Sonographic evaluation of the size of Achilles tendon: the effect of exercise and dominance of the ankle.
This study was undertaken to measure and compare the thickness and cross-sectional area of the Achilles tendon between frequent- and infrequent-exercise subjects, and between the dominant and nondominant ankles in an asymptomatic Chinese population. Interobserver variability in the measurement of the size of Achilles tendon was also evaluated. High-resolution ultrasound (US) examination of Achilles tendons was performed in 40 healthy subjects (20 who frequently exercised, had exercise at least 3 days per week and at least 2 h per session; and 20 who infrequently exercised); their age range was 19 to 25 years. The thickness and cross-sectional area of the Achilles tendons were measured in a transverse scan at the level of medial malleolus. For each subject, the Achilles tendons were measured by five operators to evaluate the interobserver variability in the measurements. The mean thickness and cross-sectional areas of the Achilles tendon in a healthy Chinese population are 5.23 mm(2) and 56.91 mm(2)(2), respectively. The mean thickness of the Achilles tendon of frequent-exercise subjects (dominant ankle 5.43 mm, nondominant ankle 5.38 mm) was significantly greater than that of infrequent-exercise subjects (dominant ankle 5.08 mm, nondominant ankle 5.04 mm) (p < 0.05). The cross-sectional area of the tendons was also larger in frequent-exercise subjects but, whereas a significant result was found in dominant ankles (frequent-exercise subjects 60.46 mm(2)(2), infrequent-exercise subjects 54.71 mm(2)(2)) (p < 0.05), this was not the case for the nondominant ankles (frequent-exercise subjects 57.09 mm(2)(2), infrequent-exercise subjects 55.4 mm(2)(2)) (p > 0.05). In both frequent- and infrequent-exercise subjects, there was no significant difference in the mean thickness and cross-sectional area of Achilles tendon between dominant and nondominant ankles (p > 0.05). There was a high reproducibility in the sonographic measurement of the thickness (68%) and cross-sectional area (81%) of Achilles tendons. Results suggested that exercise would cause increase in the thickness and cross-sectional area of Achilles tendon. Interobserver variability is not significant in the sonographic measurement of Achilles tendons.


Humoral immune responses to T cell tropic retrovirus simian T lymphotropic virus type III in monkeys with experimentally induced acquired immune deficiency-like syndrome.
The T cell tropic retrovirus of macaque monkeys simian T lymphotropic virus type III (STLV-III) has morphologic, growth, and antigenic properties indicating that it is related to human T cell lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV), the etiologic agent of the acquired immune deficiency syndrome (AIDS) of humans. STLV-III has recently been shown to induce an AIDS-like disease in macaque monkeys. In this study the humoral immune responses of six experimentally infected monkeys have been characterized to determine whether certain parameters of the antibody response to the virus might be predictive of the clinical outcome of this infection. Two distinct patterns of antibody responses were found. Four animals that died within 160 d of inoculation developed low titer anti-STLV-III antibody responses that recognized only the viral envelope protein, and progressive declines in total plasma IgG levels and absolute peripheral blood T4 lymphocyte numbers. The two animals that lived longer (one died at 352 d, the other remains alive at 430 d) developed high titer anti-STLV-III antibody responses that recognized both viral envelope and core proteins, increases in total plasma IgG, and a later decrease in number of peripheral blood T4 lymphocytes. Interestingly, the single animal that has remained clinically healthy after infection was the only one to develop detectable STLV-III neutralizing antibodies.


Hemifacial Spasm in Mucopolysaccharidosis Type VI (Maroteaux-Lamy Syndrome).
Hemifacial spasms are involuntary contractions of the muscles of one side of the face. Here, we report a 5-year-old female patient with a significant past medical history, including mucopolysaccharidosis type VI, who also presented with hemifacial spasms. Further investigations showed narrowing of skull foramina and hydrocephalus, but no compression of the facial nerve. Carbamazepine was given to the patient, which caused cessation of the spasms. Currently, there is no literature discussing the relationship between hemifacial spasms and mucopolysaccharidosis type VI; we have proposed that mucopolysaccharide deposits or dural thickening may contribute to nerve irritation.


Impact of a regionalised clinical cardiac support network on mortality among rural patients with myocardial infarction.
To evaluate the impact of the regionalised Integrated Cardiovascular Clinical Network (ICCNet) on 30-day mortality among patients with myocardial infarction (MI) in an Australian rural setting. An integrated cardiac support network incorporating standardised risk stratification, point-of-care troponin testing and cardiologist-supported decision making was progressively implemented in non-metropolitan areas of South Australia from 2001 to 2008. Hospital administrative data and statewide death records from 1 July 2001 to 30 June 2010 were used to evaluate outcomes for patients diagnosed with MI in rural and metropolitan hospitals. Risk-adjusted 30-day mortality. 29 623 independent contiguous episodes of MI were identified. The mean predicted 30-day mortality was lower among rural patients compared with metropolitan patients, while actual mortality rates were higher (30-day mortality: rural, 705/5630 [12.52%] v metropolitan, 2140/23 993 [8.92%]; adjusted odds ratio [OR], 1.46; 95% CI, 1.33-1.60; P< 0.001). After adjustment for temporal improvement in MI outcome, availability of immediate cardiac support was associated with a 22% relative odds reduction in 30-day mortality (OR, 0.78; 95% CI, 0.65-0.93; P= 0.007). A strong association between network support and transfer of patients to metropolitan hospitals was observed (before ICCNet, 1102/2419 [45.56%] v after ICCNet, 2100/3211 [65.4%]; P< 0.001), with lower mortality observed among transferred patients. Cardiologist-supported remote risk stratification, management and facilitated access to tertiary hospital-based early invasive management are associated with an improvement in 30-day mortality for patients who initially present to rural hospitals and are diagnosed with MI. These interventions closed the gap in mortality between rural and metropolitan patients in South Australia.


Hydrogen-supplemented drinking water protects cardiac allografts from inflammation-associated deterioration.
Recent evidence suggests that molecular hydrogen has therapeutic value for disease states that involve inflammation. We hypothesized that drinking hydrogen-rich water (HW) daily would protect cardiac and aortic allograft recipients from inflammation-associated deterioration. Heterotopic heart transplantation with short-course tacrolimus immunosuppression and orthotopic aortic transplantation were performed in allogeneic rat strains. HW was generated either by bubbling hydrogen gas through tap water (Bu-HW) or via chemical reaction using a magnesium stick [Mg + 2H(2) O → Mg (OH)(2) + H(2) ] immersed in tap water (Mg-HW). Recipients were given either regular water (RW), Mg-HW, Bu-HW, or Mg-HW that had been subsequently degassed (DW). Graft survival was assessed by daily palpation for a heartbeat. Drinking Mg-HW or Bu-HW was remarkably effective in prolonging heart graft survival and reducing intimal hyperplasia in transplanted aortas as compared with grafts treated with RW or DW. Furthermore, T cell proliferation was significantly inhibited in the presence of hydrogen in vitro, accompanied by less production of interleukin-2 and interferon-γ. Hydrogen treatment was also associated with increased graft ATP levels and increased activity of the enzymes in mitochondrial respiratory chain. Drinking HW prolongs survival of cardiac allografts and reduces intimal hyperplasia of aortic allografts.


Feasibility Study of the Permeability and Uptake of Mesoporous Silica Nanoparticles across the Blood-Brain Barrier.
Drug delivery into the brain is impeded by the blood-brain-barrier (BBB) that filters out the vast majority of drugs after systemic administration. In this work, we assessed the transport, uptake and cytotoxicity of promising drug nanocarriers, mesoporous silica nanoparticles (MSNs), in in vitro models of the BBB. RBE4 rat brain endothelial cells and Madin-Darby canine kidney epithelial cells, strain II, were used as BBB models. We studied spherical and rod-shaped MSNs with the following modifications: bare MSNs and MSNs coated with a poly(ethylene glycol)-poly(ethylene imine) (PEG-PEI) block copolymer. In transport studies, MSNs showed low permeability, whereas the results of the cellular uptake studies suggest robust uptake of PEG-PEI-coated MSNs. None of the MSNs showed significant toxic effects in the cell viability studies. While the shape effect was detectable but small, especially in the real-time surface plasmon resonance measurements, coating with PEG-PEI copolymers clearly facilitated the uptake of MSNs. Finally, we evaluated the in vivo detectability of one of the best candidates, i.e. the copolymer-coated rod-shaped MSNs, by two-photon in vivo imaging in the brain vasculature. The particles were clearly detectable after intravenous injection and caused no damage to the BBB. Thus, when properly designed, the uptake of MSNs could potentially be utilized for the delivery of drugs into the brain via transcellular transport.


Nuclear EGFR and Integrator/Super Elongation Complex concurrently binds to Immediate Early Genes for gene transactivation.
The gene transactivation function of nuclear EGFR (nEGFR) has been studied by investigating the genomic co-occupancies of nEGFR and RNA Polymerase II (RNAPII). However, due to RNAPII pausing, the co-recruitment of RNAPII and nEGFR does not necessarily represent productive transactivation. In this study, we integrated gatekeepers of productive transcriptional elongation such as Integrator and Super Elongation Complex (SEC) to interrogate the function of nEGFR. By analyzing publicly available ChIP-seq and RNA-seq data, we aims to 1) explore the function of nEGFR, 2) unravel nEGFR target genes, and 3) discuss potential mechanisms of nEGFR chromatin recruitment. EGF treatment in HeLa cells instigated chromatin recruitment of nEGFR, ERK, RNAPII, Integrator, and SEC in a cluster of 61 EGF-responsive genes. The function of nEGFR was identified as gene-activating rather than gene-repressing. Within the cluster of EGF-responsive genes, nEGFR targeted eleven Immediate Early Genes (IEGs) - JUN, EGR1, JUNB, IER2, KLF2, FOS, FOSL1, RHOB, CCNL1, DUSP2, and DUSP5, which up-regulated >2-fold after EGF stimulation. The promoter of these target genes commonly harbors AT-rich minimal consensus sequences for nEGFR binding. In addition, TCGA data analysis demonstrated positive correlations between EGFR and JUN/FOSL1/RHOB expressions, as well as clinical correlations in specific cancer types. To our knowledge, this is the first study to compare the genome-wide distribution of nEGFR versus Integrator and SEC, providing novel insight into supporting the gene-activating function of nEGFR. We revealed a panel of eleven nEGFR target genes, which concurrently recruited nEGFR, RNAPII, Integrator, and SEC for productive transcriptional elongation.


