Evaluating the effects of the novel GLP-1 analogue liraglutide in Alzheimer's disease: study protocol for a randomised controlled trial (ELAD study).
Liraglutide is a glucagon-like peptide-1 (GLP-1) analogue currently approved for type 2 diabetes and obesity. Preclinical evidence in transgenic models of Alzheimer's disease suggests that liraglutide exerts neuroprotective effects by reducing amyloid oligomers, normalising synaptic plasticity and cerebral glucose uptake, and increasing the proliferation of neuronal progenitor cells. The primary objective of the study is to evaluate the change in cerebral glucose metabolic rate after 12 months of treatment with liraglutide in participants with Alzheimer's disease compared to those who are receiving placebo. ELAD is a 12-month, multi-centre, randomised, double-blind, placebo-controlled, phase IIb trial of liraglutide in participants with mild Alzheimer's dementia. A total of 206 participants will be randomised to receive either liraglutide or placebo as a daily injection for a year. The primary outcome will be the change in cerebral glucose metabolic rate in the cortical regions (hippocampus, medial temporal lobe, and posterior cingulate) from baseline to follow-up in the treatment group compared with the placebo group. The key secondary outcomes are the change from baseline to 12 months in z scores for clinical and cognitive measures (Alzheimer's Disease Assessment Scale-Cognitive Subscale and Executive domain scores of the Neuropsychological Test Battery, Clinical Dementia Rating Sum of Boxes, and Alzheimer's Disease Cooperative Study-Activities of Daily Living) and the incidence and severity of treatment-emergent adverse events or clinically important changes in safety assessments. Other secondary outcomes are 12-month change in magnetic resonance imaging volume, diffusion tensor imaging parameters, reduction in microglial activation in a subgroup of participants, reduction in tau formation and change in amyloid levels in a subgroup of participants measured by tau and amyloid imaging, and changes in composite scores using support machine vector analysis in the treatment group compared with the placebo group. Alzheimer's disease is a leading cause of morbidity worldwide. As available treatments are only symptomatic, the search for disease-modifying therapies is a priority. If the ELAD trial is successful, liraglutide and GLP-1 analogues will represent an important class of compounds to be further evaluated in clinical trials for Alzheimer's treatment. ClinicalTrials.gov, NCT01843075 . Registration 30 April 2013.


Does paid vacation leave protect against depression among working Americans? A national longitudinal fixed effects analysis.
Objectives The United States is the only advanced economy globally that does not guarantee its workers paid vacation leave. Although empirical studies have linked paid vacation leave to happiness and stress, no study has investigated the association between paid vacation leave and depression. Using a nationally-representative longitudinal sample of 3380 working men and women aged 45-52 years from the National Longitudinal Survey of Youth 1979, this study explored whether paid vacation leave may protect against depression. Methods Multivariate linear and logistic regression models were employed to estimate the impacts of the number of annual paid vacation days of leave measured at age 40 on depression measured using the 7-item Center for Epidemiologic Studies Depression Scale short form (CES-D-SF) scale at age 50. Models were adjusted for demographic and socioeconomic factors, physical health, weekly hours, and individual fixed effects. Results For every ten additional days of paid vacation leave, the odds of depression in women was 29% lower [odds ratio (OR) 0.71, 95% confidence interval (CI) 0.55-0.92, P=0.01); there was no association in men. Linear regression models showed no association in either men or women. For every 10 days of paid leave, the odds of depression were 36% lower in White women and 38% lower in women with ≥2 children. Conclusions This study provides the first evidence on the linkage between paid vacation leave and depression, and supports a protective effect in White women with ≥2 children. Should this association be truly causal, and assuming a uniform effect across all ages in working adult women, the results from this study would suggest that a hypothetical increase in the average number of days of paid vacation leave of 10 days could avoid an estimated 568 442 cases of depression in women each year and lead to a cost savings of US$2.94 billion annually. Policies that mandate paid vacation leave may have marked positive impacts on the population health and economic burden of depression among working women in the USA.


Personalized Medicine Approaches in Prostate Cancer Employing Patient Derived 3D Organoids and Humanized Mice.
Prostate cancer (PCa) is the most common malignancy and the second most common cause of cancer death in Western men. Despite its prevalence, PCa has proven very difficult to propagate in vitro. PCa represents a complex organ-like multicellular structure maintained by the dynamic interaction of tumoral cells with parenchymal stroma, endothelial and immune cells, and components of the extracellular matrix (ECM). The lack of PCa models that recapitulate this intricate system has hampered progress toward understanding disease progression and lackluster therapeutic responses. Tissue slices, monolayer cultures and genetically engineered mouse models (GEMM) fail to mimic the complexities of the PCa microenvironment or reproduce the diverse mechanisms of therapy resistance. Moreover, patient derived xenografts (PDXs) are expensive, time consuming, difficult to establish for prostate cancer, lack immune cell-tumor regulation, and often tumors undergo selective engraftments. Here, we describe an interdisciplinary approach using primary PCa and tumor initiating cells (TICs), three-dimensional (3D) tissue engineering, genetic and morphometric profiling, and humanized mice to generate patient-derived organoids for examining personalized therapeutic responses in vitro and in mice co-engrafted with a human immune system (HIS), employing adaptive T-cell- and chimeric antigen receptor- (CAR) immunotherapy. The development of patient specific therapies targeting the vulnerabilities of cancer, when combined with antiproliferative and immunotherapy approaches could help to achieve the full transformative power of cancer precision medicine.


The structure of the RlmB 23S rRNA methyltransferase reveals a new methyltransferase fold with a unique knot.
In Escherichia coli, RlmB catalyzes the methylation of guanosine 2251, a modification conserved in the peptidyltransferase domain of 23S rRNA. The crystal structure of this 2'O-methyltransferase has been determined at 2.5 A resolution. RlmB consists of an N-terminal domain connected by a flexible extended linker to a catalytic C-terminal domain and forms a dimer in solution. The C-terminal domain displays a divergent methyltransferase fold with a unique knotted region, and lacks the classic AdoMet binding site features. The N-terminal domain is similar to ribosomal proteins L7 and L30, suggesting a role in 23S rRNA recognition. The conserved residues in this novel family of 2'O-methyltransferases cluster in the knotted region, suggesting the location of the catalytic and AdoMet binding sites.


Low incidence of clinically apparent thromboembolism in Korean patients with multiple myeloma treated with thalidomide.
The frequency of thromboembolic events (TE) in Caucasian patients with multiple myeloma (MM) receiving thalidomide as the initial treatment has been reported to be 10~58% without prophylactic anticoagulation. Korean MM patients treated with thalidomide were studied to determine the frequency of TE and associated risk factors. A retrospective medical record review of the Korean MM registry from 25 centers in Korea between 2003 and 2007 was performed. We assessed the incidence of arterial and venous TE and the associated clinical parameters. Three hundred and sixty MM patients (median age 61 years, range 32-88 years) received thalidomide treatment. Fourteen patients (3.9%) developed TE: 12 had venous and two had arterial locations. The sites for the venous TE included lungs (seven), lower extremities (four), upper extremities (one), and neck (one). Arterial TE developed in cerebral and peripheral arteries each. No single clinical parameter such as prerequisite for the metabolic syndrome, disease status, and treatment regimen were predictive for the development of TE. The frequency of TE in patients who received thalidomide as initial therapy (7/155) was not different from those who received thalidomide for progressive or relapsed disease (7/205, p = 0.592). The frequency of TE during thalidomide treatment in Korean patients with MM was low. No significant clinical factor was found to be a risk factor. The subgroup requiring thromboprophylaxis among the Korean patients with MM, receiving thalidomide, needs to be clarified.


