed significant heterogeneity among cluster groups of participants on neighborhood sense of community, social support, ethnic identity, and lower levels of drug use. Findings support the factor structure of the SPCS-Y and its use to measure empowerment among girls of color. Results contribute significantly to the field of social work and encourages the importance of promoting strengths-based approaches among girls of color. Implications for social work practice with girls of color are conveyed in light of the need for reliable measurement tools for practitioners.


The expression and distribution of Wnt and Wnt receptor mRNAs during early sea urchin development.
The protein beta-catenin plays a critically important role in establishing axial polarity during early animal development. In many organisms, beta-catenin is degraded preferentially on one side of the cleavage stage embryo. On the opposite side of the embryo, beta-catenin is stabilized and accumulates in the nucleus, where it functions in concert with members of the LEF/TCF family to activate the transcription of diverse target genes. Genes that are activated by beta-catenin play an essential role in the specification of endomesoderm and in the establishment of key signaling centers in the early embryo. In several organisms, the asymmetric distribution of maternal components of the canonical Wnt pathway has been shown to be responsible for the polarized stabilization of beta-catenin. In this study, we identified all Wnt and Wnt receptor mRNAs that are present in unfertilized sea urchin eggs and early embryos and analyzed their distributions along the primary (AV) axis. Our findings indicate that the asymmetric distribution of a maternal Wnt or Wnt receptor mRNA is unlikely to be a primary determinant of the polarized stabilization of beta-catenin along the AV axis. This contrasts sharply with findings in other organisms and points to remarkable evolutionary flexibility in the molecular mechanisms that underlie this otherwise very highly conserved patterning process.


Recycling attitudes and behavior among a clinic-based sample of low-income Hispanic women in southeast Texas.
We examined attitudes and behavior surrounding voluntary recycling in a population of low-income Hispanic women. Participants (N = 1,512) 18-55 years of age completed a self-report survey and responded to questions regarding household recycling behavior, recycling knowledge, recycling beliefs, potential barriers to recycling (transportation mode, time), acculturation, demographic characteristics (age, income, employment, marital status, education, number of children, birth country), and social desirability. Forty-six percent of participants (n = 810) indicated that they or someone else in their household recycled. In a logistic regression model controlling for social desirability, recycling behavior was related to increased age (P<0.05), lower acculturation (P<0.01), knowing what to recycle (P<0.01), knowing that recycling saves landfill space (P<0.05), and disagreeing that recycling takes too much time (P<0.001). A Sobel test revealed that acculturation mediated the relationship between recycling knowledge and recycling behavior (P<0.05). We offer new information on recycling behavior among Hispanic women and highlight the need for educational outreach and intervention strategies to increase recycling behavior within this understudied population.


Effect of uvulopalatopharyngoplasty on retropalatal region.
Flexible optic laryngoscopy (FOL) allows us to visualize the obstructive sleep apnea (OSA)-related airway passages. Retropalatal region is a part of upper airway contributing to the OSA. We aimed to demonstrate the changes in the retropalatal surface area (RPSA) after submucosal uvulopalatopharyngoplasty (smUPPP) in an attempt to exhibit enlargement as a predictor of surgical treatment. It is a prospective, case-control study and conducted at the Otolaryngology department in a university hospital. Twenty patients with OSA who underwent smUPPP were prospectively evaluated. Pre and postoperative respiratory disturbance index (RDI) and RPSA measurements were studied. Retropalatal region videos were obtained at the base of uvula during FOL. Photographic images were captured at the same level of a virtual horizontal line passing through the base of uvula posteriorly to measure RPSAs using AutoCad2004. RDI levels, RPSA measurements were compared using paired t test. Twenty patients underwent smUPPP. There were 17 (85 %) male and 3 (15 %) female. The mean age was 37 years. The RPSA measurements were between 18.41 and 144.102 (mean 63.39) preoperatively. The RPSA measurements were between 83.784 and 255.463 (mean 143.87) postoperatively. The RPSA measurements were significantly enlarged postoperatively (p < 0.0005). The mean RPSA increased from 63.39 ± 29.3 to 143.82 ± 57.8. The mean RDI decreased from 22.95 ± 19.2 to 9.0 ± 8.2 (p = 0.011). RPSA increases in smUPPP patients postoperatively, evidencing enlarged retropalatal region. Decreased RDI levels indicate amelioration in OSA. RPSA measurements can be used to predict UPPP surgical treatment success.


Associations between Yogurt Consumption and Weight Gain and Risk of Obesity and Metabolic Syndrome: A Systematic Review.
The role of yogurt consumption in the risk of developing overweight, obesity, or metabolic syndrome has been the subject of epidemiologic studies over the last 10 y. A comprehensive literature search on MEDLINE and ISI Web of Knowledge from 1966 through June 2016 was conducted to examine the relation between yogurt consumption and weight gain, as well as the risk of overweight, obesity, or metabolic syndrome, in prospective cohort studies. Ten articles met all the inclusion criteria and were included in our systematic review. Of the 10 cohort studies, 3 analyzed the relation between yogurt consumption and the risk of overweight or obesity, 8 analyzed changes in waist circumference or weight changes, 3 studied the association with the risk of developing metabolic syndrome, and 1 studied the probability of abdominal obesity reversion. Although an inverse association between yogurt consumption and the risk of developing overweight or obesity was not fully consistent or always statistically significant, all studies but one showed in their point estimates inverse associations between yogurt consumption and changes in waist circumference, changes in weight, risk of overweight or obesity, and risk of metabolic syndrome during follow-up, although not all estimates were statistically significant (2 studies). Prospective cohort studies consistently suggested that yogurt consumption may contribute to a reduction in adiposity indexes and the risk of metabolic syndrome. Therefore, there is a need for more prospective studies and high-quality randomized clinical trials to confirm this apparent inverse association.


B-50, the growth associated protein-43: modulation of cell morphology and communication in the nervous system.
The growth-associated protein B-50 (GAP-43) is a presynaptic protein. Its expression is largely restricted to the nervous system. B-50 is frequently used as a marker for sprouting, because it is located in growth cones, maximally expressed during nervous system development and re-induced in injured and regenerating neural tissues. The B-50 gene is highly conserved during evolution. The B-50 gene contains two promoters and three exons which specify functional domains of the protein. The first exon encoding the 1-10 sequence, harbors the palmitoylation site for attachment to the axolemma and the minimal domain for interaction with G0 protein. The second exon contains the "GAP module", including the calmodulin binding and the protein kinase C phosphorylation domain which is shared by the family of IQ proteins. Downstream sequences of the second and non-coding sequences in the third exon encode species variability. The third exon also contains a conserved domain for phosphorylation by casein kinase II. Functional interference experiments using antisense oligonucleotides or antibodies, have shown inhibition of neurite outgrowth and neurotransmitter release. Overexpression of B-50 in cells or transgenic mice results in excessive sprouting. The various interactions, specified by the structural domains, are thought to underlie the role of B-50 in synaptic plasticity, participating in membrane extension during neuritogenesis, in neurotransmitter release and long-term potentiation. Apparently, B-50 null-mutant mice do not display gross phenotypic changes of the nervous system, although the B-50 deletion affects neuronal pathfinding and reduces postnatal survival. The experimental evidence suggests that neuronal morphology and communication are critically modulated by, but not absolutely dependent on, (enhanced) B-50 presence.