Podocyte-specific knockout of cyclooxygenase 2 exacerbates diabetic kidney disease.
Enhanced expression of cyclooxygenase 2 (COX2) in podocytes contributes to glomerular injury in diabetic kidney disease, but some basal level of podocyte COX2 expression might be required to promote podocyte attachment and/or survival. To investigate the role of podocyte COX2 expression in diabetic kidney disease, we deleted COX2 specifically in podocytes in a mouse model of Type 1 diabetes mellitus (Akita mice). Podocyte-specific knockout (KO) of COX2 did not affect renal morphology or albuminuria in nondiabetic mice. Albuminuria was significantly increased in wild-type (WT) and KO Akita mice compared with nondiabetic controls, and the increase in albuminuria was significantly greater in KO Akita mice compared with WT Akita mice at both 16 and 20 wk of age. At the 20-wk time point, mesangial expansion was also increased in WT and KO Akita mice compared with nondiabetic animals, and these histologic abnormalities were not improved by KO of COX2. Tubular injury was seen only in diabetic mice, but there were no significant differences between groups. Thus, KO of COX2 enhanced albuminuria and did not improve the histopathologic features of diabetic kidney disease. These data suggest that 1) KO of COX2 in podocytes does not ameliorate diabetic kidney disease in Akita mice, and 2) some basal level of podocyte COX2 expression in podocytes is necessary to attenuate the adverse effects of diabetes on glomerular filtration barrier function.


A novel E2F binding protein with Myc-type HLH motif stimulates E2F-dependent transcription by forming a heterodimer.
The human embryonal carcinoma cells NEC14 can be induced to differentiate morphologically by the addition of 10(-2) M N, N'-hexamethylene-bis-acetamide and cease to grow in several days. Transcription factors of the E2F/DP family have been shown to be closely related to the regulation of cell proliferation. To analyse cellular proteins which interact with E2F in NEC14 cells, cDNA clones encoding E2F binding proteins were isolated from a lambdaZAP II NEC14 cell library with the 32P-labeled GST (Glutathione S-transferase)-E2F-1 fusion protein as a probe. One of the clones encodes E2FBP1 which has the helix-loop-helix (HLH) motif, but lacks the basic domain and the zipper structure usually found at N- and C-terminal sides to the HLH motif, respectively. The arrangement of amino acids in the helix 1 and helix 2 regions is quite similar to those of Mxi and Mad, but different from those of E2F-1 and DP-1. Western blot analysis of the immunoprecipitates prepared with anti-E2FBP1 antibody showed that E2FBP1 associates with both E2F-1 and DP-1 in vivo. E2FBP1 alone has no DNA binding activity, but bind to the E2F site through heterodimerization with E2F-1 but not with DP-1. Although E2FBP1 lacks the transactivation domain, it stimulates E2F site-dependent transcription in cooperation with E2F-1.


Accelerometric Assessment of Postural Balance in Children: A Systematic Review.
The correct development of postural control in children is fundamental to ensure that they fully reach their psychomotor capacities. However, this capacity is one of the least studied in the clinical and academic scope regarding children. The objective of this study was to analyze the degree of implementation of accelerometry as an evaluation technique for postural control in children and how it is being used. A systematic search was conducted in PubMed, SpringerLink, SportsDiscus, Medline, Scopus, and Web of Science with the following terms: balance, postural control, children, kids, accelerometry, and accelerometer. The search generated a total of 18 articles. Two groups of studies were differentiated: those which exclusively included healthy individuals (n = 5) and those which included children with pathologies (n = 13). Accelerometry is being used in children mainly to assess the gait and static balance, as well as to identify the differences between healthy children and children with developmental disorders. Accelerometry has a discrete degree of implementation as an evaluation tool to assess postural control. It is necessary to define a systematic method for the evaluation of postural control in pediatrics, in order to delve into the development of this capacity and its alterations in different neurodevelopmental disorders.


Chiral phosphoric acid-catalyzed enantioselective three-component Povarov reaction using enecarbamates as dienophiles: highly diastereo- and enantioselective synthesis of substituted 4-aminotetrahydroquinolines.
A chiral phosphoric acid (5)-catalyzed three-component Povarov reaction of aldehydes 2, anilines 3, and enecarbamates 4 afforded cis-4-amino-2-aryl(alkyl)-1,2,3,4-tetrahydroquinolines 1 in high yields with excellent diastereoselectivities (>95%) and almost complete enantioselectivities (up to >99% ee). The reaction was applicable to a wide range of anilines bearing electron-donating (OMe) and electron-withdrawing groups (e.g., Cl, CF(3), NO(2)) and allowed, for the first time, aliphatic aldehydes to be employed in the enantioselective Povarov reaction. With β-substituted acyclic enecarbamates, 2,3,4-trisubstituted 1,2,3,4-tetrahydroquinolines with three contiguous stereogenic centers were produced in excellent diastereo- and enantioselectivities (87 to >99% ee). A detailed study of the active catalytic species allowed us to reduce the catalyst loading from 10% to 0.5% with no deterioration of enantiomeric excess. In addition, mechanistic studies allowed us to conclude unequivocally that the Povarov reaction involving enecarbamate as dienophile proceeded via a stepwise mechanism. The key role of the free NH function of the enecarbamate in the success of this transformation was demonstrated. NMR experiments indicating the catalyst-substrate interaction as well as a linear correlation between catalyst and product ee's were also documented.


A comparison of the oral application and injection routes using the onderstepoort biological products fowl typhoid vaccine, its safety, efficacy and duration of protection in commercial laying hens.
This study was undertaken to establish whether the Onderstepoort Biological Products Fowl Typhoid (OBPft) vaccine registered as an injectable vaccine was effective and safe when administered orally to commercial layers. Its efficacy and duration of protection were compared with application by intramuscular injection. Commercial brown layer hens were used as they were found to be highly susceptible to Salmonella gallinarum infections. In the vaccine safety trial birds were euthanased at timed intervals spanning 4 weeks postvaccination. Necropsies were performed and samples were taken and tested. No clinical signs or mortalities could be attributed to the OBPft vaccine nor could active shedding of the vaccine strain be detected. Slight pathological changes were noted with both routes of vaccination; however, these changes were transient, returning to normal within the observation period. The injected groups showed a better serological response with the rapid serum plate agglutination (RSPA) test than the orally vaccinated groups. In the duration of protection trial, birds were challenged at 3-8-week intervals post-vaccination. All unvaccinated birds died. Protection 8 and 16 weeks after vaccination was above 60 %,by 24 weeks after challenge, the vaccine protection was below 30 %. It was found that there was no significant difference (P < 0.05) in the protection offered by either the oral or injected route of vaccination with the OBPft vaccine.


Switching antiretroviral regimes for the treatment of HIV: safety implications.
There are multiple reasons to switch from a virologically successful antiretroviral regimen. Some of them are related to toxicity. Lately, combination antiretroviral treatment (cART) switches have often been related to drug-drug interactions which may also eventually entail safety issues as well. The purpose of this review is to analyze causes of switching between virologically successful cART regimes related to safety issues. The most relevant papers were selected and summarized. Switching cART has been a popular strategy to address safety issues throughout the antiretroviral era. The myriad of switching studies have paralleled the study and release into clinical practice of new antiretroviral drugs with different and often improved safety profiles. Most of them have been successful in improving antiretroviral toxicity while keeping HIV replication under control. However, it should be taken into account that, whenever a new drug is given, there is a possibility of new drug-related toxicity. Notwithstanding that, an increase in cART switching is foreseen, given the fact that we have a wide antiretroviral drug armamentarium and that people living with HIV are ageing and thus more prone to developing age-related co-morbidities whose therapies may entail new interactions and eventually new toxicities.


Dynamic Reorganization of Extremely Long-Range Promoter-Promoter Interactions between Two States of Pluripotency.
Serum-to-2i interconversion of mouse embryonic stem cells (mESCs) is a valuable in vitro model for early embryonic development. To assess whether 3D chromatin organization changes during this transition, we established Capture Hi-C with target-sequence enrichment of DNase I hypersensitive sites. We detected extremely long-range intra- and inter-chromosomal interactions between a small subset of H3K27me3 marked bivalent promoters involving the Hox clusters in serum-grown cells. Notably, these promoter-mediated interactions are not present in 2i ground-state pluripotent mESCs but appear upon their further development into primed-like serum mESCs. Reverting serum mESCs to ground-state 2i mESCs removes these promoter-promoter interactions in a spatiotemporal manner. H3K27me3, which is largely absent at bivalent promoters in ground-state 2i mESCs, is necessary, but not sufficient, to establish these interactions, as confirmed by Capture Hi-C on Eed(-/-) serum mESCs. Our results implicate H3K27me3 and PRC2 as critical players in chromatin alteration during priming of ESCs for differentiation.


Genetic and physical interactions between Tel2 and the Med15 Mediator subunit in Saccharomyces cerevisiae.
In budding yeast, the highly conserved Tel2 protein is part of several complexes and its main function is now believed to be in the biogenesis of phosphatidyl inositol 3-kinase related kinases. To uncover potentially novel functions of Tel2, we set out to isolate temperature-sensitive (ts) mutant alleles of TEL2 in order to perform genetic screenings. MED15/GAL11, a subunit of Mediator, a general regulator of transcription, was isolated as a suppressor of these mutants. The isolated tel2 mutants exhibited a short telomere phenotype that was partially rescued by MED15/GAL11 overexpression. The tel2-15 mutant was markedly deficient in the transcription of EST2, coding for the catalytic subunit of telomerase, potentially explaining the short telomere phenotype of this mutant. In parallel, a two-hybrid screen identified an association between Tel2 and Rvb2, a highly conserved member of the AAA+ family of ATPases further found by in vivo co-immunoprecipitation to be tight and constitutive. Transiently overproduced Tel2 and Med15/Gal11 associated together, suggesting a potential role for Tel2 in transcription. Other Mediator subunits, as well as SUA7/TFIIB, also rescued the tel2-ts mutants. Altogether, the present data suggest the existence of a novel role for Tel2, namely in transcription, possibly in cooperation with Rvb2 and involving the existence of physical interactions with the Med15/Gal11 Mediator subunit.