Immunological disorders of human pregnancy.
Three types of disorders of human pregnancy with a possible immune etiology have been discussed. Pre-eclampsia and recurrent abortion may result from a failure of a maternal immunoregulatory response to trophoblast but the evidence is incomplete and circumstantial. There are several fetal problems which may arise because of transfer of maternal allo-, iso-, or autoantibodies to the fetus. Third, maternal graft-versus-fetal-host disease occurs rarely and may present in a delayed and incomplete way as severe combined immunodeficiency.


Bifidobacteria in feces and environmental waters.
Bifidobacteria have been recommended as potential indicators of human fecal pollution in surface waters even though very little is known about their presence in nonhuman fecal sources. The objective of this research was to shed light on the occurrence and molecular diversity of this fecal indicator group in different animals and environmental waters. Genus- and species-specific 16S rRNA gene PCR assays were used to study the presence of bifidobacteria among 269 fecal DNA extracts from 32 different animals. Twelve samples from three wastewater treatment plants and 34 water samples from two fecally impacted watersheds were also tested. The species-specific assays showed that Bifidobacterium adolescentis, B. bifidum, B. dentium, and B. catenulatum had the broadest host distribution (11.9 to 17.4%), whereas B. breve, B. infantis, and B. longum were detected in fewer than 3% of all fecal samples. Phylogenetic analysis of 356 bifidobacterial clones obtained from different animal feces showed that ca. 67% of all of the sequences clustered with cultured bifidobacteria, while the rest formed a supercluster with low sequence identity (i.e., <94%) to previously described Bifidobacterium spp. The B. pseudolongum subcluster (>97% similarity) contained 53 fecal sequences from seven different animal hosts, suggesting the cosmopolitan distribution of members of this clade. In contrast, two clades containing B. thermophilum and B. boum clustered exclusively with 37 and 18 pig fecal clones, respectively, suggesting host specificity. Using species-specific assays, bifidobacteria were detected in only two of the surface water DNA extracts, although other fecal anaerobic bacteria were detected in these waters. Overall, the results suggest that the use of bifidobacterial species as potential markers to monitor human fecal pollution in natural waters may be questionable.


Proteomic analysis of oridonin-induced apoptosis in multiple myeloma cells.
Oridonin is a diterpenoid compound isolated from the medicinal herb Rabdosia rubescens, and has shown marked antitumor effects against different types of cancer. However, the definitive systematic molecular mechanism underlying the antitumor activity of oridonin in multiple myeloma remains to be elucidated. In the present study, cell viability and cytotoxicity were examined to determine the appropriate concentration for proteomic investigation. In addition, cell apoptosis was evaluated using flow cytometry and transmission electron microscopy. A proteomic investigation using a two‑dimensional electrophoresis system and mass spectrometry was performed to identify and characterize the global proteome of the apoptosis induced by oridonin. Of the proteins identified, seven were involved in the anticancer effects of oridonin. Regulation of the expression and function of target proteins, stathmin, dihydrofolate reductase and pyruvate dehydrogenase E1β, may be potential, therapeutic strategies to effectively treat multiple myeloma. These findings provide novel information on the molecular mechanisms underlying the anticancer properties of oridonin in multiple myeloma.


Diagnostic single nucleotide polymorphism analysis of factor V Leiden and prothrombin 20210G > A. A comparison of the Nanogen Eelectronic Microarray with restriction enzyme digestion and the Roche LightCycler.
Genetic thrombosis risk factors include a sequence variant in the prothrombin gene (20210G > A) and factor V Leiden (1691G > A). These single nucleotide polymorphisms can be diagnosed with restriction fragment length polymorphism (RFLP) analysis, fluorescent genotyping on the LightCycler (Roche Diagnostics, Indianapolis, IN), and microarray-based testing on the novel NanoChip electronic microarray (NanoChip Molecular Biology Workstation, Nanogen, San Diego, CA). We compared these methods for accuracy, time to results, throughput, and interpretation. Results from 789 of 800 individual amplicons analyzed on the NanoChip were in complete agreement with the other assays. Eleven were "no calls" (uninterpreted by the NanoChip system) resulting from failed polymerase chain reaction amplifications. Although the NanoChip System, when used in a low-throughput setting, requires more overall time than the LightCycler, it is nearly equivalent per genotyping call. Owing to minimal sample handling, assay results are more reliable on the NanoChip platform and on the LightCycler than with RFLP. The NanoChip assay is reliable and may be especially valuable to laboratories with a large volume of thrombophilia test requests.


Cowpea viruses: effect of single and mixed infections on symptomatology and virus concentration.
Natural multiple viral infections of cultivated cowpeas have been reported in Nigeria. In this study, three Nigerian commercial cowpea cultivars ("Olo 11", "Oloyin" and "White") and two lines from the IITA (IT86D- 719 and TVU 76) were mechanically inoculated with Cowpea aphid-borne mosaic virus (CABMV), Bean southern mosaic virus (SBMV) and Cowpea mottle virus (CMeV) singly, as well as in all possible combinations at 10, 20 and 30 days after planting (DAP). Samples of leaves or stems were collected at 10, 20 and 30 days after inoculation (DAI) and analyzed for relative virus concentration by Enzyme-Linked Immunosrbent Assay. All the cultivars and lines {CVS/L} were susceptible to the viruses but the commercial CVS showed more severe symptoms and had relatively higher viral concentration. In single virus infections, CABMV which induced the most severe symptoms had absorbance values (at 405 nm) of 0.11 to 0.46 while SBMV and CMeV which induced moderate symptoms had virus titre of 0.74 to 1.99 and 0.11 to 0.90 respectively. Plants inoculated 10 DAP had significantly higher virus concentration than those inoculated 30 DAP. In mixed infections involving CABMV (10 DAP) apical necrosis and death were observed in commercial cultivars "Olo 11" and "White". Enhancement of CMeV titers were observed in plants infected with CMeV + CABMV. Multiple viral infections of cowpeas may result in complete yield loss, hence, the availability of seeds of cultivars with a high level of multiple virus resistance is recommended as a means of control.


Functional interactions of T cells with endothelial cells: the role of CD40L-CD40-mediated signals.
CD40 is expressed on a variety of cells, including B cells, monocytes, dendritic cells, and fibroblasts. CD40 interacts with CD40L, a 30-33-kD activation-induced CD4+ T cell surface molecule. CD40L-CD40 interactions are known to play key roles in B cell activation and differentiation in vitro and in vivo. We now report that normal human endothelial cells also express CD40 in situ, and CD40L-CD40 interactions induce endothelial cell activation in vitro. Frozen sections from normal spleen, thyroid, skin, muscle, kidney, lung, or umbilical cord were studied for CD40 expression by immunohistochemistry. Endothelial cells from all tissues studied express CD40 in situ. Moreover, human umbilical vein endothelial cells (HUVEC) express CD40 in vitro, and recombinant interferon gamma induces HUVEC CD40 upregulation. CD40 expression on HUVEC is functionally significant because CD40L+ Jurkat T cells or CD40L+ 293 kidney cell transfectants, but not control cells, upregulate HUVEC CD54 (intercellular adhesion molecule-1), CD62E (E-selectin), and CD106 (vascular cell adhesion molecule-1) expression in vitro. Moreover, the kinetics of CD40L-, interleukin 1-, or tumor necrosis factor alpha-induced CD54, CD62E, and CD106 upregulation on HUVEC are similar. Finally, CD40L-CD40 interactions do not induce CD80, CD86, or major histocompatibility complex class II expression on HUVEC in vitro. These results demonstrate that CD40L-CD40 interactions induce endothelial cell activation in vitro. Moreover, they suggest a mechanism by which activated CD4+ T cells may augment inflammatory responses in vivo by upregulating the expression of endothelial cell surface adhesion molecules.