Kinetic analysis of nitroxide radical formation under oxygenated photolysis: toward quantitative singlet oxygen topology.
Reaction kinetics for two sterically hindered secondary amines with singlet oxygen have been studied in detail. A water soluble porphyrin sensitizer, 5,10,15,20-tetrakis-(4-sulfunatophenyl)-21,23H-porphyrin (TPPS), was irradiated in oxygenated aqueous solutions containing either 2,2,6,6-tetramethylpiperidin-4-one (TMPD) or 4-[N,N,N-trimethyl-ammonium]-2,2,6,6-tetramethylpiperidinyl chloride (N-TMPCl). The resulting sensitization reaction produced singlet oxygen in high yield, ultimately leading to the formation of the corresponding nitroxide free radicals (R2NO) which were detected using steady-state electron paramagnetic resonance (EPR) spectroscopy. Careful actinometry and EPR calibration curves, coupled with a detailed kinetic analysis, led to a simple and compact expression relating the nitroxide quantum yield ΦR2NO (from the doubly-integrated EPR signal intensity) to the initial amine concentration [R2NH]i. With all other parameters held constant, a plot of ΦR2NOvs. [R2NH]i gave a straight line with a slope proportional to the rate constant for nitroxide formation, kR2NO. This establishment of a rigorous quantitative relationship between the EPR signal and the rate constant provides a mechanism for quantifying singlet oxygen production as a function of its topology in heterogeneous media. Implications for in vivo assessment of singlet oxygen topology are briefly discussed.


Delivery of large genomic DNA inserts >100 kb using HSV-1 amplicons.
The principal aim of gene therapy for recessive genetic diseases is to supplement the loss of function of an endogenous gene. For the treatment of many diseases regulation of transgene expression at physiological levels, expression of multiple splice variants, and correct tissue specificity are of utmost importance for effective therapy. We therefore believe the use of a complete genomic locus, in which the native promoter and regulatory regions drive and control expression, is an elegant and effective alternative to traditional complementary DNA (cDNA) vectors utilising heterologous promoters. Viral vectors have proved, over the years, to be an effective means of gene delivery in vitro and in vivo, but the size of complete genomic loci precludes their use in most viral systems. One notable exception comprises the amplicon-type vectors based on human herpesviruses, such as the herpes simplex virus type I (HSV-1) amplicon vector. The large genome of HSV-1 (152 kb) confers upon HSV-1 amplicons a very large transgene capacity sufficient to accommodate approximately 95% of human genomic loci. The combination of the large transgene capacity, a broad range of cell tropism, and the ability to infect dividing and non-dividing cells makes HSV-1 amplicons an excellent vector system to develop for the delivery of large genomic loci. Here we review recent work which has shown that HSV-1 amplicons can be used for the delivery and expression of large genomic inserts >100 kb to cells in culture to rescue phenotypes in cellular models of genetic disease. We then discuss applications for high capacity HSV-1 amplicons in vivo and their potential to facilitate the use of large genomic inserts in gene therapy treatment regimes.


Developing a smoke free household initiative: an Indonesian case study.
Little research has focused on women's exposure to secondhand smoke (SHS) in LMICs, local perceptions of SHS risk to women and children, and women's attempts to limit exposure to tobacco smoke in their households. This paper describes a community based survey in Indonesia that investigated these issues as one step in a movement to initiate community wide household smoking bans. The survey found high levels of exposure to SHS, high levels of awareness among both women and men that SHS placed women and children at risk for illness, a very low percentage of households having indoor smoking rules, great interest on the part of women to participate in a communitywide ban, and a promising level of male smoker agreement to comply with such a ban. Women expressed a low sense of self efficacy in individually getting their husbands to quit smoking in their homes, but a strong sense of collective efficacy that husbands might agree to a well-publicized and agreed-upon community household smoking ban. Men and women expressed concern about the social risk of asking guests not to smoke in their homes without a communitywide ban and visible displays communicating their participation in this movement. The smoke free initiative described requires the participation of doctors in community education programs, and is attempting to introduce household smoking bans as a way of turning tobacco control into a family health and not just a smokers' health issue.


Bordetella parapertussis and Bordetella bronchiseptica contain transcriptionally silent pertussis toxin genes.
Pertussis toxin, the major virulence factor of Bordetella pertussis, is not produced by the closely related species Bordetella parapertussis and Bordetella bronchiseptica. It is shown here that these two species possess but do not express the complete toxin operon. Nucleotide sequencing of an EcoRI fragment of 5 kilobases comprising the regions homologous to the pertussis toxin genes shows that in this region, B. parapertussis and B. bronchiseptica are 98.5% and 96% homologous, respectively, to B. pertussis. The changes (mostly base pair substitutions) in many cases are identical in B. parapertussis and B. bronchiseptica, suggesting that these two species derive from a common ancestor. Many of the mutations common to B. parapertussis and B. bronchiseptica involve the promoter region, which becomes very inefficient. The S1 subunits of both species, when expressed in Escherichia coli, have the same ADP-ribosylating activity as the S1 subunit from B. pertussis, indicating that the mutations in the S1 gene described here do not affect its function.


The application of CRISPR technology to high content screening in primary neurons.
Axon growth is coordinated by multiple interacting proteins that remain incompletely characterized. High content screening (HCS), in which manipulation of candidate genes is combined with rapid image analysis of phenotypic effects, has emerged as a powerful technique to identify key regulators of axon outgrowth. Here we explore the utility of a genome editing approach referred to as CRISPR (Clustered Regularly Interspersed Palindromic Repeats) for knockout screening in primary neurons. In the CRISPR approach a DNA-cleaving Cas enzyme is guided to genomic target sequences by user-created guide RNA (sgRNA), where it initiates a double-stranded break that ultimately results in frameshift mutation and loss of protein production. Using electroporation of plasmid DNA that co-expresses Cas9 enzyme and sgRNA, we first verified the ability of CRISPR targeting to achieve protein-level knockdown in cultured postnatal cortical neurons. Targeted proteins included NeuN (RbFox3) and PTEN, a well-studied regulator of axon growth. Effective knockdown lagged at least four days behind transfection, but targeted proteins were eventually undetectable by immunohistochemistry in >80% of transfected cells. Consistent with this, anti-PTEN sgRNA produced no changes in neurite outgrowth when assessed three days post-transfection. When week-long cultures were replated, however, PTEN knockdown consistently increased neurite lengths. These CRISPR-mediated PTEN effects were achieved using multi-well transfection and automated phenotypic analysis, indicating the suitability of PTEN as a positive control for future CRISPR-based screening efforts. Combined, these data establish an example of CRISPR-mediated protein knockdown in primary cortical neurons and its compatibility with HCS workflows.


The Primary Care Spend Model: a systems approach to measuring investment in primary care.
Increased investment in primary care is associated with lower healthcare costs and improved population health. The allocation of scarce resources should be driven by robust models that adequately describe primary care activities and spending within a health system, and allow comparisons within and across health systems. However, disparate definitions result in wide variations in estimates of spending on primary care. We propose a new model that allows for a dynamic assessment of primary care spending (PC Spend) within the context of a system's total healthcare budget. The model articulates varied definitions of primary care through a tiered structure which includes overall spending on primary care services, spending on services delivered by primary care professionals and spending delivered by providers that can be characterised by the '4Cs' (first contact, continuous, comprehensive and coordinated care). This unifying framework allows a more refined description of services to be included in any estimate of primary care spend and also supports measurement of primary care spending across nations of varying economic development, accommodating data limitations and international health system differences. It provides a goal for best accounting while also offering guidance, comparability and assessments of how primary care expenditures are associated with outcomes. Such a framework facilitates comparison through the creation of standard definitions and terms, and it also has the potential to foster new areas of research that facilitate robust policy analysis at the national and international levels.