Odontonema cuspidatum and Psychotria punctata, Two New Hosts of Cucumber mosaic virus in the United States.
Cucumber mosaic virus (CMV) has a reported host range of 750 to 1,200 species (2,3) that includes weeds, wild plants, crops, and ornamentals. Two new CMV hosts were recently identified in Florida. In July 2011, leaves of Odontonema cuspidatum (firespike), a member of the Acanthaceae, with virus-like symptoms were sent to FDACS-DPI. Firespike is an ornamental shrub native to Mexico with evergreen ovate leaves tapering to a pointed tip. Leaf symptoms included severe leaf distortion with some subtle yellowing or mosaic on younger leaves. Pink-red crystals were seen in leaf strips stained with the nucleic acid stain Azure A, indicating a viral infection. In January 2012, leaves of Psychotria punctata (dotted wild coffee), a member of the Rubiaceae, with virus-like symptoms were sent to FDACS-DPI. Dotted wild coffee is a small exotic tropical tree found in south Florida with many tiny leaf nodules inhabited by endosymbiotic bacteria. In addition to the nodules, these leaves had many large dark green ringspots surrounded with a yellow halo. Both samples were positive for CMV when tested with ImmunoStrips and/or by conventional ELISA using CMV antiserum (Agdia, Elkhart, IN). To confirm CMV infection, reverse transcriptase (RT)-PCR on total RNA from a leaf sample of each plant species was used with previously published cucumovirus primers (1). An expected ~940 bp product was amplified from each sample and cloned into pGEM-T (Promega, Madison, WI). Ten clones from each plant species were sequenced in both directions. After removal of primer sequences, the 906 bp products were 96.3% identical with each other and showed 96.8 to 98.9% nucleotide identity with CMV sequences from Hungary, the United States, and Austria (GenBank Accession Nos. AF517802, U20668, and HQ916354, respectively). Identification of CMV infection in these two species expands the known host range and therefore the reservoir of this plant virus. This has implications for the ornamental industry in general and Florida farmers in particular. References: (1) S. K. Choi et al. J. Virol. Methods 83:67, 1999. (2) E. J. Sikora. Cucumber Mosaic Virus, Pant Disease Notes, Alabama Cooperative Extensions System, retrieved online at http://www.aces.edu/pubs/docs/A/ANR-0868/ANR-0868.pdf , 2004. (3) T. A. Zitter and J. F. Murphy. The Plant Health Instructor. DOI: 10.1094/PHI-I-2009-0518-01, 2009.


Beta-arrestin-dependent formation of beta2 adrenergic receptor-Src protein kinase complexes.
The Ras-dependent activation of mitogen-activated protein (MAP) kinase pathways by many receptors coupled to heterotrimeric guanine nucleotide binding proteins (G proteins) requires the activation of Src family tyrosine kinases. Stimulation of beta2 adrenergic receptors resulted in the assembly of a protein complex containing activated c-Src and the receptor. Src recruitment was mediated by beta-arrestin, which functions as an adapter protein, binding both c-Src and the agonist-occupied receptor. beta-Arrestin 1 mutants, impaired either in c-Src binding or in the ability to target receptors to clathrin-coated pits, acted as dominant negative inhibitors of beta2 adrenergic receptor-mediated activation of the MAP kinases Erk1 and Erk2. These data suggest that beta-arrestin binding, which terminates receptor-G protein coupling, also initiates a second wave of signal transduction in which the "desensitized" receptor functions as a critical structural component of a mitogenic signaling complex.


The impact of supplemental macular carotenoids in Alzheimer's disease: a randomized clinical trial.
Patients with Alzheimer's disease (AD) exhibit significantly less macular pigment (MP) and poorer vision when compared to control subjects. To investigate supplementation with the macular carotenoids on MP, vision, and cognitive function in patients with AD versus controls. A randomized, double-blind clinical trial with placebo and active arms. 31 AD patients and 31 age-similar control subjects were supplemented for six months with either Macushield (10 mg meso-zeaxanthin [MZ]; 10 mg lutein [L]; 2 mg zeaxanthin [Z]) or placebo (sunflower oil). MP was measured using dual-wavelength autofluorescence (Heidelberg Spectralis®). Serum L, Z, and MZ were quantified by high performance liquid chromatography. Visual function was assessed by best corrected visual acuity and contrast sensitivity (CS). Cognitive function was assessed using a battery of cognition tests, including the Cambridge Neuropsychological Test Automated Battery (CANTAB)). Subjects on the active supplement (for both AD and non-AD controls) exhibited statistically significant improvement in serum concentrations of L, Z, MZ, and MP (p < 0.001, for all) and also CS at (p = 0.039). Also, for subjects on the active supplement, paired samples t-tests exhibited four significant results (from five spatial frequencies tested) in the AD group, and two for the non-AD group, and all indicating improvements in CS. We found no significant changes in any of the cognitive function outcome variables measured (p > 0.05, for all). Supplementation with the macular carotenoids (MZ, Z, and L) benefits patients with AD, in terms of clinically meaningful improvements in visual function and in terms of MP augmentation.


Many people in Scotland now benefit from anticipatory care before they die: an after death analysis and interviews with general practitioners.
Key Information Summaries (KIS) were introduced throughout Scotland in 2013 so that anticipatory care plans written by general practitioners (GPs) could be routinely shared electronically and updated in real time, between GPs and providers of unscheduled and secondary care. We aimed to describe the current reach of anticipatory and palliative care, and to explore GPs' views on using KIS. We studied the primary care records of all patients who died in 2014 in 9 diverse Lothian practices. We identified if anticipatory or palliative care had been started, and if so how many weeks before death and which aspects of care had been documented. We interviewed 10 GPs to understand barriers and facilitating factors. Overall, 60% of patients were identified for a KIS, a median of 18 weeks before death. The numbers identified were highest for patients with cancer, with 75% identified compared with 66% of those dying with dementia/frailty and only 41% dying from organ failure. Patients were more likely to die outside hospital if they had a KIS. GPs identified professional, patient and societal challenges in identifying patients for palliative care, especially those with non-cancer diagnoses. GPs are identifying patients for anticipatory and palliative care more equitably across the different disease trajectories and earlier in the disease process than they were previously identifying patients specifically for palliative care. However, many patients still lack care planning, particularly those dying with organ failure.


Treatment of chronic non-healing ulcers using autologous platelet rich plasma: a case series.
Non-healing ulcers are a major health problem worldwide and have great impact at personal, professional and social levels, with high cost in terms of human and material resources. Recalcitrant non-healing ulcers are inevitable and detrimental to the lower limb and are a major cause of non-traumatic lower limb amputations. Application of autologous Platelet Rich Plasma (PRP) has been a major breakthrough for the treatment of non-healing and diabetic foot ulcers, as it is an easy and cost-effective method, and provides the necessary growth factors that enhance tissue healing. PRP is a conglomeration of thrombocytes, cytokines and various growth factors which are secreted by α-granules of platelets that augment the rate of natural healing process with decrease in time. The purpose of this case series was to evaluate the safety and efficacy of autologous platelet rich plasma for the treatment of chronic non-healing ulcers on the lower extremity. Autologous PRP was prepared from whole blood utilizing a rapid, intraoperative point-of-care system that works on the principle of density gradient centrifugation. Twenty Four (24) patients with non-healing ulcers of different etiologies, who met the inclusion criteria, were treated with single dose of subcutaneous PRP injections along with topical application of PRP gel under compassionate use. The mean age of the treated patients was 62.5 ± 13.53 years and they were followed-up for a period of 24 weeks. All the patients showed signs of wound healing with reduction in wound size, and the mean time duration to ulcer healing was 8.2 weeks. Also, an average five fold increase in the platelet concentrate was observed in the final PRP product obtained using the rapid point-of-care device, and the average platelet dose administered to the patients was 70.10 × 108. This case series has demonstrated the potential safety and efficacy of autologous platelet rich plasma for the treatment of chronic non-healing ulcers. NCT03026855 , Registered 4 January 2017 'Retrospectively'.


Experimental reconstruction of a highly reflecting fiber Bragg grating by using spectral regularization and inverse scattering.
We demonstrate experimentally, for the first time to our knowledge, a reconstruction of a highly reflecting fiber Bragg grating from its complex reflection spectrum by using a regularization algorithm. The regularization method is based on correcting the measured reflection spectrum at the Bragg zone frequencies and enables the reconstruction of the grating profile using the integral-layer-peeling algorithm. A grating with an approximately uniform profile and with a maximum reflectivity of 99.98% was accurately reconstructed by measuring only its complex reflection spectrum.


Microscale locomotion in a nematic liquid crystal.
Microorganisms often encounter anisotropy, for example in mucus and biofilms. We study how anisotropy and elasticity of the ambient fluid affects the speed of a swimming microorganism with a prescribed stroke. Motivated by recent experiments on swimming bacteria in anisotropic environments, we extend a classical model for swimming microorganisms, the Taylor swimming sheet, actuated by small-amplitude traveling waves in a three-dimensional nematic liquid crystal without twist. We calculate the swimming speed and entrained volumetric flux as a function of the swimmer's stroke properties as well as the elastic and rheological properties of the liquid crystal. These results are then compared to previous results on an analogous swimmer in a hexatic liquid crystal, indicating large differences in the cases of small Ericksen number and in a nematic fluid when the tumbling parameter is near the transition to a shear-aligning nematic. We also propose a novel method of swimming or pumping in a nematic fluid by passing a traveling wave of director oscillation along a rigid wall.


Comparative characterization of rep proteins from the helper-dependent adeno-associated virus type 2 and the autonomous goose parvovirus.
Adeno-associated viruses (AAVs) are nonautonomous human parvoviruses in that they are dependent on helper functions supplied by other viruses or on genotoxic stimuli for conditions permissive for replication. In the absence of helper, AAV type 2 enters latency by integration into a specific site on human chromosome 19. This feature of AAV, in combination with a lack of pathogenicity, makes AAV an attractive candidate vector for human gene therapy. Goose parvovirus (GPV) is both autonomous and pathogenic yet is highly homologous to AAV. To address the molecular bases for the different viral lifestyles, we compare the AAV and GPV nonstructural proteins, Rep78 and Rep1, respectively. We find that Rep78 and Rep1 possess several biochemical activities in common, including (i) high-affinity DNA binding for sequences that constitute the minimal DNA replication origin; (ii) nucleoside triphosphate-dependent DNA helicase activity; and (iii) origin-specific replication of double-stranded linear DNA. These experiments also establish a specific 38-bp DNA sequence as the minimal GPV DNA replication origin. It is noteworthy that although the proposed Rep binding sites of GPV and AAV are highly similar, Rep1 and Rep78 show a high degree of specificity for their respective origins, in both binding and replication assays. One significant difference was observed; with the minimal replication origin in adenovirus-uninfected extracts, Rep78-mediated replication exhibited low processivity, as previously reported. In contrast, Rep1 efficiently replicated full-length template. Overall, our studies indicate that GPV Rep1 and AAV Rep78 support a comparable mode of replication. Thus, a comparison of the two proteins provides a model system with which to determine the contribution of Rep in the regulation of dependence and autonomy at the level of DNA replication.