Better together: engineering and application of microbial symbioses.
Symbioses provide a way to surpass the limitations of individual microbes. Natural communities exemplify this in symbioses like lichens and biofilms that are robust to perturbations, an essential feature in fluctuating environments. Metabolic capabilities also expand in consortia enabling the division of labor across organisms as seen in photosynthetic and methanogenic communities. In engineered consortia, the external environment provides levers of control for microbes repurposed from nature or engineered to interact through synthetic biology. Consortia have successfully been applied to real-world problems including remediation and energy, however there are still fundamental questions to be answered. It is clear that continued study is necessary for the understanding and engineering of microbial systems that are more than the sum of their parts.


Consumption of fa cai Nostoc soup: a potential for BMAA exposure from Nostoc cyanobacteria in China?
Grown in arid regions of western China the cyanobacterium Nostoc flagelliforme--called fa cai in Mandarin and fat choy in Cantonese--is wild-harvested and used to make soup consumed during New Year's celebrations. High prices, up to $125 USD/kg, led to overharvesting in Inner Mongolia, Ningxia, Gansu, Qinghai, and Xinjiang. Degradation of arid ecosystems, desertification, and conflicts between Nostoc harvesters and Mongol herdsmen concerned the Chinese environmental authorities, leading to a government ban of Nostoc commerce. This ban stimulated increased marketing of a substitute made from starch. We analysed samples purchased throughout China as well as in Chinese markets in the United States and the United Kingdom. Some were counterfeits consisting of dyed starch noodles. A few samples from California contained Nostoc flagelliforme but were adulterated with starch noodles. Other samples, including those from the United Kingdom, consisted of pure Nostoc flagelliforme. A recent survey of markets in Cheng Du showed no real Nostoc flagelliforme to be marketed. Real and artificial fa cai differ in the presence of beta-N-methylamino-L-alanine (BMAA). Given its status as a high-priced luxury food, the government ban on collection and marketing, and the replacement of real fa cai with starch substitutes consumed only on special occasions, it is anticipated that dietary exposure to BMAA from fa cai will be reduced in the future in China.


Diet strongly influences the gut microbiota of surgeonfishes.
Intestinal tracts are among the most densely populated microbial ecosystems. Gut microbiota and their influence on the host have been well characterized in terrestrial vertebrates but much less so in fish. This is especially true for coral reef fishes, which are among the most abundant groups of vertebrates on earth. Surgeonfishes (family: Acanthuridae) are part of a large and diverse family of reef fish that display a wide range of feeding behaviours, which in turn has a strong impact on the reef ecology. Here, we studied the composition of the gut microbiota of nine surgeonfish and three nonsurgeonfish species from the Red Sea. High-throughput pyrosequencing results showed that members of the phylum Firmicutes, especially of the genus Epulopiscium, were dominant in the gut microbiota of seven surgeonfishes. Even so, there were large inter- and intraspecies differences in the diversity of surgeonfish microbiota. Replicates of the same host species shared only a small number of operational taxonomic units (OTUs), although these accounted for most of the sequences. There was a statistically significant correlation between the phylogeny of the host and their gut microbiota, but the two were not completely congruent. Notably, the gut microbiota of three nonsurgeonfish species clustered with some surgeonfish species. The microbiota of the macro- and microalgavores was distinct, while the microbiota of the others (carnivores, omnivores and detritivores) seemed to be transient and dynamic. Despite some anomalies, both host phylogeny and diet were important drivers for the intestinal microbial community structure of surgeonfishes from the Red Sea.


3D bioprinting of BM-MSCs-loaded ECM biomimetic hydrogels for in vitro neocartilage formation.
In this work we demonstrate how to print 3D biomimetic hydrogel scaffolds for cartilage tissue engineering with high cell density (>10(7) cells ml(-1)), high cell viability (85 ÷ 90%) and high printing resolution (≈100 μm) through a two coaxial-needles system. The scaffolds were composed of modified biopolymers present in the extracellular matrix (ECM) of cartilage, namely gelatin methacrylamide (GelMA), chondroitin sulfate amino ethyl methacrylate (CS-AEMA) and hyaluronic acid methacrylate (HAMA). The polymers were used to prepare three photocurable bioinks with increasing degree of biomimicry: (i) GelMA, (ii) GelMA + CS-AEMA and (iii) GelMA + CS-AEMA + HAMA. Alginate was added to the bioinks as templating agent to form stable fibers during 3D printing. In all cases, bioink solutions were loaded with bone marrow-derived human mesenchymal stem cells (BM-MSCs). After printing, the samples were cultured in expansion (negative control) and chondrogenic media to evaluate the possible differentiating effect exerted by the biomimetic matrix or the synergistic effect of the matrix and chondrogenic supplements. After 7, 14, and 21 days, gene expression of the chondrogenic markers (COL2A1 and aggrecan), marker of osteogenesis (COL1A1) and marker of hypertrophy (COL10A1) were evaluated qualitatively by means of fluorescence immunocytochemistry and quantitatively by means of RT-qPCR. The observed enhanced viability and chondrogenic differentiation of BM-MSCs, as well as high robustness and accuracy of the employed deposition method, make the presented approach a valid candidate for advanced engineering of cartilage tissue.


Mechanisms of lung endothelial barrier disruption induced by cigarette smoke: role of oxidative stress and ceramides.
The epithelial and endothelial cells lining the alveolus form a barrier essential for the preservation of the lung respiratory function, which is, however, vulnerable to excessive oxidative, inflammatory, and apoptotic insults. Whereas profound breaches in this barrier function cause pulmonary edema, more subtle changes may contribute to inflammation. The mechanisms by which cigarette smoke (CS) exposure induce lung inflammation are not fully understood, but an early alteration in the epithelial barrier function has been documented. We sought to investigate the occurrence and mechanisms by which soluble components of mainstream CS disrupt the lung endothelial cell barrier function. Using cultured primary rat microvascular cell monolayers, we report that CS induces endothelial cell barrier disruption in a dose- and time-dependent manner of similar magnitude to that of the epithelial cell barrier. CS exposure triggered a mechanism of neutral sphingomyelinase-mediated ceramide upregulation and p38 MAPK and JNK activation that were oxidative stress dependent and that, along with Rho kinase activation, mediated the endothelial barrier dysfunction. The morphological changes in endothelial cell monolayers induced by CS included actin cytoskeletal rearrangement, junctional protein zonula occludens-1 loss, and intercellular gap formation, which were abolished by the glutathione modulator N-acetylcysteine and ameliorated by neutral sphingomyelinase inhibition. The direct application of ceramide recapitulated the effects of CS, by disrupting both endothelial and epithelial cells barrier, by a mechanism that was redox and apoptosis independent and required Rho kinase activation. Furthermore, ceramide induced dose-dependent alterations of alveolar microcirculatory barrier in vivo, measured by two-photon excitation microscopy in the intact rat. In conclusion, soluble components of CS have direct endothelial barrier-disruptive effects that could be ameliorated by glutathione modulators or by inhibitors of neutral sphingomyelinase, p38 MAPK, JNK, and Rho kinase. Amelioration of endothelial permeability may alleviate lung and systemic vascular dysfunction associated with smoking-related chronic obstructive lung diseases.


Acute kidney injury due to tropical infectious diseases and animal venoms: a tale of 2 continents.
South and Southeast Asia and Latin American together comprise 46 countries and are home to approximately 40% of the world population. The sociopolitical and economic heterogeneity, tropical climate, and malady transitions characteristic of the region strongly influence disease behavior and health care delivery. Acute kidney injury epidemiology mirrors these inequalities. In addition to hospital-acquired acute kidney injury in tertiary care centers, these countries face a large preventable burden of community-acquired acute kidney injury secondary to tropical infectious diseases or animal venoms, affecting previously healthy young individuals. This article reviews the epidemiology, clinical picture, prevention, risk factors, and pathophysiology of acute kidney injury associated with tropical diseases (malaria, dengue, leptospirosis, scrub typhus, and yellow fever) and animal venom (snakes, bees, caterpillars, spiders, and scorpions) in tropical regions of Asia and Latin America, and discusses the potential future challenges due to emerging issues.