30-Day readmission after bariatric surgery in a publicly funded regionalized center of excellence system.
Avoidable readmission after surgery is a major burden on healthcare resources and is common after major surgery. Bariatric surgery is one of the most common surgical procedures in North America, and there is a paucity of strategies to prevent readmission. Strategies for prevention must first identify actual risk factors before interventions can be designed. Our objective was to evaluate the readmission rate, characteristics of readmitted patients, and factors associated with readmission. We performed a population-based cohort study that included all patients who received a Roux-en-Y gastric bypass (RYGB) or sleeve gastrectomy (SG) procedure in Ontario from April 2009 until March 2012 for the purposes of weight loss. Data were derived from the Canadian Institute for Health Information Discharge Abstract Database and Hospital Morbidity Database. Over 3 years, 5007 procedures (91.7 % RYGB, 8.1 % SG) were performed with an overall 30-day readmission rate of 6.1 %. Readmission stays of 72 h or less accounted for 83 % of the cohort. The most common reasons for readmission were: infectious complications (24.6 %), pain (16.4 %) nausea/vomiting (11.5 %), bleeding complications (11.5 %), obstruction (5.6 %). A complication during initial admission OR 2.07 (95 % CI 1.44-2.97; P value < 0.001) and a length of stay greater than 2 days OR 1.40 (95 % CI 1.07-1.84; P value = 0.013) were independent predictors of readmission within 30 days. The readmission rate after bariatric surgery in Ontario is similar to other major population-based bariatric surgery programs. Complications on initial admission and prolonged length of stay were independent predictors of readmission. Considering a large proportion of the readmissions were short term, future research into potential measures to prevent these readmissions is essential.


Altered expression of tyrosine kinases of the Src and Syk families in human T-cell leukemia virus type 1-infected T-cell lines.
During the late phase of adult T-cell leukemia/lymphoma, a severe lymphoproliferative disorder caused by human T-cell leukemia virus type 1 (HTLV-1), leukemic cells no longer produce interleukin-2. Several studies have reported the lack of the Src-like protein tyrosine kinase Lck and overexpression of Lyn and Fyn in these cells. In this report we demonstrate that, in addition to the downregulation of TCR, CD45, and Lck (which are key components of T-cell activation), HTLV-1-infected cell lines demonstrate a large increase of FynB, a Fyn isoform usually poorly expressed in T cells. Furthermore, similar to anergic T cells, Fyn is hyperactive in one of these HTLV-1-infected T-cell lines, probably as a consequence of Csk downregulation. A second family of two proteins, Zap-70 and Syk, relay the signal of T-cell activation. We demonstrate that in contrast to uninfected T cells, Zap-70 is absent in HTLV-1-infected T cells, whereas Syk is overexpressed. In searching for the mechanism responsible for FynB overexpression and Zap-70 downregulation, we have investigated the ability of the Tax and Rex proteins to modulate Zap-70 expression and the alternative splicing mechanism which gives rise to either FynB or FynT. By using Jurkat T cells stably transfected with the tax and rex genes or inducibly expressing the tax gene, we found that the expression of Rex was necessary to increase fynB expression, suggesting that Rex controls fyn gene splicing. Conversely, with the same Jurkat clones, we found that the expression of Tax but not Rex could downregulate Zap-70 expression. These results suggest that the effect of Tax and Rex must cooperate to deregulate the pathway of T-cell activation in HTLV-1-infected T cells.


The continuity and quality of primary care.
Patients who have access to different health care systems, such as Medicare-eligible veterans, may obtain services in either or both health systems. We examined whether quality of diabetes care was associated with care continuity or veterans' usual source of primary care in a retrospective cohort study of 1,867 Medicare-eligible veterans with diabetes in 2001 to 2004. Underprovision of quality of diabetes care was more common than overprovision. In adjusted analyses, veterans who relied only on Medicare fee-for-service (FFS) for primary care were more likely to be underprovided HbA1c testing than veterans who relied only on Veteran Affairs (VA) for primary care. Dual users of VA and Medicare FFS primary care were significantly more likely to be overprovided HbA1c and microalbumin testing than VA-only users. VA and Medicare providers may need to coordinate more effectively to ensure appropriate diabetes care to Medicare-eligible veterans, because VA reliance was a stronger predictor than care continuity.


Stakeholder engagement for comparative effectiveness research in cancer care: experience of the DEcIDE Cancer Consortium.
Stakeholder input is a critical component of comparative effectiveness research. To ensure that the research activities of the Developing Evidence to Inform Decisions about Effectiveness (DEcIDE) Network, supported by the Agency for Healthcare Research and Quality, translate into the greatest impact for everyday practice and policy-making in cancer, we were tasked with soliciting stakeholder input regarding priority areas in cancer-related comparative effectiveness research for the DEcIDE Cancer Consortium. Given the increasing emphasis on stakeholder engagement in research, many investigators are facing a similar task, yet there is limited literature to guide such efforts, particularly in cancer care. To help fill this gap, we present our approach to operationalizing stakeholder engagement and discuss it in the context of other recent developments in the area. We describe challenges encountered in convening stakeholders from multiple vantage points to prioritize topics and strategies used to mitigate these barriers. We offer several recommendations regarding how to best solicit stakeholder input to inform comparative effectiveness research in cancer care. These recommendations can inform other initiatives currently facing the challenges of engaging stakeholders in priority setting for cancer.


Switch from sexual to parthenogenetic reproduction in a zebra shark.
Parthenogenesis is a natural form of asexual reproduction in which embryos develop in the absence of fertilisation. Most commonly found in plants and invertebrate organisms, an increasing number of vertebrate species have recently been reported employing this reproductive strategy. Here we use DNA genotyping to report the first demonstration of an intra-individual switch from sexual to parthenogenetic reproduction in a shark species, the zebra shark Stegostoma fasciatum. A co-housed, sexually produced daughter zebra shark also commenced parthenogenetic reproduction at the onset of maturity without any prior mating. The demonstration of parthenogenesis in these two conspecific individuals with different sexual histories provides further support that elasmobranch fishes may flexibly adapt their reproductive strategy to environmental circumstances.