Ultrastructural transmission electron microscopy (TEM) study of hybrid layers formed beneath a one-bottle adhesive system using the total-etch technique and a self-etching system.
To compare, using transmission electron microscopy (TEM), the ultrastructure of the hybrid layers formed beneath a one-bottle adhesive system using the total-etch technique with that of a self-etching system. Occlusal cavity preparations were made in vivo in 18 human premolars and randomly appointed to three groups (n = 6), according to the following bonding procedures: (1) OptiBond Solo, a single-bottle adhesive system, was applied following 15 seconds etching with 37% phosphoric acid and rinsing; (2) Clearfil Liner Bond 2V, a self-etching adhesive system was applied; and (3) no bonding material was used, with the untreated smear layer acting as a control. The cavities were then filled with resin composite restorations. In group 1, dense resin tags obturated the tubules. A layer of inorganic silicon microgranules formed at the top of the interdiffusion zone, and below this was a zone of loosely arranged collagen fibers. Toward the base, there was a more dense accumulation of hydroxyapatite crystals. In group 2, dense resin tags obturated the tubular orifices. Collagen fibers were densely compacted within monomer material. Toward the base, hydroxyapatite crystals were observed between collagen bundles. In group 3, a rough fragmented smear layer covered the dentinal floor and occluded the tubules. Irregularities occurred in the coronal zone of the hybrid layer in both systems. In the one-bottle system, this was attributed to the etching technique, while in the case of the self-etching system, this was attributed to the resin materials. These irregularities are potential sites for debonding in the clinical situation.


Australia antigen and hepatitis in Accra, Ghana.
Viral hepatitis in young adults in Accra, Ghana, is associated with Australia antigen (H.A.A.). Sera from 85 patients in hospital with viral hepatitis were available for determinations of H.A.A. Of the 16 patients whose serum was obtained within the first week of symptoms, 15 were positive. The only factor related to finding H.A.A. was the time between onset of symptoms and the collection of the serum sample. Persistence of H.A.A. was associated with persistence of jaundice in men but not in women. Previous epidemiological studies in Accra found no evidence for parenteral transmission of viral hepatitis and showed a shanty-town predilection pointing to faecal-oral transmission. It thus seems that H.A.A.-associated hepatitis is transmitted in West Africa either faecal-orally or by shanty-town associated arthropods. The finding that H.A.A. hepatitis is the usual hepatitis in young adults in Accra is in accord with the high prevalence of H.A.A. elsewhere in the general population in Africa and may be related to the high rate of cirrhosis and hepatoma in Africa.


Healthcare personnel exposure in an emergency department during influenza season.
Healthcare personnel are at high risk for exposure to influenza by direct and indirect contact, droplets and aerosols, and by aerosol generating procedures. Information on air and surface influenza contamination is needed to assist in developing guidance for proper prevention and control strategies. To understand the vulnerabilities of healthcare personnel, we measured influenza in the breathing zone of healthcare personnel, in air and on surfaces within a healthcare setting, and on filtering facepiece respirators worn by healthcare personnel when conducting patient care. Thirty participants were recruited from an adult emergency department during the 2015 influenza season. Participants wore personal bioaerosol samplers for six hours of their work shift, submitted used filtering facepiece respirators and medical masks and completed questionnaires to assess frequency and types of interactions with potentially infected patients. Room air samples were collected using bioaerosol samplers, and surface swabs were collected from high-contact surfaces within the adult emergency department. Personal and room bioaerosol samples, surface swabs, and filtering facepiece respirators were analyzed for influenza A by polymerase chain reaction. Influenza was identified in 42% (53/125) of personal bioaerosol samples, 43% (28/ 96) of room bioaerosol samples, 76% (23/30) of pooled surface samples, and 25% (3/12) of the filtering facepiece respirators analyzed. Influenza copy numbers were greater in personal bioaerosol samples (17 to 631 copies) compared to room bioaerosol samples (16 to 323 copies). Regression analysis suggested that the amount of influenza in personal samples was approximately 2.3 times the amount in room samples (Wald χ2 = 16.21, p<0.001). Healthcare personnel may encounter increased concentrations of influenza virus when in close proximity to patients. Occupations that require contact with patients are at an increased risk for influenza exposure, which may occur throughout the influenza season. Filtering facepiece respirators may become contaminated with influenza when used during patient care.


Factors Impacting Transgender Patients' Discomfort with Their Family Physicians: A Respondent-Driven Sampling Survey.
Representing approximately 0.5% of the population, transgender (trans) persons in Canada depend on family physicians for both general and transition-related care. However, physicians receive little to no training on this patient population, and trans patients are often profoundly uncomfortable and may avoid health care. This study examined factors associated with patient discomfort discussing trans health issues with a family physician in Ontario, Canada. 433 trans people age 16 and over were surveyed using respondent-driven sampling for the Trans PULSE Project; 356 had a family physician. Weighted logistic regression models were fit to produce prevalence risk ratios (PRRs) via average marginal predictions, for transmasculine (n = 184) and transfeminine (n = 172) trans persons. Among the 83.1% (95% CI = 77.4, 88.9) of trans Ontarians who had a family physician, approximately half reported discomfort discussing trans health issues. 37.2% of transmasculine and 38.1% of transfeminine persons reported at least one trans-specific negative experience. In unadjusted analysis, sociodemographics did not predict discomfort, but those who planned to medically transition sex, but had not begun, were more likely to report discomfort (transmasculine: PRR = 2.62 (95% CI = 1.44, 4.77); transfeminine: PRR = 1.85 (95% CI = 1.08, 3.15)). Adjusted for other factors, greater perceived physician knowledge about trans issues was associated with reduced likelihood of discomfort, and previous trans-specific negative experiences with a family physician with increased discomfort. Transfeminine persons who reported three or more types of negative experiences were 2.26 times as likely, and transmasculine persons 1.61 times as likely, to report discomfort. In adjusted analyses, sociodemographic associations differed by gender, with being previously married or having higher education associated with increased risk of discomfort among transfeminine persons, but decreased risk among transmasculine persons. Within this transgender population, discomfort in discussing trans health issues with a family physician was common, presenting a barrier to accessing primary care despite having a regular family physician and "universal" health insurance.


2-deoxyglucose transiently inhibits yeast AMPK signaling and triggers glucose transporter endocytosis, potentiating the drug toxicity.
2-deoxyglucose is a glucose analog that impacts many aspects of cellular physiology. After its uptake and its phosphorylation into 2-deoxyglucose-6-phosphate (2DG6P), it interferes with several metabolic pathways including glycolysis and protein N-glycosylation. Despite this systemic effect, resistance can arise through strategies that are only partially understood. In yeast, 2DG resistance is often associated with mutations causing increased activity of the yeast 5'-AMP activated protein kinase (AMPK), Snf1. Here we focus on the contribution of a Snf1 substrate in 2DG resistance, namely the alpha-arrestin Rod1 involved in nutrient transporter endocytosis. We report that 2DG triggers the endocytosis of many plasma membrane proteins, mostly in a Rod1-dependent manner. Rod1 participates in 2DG-induced endocytosis because 2DG, following its phosphorylation by hexokinase Hxk2, triggers changes in Rod1 post-translational modifications and promotes its function in endocytosis. Mechanistically, this is explained by a transient, 2DG-induced inactivation of Snf1/AMPK by protein phosphatase 1 (PP1). We show that 2DG-induced endocytosis is detrimental to cells, and the lack of Rod1 counteracts this process by stabilizing glucose transporters at the plasma membrane. This facilitates glucose uptake, which may help override the metabolic blockade caused by 2DG, and 2DG export-thus terminating the process of 2DG detoxification. Altogether, these results shed a new light on the regulation of AMPK signaling in yeast and highlight a remarkable strategy to bypass 2DG toxicity involving glucose transporter regulation.


miR-296-3p, miR-298-5p and their downstream networks are causally involved in the higher resistance of mammalian pancreatic α cells to cytokine-induced apoptosis as compared to β cells.
The molecular bases of mammalian pancreatic α cells higher resistance than β to proinflammatory cytokines are very poorly defined. MicroRNAs are master regulators of cell networks, but only scanty data are available on their transcriptome in these cells and its alterations in diabetes mellitus. Through high-throughput real-time PCR, we analyzed the steady state microRNA transcriptome of murine pancreatic α (αTC1-6) and β (βTC1) cells: their comparison demonstrated significant differences. We also characterized the alterations of αTC1-6 cells microRNA transcriptome after treatment with proinflammatory cytokines. We focused our study on two microRNAs, miR-296-3p and miR-298-5p, which were: (1) specifically expressed at steady state in αTC1-6, but not in βTC1 or INS-1 cells; (2) significantly downregulated in αTC1-6 cells after treatment with cytokines in comparison to untreated controls. These microRNAs share more targets than expected by chance and were co-expressed in αTC1-6 during a 6-48 h time course treatment with cytokines. The genes encoding them are physically clustered in the murine and human genome. By exploiting specific microRNA mimics, we demonstrated that experimental upregulation of miR-296-3p and miR-298-5p raised the propensity to apoptosis of transfected and cytokine-treated αTC1-6 cells with respect to αTC1-6 cells, treated with cytokines after transfection with scramble molecules. Both microRNAs control the expression of IGF1Rβ, its downstream targets phospho-IRS-1 and phospho-ERK, and TNFα. Our computational analysis suggests that MAFB (a transcription factor exclusively expressed in pancreatic α cells within adult rodent islets of Langerhans) controls the expression of miR-296-3p and miR-298-5p. Altogether, high-throughput microRNA profiling, functional analysis with synthetic mimics and molecular characterization of modulated pathways strongly suggest that specific downregulation of miR-296-3p and miR-298-5p, coupled to upregulation of their targets as IGF1Rβ and TNFα, is a major determinant of mammalian pancreatic α cells resistance to apoptosis induction by cytokines.