Diagnoses based on the Research Diagnostic Criteria for Temporomandibular Disorders in a biracial population of young women.
To compare the clinical characteristics of diagnostic subtypes of temporomandibular disorders (TMD) based on the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) in terms of physical findings (Axis I) and psychosocial findings (Axis II) among Caucasian and African American young women. An ancillary goal was to assess the value of using self-reported TMD pain as a screening tool compared to RDC/TMD examinations. A biracial community sample (n = 830) of young women 19 to 23 years old was screened for facial pain with the Chronic Pain Grade questionnaire. Patients were considered to be putative cases of TMD if they reported facial pain present within the last 6 months; putative controls had no facial pain history or jaw symptoms. Women with facial pain more than 6 months ago and jaw symptoms (jaw symptom-past pain, JSPP group) were added. 129 women were clinically examined for TMD diagnosis for final confirmation of case-control status. 41 of 43 Caucasian and 11 of 18 African American putative cases were confirmed as cases; 9 of 27 Caucasians, but 0 of 17 African Americans from the JSPP group were confirmed as cases. All 24 putative controls were confirmed as controls. Based on RDC/TMD Axis I, 80% of 61 cases were muscle-related diagnoses, 33% as disc-related diagnoses, and 48% as arthralgia/arthritis/arthrosis. Based on Axis I, there were no significant differences in diagnoses between African American and Caucasian women. Based on Axis II, cases had significantly greater depression (P = .002) and somatization with pain (P < .001) than controls as expected. African Americans had significantly greater somatization with pain than Caucasians (P = .020). There were no other significant racial differences. Among young women reporting facial pain, clinical TMD subtypes, pain impact, treatment utilization, and additional characteristics other than somatization with pain were similar between races. A high percentage of these young non-clinical cases presented severe depression and somatization.


Production of IGF-I and IGF binding proteins by retinal cells in vitro.
Insulin-like growth factor I (IGF-I) and its associated binding proteins (IGFBPs) have been identified in retinal tissues but their precise cellular origin remains unclear. The aim of this study was to examine the ability of three retinal cell types (microvascular endothelial cells, pericytes, and pigment epithelial cells) and a non-retinal cell type (Tenon's fibroblasts) to produce IGF-I and cell specific IGFBPs in vitro. Using a radioimmunoassay we demonstrated that all four cell types produce IGF-I and that this production continued over a 72 hour period. In addition, western ligand blotting revealed that each cell type produced at least one IGFBP and that each cell type produced a different IGFBP profile. Endothelial cells produced a 24 kDa band only, pericytes produced both a 24 kDa and a 34 kDa band, retinal pigment epithelial cells produced a 38-41 kDa band, while fibroblasts produced both a 24 kDa and a 30 kDa band. Laser scanning densitometry demonstrated that in the majority of experiments the IGFBPs accumulated in the culture medium over a 72 hour period. Neither IGF-I nor IGFBPs were observed in cell lysates indicating de novo synthesis and secretion in vitro. These results suggest an autocrine/paracrine function for IGF-I and its associated binding proteins which may play a significant role in both retinal physiology and disease.


Prevalence and analysis of Renibacterium salmoninarum infection among juvenile Chinook salmon Oncorhynchus tshawytscha in North Puget Sound.
Renibacterium salmoninarum causes bacterial kidney disease (BKD), a chronic and sometimes fatal disease of salmon and trout that could lower fitness in populations with high prevalences of infection. Prevalence of R. salmoninarum infection among juvenile Chinook salmon Oncorhynchus tshawytscha inhabiting neritic marine habitats in North Puget Sound, Washington, USA, was assessed in 2002 and 2003. Fish were collected by monthly surface trawl at 32 sites within 4 bays, and kidney infections were detected by a quantitative fluorescent antibody technique (qFAT). The sensitivity of the qFAT was within an order of magnitude of the quantitative real-time PCR (qPCR) sensitivity. Prevalence of infection was classified by fish origin (marked/hatchery vs. unmarked/likely natural spawn), month of capture, capture location and stock origin. The highest percentages of infected fish (63.5 to 63.8%) and the greatest infection severity were observed for fish collected in Bellingham Bay. The lowest percentages were found in Skagit Bay (11.4 to 13.5%); however, there was no difference in prevalence between marked and unmarked fish among the capture locations. The optimal logistic regression model of infection probabilities identified the capture location of Bellingham Bay as the strongest effect, and analysis of coded wire tagged (CWT) fish revealed that prevalence of infection was associated with the capture location and not with the originating stock. These results suggest that infections can occur during the early marine life stages of Chinook salmon that may be due to common reservoirs of infection or horizontal transmission among fish stocks.


The prognosis of primary lymphedema of the lower limbs.
Primary lymphedema in most patients can be managed conservatively, but, in a few, severe lymphedema develops that requires surgical reduction of the limb. The five- to 20-year course of the disease has been studied in 372 patients. The eventual extent of the disease could be assessed within the first year. Further spreading of the lymphedema in the same limb occurred in only 7% of the patients after the first year and in less than 1% after five years. Only 9% of the patients suffered lymphedema of the other leg at a later time. Patients without pelvic lymphatic obstruction had a good prognosis even if the distal lymphatics were obliterated, but 32% of those with proximal obstructive hypoplasia had severe lymphedema that required surgical reduction. In this latter group, men and women were affected equally.


Cocaine Use and Risk of Ischemic Stroke in Young Adults.
Although case reports have long identified a temporal association between cocaine use and ischemic stroke (IS), few epidemiological studies have examined the association of cocaine use with IS in young adults, by timing, route, and frequency of use. A population-based case-control study design with 1090 cases and 1154 controls was used to investigate the relationship of cocaine use and young-onset IS. Stroke cases were between the ages of 15 and 49 years. Logistic regression analysis was used to evaluate the association between cocaine use and IS with and without adjustment for potential confounders. Ever use of cocaine was not associated with stroke with 28% of cases and 26% of controls reporting ever use. In contrast, acute cocaine use in the previous 24 hours was strongly associated with increased risk of stroke (age-sex-race adjusted odds ratio, 6.4; 95% confidence interval, 2.2-18.6). Among acute users, the smoking route had an adjusted odds ratio of 7.9 (95% confidence interval, 1.8-35.0), whereas the inhalation route had an adjusted odds ratio of 3.5 (95% confidence interval, 0.7-16.9). After additional adjustment for current alcohol, smoking use, and hypertension, the odds ratio for acute cocaine use by any route was 5.7 (95% confidence interval, 1.7-19.7). Of the 26 patients with cocaine use within 24 hours of their stroke, 14 reported use within 6 hours of their event. Our data are consistent with a causal association between acute cocaine use and risk of early-onset IS.


Alcohol mixed with energy drinks: are there associated negative consequences beyond hazardous drinking in college students?
The consumption of alcohol mixed with energy drinks (AmED) is prevalent among college students as is hazardous drinking, a drinking pattern that places one at risk for alcohol-related harm. The present study, therefore, examined associations between AmED use, hazardous drinking, and alcohol-related consequences in college students. Based on a probability sample conducted in 2010, participants were 606 undergraduate students aged 18-25. AmED consumption included lifetime and past year use. Hazardous drinking and alcohol-related consequences were measured during the past year. Point prevalence was used to estimate rates of AmED use, and chi-square, ANOVA, and logistic regression were used to examine associations between AmED use, hazardous drinking, and alcohol-related consequences. Lifetime and past year AmED use prevalence rates were 75.2% and 64.7%, respectively. Hazardous drinkers who engaged in AmED use were significantly more likely than past year hazardous drinkers who did not engage in AmED use to have had unprotected sex (OR=2.35, CI 1.27-4.32). AmED use appears to be highly prevalent among college students, and AmED use may confer additional risk for unprotected sex beyond hazardous drinking. Unprotected sex has implications for public health, and students who drink hazardously and consume AmED may be at greater risk.