Whole-abdominal radiation in endometrial carcinoma: an analysis of toxicity, patterns of recurrence, and survival.
The purpose of this study was to determine the toxicity, patterns of recurrence, and survival in high-risk endometrial cancer patients treated with whole-abdominal radiation. All patients with endometrial cancer treated at Northwestern University since 1994 and at Rush University since 1993 were retrospectively reviewed. Patients believed to be at high risk for intra-abdominal recurrence and who received whole-abdominal radiation were reviewed for this study. A total of 30 patients completed whole-abdominal radiation (WAR) and were available for study. The mean and median follow-up was 2.3 and 2.1 years, respectively, with a range of 0.13 to 6.1 years. Seventy-eight percent of the cohort received surgical staging with bilateral salpingo-oophorectomy/total abdominal hysterectomy/lymph node sampling. Forty-seven percent of the patients were found to have serous histology as a component of their tumor. Surgical staging results included 19% stage 1B, 4% stage IC, 8% stage IIB, 37% stage IIIA, 26% stage IIIC, and 7% stage IVB. Two patients had gross residual disease at the completion of surgery. Megestrol acetate (Megace) was used as an adjuvant treatment in 37% of patients, and no cases received initial chemotherapy. All patients received WAR with a mean total dose and dose per fraction of 2620 and 143 cGy, respectively. Twenty-two percent of patients received a para-aortic boost. The mean total pelvic dose was 4956 cGy. Seventy percent of patients received a vaginal cuff boost. Eight percent of patients had grade 3 acute gastrointestinal morbidity, and 4% had grade 4 acute gastrointestinal morbidity. No other grade 3 or greater acute or long-term morbidity was noted. At last follow-up, seven (23%) patients had experienced recurrence. The pattern of first recurrence was 0% in the vaginal cuff, 3% other vaginal, 7% pelvic, 7% upper abdominal, 3% lung, 7% bone, and 7% para-aortic lymph nodes. Ultimate recurrences were similar. At last follow-up, 77% patients had no evidence of disease, 13% were alive with disease, and 10% had died of disease. Utilizing a conservatrive total whole-abdominal radiation dose and limited para-aortic nodal boost resulted in very tolerable treatments. The patterns of recurrence and survival in this early report are encouraging.


Discriminative Brain Effective Connectivity Analysis for Alzheimer's Disease: A Kernel Learning Approach upon Sparse Gaussian Bayesian Network.
Analyzing brain network from neuroimages is becoming a promising approach in identifying novel connectivity-based biomarkers for the Alzheimer's disease (AD). In this regard, brain "effective connectivity" analysis, which studies the causal relationship among brain regions, is highly challenging and of many research opportunities. Most of the existing works in this field use generative methods. Despite their success in data representation and other important merits, generative methods are not necessarily discriminative, which may cause the ignorance of subtle but critical disease-induced changes. In this paper, we propose a learning-based approach that integrates the benefits of generative and discriminative methods to recover effective connectivity. In particular, we employ Fisher kernel to bridge the generative models of sparse Bayesian network (SBN) and the discriminative classifiers of SVMs, and convert the SBN parameter learning to Fisher kernel learning via minimizing a generalization error bound of SVMs. Our method is able to simultaneously boost the discrimination power of both the generative SBN models and the SBN-induced SVM classifiers via Fisher kernel. The proposed method is tested on analyzing brain effective connectivity for AD from ADNI data. It demonstrates significant improvements over the state-of-the-art: classification accuracy increased above 10% by our SBN models, and above 16% by our SBN-induced SVM classifiers with a simple feature selection.


Localization of Apaf1 gene expression in the early development of the mouse by means of in situ reverse transcriptase-polymerase chain reaction.
Apoptosis is an essential ubiquitous process that controls the duration of the life span of cells, thus playing a crucial role in morphogenetic, histogenetic, and phylogenetic developmental processes. Apaf1 (apoptosis protease activating factor 1) is one of the central mediators of the intrinsic apoptotic pathway and a part of the apoptosome, which activates procaspase-3 and promotes cell death. Gene knockout of Apaf1 in mice leads to late embryonic lethality with malformations such as the persistence of interdigital webs and hyperplasia of brain and retina. Therefore, Apaf1 is generally believed to play a crucial role in developmental apoptosis and have a widespread expression. However, its pattern of expression in early development remains unknown. To specify whether Apaf1 indeed plays this key role, we investigated the pattern of gene expression for Apaf1 in mouse embryos on day 7, 9, and 12 of development. Our results show, that gene expression for Apaf1 first occurs within the embryo between day 7 and 9 of development, becoming more widespread toward day 12 and then includes structures, such as yolk sac, mesenchyme, cartilage, heart anlage, otic vesicle, peridermis, and anlagen of the spinal ganglia and vertebral bodies. Our results also show that gene expression for Apaf1 is not ubiquitous in early mouse development. This finding indicates that cell death processes are independent of or less dependent on Apaf1 during this time. Of interest, an active gene expression for Apaf1 is also present in organ anlagen such as heart or intestine, in which no obvious phenotype is seen after Apaf1 deletion. This finding suggests a possible role for Apaf1 in such anlagen as a putative alternative compensatory pathway, which could be switched on in the case of defects in the mediators that are normally involved in such organs.


Intraoperative skeletal traction in the dog: a cadaveric study.
A standard skeletal traction technique was applied to each major segment of the appendicular skeleton of ten fresh dog cadavers. Opposition points and anchorage points for the application of traction were determined for each skeletal segment. Traction was exerted by means of a micrometric traction stand, connected to the limb by bands or a stirrup. Traction was applied to the antebrachium and the tibia through nylon traction bands anchored to the metacarpus and metatarsus, respectively. A traction stirrup applied to the condylar region was used as the anchorage point to load the humerus and femur. Once a peak force of 25 kg weight was achieved, the load was monitored for half an hour to check for any variation. After that, each skeletal segment was osteotomized in the mid-diaphyseal region, and evaluated for any angular malalignment due to a mismatch between the axis of the bone and the applied loading. Any ensuing angular malalignment was successfully corrected by manoeuvres using the traction stand. The technique used in this study to perform intraoperative skeletal traction proved to be reliable and consistent for each segment of the appendicular skeleton.


Iniferter-mediated grafting of molecularly imprinted polymers on porous silica beads for the enantiomeric resolution of drugs.
A surface-imprinted chiral stationary phase for the enantiomeric resolution of the antidepressant drug, citalopram, is presented in this work. N, N'-diethylaminodithiocarbamoylpropyl(trimethoxy)silane has been used as silane iniferter for the surface functionalization of the solid silica support. A molecularly imprinted polymer thin film, in the nm scale, was then grafted on the silanized silica using itaconic acid as the functional monomer and ethylene glycol dimethacrylate as the cross-linker in the presence of the template S-citalopram. The total monomer amount was calculated to obtain the desired thickness. Non-imprinted stationary phases were prepared similarly in the absence of S-citalopram. Characterization of the materials was carried out by scanning electron microscopy, thermogravimetric analysis, elemental analysis and Fourier transform infrared spectroscopy. Stationary phases have been applied to the chromatographic separation of the target. Conditions for best chromatographic resolution of the enantiomers were optimized, and it was found that a mobile phase consisting of a mixture of formate buffer (40 mM, pH 3) and acetonitrile (30:70 v/v) at 40 °C provided best results. Binding behaviour of the developed material was finally assessed by batch rebinding experiments. The obtained binding isotherm was fitted to different binding models being the Freundlich-Langmuir model, the one that best fitted the experimental data. The developed material has shown high selectivity for the target enantiomer, and the stationary phase could be undoubtedly exploited for chiral separation of the drug.