Designing for Motivation, Engagement and Wellbeing in Digital Experience.
Research in psychology has shown that both motivation and wellbeing are contingent on the satisfaction of certain psychological needs. Yet, despite a long-standing pursuit in human-computer interaction (HCI) for design strategies that foster sustained engagement, behavior change and wellbeing, the basic psychological needs shown to mediate these outcomes are rarely taken into account. This is possibly due to the lack of a clear model to explain these needs in the context of HCI. Herein we introduce such a model: Motivation, Engagement and Thriving in User Experience (METUX). The model provides a framework grounded in psychological research that can allow HCI researchers and practitioners to form actionable insights with respect to how technology designs support or undermine basic psychological needs, thereby increasing motivation and engagement, and ultimately, improving user wellbeing. We propose that in order to address wellbeing, psychological needs must be considered within five different spheres of analysis including: at the point of technology adoption, during interaction with the interface, as a result of engagement with technology-specific tasks, as part of the technology-supported behavior, and as part of an individual's life overall. These five spheres of experience sit within a sixth, society, which encompasses both direct and collateral effects of technology use as well as non-user experiences. We build this model based on existing evidence for basic psychological need satisfaction, including evidence within the context of the workplace, computer games, and health. We extend and hone these ideas to provide practical advice for designers along with real world examples of how to apply the model to design practice.


Molecular genetics of mouse serotonin neurons across the lifespan.
New molecular genetics approaches have been developed over the past several years to study brain serotonin (5-HT) neuron development and the roles of 5-HT neurons in behavior and physiology. These approaches were enabled by manipulation of the gene encoding the Pet-1 ETS transcription factor whose expression in the brain is restricted to developing and adult 5-HT neurons. Targeting of the Pet-1 gene led to the development of a mouse line with a severe and stable deficiency of embryonic 5-HT-synthesizing neurons. The Pet-1 transcription regulatory region has been used to create several new 5-HT neuron-type transgenic tools that have greatly increased the experimental accessibility of the small number of brain 5-HT neurons. Permanent and specific marking of 5-HT neurons with Pet-1-based transgenic tools have now been used for flow cytometry, whole cell electrophysiological recordings, progenitor fate mapping, and live time lapse imaging of these neurons. Additional tools provide multiple strategies for conditional temporal targeting of gene expression in 5-HT neurons at different stages of life. Pet-1-based approaches have led to advances in understanding the role of 5-HT neurons in respiration, thermoregulation, emotional behaviors, maternal behavior, and the mechanism of antipsychotic drug actions. In addition, these approaches have begun to reveal the molecular basis of 5-HT neuron heterogeneity and the transcriptional mechanisms that direct 5-HT neuron-type identity, maturation, and maintenance.


Efficacy and safety of gabapentin for treatment of postherpetic neuralgia: a meta-analysis of randomized controlled trials.
Postherpetic neuralgia (PHN) is a common type of neuropathic pain occurring after resolution of herpes zoster rash. Although gabapentin is a widely used treatment, some disagreements exist about its efficacy and safety. Meta-analysis was performed to better evaluate the efficacy and safety of gabapentin for management of PHN. Randomized, double-blind, placebo-controlled trials of gabapentin to treat PHN were identified by searching MEDLINE, EMBASE, and CENTRAL databases. Searches were restricted to studies published in English. Seven trials involving a total of 2039 participants were identified. Pooled analysis showed that gabapentin reduced PHN-related pain significantly more than placebo (mean difference, MD=-0.89, 95% CI -1.58 to -0.18, P<0.001). Gabapentin reduced pain below baseline by at least 50% in significantly more patients than did placebo (RR=1.59, 95% CI 1.35 to 1.88, P<0.001). Gabapentin was significantly more likely than placebo to lead patients to rate their global impression of change as "much improved" or "very much improved" (RR=1.82, 95% CI 1.41 to 2.35, P=0.003). Gabapentin also improved sleep quality significantly more than did placebo (MD=-0.62, 95% CI -0.67 to -0.57, P<0.001). On the other hand, patients given gabapentin were significantly more likely to experience dizziness, somnolence, peripheral edema, ataxia or gait disturbance and diarrhea. Subgroup analysis on formulation of gabapentin showed that gabapentin enacarbil had similar efficacy of pain relief with other formulations while it may be superior to others in term of compliance and safety. This meta-analysis indicates that gabapentin is an effective and well-tolerated treatment for patients with PHN.


Knowledge Level on Administration of Chemotherapy through Peripheral and Central Venous Catheter among Oncology Nurses.
The aim of this study is to determine the knowledge levels of oncology nurses about peripheral and central venous catheter during their chemotherapy administration. Data collection of this descriptive study was started on April 15, 2015-July 15, 2015. The data presented in this summary belong to 165 nurses. Data were collected with data collection form including questions related to sociodemographic qualifications and knowledge levels of nurses. Data collection forms were E-mailed to the members of Turkish Oncology Nursing Society. Data presented with numbers, percentages, and mean ± standard deviation. The mean age of nurses was 33.60 ± 7.34 years and mean duration for oncology nursing experience was 2.65 ± 0.91 years. Nurses had correct information about the importance of selecting peripheral venous catheter and choosing the placement area for chemotherapy administration (63.6%), control of catheter before the administration (93.9%), influence of chemotherapeutic agent on length of catheter (40.6%), and management of extravasation (75.7%). Nurses also had correct information about the first use of port catheter (67.3%) and checking the catheter whether it is working properly or not (75.8%). In General, nurses' level of knowledge related to catheter is 50% and higher. It is recommended to increase the knowledge of nurses about evidence-based information for catheter care as a step to safe chemotherapy practice.


Synthesis of structurally well-defined and liquid-phase-processable graphene nanoribbons.
The properties of graphene nanoribbons (GNRs) make them good candidates for next-generation electronic materials. Whereas 'top-down' methods, such as the lithographical patterning of graphene and the unzipping of carbon nanotubes, give mixtures of different GNRs, structurally well-defined GNRs can be made using a 'bottom-up' organic synthesis approach through solution-mediated or surface-assisted cyclodehydrogenation reactions. Specifically, non-planar polyphenylene precursors were first 'built up' from small molecules, and then 'graphitized' and 'planarized' to yield GNRs. However, fabrication of processable and longitudinally well-extended GNRs has remained a major challenge. Here we report a bottom-up solution synthesis of long (>200 nm) liquid-phase-processable GNRs with a well-defined structure and a large optical bandgap of 1.88 eV. Self-assembled monolayers of GNRs can be observed by scanning probe microscopy, and non-contact time-resolved terahertz conductivity measurements reveal excellent charge-carrier mobility within individual GNRs. Such structurally well-defined GNRs may prove useful for fundamental studies of graphene nanostructures, as well as the development of GNR-based nanoelectronics.


Supplementation with Whey Protein, Omega-3 Fatty Acids and Polyphenols Combined with Electrical Muscle Stimulation Increases Muscle Strength in Elderly Adults with Limited Mobility: A Randomized Controlled Trial.
Age-related sarcopenia is a progressive and generalized skeletal muscle disorder associated with adverse outcomes. Herein, we evaluate the effects of a combination of electrical muscle stimulation (EMS) and a whey-based nutritional supplement (with or without polyphenols and fish oil-derived omega-3 fatty acids) on muscle function and size. Free-living elderly participants with mobility limitations were included in this study. They received 2 sessions of EMS per week and were randomly assigned to ingest an isocaloric beverage and capsules for 12 weeks: (1) carbohydrate + placebo capsules (CHO, n = 12), (2) whey protein isolate + placebo capsules (WPI, n = 15) and (3) whey protein isolate + bioactives (BIO) capsules containing omega-3 fatty acids, rutin, and curcumin (WPI + BIO, n = 10). The change in knee extension strength was significantly improved by 13% in the WPI + BIO group versus CHO on top of EMS, while WPI alone did not provide a significant benefit over CHO. On top of this, there was the largest improvement in gait speed (8%). The combination of EMS and this specific nutritional intervention could be considered as a new approach for the prevention of sarcopenia but more work is needed before this approach should be recommended. This trial was registered at the Japanese University Hospital Medical Information Network (UMIN) clinical trial registry (UMIN000008382).


A quasi-experimental study on the effect of health and food safety training intervention on restaurant food handlers during the COVID-19 pandemic.
The restaurant business has turned into a dynamic and ever-growing industry. So, food safety must be a priority for these establishments, especially during the COVID-19 pandemic. The aim of this study was to determine the effect of training intervention on the health and food safety knowledge, attitude, and self-reported practice (KAP) of restaurant food handlers during the COVID-19 pandemic. This quasi-experimental study was conducted on 159 restaurant food handlers in Tehran, Iran. The training intervention was developed based on the latest global guidelines. The KAP of the subjects was measured before and after the training. Fisher's exact test, paired t test, and repeated measures ANOVA were used for statistical analysis. Data analysis was done using the IBM_SPSS software. The total knowledge scores of participants were low (17.6%), moderate (35.2%), and good (47.2%) before training, which were changed to 5% (low), 23.9% (moderate), and 71.1% (good) after training. The total pretraining attitude scores were 0.6, 77.4, 18.2, and 3.8% that were changed to 0% (strongly negative), 49.1% (negative), 33.3% (positive), and 17.6% (strongly positive), respectively. Also, the self-reported practice scores of the participants before training were 1.3, 56, and 42.7 that were changed to 0% (weak), 26.4% (acceptable), and 73.6% (desirable) after the intervention, respectively. Paired t test results showed a statistically significant increase in all scores. The interaction of training with age and education was statistically significant in increasing the knowledge and attitude scores of the participants by the repeated measures ANOVA. Improving the KAP of food handlers by health and food safety training can improve the status of restaurants and minimize the outbreak of pandemic diseases, including COVID-19, which is an effective step in community health. Thus, it is an urgent need for policymakers to design an online system of continuous food safety training for food handlers.


Computed tomography and magnetic resonance imaging of the equine head.
Horses are frequently presented with clinical conditions warranting diagnostic imaging of the head. Unfortunately, evaluation of the equine head can be difficult using conventional radiography. The anatomic complexity and superimposition of the osseous, dental, and soft tissue structures complicate radiographic interpretation and diagnoses. Multiple radiographic projections may be necessary to image specific regions, and side-to-side comparison is often essential.


Marsilea vestita: Conversion of the Water Form to the Land Form by Darkness and by Far-Red Light.
A period in darkness or in continuous far-red light can cause the ferns of the species Marsilea vestita Hook. and Grev. to develop as a land form in a medium which normally allows only the water form to develop. Far-red light is more effective than darkness in causing this conversion. The difference in the quality of light reaching plants growing in water or on land may cause this conversion to occur in nature.