Coupled double triplet state in singlet fission.
The highly unusual state, 1(TT), is a coupled, double triplet state that has recently garnered significant attention. This multiexcitonic state can be formed by a quantum transition from a single-photon bright state in a variety of organic semiconducting materials. 1(TT)'s transient nature and similarity to independent triplets, however, has led to significant difficulties in characterization and prediction of its properties. Recent progress describing 1(TT) from theory and experiment are breaking through these difficulties, and have greatly advanced our comprehension of this state. Starting from the early description of 1(TT) in polyenes, this perspective discusses formation mechanisms, spectroscopic signatures, and the scope of intertriplet interactions. When employing singlet fission to generate charge carriers in a solar cell, 1(TT) has a central role. Due to the variety of coupling strengths between triplet states in 1(TT) amongst different chromophores, two different strategies are discussed to enable efficient charge carrier extraction. Continued growth in our understanding of 1(TT) may lead to control over complex quantum states for intriguing applications beyond high-efficiency, organic solar cells.


Scaffold-Constrained Molecular Generation.
One of the major applications of generative models for drug discovery targets the lead-optimization phase. During the optimization of a lead series, it is common to have scaffold constraints imposed on the structure of the molecules designed. Without enforcing such constraints, the probability of generating molecules with the required scaffold is extremely low and hinders the practicality of generative models for de novo drug design. To tackle this issue, we introduce a new algorithm, named SAMOA (Scaffold Constrained Molecular Generation), to perform scaffold-constrained in silico molecular design. We build on the well-known SMILES-based Recurrent Neural Network (RNN) generative model, with a modified sampling procedure to achieve scaffold-constrained generation. We directly benefit from the associated reinforcement learning methods, allowing to design molecules optimized for different properties while exploring only the relevant chemical space. We showcase the method's ability to perform scaffold-constrained generation on various tasks: designing novel molecules around scaffolds extracted from SureChEMBL chemical series, generating novel active molecules on the Dopamine Receptor D2 (DRD2) target, and finally, designing predicted actives on the MMP-12 series, an industrial lead-optimization project.


Melanoma biomolecules: independently identified but functionally intertwined.
The majority of patients diagnosed with melanoma present with thin lesions and generally these patients have a good prognosis. However, 5% of patients with early melanoma (<1 mm thick) will have recurrence and die within 10 years, despite no evidence of local or metastatic spread at the time of diagnosis. Thus, there is a need for additional prognostic markers to help identify those patients that may be at risk of recurrent disease. Many studies and several meta-analyses have compared gene and protein expression in melanocytes, naevi, primary, and metastatic melanoma in an attempt to find informative prognostic markers for these patients. However, although a large number of putative biomarkers have been described, few of these molecules are informative when used in isolation. The best approach is likely to involve a combination of molecules. We believe one approach could be to analyze the expression of a group of interacting proteins that regulate different aspects of the metastatic pathway. This is because a primary lesion expressing proteins involved in multiple stages of metastasis may be more likely to lead to secondary disease than one that does not. This review focuses on five putative biomarkers - melanoma cell adhesion molecule (MCAM), galectin-3 (gal-3), matrix metalloproteinase 2 (MMP-2), chondroitin sulfate proteoglycan 4 (CSPG4), and paired box 3 (PAX3). The goal is to provide context around what is known about the contribution of these biomarkers to melanoma biology and metastasis. Although each of these molecules have been independently identified as likely biomarkers, it is clear from our analyses that each are closely linked with each other, with intertwined roles in melanoma biology.


Frequency self-doubling optical parametric amplification: noncollinear red-green-blue light-source generation based on a hexagonally poled lithium tantalate.
Simultaneous generation of noncollinear red, green, and blue light from a single hexagonally poled lithium tantalate is reported. It results from the frequency self-doubling optical parametric amplification process, a process of second-order harmonic generation cascaded optical parametric amplification in a single-pass setup. The temperature and spectrum detuning characters of each cascaded quasi-phase-matching process are studied. This unique red-green-blue light source has potential applications in laser display and other laser industries.


Fanconi anemia with sun-sensitivity caused by a Xeroderma pigmentosum-associated missense mutation in XPF.
Fanconi anemia (FA) is an inherited genomic instability disorder with congenital and developmental abnormalities, bone marrow failure and predisposition to cancer early in life, and cellular sensitivity to DNA interstrand crosslinks. A fifty-one-year old female patient, initially diagnosed with FA in childhood on the basis of classic features and increased chromosomal breakage, and remarkable sun-sensitivity is described. She only ever had mild haematological abnormalities and no history of malignancy. To identify and characterise the genetic defect in this lady, who is one of the oldest reported FA patients, we used whole-exome sequencing for identification of causative mutations, and functionally characterized the cellular phenotype. Detection of the novel splice site mutation c.793-2A > G and the previously described missense mutation c.1765C > T (p.Arg589Trp) in XPF/ERCC4/FANCQ assign her as the third individual of complementation group FA-Q. Ectopic expression of wildtype, but not mutant, XPF/ERCC4/FANCQ, in patient-derived fibroblasts rescued cellular resistance to DNA interstrand-crosslinking agents. Patient derived FA-Q cells showed impaired nuclear excision repair capacity. However, mutated XPF/ERCC4/FANCQ protein in our patient's cells, as in the two other patients with FA-Q, was detectable on chromatin, in contrast to XP-F cells, where missense-mutant protein failed to properly translocate to the nucleus. Patients with FA characteristics and UV sensitivity should be tested for mutations in XPF/ERCC4/FANCQ. The missense mutation p.Arg589Trp was previously detected in patients diagnosed with Xeroderma pigmentosum or Cockayne syndrome. Hence, phenotypic manifestations associated with this XPF/ERCC4/ FANCQ mutation are highly variable.


Structure-Guided Optimization of HIV Integrase Strand Transfer Inhibitors.
Integrase mutations can reduce the effectiveness of the first-generation FDA-approved integrase strand transfer inhibitors (INSTIs), raltegravir (RAL) and elvitegravir (EVG). The second-generation agent, dolutegravir (DTG), has enjoyed considerable clinical success; however, resistance-causing mutations that diminish the efficacy of DTG have appeared. Our current findings support and extend the substrate envelope concept that broadly effective INSTIs can be designed by filling the envelope defined by the DNA substrates. Previously, we explored 1-hydroxy-2-oxo-1,2-dihydro-1,8-naphthyridine-3-carboxamides as an INSTI scaffold, making a limited set of derivatives, and concluded that broadly effective INSTIs can be developed using this scaffold. Herein, we report an extended investigation of 6-substituents as well the first examples of 7-substituted analogues of this scaffold. While 7-substituents are not well-tolerated, we have identified novel substituents at the 6-position that are highly effective, with the best compound (6p) retaining better efficacy against a broad panel of known INSTI resistant mutants than any analogues we have previously described.


Benchmark simulation model no 2: general protocol and exploratory case studies.
Over a decade ago, the concept of objectively evaluating the performance of control strategies by simulating them using a standard model implementation was introduced for activated sludge wastewater treatment plants. The resulting Benchmark Simulation Model No 1 (BSM1) has been the basis for a significant new development that is reported on here: Rather than only evaluating control strategies at the level of the activated sludge unit (bioreactors and secondary clarifier) the new BSM2 now allows the evaluation of control strategies at the level of the whole plant, including primary clarifier and sludge treatment with anaerobic sludge digestion. In this contribution, the decisions that have been made over the past three years regarding the models used within the BSM2 are presented and argued, with particular emphasis on the ADM1 description of the digester, the interfaces between activated sludge and digester models, the included temperature dependencies and the reject water storage. BSM2-implementations are now available in a wide range of simulation platforms and a ring test has verified their proper implementation, consistent with the BSM2 definition. This guarantees that users can focus on the control strategy evaluation rather than on modelling issues. Finally, for illustration, twelve simple operational strategies have been implemented in BSM2 and their performance evaluated. Results show that it is an interesting control engineering challenge to further improve the performance of the BSM2 plant (which is the whole idea behind benchmarking) and that integrated control (i.e. acting at different places in the whole plant) is certainly worthwhile to achieve overall improvement.