Pharmacokinetics of dihydroartemisinin and piperaquine in pregnant and nonpregnant women with uncomplicated falciparum malaria.
Dihydroartemisinin-piperaquine is a fixed-dose artemisinin-based combination treatment. Some antimalarials have altered pharmacokinetics in pregnancy. Pregnant women in the 2nd or 3rd trimester and matched nonpregnant women with uncomplicated falciparum malaria were treated with a total of 6.4 mg/kg of body weight dihydroartemisinin and 51.2 mg/kg piperaquine once daily for 3 days. Venous blood samples were drawn at prespecified time points over 9 weeks. Plasma dihydroartemisinin and piperaquine concentrations were analyzed by liquid chromatography-mass spectrometry. Piperaquine and dihydroartemisinin pharmacokinetics were well described. There were no significant differences in total piperaquine exposure (P = 0.80) or drug exposure during the terminal elimination phase (72 h to infinity) (P = 0.64) between the two groups. The apparent volume of distribution of piperaquine was significantly smaller (602 liters/kg versus 877 liters/kg) in pregnant women than in nonpregnant women (P = 0.0057), and the terminal elimination half-life was significantly shorter (17.8 days versus 25.6 days; P = 0.0023). Dihydroartemisinin exposure after the first dose was significantly lower (844 h × ng/ml versus 1,220 h × ng/ml, P = 0.0021) in pregnant women, but there were no significant differences in total dihydroartemisinin exposure or maximum concentrations between the two groups. There were no significant differences in any pharmacokinetic parameters between the second and third trimester. These results obtained through noncompartmental analysis suggest that in the treatment of falciparum malaria, there are no clinically important differences in the pharmacokinetics of dihydroartemisinin or piperaquine between pregnant and nonpregnant women. However, a more detailed analysis using population pharmacokinetic modeling is needed to fully investigate the differences found for some of the pharmacokinetic parameters, such as the terminal half-life.


A 4-dimensional representation of antennal lobe output based on an ensemble of characterized projection neurons.
A central problem facing studies of neural encoding in sensory systems is how to accurately quantify the extent of spatial and temporal responses. In this study, we take advantage of the relatively simple and stereotypic neural architecture found in invertebrates. We combine standard electrophysiological techniques, recently developed population analysis techniques, and novel anatomical methods to form an innovative 4-dimensional view of odor output representations in the antennal lobe of the moth Manduca sexta. This novel approach allows quantification of olfactory responses of characterized neurons with spike time resolution. Additionally, arbitrary integration windows can be used for comparisons with other methods such as imaging. By assigning statistical significance to changes in neuronal firing, this method can visualize activity across the entire antennal lobe. The resulting 4-dimensional representation of antennal lobe output complements imaging and multi-unit experiments yet provides a more comprehensive and accurate view of glomerular activation patterns in spike time resolution.


HOXB13 mediates tamoxifen resistance and invasiveness in human breast cancer by suppressing ERα and inducing IL-6 expression.
Most breast cancers expressing the estrogen receptor α (ERα) are treated successfully with the receptor antagonist tamoxifen (TAM), but many of these tumors recur. Elevated expression of the homeodomain transcription factor HOXB13 correlates with TAM-resistance in ERα-positive (ER+) breast cancer, but little is known regarding the underlying mechanism. Our comprehensive evaluation of HOX gene expression using tiling microarrays, with validation, showed that distant metastases from TAM-resistant patients also displayed high HOXB13 expression, suggesting a role for HOXB13 in tumor dissemination and survival. Here we show that HOXB13 confers TAM resistance by directly downregulating ERα transcription and protein expression. HOXB13 elevation promoted cell proliferation in vitro and growth of tumor xenografts in vivo. Mechanistic investigations showed that HOXB13 transcriptionally upregulated interleukin (IL)-6, activating the mTOR pathway via STAT3 phosphorylation to promote cell proliferation and fibroblast recruitment. Accordingly, mTOR inhibition suppressed fibroblast recruitment and proliferation of HOXB13-expressing ER+ breast cancer cells and tumor xenografts, alone or in combination with TAM. Taken together, our results establish a function for HOXB13 in TAM resistance through direct suppression of ERα and they identify the IL-6 pathways as mediator of disease progression and recurrence.


Syntheses, structural characterizations, and optical and electrochemical properties of directly fused diporphyrins.
Directly fused diporphyrins display the extensive pi conjugation as evinced by highly perturbed electronic absorption spectra as well as lowered and largely split first oxidation potentials. Such diporphyrins prepared include meso-beta doubly linked diporphyrins 7, meso-meso beta-beta beta-beta triply linked diporphyrins 8, and meso-meso beta-beta doubly linked diporphyrins 9. Oxidation of 5,15-diaryl-substituted and 5,10,15-triaryl-substituted Ni(II)-, Cu(II)-, and Pd(II)-porphyrins with tris(4-bromophenyl)aminium hexachloroantimonate (BAHA) in CHCl(3) afforded 7, and triply linked Cu(II)-diporphyrins 8a and 8g were respectively prepared by the oxidation of meso-meso singly linked Cu(II)-diporphyrins 5c and 5f with BAHA. Meso-meso beta-beta doubly linked Ni(II)-diporphyrin 9a was isolated along with triply linked Ni(II)-diporphyrin 8e from the similar oxidation of meso-meso singly linked Ni(II)-diporphyrin 5a. Doubly linked diporphyrins 7 and 9a both exhibit significantly perturbed electronic absorption spectra, in which the Soret-like bands are largely split at around 405-418 and 500-616 nm and the Q-bandlike absorption bands are substantially intensified and red-shifted at 748-820 nm, probably as a consequence of symmetry lowering. Triply linked diporphyrins 8 display more strongly perturbed electronic absorption spectra with split Soret-like bands at 408-419 and 567-582 nm and Q-bandlike absorption bands reaching far-infrared region. Structures of three types of fused diporphyrins 7b and 7c, 8g and 8j, and 9a have been unambiguously determined by X-ray crystallography to be nearly coplanar. Both the triply linked diporphyrins 8g and 8j exhibit very flat structures, whereas the doubly linked diporphyrins 7b and 7c exhibit ruffled structures. The doubly linked diporphyrin 9a shows a helically twisted conformation with larger ruffling toward the opposite directions and has been actually separated into two enantiomers, which display strong Cotton effects in the CD spectra. The first oxidation potentials (E(OX1)) decrease in the order of 5 > 7 > or = 9 > 8, indicating lift-up of HOMO orbital in this order, and split potential differences DeltaE = E(OX1) - E(OX2), in turn, increase in the reverse order of 5 < 7< or = 9 < 8. The (1)H NMR spectra have indicated that the aromatic porphyrin ring current becomes weakened in the order of 5 > 7 > 8. Collectively, the electronic interactions between the diporphyrins have been concluded to increase in the other of 5 << 7 < or = 9 < 8.


Biological preservation of plant derived animal feed with antifungal microorganisms: safety and formulation aspects.
During storage of moist animal feed, growth of detrimental fungi causing spoilage, or being mycotoxigenic or pathogenic, is a severe problem. Addition of biopreservative yeasts or lactic acid bacteria can significantly reduce this problem. However, their use requires several careful considerations. One is the safety to the animal, humans and the environment, tightly connected to legal aspects and the need for pre-market authorisation when supplementing feed with microorganisms. Although both yeasts and lactic acid bacteria are considered comparatively safe organisms due to low production of toxic metabolites, it is of great importance to understand the mechanisms behind the biopreservative abilities. Another important issue concerns practical aspects, such as the economic production of large amounts of the organisms and the development of a suitable formulation giving the organisms a long shelf life. These aspects are discussed and a recommendation of this review is that both safety and formulation aspects of a specific microbe should be considered at an early stage in the selection of new organisms with biopreservation potential.