Agonist-stimulated cytoskeletal reorganization and signal transduction at focal adhesions in vascular smooth muscle cells require c-Src.
Thrombin and angiotensin II (angII) have trophic properties as mediators of vascular remodeling. Focal adhesions and actin cytoskeleton are involved in cell growth, shape, and movement and may be important in vascular remodeling. To characterize mechanisms by which thrombin and angII modulate vessel structure, we studied the effects of these G protein-coupled receptor ligands on focal adhesions in vascular smooth muscle cells (VSMCs). Both thrombin and angII stimulated bundling of actin filaments to form stress fibers, assembly of focal adhesions, and protein tyrosine phosphorylation at focal adhesions, such as p130Cas, paxillin, and tensin. To test whether c-Src plays a critical role in focal adhesion rearrangement, we analyzed cells with altered c-Src activity by retroviral transduction of wild-type (WT) and kinase-inactive (KI) c-Src into rat VSMCs, and by use of VSMCs from WT (src+/+) and Src-deficient (src-/-) mice. Tyrosine phosphorylation of Cas, paxillin, and tensin were markedly decreased in VSMCs expressing KI-Src and in src-/- VSMCs. Expression of KI-Src did not inhibit stress fiber formation by thrombin. Surprisingly, actin bundling was markedly decreased in VSMCs from src-/- mice both basally and after thrombin stimulation, compared with src+/+ mice. We also studied the effect of KI-Src and WT-Src on VSMC spreading. Expression of KI-Src reduced the rate of VSMC spreading on collagen, whereas WT-Src enhanced cell spreading. In conclusion, c-Src plays a critical role in agonist-stimulated cytoskeletal reorganization and signal transduction at focal adhesions in VSMCs. c-Src kinase activity is required for the cytoskeletal turnover that occurs in cell spreading, whereas c-Src appears to regulate actin bundling via a kinase-independent mechanism.


Immunological study of hydatidosis. I. Evaluation of immunoelectrodiffusion tests and enzyme-linked immunoelectrodiffusion assay (ELIEDA) in human hydatidosis.
The comparative sensitivity, specificity and rapidity of immunoelectrodiffusion (IED) on cellulose acetate membranes and of immunoelectrophoresis (IEP) on agarose gel, were evaluated in the diagnosis of hydatidosis. Pooled crude fluid from several cattle hydatis cysts was used as antigen in tests on 1,750 non-hydatis and 400 hydatis sera obtained from patients with hepatic, pulmonary, splenic, peritoneal and cerebral hydatidosis before and after surgery. Coupled to a specific human reference serum for arc 5, IED shows a specificity comparable to that of IEP but its sensitivity is slightly higher and the amount of antigen needed is very small. The appearance of a typical "gloved finger" pattern in sera from patients with ruptured cysts emphasizes the interest of quick results (3 hours) obtained by this method. In order to increase the sensitivity of IED and to define the class of immunoglobulins involved in the antigen-antibody reaction, we have coupled this method to an enzymatic technique. The immune complexes precipitated by IED were treated with peroxidase-labelled antibodies specific to each class of human immunoglobulins. The specificity of this enzyme-linked immunoelectrodiffusion assay (ELIEDA) permits one to follow the immunologic evolution of hydatidosis and to identify IgM in ruptured hepatic cysts and IgA in pulmonary cysts.


Nature of variability in saccades.
We studied the variability in saccades by comparing the peak velocities of saccades with the same target amplitude made with different actual amplitudes. We tested three hypotheses: the pulse-height noise hypothesis (peak velocity and amplitude vary proportionally), the localization noise hypothesis (variability in amplitude and peak velocity lie along the main sequence), and the independent noise hypothesis (variability in amplitude and peak velocity are independent). We measured eye orientation in two experiments by a scleral coil and a video system. Surprisingly, the main source of variability of saccades depended on the measurement system used. A combination of localization noise and independent noise best describes the data obtained by the video system. The independent noise (e.g., measurement inaccuracy) was the main source of variability. For the scleral coils, the variability was considerably larger than for the less accurate video system. The pulse-height noise hypothesis best describes this additional variability. Therefore we conclude that pulse-height noise is the main source of variability in saccades measured with scleral coils. We discuss the influence of scleral coils on saccade generation and suggest that a change in motor strategy due to the discomfort of wearing the coils might be the cause of the increased variability.


Active convulsive epilepsy in a rural district of Kenya: a study of prevalence and possible risk factors.
Few large-scale studies of epilepsy have been done in sub-Saharan Africa. We aimed to estimate the prevalence of, treatment gap in, and possible risk factors for active convulsive epilepsy in Kenyan people aged 6 years or older living in a rural area. We undertook a three-phase screening survey of 151,408 individuals followed by a nested community case-control study. Treatment gap was defined as the proportion of cases of active convulsive epilepsy without detectable amounts of antiepileptic drugs in blood. Overall prevalence of active convulsive epilepsy was 2.9 per 1000 (95% CI 2.6-3.2); after adjustment for non-response and sensitivity, prevalence was 4.5 per 1000 (4.1-4.9). Substantial heterogeneity was noted in prevalence, with evidence of clustering. Treatment gap was 70.3% (65.9-74.5), with weak evidence of a difference by sex and area. Adjusted odds of active convulsive epilepsy for all individuals were increased with a family history of non-febrile convulsions (odds ratio 3.3, 95% CI 2.4-4.7; p<0.0001), family history of febrile convulsions (14.6, 6.3-34.1; p<0.0001), history of both seizure types (7.3, 3.3-16.4; p<0.0001), and previous head injury (4.1, 2.1-8.1; p<0.0001). Findings of multivariable analyses in children showed that adverse perinatal events (5.7, 2.6-12.7; p<0.0001) and the child's mother being a widow (5.1, 2.4-11.0; p<0.0001) raised the odds of active convulsive epilepsy. Substantial heterogeneity exists in prevalence of active convulsive epilepsy in this rural area in Kenya. Assessment of prevalence, treatment use, and demographic variation in screening response helped to identify groups for targeted interventions. Adverse perinatal events, febrile illness, and head injury are potentially preventable associated factors for epilepsy in this region.


Determinants of the risk of dying of HIV/AIDS in a rural South African community over the period of the decentralised roll-out of antiretroviral therapy: a longitudinal study.
Antiretroviral treatment (ART) has significantly reduced HIV mortality in South Africa. The benefits have not been experienced by all groups. Here we investigate the factors associated with these inequities. This study was located in a rural South African setting and used data collected from 2007 to 2010, the period when decentralised ART became available. Approximately one-third of the population were of Mozambican origin. There was a pattern of repeated circular migration between urban areas and this community. Survival analysis models were developed to identify demographic, socioeconomic, and spatial risk factors for HIV mortality. Among the study population of 105,149 individuals, there were 2,890 deaths. The HIV/TB mortality rate decreased by 27% between 2007-2008 and 2009-2010. For other causes of death, the reduction was 10%. Bivariate analysis found that the HIV/TB mortality risk was lower for: those living within 5 km of the Bhubezi Community Health Centre; women; young adults; in-migrants with a longer period of residence; permanent residents; and members of households owning motorised transport, holding higher socioeconomic positions, and with higher levels of education. Multivariate modelling showed, in addition, that those with South Africa as their country of origin had an increased risk of HIV/TB mortality compared to those with Mozambican origins. For males, those of South African origin, and recent in-migrants, the risk of death associated with HIV/TB was significantly greater than that due to other causes. In this community, a combination of factors was associated with an increased risk of dying of HIV/TB over the period of the roll-out of ART. There is evidence for the presence of barriers to successful treatment for particular sub-groups in the population, which must be addressed if the recent improvements in population-level mortality are to be maintained.


Current state of educational compensation in academic neurology: Results of a US national survey.
In the current medical climate, medical education is at risk of being de-emphasized, leading to less financial support and compensation for faculty. A rise in compensation plans that reward clinical or research productivity fails to incentivize and threatens to erode the educational missions of our academic institutions. Aligning compensation with the all-encompassing mission of academic centers can lead to increased faculty well-being, clinical productivity, and scholarship. An anonymous survey developed by members of the A.B. Baker Section on Neurologic Education was sent to the 133 chairs of neurology to assess the type of compensation faculty receive for teaching efforts. Seventy responses were received, with 59 being from chairs. Key results include the following: 36% of departments offered direct compensation; 36% did not; residency program directors received the most salary support at 36.5% full-time equivalent; and administrative roles had greatest weight in determining academic compensation. We believe a more effective, transparent system of recording and rewarding faculty for their educational efforts would encourage faculty to teach, streamline promotions for clinical educators, and strengthen undergraduate and graduate education in neurology.


Genetic profile of ischemic cerebrovascular disease and carotid stenosis.
Carotid artery stenosis (CS) is a major risk factor for ischemic cerebrovascular disease (ICVD) and is therefore of interest in genetic investigating. Here we report the distribution of 100 polymorphisms in 47 suspected susceptibility genes for ICVD and its risk factors. Previously published markers in suspected susceptibility genes were genotyped in ICVD patients and controls (928/602). Genotyping was performed using multiplex polymerase chain reaction (PCR) and linear immobilized probe array assays. ICVD cases were subtyped according to Trial of Org 10172 in Acute Stroke Treatment (TOAST) or subdivided into CS and non-CS patients by ultrasonography in a separate analysis. Three polymorphisms located in the lipoprotein lipase (LPL), angiotensinogen (AGT) and guanine nucleotide-binding protein beta-3 (GNB3) genes were significantly associated with ICVD after correction for age and gender. The strongest association was found for the protective LPL Ser447Term polymorphism. All the significant markers showed varying frequencies in different subphenotypes of ICVD. Factor VII, apolipoprotein E and two renin polymorphisms were differentially frequent in patients with evidence of CS compared with non-CS patients. We have found that some previously described susceptibility polymorphisms are weakly associated with ICVD and that subdivision of patients into CS and non-CS groups may help to identify new candidate polymorphisms.


Mortality from diabetes and ischaemic heart disease in textile workers.
To explore explanations for elevated mortality from diabetes among male garment manufacturers and repairers in England and Wales during 1979-1990, analysis was extended by 10 years, looking also at other textile workers and at deaths from ischaemic heart disease (IHD). Data on some 3.5 million deaths were used to compute proportional mortality ratios (PMRs) for diabetes and IHD, standardised for age and social class, in 10 textile-associated job groups, with additional analyses by place of birth for 1993-2000. Male mortality from diabetes was elevated in nine of the 10 textile jobs, with overall PMRs of 147 (95% CI 131 to 165) during 1979-90 and 170 (95% CI 144 to 199) during 1991-2000. Proportional mortality from IHD was also consistently high. Female mortality from both diseases was close to that for other occupations. In both sexes, mortality from diabetes and IHD was increased among people born in the Indian subcontinent (PMRs 353 and 139 in men; 262 and 130 in women). In men, the proportion of deceased textile workers born in the Indian subcontinent (11.4%) was much higher than for all occupations (1.8%), but not in women (1.1% vs 0.7%). PMRs for male textile workers standardised for place of birth were lower but still significantly elevated (133, 95% CI 110 to 159 for diabetes; 109, 95% CI 105 to 114 for IHD). No obvious occupational hazard explains the increased risk specific to men across a wide range of textile occupations. One possible explanation is uncontrolled residual confounding related to place of birth. This could be tested through suitably designed morbidity surveys.