Cathodic electrochemical activation of Co3O4 nanoarrays: a smart strategy to significantly boost the hydrogen evolution activity.
Co(hydro)oxides show unsatisfactory catalytic activity for the hydrogen evolution reaction (HER) in alkaline media, and it is thus highly desirable but still remains a challenge to design and develop Co(hydro)oxide derived materials as superb hydrogen-evolving catalysts using a facile, rapid and less energy-intensive method. Here, we propose a cathodic electrochemical activation strategy toward greatly boosted HER activity of a Co3O4 nanoarray via room-temperature cathodic polarization in sodium hypophosphite solution. After activation, the overpotential significantly decreases from 260 to 73 mV to drive a geometrical catalytic current density of 10 mA cm-2 in 1.0 M KOH. Notably, this activated electrode also shows strong long-term electrochemical durability with the retention of its catalytic activity at 100 mA cm-2 for at least 40 h.


Comparison of surgical outcomes between simple posterior layer advancement of lower eyelid retractors and combination with a lateral tarsal strip procedure for involutional entropion in a Japanese population.
To compare the effectiveness of simple posterior layer advancement of the lower eyelid retractor (LER) and combination surgery of posterior layer advancement of the LER with a lateral tarsal strip procedure for involutional lower eyelid entropion in a Japanese population. 46 eyelids of 37 patients with horizontal laxity (Group A) and 47 eyelids of 42 patients without horizontal laxity (Group B) that underwent simple posterior layer advancement of the LER, and 47 eyelids of 37 patients with horizontal laxity that had the combination surgery (Group C) were retrospectively reviewed. All eyelids were judged as successfully corrected without recurrence in Groups B and C. On the other hand, although 42 eyelids (91.3%) were successfully altered in Group A, four eyelids (8.7%) showed recurrence at a mean period of 7.6 months postoperatively. Surgical success rates in Groups B and C tended to be higher than in Group A (p=0.056). The combination surgery in Group C and simple posterior layer advancement of the LER in Group B provided complete surgical success. The present study suggests the importance of preoperative evaluation of horizontal laxity, allowing surgeons to perform the least amount of surgery to achieve success.


Lateralization of motor imagery following stroke.
Motor imagery may activate the primary motor cortex (M1) and promote functional recovery following stroke. We investigated whether the hemisphere affected by stroke affects performance and M1 activity during motor imagery. Twelve stroke patients (6 left, 6 right hemisphere) and eight healthy age-matched adults participated. Experiment 1 assessed the speed and ease of actual and imagined motor performance. Experiment 2 measured corticomotor excitability during imagined movement of each hand separately, and both hands together, using transcranial magnetic stimulation. For control participants, imagined movements were performed more slowly than actual movements, and right-hand MEPs were facilitated when they imagined moving their right hand or both hands together. Patients reported being able to imagine movements with either hand, despite no measurable facilitation of MEPs in the stroke-affected hand. In left hemisphere patients, MEPs were facilitated in the left hand during imagery of the right hand and both hands together. In right hemisphere patients, motor imagery did not facilitate MEPs in either hand. Motor imagery does not appear to facilitate the ipsilesional M1 following stroke. Motor imagery may play a role in rehabilitating movement planning, but its role in directly facilitating corticomotor output appears limited.


Production of a monoclonal antibody reactive with human dendritic reticulum cells and its use in the immunohistological analysis of lymphoid tissue.
A murine monoclonal antibody (designated R4/23) which reacts strongly with human dendritic reticulum cells (DRC) is described. Immunoperoxidase staining of tissue cryostat sections revealed that this antibody reacts strongly with DRC in lymphoid follicles (both primary and secondary), and also weakly with marginal zone splenic B cells and with some peripheral follicular mantle B lymphocytes in lymph node cortical follicles. The value of antibody R4/23 is that it allows the distribution of DRC in reactive and neoplastic lymphoid tissue to be clearly delineated. Of particular interest is the fact that all cases of follicular lymphoma of germinal centre cell origin are consistently accompanied by a proliferation of DRC, even when the neoplasm is present in non-lymphoid tissue--for example, in the kidney. In contrast, DRC in B cell lymphomas of non-germinal centre origin are partially or totally obliterated.


Identifying targets of multiple co-regulating transcription factors from expression time-series by Bayesian model comparison.
Complete transcriptional regulatory network inference is a huge challenge because of the complexity of the network and sparsity of available data. One approach to make it more manageable is to focus on the inference of context-specific networks involving a few interacting transcription factors (TFs) and all of their target genes. We present a computational framework for Bayesian statistical inference of target genes of multiple interacting TFs from high-throughput gene expression time-series data. We use ordinary differential equation models that describe transcription of target genes taking into account combinatorial regulation. The method consists of a training and a prediction phase. During the training phase we infer the unobserved TF protein concentrations on a subnetwork of approximately known regulatory structure. During the prediction phase we apply Bayesian model selection on a genome-wide scale and score all alternative regulatory structures for each target gene. We use our methodology to identify targets of five TFs regulating Drosophila melanogaster mesoderm development. We find that confident predicted links between TFs and targets are significantly enriched for supporting ChIP-chip binding events and annotated TF-gene interations. Our method statistically significantly outperforms existing alternatives. Our results show that it is possible to infer regulatory links between multiple interacting TFs and their target genes even from a single relatively short time series and in presence of unmodelled confounders and unreliable prior knowledge on training network connectivity. Introducing data from several different experimental perturbations significantly increases the accuracy.


Mechanism of activation of protein kinase JAK2 by the growth hormone receptor.
Signaling from JAK (Janus kinase) protein kinases to STAT (signal transducers and activators of transcription) transcription factors is key to many aspects of biology and medicine, yet the mechanism by which cytokine receptors initiate signaling is enigmatic. We present a complete mechanistic model for activation of receptor-bound JAK2, based on an archetypal cytokine receptor, the growth hormone receptor. For this, we used fluorescence resonance energy transfer to monitor positioning of the JAK2 binding motif in the receptor dimer, substitution of the receptor extracellular domains with Jun zippers to control the position of its transmembrane (TM) helices, atomistic modeling of TM helix movements, and docking of the crystal structures of the JAK2 kinase and its inhibitory pseudokinase domain with an opposing kinase-pseudokinase domain pair. Activation of the receptor dimer induced a separation of its JAK2 binding motifs, driven by a ligand-induced transition from a parallel TM helix pair to a left-handed crossover arrangement. This separation leads to removal of the pseudokinase domain from the kinase domain of the partner JAK2 and pairing of the two kinase domains, facilitating trans-activation. This model may well generalize to other class I cytokine receptors.