Early initiation of prophylactic anticoagulation for prevention of COVID-19 mortality: a nationwide cohort study of hospitalized patients in the United States.
Deaths among patients with coronavirus disease 2019 (COVID-19) are partially attributed to venous thromboembolism and arterial thromboses. Anticoagulants prevent thrombosis formation, possess anti-inflammatory and anti-viral properties, and may be particularly effective for treating patients with COVID-19. To evaluate whether initiation of prophylactic anticoagulation within 24 hours of admission is associated with decreased risk of death among patients hospitalized with COVID-19. Observational cohort study. Nationwide cohort of patients receiving care in the Department of Veterans Affairs, the largest integrated healthcare system in the United States. All patients hospitalized with laboratory-confirmed SARS-CoV-2 infection March 1 to July 31, 2020, without a history of therapeutic anticoagulation. Prophylactic doses of subcutaneous heparin, low-molecular-weight heparin, or direct oral anticoagulants. 30-day mortality. Secondary outcomes: inpatient mortality and initiating therapeutic anticoagulation. Of 4,297 patients hospitalized with COVID-19, 3,627 (84.4%) received prophylactic anticoagulation within 24 hours of admission. More than 99% (n=3,600) received subcutaneous heparin or enoxaparin. We observed 622 deaths within 30 days of admission, 513 among those who received prophylactic anticoagulation. Most deaths (510/622, 82%) occurred during hospitalization. In inverse probability of treatment weighted analyses, cumulative adjusted incidence of mortality at 30 days was 14.3% (95% CI 13.1-15.5) among those receiving prophylactic anticoagulation and 18.7% (95% CI 15.1-22.9) among those who did not. Compared to patients who did not receive prophylactic anticoagulation, those who did had a 27% decreased risk for 30-day mortality (HR 0.73, 95% CI 0.66-0.81). Similar associations were found for inpatient mortality and initiating therapeutic anticoagulation. Quantitative bias analysis demonstrated that results were robust to unmeasured confounding (e-value lower 95% CI 1.77). Results persisted in a number of sensitivity analyses. Early initiation of prophylactic anticoagulation among patients hospitalized with COVID-19 was associated with a decreased risk of mortality. These findings provide strong real-world evidence to support guidelines recommending the use of prophylactic anticoagulation as initial therapy for COVID-19 patients upon hospital admission.


The effect of silica coating on the resin bond to the intaglio surface of Procera AllCeram restorations.
This study investigated the influence of a silica-coating method on the resin bond of two different resin composite cements to the intaglio surface of Procera AllCeram densely sintered, high-purity, alumina ceramic restorations after long-term storage and thermocycling. Densely sintered alumina ceramic specimens were fabricated with the intaglio surface of the Procera AlICeram coping and randomly divided into five adhesive groups (100 total specimens). Resin composite cylinders were bonded either to the untreated or to the tribochemical silica/silane-coated ceramic surface with either a conventional Bis-GMA resin cement or a resin composite containing an adhesive phosphate monomer (Panavia 21) in combination with their corresponding bonding/silane coupling agents. Panavia was also used without silanization to the untreated ceramic surface (control). Subgroups of 10 specimens were stored in distilled water for either 3 (baseline) or 180 days prior to shear bond strength testing. The 180-day samples were subjected to repeated thermocycling for a total of 12,000 cycles. Data were analyzed with one-way analysis of variance and Tukey's multiple comparison. Silica coating significantly increased overall bond strength to Procera AllCeram. RelyX ARC and silica coating revealed the highest bond strength at baseline. Long-term storage and thermocycling significantly decreased overall bond strength. Two groups revealed the significantly highest bond strength values after artificial aging: Panavia 21 with its silane/bonding agent to the original ceramic surface and Panavia 21 to the silica-coated ceramic surface. The use of a resin composite containing an adhesive phosphate monomer either in combination with a silane coupling/bonding agent or after tribochemical silica/silane coating revealed the highest long-term shear bond strength to the intaglio surface of Procera AllCeram restorations.


Oil type and temperature dependent biodegradation dynamics - Combining chemical and microbial community data through multivariate analysis.
This study investigates a comparative multivariate approach for studying the biodegradation of chemically dispersed oil. The rationale for this approach lies in the inherent complexity of the data and challenges associated with comparing multiple experiments with inconsistent sampling points, with respect to inferring correlations and visualizing multiple datasets with numerous variables. We aim to identify novel correlations among microbial community composition, the chemical change of individual petroleum hydrocarbons, oil type and temperature by creating modelled datasets from inconsistent sampling time points. Four different incubation experiments were conducted with freshly collected Norwegian seawater and either Grane and Troll oil dispersed with Corexit 9500. Incubations were conducted at two different temperatures (5 °C and 13 °C) over a period of 64 days. PCA analysis of modelled chemical datasets and calculated half-lives revealed differences in the biodegradation of individual hydrocarbons among temperatures and oil types. At 5 °C, most n-alkanes biodegraded faster in heavy Grane oil compared to light Troll oil. PCA analysis of modelled microbial community datasets reveal differences between temperature and oil type, especially at low temperature. For both oils, Colwelliaceae and Oceanospirillaceae were more prominent in the colder incubation (5 °C) than the warmer (13 °C). Overall, Colwelliaceae, Oceanospirillaceae, Flavobacteriaceae, Rhodobacteraceae, Alteromonadaceae and Piscirickettsiaceae consistently dominated the microbial community at both temperatures and in both oil types. Other families known to include oil-degrading bacteria were also identified, such as Alcanivoracaceae, Methylophilaceae, Sphingomonadaceae and Erythrobacteraceae, but they were all present in dispersed oil incubations at a low abundance (< 1%). In the current study, our goal was to introduce a comparative multivariate approach for studying the biodegradation of dispersed oil, including curve-fitted models of datasets for a greater data resolution and comparability. By applying these approaches, we have shown how different temperatures and oil types influence the biodegradation of oil in incubations with inconsistent sampling points. Clustering analysis revealed further how temperature and oil type influence single compound depletion and microbial community composition. Finally, correlation analysis of degraders community, with single compound data, revealed complexity beneath usual abundance cut-offs used for microbial community data in biodegradation studies.


A quantum dot single-photon turnstile device.
Quantum communication relies on the availability of light pulses with strong quantum correlations among photons. An example of such an optical source is a single-photon pulse with a vanishing probability for detecting two or more photons. Using pulsed laser excitation of a single quantum dot, a single-photon turnstile device that generates a train of single-photon pulses was demonstrated. For a spectrally isolated quantum dot, nearly 100% of the excitation pulses lead to emission of a single photon, yielding an ideal single-photon source.


Effects of glutamine on adhesion molecule expression and leukocyte transmigration in endothelial cells exposed to arsenic.
This study evaluated whether glutamine (GLN) concentration was related to endothelial surface molecule expression and the migration of polymorphonuclear neutrophils (PMNs) through endothelial cells (ECs) stimulated by arsenic. Human umbilical vein endothelial cells (HUVECs) and PMNs were treated with different GLN concentrations (0, 300, 600 and 1000 microM) for 24 h. After that, we stimulated HUVECs for 3 h with 0.5 microM arsenic, and PMNs were allowed to transmigrate to ECs for 2 h. HUVEC surface expressions of cell adhesion molecules and integrin (CD11b) and interleukin (IL)-8 receptor expressions on PMNs were measured. The transendothelial migration of PMNs was also analyzed. The results showed that cell adhesion molecule (CAM) and integrin expressions in arsenic groups were higher than in those without arsenic. Among the arsenic groups, the expression of CAMs on ECs and CD11b, and IL-8 receptor on PMNs was lowest with 0 microM compared with the other GLN concentrations. Vascular CAM-1 on ECs and CD11b on PMN expression were higher with 300 microM than with 600 and 1000 microM GLN. IL-8 secretions from ECs and PMNs were higher with 300 muM than with 600 and 1000 microM GLN, and this was consistent with the expression of the IL-8 receptor on PMNs. Polymorphonuclear neutrophil transmigration was significantly higher with 300 muM GLN than with other GLN concentrations. These results suggest that ECs and PMNs were activated after arsenic stimulation. Cell adhesion molecule expressions on ECs and PMNs were suppressed in the absence of GLN. A low GLN concentration comparable to catabolic conditions resulted in higher adhesion molecule expression and greater transendothelial migration of neutrophils. Glutamine administration at levels similar to or higher than physiological concentrations reduced IL-8 and adhesion molecule expression; PMN transmigration was also decreased after stimulation with arsenic.