Genome-Wide Prediction of Metabolic Enzymes, Pathways, and Gene Clusters in Plants.
Plant metabolism underpins many traits of ecological and agronomic importance. Plants produce numerous compounds to cope with their environments but the biosynthetic pathways for most of these compounds have not yet been elucidated. To engineer and improve metabolic traits, we need comprehensive and accurate knowledge of the organization and regulation of plant metabolism at the genome scale. Here, we present a computational pipeline to identify metabolic enzymes, pathways, and gene clusters from a sequenced genome. Using this pipeline, we generated metabolic pathway databases for 22 species and identified metabolic gene clusters from 18 species. This unified resource can be used to conduct a wide array of comparative studies of plant metabolism. Using the resource, we discovered a widespread occurrence of metabolic gene clusters in plants: 11,969 clusters from 18 species. The prevalence of metabolic gene clusters offers an intriguing possibility of an untapped source for uncovering new metabolite biosynthesis pathways. For example, more than 1,700 clusters contain enzymes that could generate a specialized metabolite scaffold (signature enzymes) and enzymes that modify the scaffold (tailoring enzymes). In four species with sufficient gene expression data, we identified 43 highly coexpressed clusters that contain signature and tailoring enzymes, of which eight were characterized previously to be functional pathways. Finally, we identified patterns of genome organization that implicate local gene duplication and, to a lesser extent, single gene transposition as having played roles in the evolution of plant metabolic gene clusters.


Neural Dynamics of Serial Dependence in Numerosity Perception.
Serial dependence-an attractive perceptual bias whereby a current stimulus is perceived to be similar to previously seen ones-is thought to represent the process that facilitates the stability and continuity of visual perception. Recent results demonstrate a neural signature of serial dependence in numerosity perception, emerging very early in the time course during perceptual processing. However, whether such a perceptual signature is retained after the initial processing remains unknown. Here, we address this question by investigating the neural dynamics of serial dependence using a recently developed technique that allowed a reactivation of hidden memory states. Participants performed a numerosity discrimination task during EEG recording, with task-relevant dot array stimuli preceded by a task-irrelevant stimulus inducing serial dependence. Importantly, the neural network storing the representation of the numerosity stimulus was perturbed (or pinged) so that the hidden states of that representation can be explicitly quantified. The results first show that a neural signature of serial dependence emerges early in the brain signals, starting soon after stimulus onset. Critical to the central question, the pings at a later latency could successfully reactivate the biased representation of the initial stimulus carrying the signature of serial dependence. These results provide one of the first pieces of empirical evidence that the biased neural representation of a stimulus initially induced by serial dependence is preserved throughout a relatively long period.


The impact of the congestion charging scheme on air quality in London. Part 2. Analysis of the oxidative potential of particulate matter.
There is growing scientific consensus that the ability of inhaled particulate matter (PM*) to elicit oxidative stress both at the air-lung interface and systemically might underpin many of the acute and chronic respiratory and cardiovascular responses observed in exposed populations. In the current study (which is part two of a two-part HEI study of a congestion charging scheme [CCS] introduced in London, United Kingdom, in 2003), we tested the hypothesis that the reduction in vehicle numbers and changes in traffic composition resulting from the introduction of the CCS would result in decreased concentrations of traffic-specific emissions, both from vehicle exhaust and other sources (brake wear and tire wear), and an associated reduction in the oxidative potential of PM with an aerodynamic diameter < or = 10 microm (PM10). To test this hypothesis, we obtained, extracted, and analyzed tapered element oscillating microbalance (TEOM) PM10 filters from six monitoring sites within, bordering, or outside the area of the congestion charging zone (CCZ) for the 3 years before and after the introduction of the scheme. In addition, from January 2005, TEOM PM10 filters were obtained from an additional 10 sites outside the zone in order to perform the first-ever assessment of within-city spatial variability in the oxidative potential of PM10. Although London's PM10 was found to have remarkably high oxidative potential, it varied markedly between the studied sites, with evidence of increased potential at roadside locations compared with urban background locations. This difference appeared to reflect increased concentrations of copper (Cu), barium (Ba), and bioavailable iron (Fe) in PM10 collected at the roadside sites. PM10's oxidative potential after the introduction of the CCS did not change at the one urban background site within the zone. Yet compositional changes in PM10 were noted at the same site, including significant decreases in Cu and zinc (Zn) content, probably reflecting brake and tire wear (compared with increases in these metals at all sites outside the zone in the 3 years since the scheme's introduction). This pattern of results is consistent with observations of increased vehicle use throughout London in recent years and decreases in the number of vehicles entering the zone since the scheme's introduction.


Nerve transfers in tetraplegia I: Background and technique.
The recovery of hand function is consistently rated as the highest priority for persons with tetraplegia. Recovering even partial arm and hand function can have an enormous impact on independence and quality of life of an individual. Currently, tendon transfers are the accepted modality for improving hand function. In this procedure, the distal end of a functional muscle is cut and reattached at the insertion site of a nonfunctional muscle. The tendon transfer sacrifices the function at a lesser location to provide function at a more important location. Nerve transfers are conceptually similar to tendon transfers and involve cutting and connecting a healthy but less critical nerve to a more important but paralyzed nerve to restore its function. We present a case of a 28-year-old patient with a C5-level ASIA B (international classification level 1) injury who underwent nerve transfers to restore arm and hand function. Intact peripheral innervation was confirmed in the paralyzed muscle groups corresponding to finger flexors and extensors, wrist flexors and extensors, and triceps bilaterally. Volitional control and good strength were present in the biceps and brachialis muscles, the deltoid, and the trapezius. The patient underwent nerve transfers to restore finger flexion and extension, wrist flexion and extension, and elbow extension. Intraoperative motor-evoked potentials and direct nerve stimulation were used to identify donor and recipient nerve branches. The patient tolerated the procedure well, with a preserved function in both elbow flexion and shoulder abduction. Nerve transfers are a technically feasible means of restoring the upper extremity function in tetraplegia in cases that may not be amenable to tendon transfers.


Genetic Variation in the 3'-Untranslated Region of NBN Gene Is Associated with Gastric Cancer Risk in a Chinese Population.
NBN plays a crucial role in carcinogenesis as a core component for both homologous recombination (HR) and non-homologous end-joining (NHEJ) DNA double-strand breaks (DSBs) repair pathways. Genetic variants in the NBN gene have been associated with multiple cancers risk, suggesting pleiotropic effect on cancer. We hypothesized that genetic variants in the NBN gene may modify the risk of gastric cancer. To test this hypothesis, we evaluated the association between four potentially functional single nucleotide polymorphisms in NBN and gastric cancer risk in a case-control study of 1,140 gastric cancer cases and 1,547 controls in a Chinese population. We found that the A allele of rs10464867 (G>A) was significantly associated with a decreased risk of gastric cancer (odds ratio [OR] = 0.81, 95% confidence interval [95% CI] = 0.71-0.94; P = 4.71×10-3). Furthermore, the association between A allele of rs10464867 and decreased risk of gastric cancer was more significantly in elder individuals (per-allele OR = 0.72[0.59-0.88], P = 1.07×10-3), and male individuals (per-allele OR = 0.73[0.62-0.87], P = 3.68×10-4). We further conducted a haplotype analysis and identified that the NBN Ars10464867Grs14448Grs1063053 haplotype conferred stronger protective effect on gastric cancer (OR = 0.76[0.65-0.89], P = 6.39×10-4). In summary, these findings indicate that genetic variants at NBN gene may contribute to gastric cancer susceptibility and may further advance our understanding of NBN gene in cancer development.


Nanoparticulate flurbiprofen reduces amyloid-β42 generation in an in vitro blood-brain barrier model.
The amyloid-β42 (Aβ42) peptide plays a crucial role in the pathogenesis of Alzheimer's disease (AD), the most common neurodegenerative disorder affecting the elderly. Over the past years, several approaches and compounds developed for the treatment of AD have failed in clinical studies, likely in part due to their low penetration of the blood-brain barrier (BBB). Since nanotechnology-based strategies offer new possibilities for the delivery of drugs to the brain, this technique is studied intensively for the treatment of AD and other neurological disorders. The Aβ42 lowering drug flurbiprofen was embedded in polylactide (PLA) nanoparticles by emulsification-diffusion technique and their potential as drug carriers in an in vitro BBB model was examined. First, the cytotoxic potential of the PLA-flurbiprofen nanoparticles on endothelial cells and the cellular binding and uptake by endothelial cells was studied. Furthermore, the biological activity of the nanoparticulate flurbiprofen on γ-secretase modulation as well as its in vitro release was examined. Furthermore, the protein corona of the nanoparticles was studied as well as their ability to transport flurbiprofen across an in vitro BBB model. PLA-flurbiprofen nanoparticles were endocytosed by endothelial cells and neither affected the vitality nor barrier function of the endothelial cell monolayer. The exposure of the PLA-flurbiprofen nanoparticles to human plasma occurred in a rapid protein corona formation, resulting in their decoration with bioactive proteins, including apolipoprotein E. Furthermore, luminally administered PLA-flurbiprofen nanoparticles in contrast to free flurbiprofen were able to modulate γ-secretase activity by selectively decreasing Aβ42 levels in the abluminal compartment of the BBB model. In this study, we were able to show that flurbiprofen can be transported by PLA nanoparticles across an in vitro BBB model and most importantly, the transported flurbiprofen modulated γ-secretase activity by selectively decreasing Aβ42 levels. These results demonstrate that the modification of drugs via embedding in nanoparticles is a promising tool to facilitate drug delivery to the brain, which enables future development for the treatment of neurodegenerative disorders like AD.