The Health Information National Trends Survey (HINTS): A Resource for Consumer Engagement and Health Communication Research.
The contemporary healthcare system can help improve health literacy outcomes in two ways: first, by nurturing the skills and motivations needed for patients to be actively engaged in their own health and healthcare decisions; and, second, by creating a prepared and proactive healthcare system that adapts to patients' capacities and needs in efficacious ways. In 2001, the National Cancer Institute launched the Health Information National Trends Survey (HINTS) as a way for researchers and planners to understand how the public is interacting with a rapidly changing health information environment. Original iterations of the HINTS national probability sampling strategies took place on a biennial basis, but in subsequent years the protocol moved to a yearly administration. This yields a rich resource of cross-sectional, national surveillance data to evaluate for trends across and within vulnerable populations. Sixteen studies are presented from the published literature to illustrate how HINTS data were used to explore constructs of direct interest to health literacy researchers. Suggestions are given for how this ongoing public surveillance mechanism can be used: (a) to provide a sentinel view of how the public is interacting with information in the environment to address their health needs; (b) to generate research questions and hypotheses for further exploration using complementary methodologies; and


Non-linear spelling in writing after a pure cerebellar lesion.
The most common deficits in processing written language result from damage to the graphemic buffer system and refer to semantic and lexical problems or difficulties in phoneme-graphene conversion. However, a writing disorder that has not yet been studied in depth is the non-linear spelling phenomenon. Indeed, although some cases have been described, no report has exhaustively explained the cognitive mechanism and the anatomical substrates underlying this process. In the present study, we analyzed the modality of non-linear writing in a patient affected by a focal cerebellar lesion, who presented with an alteration of the normal trend to write the order of the letters. Based on this evidence, we analyzed the functional connectivity between the cerebellum and the brain network that subtends handwriting and demonstrated how the cerebellar lesion of the patient affected the connections between the cerebellum and cortical areas that support the anatomical system of writing. This is the first report of non-linear spelling in a patient with a lesion outside the fronto-parietal network, specifically with a focal cerebellar lesion. We propose that non-linear writing can be interpreted in view of the role of the cerebellum in timing and sequential processing. Thus, considering the current functional connectivity data, we hypothesize that the cerebellum might be relevant in the mechanism that allows the correct activation timing of letters within a string and placement of the letters in a specific sequential writing order.


Mitochondrial DNA deletions induce the adenosine monophosphate-activated protein kinase energy stress pathway and result in decreased secretion of some proteins.
Mitochondrial DNA (mtDNA) deletions occur sporadically in zygotic and somatic tissues and reach their highest concentration in substantia nigra. Previously, we noted the increase of the adenosine monophosphate (AMP)-activated protein kinase (AMPK) transcript by microarray in multiple cells and tissues bearing deletions. In this work, we demonstrate that the induction of AMPK transcript is dependent on deletions by quantitative polymerase chain reaction, and also demonstrate a deficiency in adenosine triphosphate (ATP) synthesis in the same cells. Consistent with AMPK induction, its known targets SREBF1 (sterol regulatory element binding protein-1) and ATG12 were inhibited and induced, respectively. AMPK induction is known to decrease secretory processes in some cells, and the secretion of both osteoprotegerin (OPG) and fibronectin (FN) proteins to the extracellular space was significantly deficient. Deletions caused a defect in the adenosine diphosphate (ADP)-ribosylation factor-like 2 (ARL2) transcript, which is known to be important in secretion and interacts with protein phosphatase 2A (PP2A) and thus AMPK. The deletion-dependent dysfunctions occurred even in cells bearing less than 30% deletions, suggesting that the concept of a high biological 'threshold' for deletions should be further revised downward. The defects in ATP synthesis, induction of the AMPK and SREBF1 transcripts, and decreased expression of ARL2 and secretion of OPG and FN were recapitulated by low doses of rotenone, demonstrating that they were a specific consequence of electron transport chain inhibition. Thus, mtDNA deletions result in cellular energy depletion, which causes the induction of AMPK and its regulated targets, and inhibit secretion of some proteins. We integrate these observations into a pathophysiological model for how mitochondrial deletions cause disease.


Evaluating the appropriateness of electronic information resources for learning.
Current US medical students have begun to rely on electronic information repositories-such as UpToDate, AccessMedicine, and Wikipedia-for their pre-clerkship medical education. However, it is unclear whether these resources are appropriate for this level of learning due to factors involving information quality, level of evidence, and the requisite knowledgebase. This study evaluated appropriateness of electronic information resources from a novel perspective: amount of mental effort learners invest in interactions with these resources and effects of the experienced mental effort on learning. Eighteen first-year medical students read about three unstudied diseases in the above-mentioned resources (a total of fifty-four observations). Their eye movement characteristics (i.e., fixation duration, fixation count, visit duration, and task-evoked pupillary response) were recorded and used as psychophysiological indicators of the experienced mental effort. Post reading, students' learning was assessed with multiple-choice tests. Eye metrics and test results constituted quantitative data analyzed according to the repeated Latin square design. Students' perceptions of interacting with the information resources were also collected. Participants' feedback during semi-structured interviews constituted qualitative data and was reviewed, transcribed, and open coded for emergent themes. Compared to AccessMedicine and Wikipedia, UpToDate was associated with significantly higher values of eye metrics, suggesting learners experienced higher mental effort. No statistically significant difference between the amount of mental effort and learning outcomes was found. More so, descriptive statistical analysis of the knowledge test scores suggested similar levels of learning regardless of the information resource used. Judging by the learning outcomes, all three information resources were found appropriate for learning. UpToDate, however, when used alone, may be less appropriate for first-year medical students' learning as it does not fully address their information needs and is more demanding in terms of cognitive resources invested.


Protease-activated receptor (PAR) 1 and PAR4 differentially regulate factor V expression from human platelets.
With the recent interest of protease-activated receptors (PAR) 1 and PAR4 as possible targets for the treatment of thrombotic disorders, we compared the efficacy of protease-activated receptor (PAR)1 and PAR4 in the generation of procoagulant phenotypes on platelet membranes. PAR4-activating peptide (AP)-stimulated platelets promoted thrombin generation in plasma up to 5 minutes earlier than PAR1-AP-stimulated platelets. PAR4-AP-mediated factor V (FV) association with the platelet surface was 1.6-fold greater than for PAR1-AP. Moreover, PAR4 stimulation resulted in a 3-fold greater release of microparticles, compared with PAR1 stimulation. More robust FV secretion and microparticle generation with PAR4-AP was attributable to stronger and more sustained phosphorylation of myosin light chain at serine 19 and threonine 18. Inhibition of Rho-kinase reduced PAR4-AP-mediated FV secretion and microparticle generation to PAR1-AP-mediated levels. Thrombin generation assays measuring prothrombinase complex activity demonstrated 1.5-fold higher peak thrombin levels on PAR4-AP-stimulated platelets, compared with PAR1-AP-stimulated platelets. Rho-kinase inhibition reduced PAR4-AP-mediated peak thrombin generation by 25% but had no significant effect on PAR1-AP-mediated thrombin generation. In conclusion, stimulation of PAR4 on platelets leads to faster and more robust thrombin generation, compared with PAR1 stimulation. The greater procoagulant potential is related to more efficient FV release from intracellular stores and microparticle production driven by stronger and more sustained myosin light chain phosphorylation. These data have implications about the role of PAR4 during hemostasis and are clinically relevant in light of recent efforts to develop PAR antagonists to treat thrombotic disorders.


Evaluation of drug release kinetics from ibuprofen matrix tablets using HPMC.
The aim of this study is to develop a once-daily sustained release matrix tablet of ibuprofen using hydroxypropyl methylcellulose (HPMC) as release controlling factor and to evaluate drug release parameters as per various release kinetic models. In order to achieve required sustained release profile tablets were directly compressed using Avicel pH 101 and Magnesium stearate. The formulated tablets were also characterized by physical and chemical parameters and results were found in acceptable limits. Different dissolution models were applied to drug release data in order to evaluate release mechanisms and kinetics. Criteria for selecting the most appropriate model was based on linearity (coefficient of correlation). The drug release data fit well to the Higuchi expression. Drug release mechanism was found as a complex mixture of diffusion, swelling and erosion.