Two-step conversion of carboxylic esters into distally fluorinated ketones via ring cleavage of cyclopropanol intermediates: application of sulfinate salts as fluoroalkylating reagents.
Tertiary cyclopropanols easily available from carboxylic esters have been used in the synthesis of distally fluorinated ketones. Cyclopropane ring cleavage reactions in methanol with aqueous tert-butyl hydroperoxide in the presence of a copper(ii) acetate catalyst and sodium triflinate (Langlois reagent) afford β-trifluoromethyl ketones in 16-74% isolated yields. Sodium triflinate serves as a precursor of reactive trifluoromethyl copper species, enabling ring-opening trifluoromethylation, as evidenced by mechanistic studies. We also demonstrate here that other sulfinate salts, such as sodium 1,1-difluoroethanesulfinate, sodium 2-(4-bromophenyl)-1,1-difluoroethanesulfinate and sodium 1-(trifluoromethyl)cyclopropanesulfinate, can be used as fluoroalkylation reagents, resulting in the corresponding fluorinated ketones.


Construction, characterization, and preliminary BAC-end sequence analysis of a bacterial artificial chromosome library of the tea plant (Camellia sinensis).
We describe the construction and characterization of a publicly available BAC library for the tea plant, Camellia sinensis. Using modified methods, the library was constructed with the aim of developing public molecular resources to advance tea plant genomics research. The library consists of a total of 401,280 clones with an average insert size of 135 kb, providing an approximate coverage of 13.5 haploid genome equivalents. No empty vector clones were observed in a random sampling of 576 BAC clones. Further analysis of 182 BAC-end sequences from randomly selected clones revealed a GC content of 40.35% and low chloroplast and mitochondrial contamination. Repetitive sequence analyses indicated that LTR retrotransposons were the most predominant sequence class (86.93%-87.24%), followed by DNA retrotransposons (11.16%-11.69%). Additionally, we found 25 simple sequence repeats (SSRs) that could potentially be used as genetic markers.


Treatment of onyalai with prednisolone, intravenous gammaglobulin and ascorbic acid. A prospective clinical trial.
Forty-two patients in the acute phase of onyalai were admitted to hospital and randomly divided into three groups of 14 to receive prednisolone 3 mg/kg/d for 1 week, intravenous gammaglobulin 100 mg/kg on 2 successive days, or ascorbic acid 1000 mg 3 times a day for 1 week. The groups were comparable with regard to sex and age distribution, initial platelet counts and initial haemoglobin values. No statistical difference was observed between the three treatment groups with regard to changes in the platelet counts during the period of observation. One patient died from haemorrhagic shock. The low mortality rate in this series suggests that admission to hospital and early correction of blood loss are important factors in reducing the mortality rate associated with onyalai.


The nature of geometric frustration in the Kob-Andersen mixture.
Geometric frustration is an approach to the glass transition based upon the consideration of locally favoured structures (LFS), which are geometric motifs which minimise the local free energy. Geometric frustration proposes that a transition to a crystalline state is frustrated because these LFS do not tile space. However, this concept is based on icosahedra which are not always the LFS for a given system. The LFS of the popular Kob-Andersen (KA) model glassformer are the bicapped square antiprism, which does tile space. Such a LFS-crystal is indeed realised in the Al2Cu structure, which is predicted to be a low energy state for the KA model with a 2:1 composition. We, therefore, hypothesise that upon changing the composition in the KA model towards 2:1, geometric frustration may be progressively relieved, leading to larger and larger domains of LFS which would ultimately correspond to the Al2Cu crystal. Remarkably, rather than an increase, upon changing composition we find a small decrease in the LFS population, and the system remains impervious to nucleation of LFS crystals. We suggest that this may be related to the composition of the LFS, as only a limited subset is compatible with the crystal. We further demonstrate that the Al2Cu crystal will grow from a seed in the KA model with 2:1 composition and identify the melting temperature to be 0.447(2).


Labor Complicated by Cervical Stenosis Following a Laser Cone Biopsy.
A 34-year-old lady, with three previous spontaneous vaginal deliveries, presented in labor at 37 + 0 weeks' gestation. Following the birth of her third child, she underwent a laser cone biopsy for cervical intraepithelial neoplasia (CIN) 3. Despite sustained regular contractions and augmentation with Syntocinon, progressive cervical dilatation beyond 1 cm failed to occur in this multiparous lady. A delayed diagnosis of cervical stenosis was made. She eventually underwent a cesarean section where her internal cervical os was found to be thin and fibrotic. This case describes an uncommon occurrence of cervical stenosis presenting in labor and seeks to increase awareness of this condition so as to allow preemptive counselling of similar patients, early recognition in a labor with poor progress and a swift, better-informed decision to deliver via an emergency cesarean section. We review the existing literature on cervical stenosis following cervical conization and analyze the various definitions available.


Identification and functional analysis of type III effector proteins in Mesorhizobium loti.
Mesorhizobium loti MAFF303099, a microsymbiont of the model legume Lotus japonicus, possesses a cluster of genes (tts) that encode a type III secretion system (T3SS). In the presence of heterologous nodD from Rhizobium leguminosarum and a flavonoid naringenin, we observed elevated expression of the tts genes and secretion of several proteins into the culture medium. Inoculation experiments with wild-type and T3SS mutant strains revealed that the presence of the T3SS affected nodulation at a species level within the Lotus genus either positively (L. corniculatus subsp. frondosus and L. filicaulis) or negatively (L. halophilus and two other species). By inoculating L. halophilus with mutants of various type III effector candidate genes, we identified open reading frame mlr6361 as a major determinant of the nodulation restriction observed for L. halophilus. The predicted gene product of mlr6361 is a protein of 3,056 amino acids containing 15 repetitions of a sequence motif of 40 to 45 residues and a shikimate kinase-like domain at its carboxyl terminus. Homologues with similar repeat sequences are present in the hypersensitive-response and pathogenicity regions of several plant pathogens, including strains of Pseudomonas syringae, Ralstonia solanacearum, and Xanthomonas species. These results suggest that L. halophilus recognizes Mlr6361 as potentially pathogen derived and subsequently halts the infection process.