The relationship between high-fat dairy consumption and obesity, cardiovascular, and metabolic disease.
To comprehensively review the data on the relationship between the consumption of dairy fat and high-fat dairy foods, obesity, and cardiometabolic disease. We have conducted a systematic literature review of observational studies on the relationship between dairy fat and high-fat dairy foods, obesity, and cardiometabolic disease. We have integrated these findings with data from controlled studies showing effects of several minor dairy fatty acids on adiposity and cardiometabolic risk factors, and data on how bovine feeding practices influence the composition of dairy fat. In 11 of 16 studies, high-fat dairy intake was inversely associated with measures of adiposity. Studies examining the relationship between high-fat dairy consumption and metabolic health reported either an inverse or no association. Studies investigating the connection between high-fat dairy intake and diabetes or cardiovascular disease incidence were inconsistent. We discuss factors that may have contributed to the variability between studies, including differences in (1) the potential for residual confounding; (2) the types of high-fat dairy foods consumed; and (3) bovine feeding practices (pasture- vs. grain-based) known to influence the composition of dairy fat. The observational evidence does not support the hypothesis that dairy fat or high-fat dairy foods contribute to obesity or cardiometabolic risk, and suggests that high-fat dairy consumption within typical dietary patterns is inversely associated with obesity risk. Although not conclusive, these findings may provide a rationale for future research into the bioactive properties of dairy fat and the impact of bovine feeding practices on the health effects of dairy fat.


Association between Cigarette Smoking and Histotype-Specific Epithelial Ovarian Cancer: A Review of Epidemiologic Studies.
Many studies have demonstrated that smoking can influence ovarian cancer risk and survival; however, the number of studies investigating this relationship according to histologic subtypes is limited. We conducted a review of epidemiologic research that assessed the role of smoking on ovarian cancer risk and survival after diagnosis, specifically capturing studies that discerned between various histologic subtypes of this disease. In the majority of studies, current smoking was associated with increased risk of mucinous cancer. There was also evidence of a decreased risk of clear cell and endometrioid histotypes. No significant association was observed between cigarette smoking and serous cancer. In the studies investigating the relationship between smoking and survival, all the studies reported an increased risk of mortality associated with smoking. Smoking appeared to be a risk factor for both ovarian cancer risk and mortality. Future studies need to investigate further a potential link between smoking and ovarian cancer by having a better assessment of exposure to smoking and having a larger number of participants with the ability to detect associations within rare histotypes.


Measurement of cerebral blood volume in mouse brain regions using micro-computed tomography.
Micro-computed tomography (micro-CT) is an X-ray imaging technique that can produce detailed 3D images of cerebral vasculature. This paper describes the development of a novel method for using micro-CT to measure cerebral blood volume (CBV) in the mouse brain. As an application of the methodology, we test the hypotheses that differences in CBV exist over anatomical brain regions and that high energy demanding primary sensory regions of the cortex have locally elevated CBV, which may reflect a vascular specialization. CBV was measured as the percentage of tissue space occupied by a radio-opaque silicon rubber that fills the vasculature. To ensure accuracy of the CBV measurements, several innovative refinements were made to standard micro-CT specimen preparation and analysis procedures. Key features of the described method are vascular perfusion under controlled pressure, registration of the micro-CT images to an MRI anatomical brain atlas and re-scaling of micro-CT intensities to CBV units with selectable exclusion of major vessels. Histological validation of the vascular perfusion showed that the average percentage of vessels filled was 93+/-3%. Comparison of thirteen brain regions in nine mice revealed significant differences in CBV between regions (p<0.0001) while cortical maps showed that primary visual and auditory areas have higher CBV than primary somatosensory areas.


Towards a neurobiological understanding of pain in neurofibromatosis type 1: mechanisms and implications for treatment.
Neurofibromatosis type 1 (NF1) is the most common of a group of rare diseases known by the term, "Neurofibromatosis," affecting 1 in 3000 to 4000 people. NF1 patients present with, among other disease complications, café au lait patches, skin fold freckling, Lisch nodules, orthopedic complications, cutaneous neurofibromas, malignant peripheral nerve sheath tumors, cognitive impairment, and chronic pain. Although NF1 patients inevitably express pain as a debilitating symptom of the disease, not much is known about its manifestation in the NF1 disease, with most current information coming from sporadic case reports. Although these reports indicate the existence of pain, the molecular signaling underlying this symptom remains underexplored, and thus, we include a synopsis of the literature surrounding NF1 pain studies in 3 animal models: mouse, rat, and miniswine. We also highlight unexplored areas of NF1 pain research. As therapy for NF1 pain remains in various clinical and preclinical stages, we present current treatments available for patients and highlight the importance of future therapeutic development. Equally important, NF1 pain is accompanied by psychological complications in comorbidities with sleep, gastrointestinal complications, and overall quality of life, lending to the importance of investigation into this understudied phenomenon of NF1. In this review, we dissect the presence of pain in NF1 in terms of psychological implication, anatomical presence, and discuss mechanisms underlying the onset and potentiation of NF1 pain to evaluate current therapies and propose implications for treatment of this severely understudied, but prevalent symptom of this rare disease.


Projection subtypes of rat neostriatal matrix cells revealed by intracellular injection of biocytin.
Individual neostriatal-matrix spiny neurons were stained intracellularly with biocytin after intracellular recording in vivo, and their axons were traced into the globus pallidus (GP), entopeduncular nucleus (EP), and/or substantia nigra (SN). The locations of the neurons within the matrix compartment of the neostriatum (NS) were established by immunocytochemical counterstaining of sections containing the cell bodies using antibodies for calbindin-D28K. This allowed nearly complete visualization of the axonal projections of single NS neurons. On the basis of their intrastriatal axonal arborizations, matrix spiny neurons could be divided into 2 types. One type, which was the more common, had local axonal arborizations restricted to the region of the dendritic field, often with axon collaterals arborizing within the dendritic field of the cells of origin. A second, less common, cell type in the matrix had local axon collaterals distributed widely in the NS. Among matrix neurons with restricted local collateral fields, 3 subtypes could be distinguished on the basis of their efferent axonal projections. Type I cells projected only to the GP. Type IIa cells projected to the GP, EP, and SN pars reticulata. Type IIb cells projected to the GP and SN but not to the EP. The shapes and densities of the GP arborizations varied in the 3 cell types, with the cells projecting only to the GP (type I) projecting more heavily and filling a larger volume there than type II cells. The dendrites and intrastriatal axon collaterals of 3 subtypes were similar in morphology. The class of matrix spiny neurons with intrastriatal axon collaterals distributed widely in the NS were observed to project to the GP. Projections beyond the GP were not identified for this cell type, but could not be ruled out. Somatodendritic morphologies of neurons did not differ according to the projection site. These results demonstrate that NS matrix spiny cells are more heterogeneous in their efferent projection patterns than previously suspected on the basis of retrograde axonal tracing and immunocytochemical studies. As predicted by those previous studies, there is a class of matrix neurons that projects only to the GP. Presumably, these cells contain enkephalin. Cells projecting to the SN and EP, and so presumably containing substance P, give off a small projection to the GP, as well, and differ in their collateralization patterns within the 3 major target nuclei.


An extensive circuitry for cell wall regulation in Candida albicans.
Protein kinases play key roles in signaling and response to changes in the external environment. The ability of Candida albicans to quickly sense and respond to changes in its environment is key to its survival in the human host. Our guiding hypothesis was that creating and screening a set of protein kinase mutant strains would reveal signaling pathways that mediate stress response in C. albicans. A library of protein kinase mutant strains was created and screened for sensitivity to a variety of stresses. For the majority of stresses tested, stress response was largely conserved between C. albicans, Saccharomyces cerevisiae, and Schizosaccharomyces pombe. However, we identified eight protein kinases whose roles in cell wall regulation (CWR) were not expected from functions of their orthologs in the model fungi Saccharomyces cerevisiae and Schizosaccharomyces pombe. Analysis of the conserved roles of these protein kinases indicates that establishment of cell polarity is critical for CWR. In addition, we found that septins, crucial to budding, are both important for surviving and are mislocalized by cell wall stress. Our study shows an expanded role for protein kinase signaling in C. albicans cell wall integrity. Our studies suggest that in some cases, this expansion represents a greater importance for certain pathways in cell wall biogenesis. In other cases, it appears that signaling pathways have been rewired for a cell wall integrity response.


Photophysical dynamics of the efficient emission and photosensitization of [Ir(pqi)2(NN)]+ complexes.
The photophysical dynamics of three complexes in the highly-emissive [Ir(pqi)2(NN)]+ series were investigated aiming at unique photophysical features and applications in light-emitting and singlet oxygen sensitizing research fields. Rational elucidation and Franck-Condon analyses of the observed emission spectra in nitrile solutions at 298 and 77 K reveal the true emissive nature of the lowest-lying triplet excited state (T1), consisting of a hybrid 3MLCT/LCIr(pqi)→pqi state. Emissive deactivations from T1 occur mainly by very intense, yellow-orange phosphorescence with high quantum yields and radiative rates. The emission nature experimentally verified is corroborated by theoretical calculations (TD-DFT), with T1 arising from a mixing of several transitions induced by the spin-orbit coupling, majorly ascribed to 3MLCT/LCIr(pqi)→pqi and increasing contributions of 3MLCT/LLCTIr(pqi)→NN. The microsecond-lived emission of T1 is rapidly quenched by molecular oxygen, with an efficient generation of singlet oxygen. Our findings show that the photophysics of [Ir(pqi)2(NN)][PF6] complexes is suitable for many applications, from the active layer of electroluminescent devices to photosensitizers for photodynamic therapy and theranostics.


Spatio-temporal reconstruction of air temperature maps and their application to estimate rice growing season heat accumulation using multi-temporal MODIS data.
The accumulation of thermal time usually represents the local heat resources to drive crop growth. Maps of temperature-based agro-meteorological indices are commonly generated by the spatial interpolation of data collected from meteorological stations with coarse geographic continuity. To solve the critical problems of estimating air temperature (T(a)) and filling in missing pixels due to cloudy and low-quality images in growing degree days (GDDs) calculation from remotely sensed data, a novel spatio-temporal algorithm for T(a) estimation from Terra and Aqua moderate resolution imaging spectroradiometer (MODIS) data was proposed. This is a preliminary study to calculate heat accumulation, expressed in accumulative growing degree days (AGDDs) above 10 °C, from reconstructed T(a) based on MODIS land surface temperature (LST) data. The verification results of maximum T(a), minimum T(a), GDD, and AGDD from MODIS-derived data to meteorological calculation were all satisfied with high correlations over 0.01 significant levels. Overall, MODIS-derived AGDD was slightly underestimated with almost 10% relative error. However, the feasibility of employing AGDD anomaly maps to characterize the 2001-2010 spatio-temporal variability of heat accumulation and estimating the 2011 heat accumulation distribution using only MODIS data was finally demonstrated in the current paper. Our study may supply a novel way to calculate AGDD in heat-related study concerning crop growth monitoring, agricultural climatic regionalization, and agro-meteorological disaster detection at the regional scale.