Multiple membrane transport systems for the uptake of folate-based thymidylate synthase inhibitors.
N10-Propargyl-5,8-dideazafolic acid (CB3717) and 2-desamino-2-methyl-N10-propargyl-5,8-dideazafolic acid (ICI-198,583) are potent folate-based inhibitors of thymidylate synthase. We studied the membrane transport and the growth-inhibitory effects of the two thymidylate synthase inhibitors on human CCRF-CEM leukemia cells with different transport properties for folic acid, reduced folates, and methotrexate (MTX). Membrane transport of [3H]ICI-198,583 can proceed via the high affinity/low capacity reduced folate carrier as supported by findings that (a) uptake of [3H]ICI-198,583 was significantly impaired in CEM cells which have a transport defect for MTX, (b) variants of CEM cells which overproduce the reduced folate carrier system showed a concomitant increase in the uptake of [3H]ICI-198,583 as for [3H]MTX, (c) MTX inhibited transport of [3H]ICI-198,583, and (d) uptake of [3H]ICI-198,583 was inhibited after treatment of CEM cells with an N-hydroxysuccinimide ester of MTX, which is a potent inhibitor of MTX transport. However, a membrane-associated folate-binding protein (FBP) offers another route for entry of CB3717 and ICI-198,583. CEM-FBP cells that have an elevated amount of FBP and do not have a functional reduced folate carrier were 640- and 61-fold more sensitive to CB3717 and ICI-198,583, respectively, compared to control CEM cells expressing the reduced folate/MTX carrier. This high sensitivity was related to a high affinity of the FBP for CB3717 and ICI-198,583 (Kd 2-3 nM), which is only 3-fold lower than for folic acid (Kd 1 nM) but significantly higher than for MTX (Kd 100 nM). Furthermore, after incubation of CEM-FBP cells for 24 h at 10 nM [3H]ICI-198,583, the high affinity binding of the FBP for ICI-198,583 allowed a 600-fold concentrative uptake of [3H]ICI-198,583 and its conversion to polyglutamate forms. These results indicate that multiple folate transport systems may be involved in the uptake of folate-based thymidylate synthase inhibitors.


Effects of protein-rich supplementation and nandrolone on bone tissue after a hip fracture.
Osteoporosis is a major health problem worldwide. Low weight is a major risk factor for low bone mass and fractures. The aim of this study was to investigate the effects on bone tissue of protein-rich supplementation alone or in combination with nandrolone decanoate in lean elderly women after a hip fracture. Sixty elderly women with BMI <24 kg/m(2) admitted to hospital due to a femoral neck fracture were randomised to a control group, to receive a protein-rich formula or to receive the same formula with an addition of nandrolone decanoate for 6 months. All patients received additional calcium and vitamin D. The effects after 6 and 12 months were measured by means of bone mineral density (BMD) using dual-energy X-ray absorptiometry (DXA), and with biochemical bone markers. Osteocalcin and C-terminal telopeptide of collagen-1 (CTX) were used to estimate bone formation and bone resorption, respectively. The analyses showed an increase in total body BMD at 6 and 12 months in patients who received protein-rich supplementation. Nandrolone decanoate did not appear to have any additional effect on BMD. Osteocalcin increased in all groups while no significant changes were found for CTX. The overall results of the study indicated that protein-rich supplementation given to lean elderly female hip fracture patients increased the total body BMD.


Dosing strategies for antiepileptic drugs: from a standard dose for all to individualised treatment by implementation of therapeutic drug monitoring.
This review focuses on the evolution of approaches to dosing of antiepileptic drugs (AEDs) in clinical practice through history. There has been a shift in the view of treatment of epilepsy, from "one dose fits all patients" in the early days to individualisation of treatment. Over the past 50 years, our knowledge of pharmacological variability of AEDs has markedly increased through implementation of therapeutic drug monitoring (TDM). The use of TDM has demonstrated extensive pharmacokinetic variability for AEDs and a need to individualise the treatment for an optimal outcome. Factors that contribute to pharmacokinetic variability include external factors (including food and comedication), physiological factors (gender, age, and pregnancy), pathological conditions (organ dysfunction), and genetic factors (polymorphisms in metabolising enzymes). Patient groups of children, pregnant women, and the elderly, in whom the most extensive pharmacokinetic changes occur, need special attention and close follow-up of treatment. Patients with complicated and changing combination treatments are also vulnerable. Therapeutic drug monitoring may be particularly helpful in such situations. There are also challenges regarding the use and misuse of therapeutic drug monitoring, such as the use of drug monitoring without a clear indication, misinterpretation of the reference range, and erroneous sampling times.


Wideband tunable optoelectronic oscillator based on the deamplification of stimulated Brillouin scattering.
A wideband tunable optoelectronic oscillator (OEO) based on the deamplification of stimulated Brillouin scattering (SBS) is proposed and experimentally demonstrated. A tunable single passband microwave photonic filter (MPF) utilizing phase modulation and SBS deamplification is used to realize the tunability of the OEO. Theoretical analysis of the MPF and phase noise performance of the OEO are presented. The frequency response of the MPF is determined by the + 1st sideband attenuation due to SBS deamplification and phase shift difference between the two sidebands due to chromatic dispersion and SBS. The close-in (< 1 MHz) phase noise of the proposed OEO is shown to be dominated by the laser frequency noise via phase shift of SBS. The conversion of the laser frequency noise to the close-in phase noise of the proposed OEO is effectively reduced compared with the OEO based on amplification by SBS. Tunable 7 to 40 GHz signals are experimentally obtained. The single-sideband (SSB) phase noise at 10 kHz offset is -128 dBc/Hz for 10.30 GHz signal. Compared with the OEO based on SBS amplification, the proposed OEO can achieve a phase noise performance improvement beyond 20 dB at 10 kHz offset. The maximum frequency and power drifts at 10.69 GHz are within 1 ppm and 1.4 dB during 1000 seconds, respectively. To achieve better close-in phase noise performance, lower frequency noise laser and higher pump power are preferred. The experimental results agree well with the theoretical models.


Neither per, nor tim1, nor cry2 alone are essential components of the molecular circadian clockwork in the Madeira cockroach.
Circadian clocks control rhythms in physiology and behavior entrained to 24 h light-dark cycles. Despite of conserved general schemes, molecular circadian clockworks differ between insect species. With RNA interference (RNAi) we examined an ancient circadian clockwork in a basic insect, the hemimetabolous Madeira cockroach Rhyparobia maderae. With injections of double-stranded RNA (dsRNA) of cockroach period (Rm´per), timeless 1 (Rm´tim1), or cryptochrome 2 (Rm´cry2) we searched for essential components of the clock´s core negative feedback loop. Single injections of dsRNA of each clock gene into adult cockroaches successfully and permanently knocked down respective mRNA levels within ~two weeks deleting daytime-dependent mRNA rhythms for Rm´per and Rm´cry2. Rm´perRNAi or Rm´cry2RNAi affected total mRNA levels of both genes, while Rm´tim1 transcription was independent of both, also keeping rhythmic expression. Unexpectedly, circadian locomotor activity of most cockroaches remained rhythmic for each clock gene knockdown employed. It expressed weakened rhythms and unchanged periods for Rm´perRNAi and shorter periods for Rm´tim1RNAi and Rm´cry2RNAi.As a hypothesis of the cockroach´s molecular clockwork, a basic network of switched differential equations was developed to model the oscillatory behavior of clock cells expressing respective clock genes. Data were consistent with two synchronized main groups of coupled oscillator cells, a leading (morning) oscillator, or a lagging (evening) oscillator that couple via mutual inhibition. The morning oscillators express shorter, the evening oscillators longer endogenous periods based on core feedback loops with either PER, TIM1, or CRY2/PER complexes as dominant negative feedback of the clockwork. We hypothesize that dominant morning oscillator cells with shorter periods express PER, but not CRY2, or TIM1 as suppressor of clock gene expression, while two groups of evening oscillator cells with longer periods either comprise TIM1 or CRY2/PER suppressing complexes. Modelling suggests that there is an additional negative feedback next to Rm´PER in cockroach morning oscillator cells.