Polyclonal rearrangements of the T-cell receptor beta-chain in fatal angioimmunoblastic lymphadenopathy.
Genomic rearrangement of germline T-cell antigen receptor (TcR) and immunoglobulin (Ig) genes was studied by Southern blot analysis in seven patients with angioimmunoblastic lymphadenopathy (AILD). In three cases clinically suspected of transformation into malignant lymphoma, hybridization with the TcR beta probe showed markedly dimished intensity in the 11.5 kb germline band after Eco RI digestion and normal germline configuration after Hind III and Bam HI digestion, indicating polyclonal T cell rearrangements. A clonal rearrangement of the TcR beta gene was detected in only one case at initial biopsy. No monoclonal rearrangement of Ig genes was observed. These data show that in some cases of AILD disease progression is indicated by polyclonal TcR rearrangements and not by outgrowth of a malignant clone, supporting the concept of AILD as an immunoregulatory disorder.


Differences in the quality of breast cancer care among vulnerable populations.
It is unknown whether differences in the quality of breast cancer care among women from racial and ethnic minority groups, the elderly, and rural areas have changed over time across the continuum of care. The linked Surveillance, Epidemiology, and End Results-Medicare database identified 22,701 women ages 66-79 years diagnosed with early stage breast cancer from 1992-1999. Multiple breast cancer processes of care were measured, including breast-conserving surgery, radiation therapy, documentation of estrogen receptor status, surveillance mammography, and a combined measure of "adequate care". African-American and Hispanic women were significantly less likely to receive adequate care than White women in unadjusted comparisons (54.7% and 58.0% vs. 68.4% for African-American and Hispanic vs. White women) and adjusted comparisons (adjusted odds ratio [AOR] 0.67; 95% confidence interval [95% CI] 0.59-0.76, and AOR 0.77; 95% CI 0.66-0.90 for African-American and Hispanic women, respectively). The proportion of Asian/Pacific Islander women receiving adequate care was similar to White women. When considering only women diagnosed with breast cancer from 1997-1999, African-American women remained less likely than White women to receive adequate care (AOR 0.63; 95% CI 0.50-0.79). Women ages 75-79 years were less likely to receive adequate care compared with women ages 66-69 years (AOR 0.74; 95% CI 0.69-0.80), and women from rural (vs. metropolitan) areas were less likely to receive adequate care (AOR 0.81; 95% CI 0.73-0.89). The quality of breast cancer care is lower among vulnerable populations across the continuum of care, and many of these differences have not improved in more recent years.


[Reinterventions in anaplastic cancers and sarcomas of the thyroid gland].
In the years 1979-1988, operations were performed on 41 anaplastic carcinomas and 11 sarcomas of the thyroid gland. Out of these cases reoperation was indicated in 18 patients (34.6%): in 11 cases only a palliative tracheotomy could be done (group I), whilst in the remaining 7 patients surgical removal of the recurrent tumour was possible (group II). Analysis of group II patients revealed that the initial surgical intervention had been a macroscopically radical operation. The difference between the survival rates of both groups was highly significant (group I 64 days, group II 412 days mean survival). Despite the fatal prognosis of these highly malignant tumours, we conclude that reoperation for the removal of a recurrent tumour is advisable in those patients in whom the initial operation was a radical excision. Longer survival rates and improved quality of life were achieved. If the initial operation was merely palliative, then only a tracheotomy is the likely procedure in case of recurrence.


Chronic antidepressant exposure enhances 5-hydroxytryptamine7 receptor-mediated cyclic adenosine monophosphate accumulation in rat frontocortical astrocytes.
In rat frontal cortical astrocytes, 5-hydroxytryptamine (5-HT), 5-carboxamidotryptamine and 5-methoxytryptamine elicited a concentration-dependent increase of cyclic AMP accumulation, with EC50 values of 137 +/- 7 nM (5-HT), 10.4 +/- 3.9 nM (5-carboxamidotryptamine) and 57.8 +/- 13.6 nM (5-methoxytryptamine). Accumulation of cyclic AMP stimulated by 5-HT (10 microM) was inhibited by methiothepin (IC50 = 21 +/- 13 nM) but not by pindolol, SCH 23390, ICS 205-930 or GR 113808. These pharmacological effects on cyclic AMP formation in astrocytes are consistent with those observed in 5-HT7 receptor binding studies. Indeed, reverse transcription-polymerase chain reaction analysis revealed the presence of 5-HT7 mRNA in astrocytes. Chronic exposure of astrocytes to mianserin (0.05-4 microM) or amitriptyline (1-8 microM) for 3 days produced a concentration-dependent enhancement of 5-HT-stimulated cyclic AMP accumulation, with EC50 values of 0.47 +/- 0.04 microM (mianserin) and 2.5 +/- 0.3 microM (amitriptyline). The enhancement of cyclic AMP accumulation was time dependent, reaching maximal levels within 2 days. Mianserin exposure (2 microM, for 3 days) resulted in a comparable enhancement of cyclic AMP accumulation mediated by 5-carboxamidotryptamine and 5-methoxytryptamine, whereas this treatment had no effect on cyclic AMP production induced by G protein-specific stimulators and catalytic subunit-selective stimulators. The mianserin-induced enhancement of 5-HT-stimulated cyclic AMP accumulation was decreased by methiothepin (IC50 = 15 +/- 8 nM) and significantly attenuated by pretreatment with 5-HT7 receptor antisense oligonucleotides, suggesting that chronic mianserin exposure produces an increase in 5-HT7 receptor activity. Chronic exposure to maprotiline, setiptiline or clomipramine (5 microM, for 3 days) mimicked the effect of mianserin. The enhancement of 5-HT7 receptor activity after chronic antidepressant exposure may be a mechanism underlying the therapeutic effects.


Apamin inhibits TNF-α- and IFN-γ-induced inflammatory cytokines and chemokines via suppressions of NF-κB signaling pathway and STAT in human keratinocytes.
Atopic dermatitis (AD) is identified by an increase in infiltrations of several inflammatory cells including type 2 helper (Th2) lymphocytes. Th2-related chemokines such as thymus and activation-regulated chemokine (TARC/CCL17) and macrophage-derived chemokine (MDC/CCL22), and pro-inflammatory cytokines including interleukin (IL)-1β and IL-6 are considered to play a crucial role in AD. Tumor necrosis factor (TNF)-α- and interferon (IFN)-γ induce the inflammatory condition through production of TARC, MDC, IL-1β and IL-6, and activations of related transcription factors, such as nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and signal transducer and activator of transcription (STAT) in keratinocytes. Apamin, a peptide component of bee venom, has been reported its beneficial activities in various diseases. However, anti-inflammatory effects of apamin on inflammatory condition in keratinocytes have not been explored. Therefore, the present study aimed to demonstrate the anti-inflammatory effect of apamin on TNF-α- and IFN-γ-induced inflammatory condition in keratinocytes. HaCaT was used as human keratinocytes cell line. Cell Counting Kit-8 was performed to measure a cytotoxicity of apamin. The effects of apamin on TNF-α-/IFN-γ-induced inflammatory condition were determined by real-time PCR and Western blot analysis. Further, NF-κB signaling pathways, STAT1, and STAT3 were analyzed by Western blot and immunofluorescence. Apamin ameliorated the inflammatory condition through suppression of Th2-related chemokines and pro-inflammatory cytokines. Further, apamin down-regulated the activations of NF-κB signaling pathways and STATs in HaCaT cells. These results suggest that apamin has therapeutic effect on AD through improvement of inflammatory condition.


Encoding of emotional memories depends on amygdala and hippocampus and their interactions.
We have studied patients with variable degrees of left hippocampal and amygdala pathology who performed a verbal encoding task during functional magnetic resonance imaging (fMRI) to assess the impact of pathology on emotional-memory performance and encoding-evoked activity. The severity of left hippocampal pathology predicted memory performance for neutral and emotional items alike, whereas the severity of amygdala pathology predicted memory performance for emotional items alone. Encoding-related hippocampal activity for successfully remembered emotional items correlated with the degree of left amygdala pathology. Conversely, amygdala-evoked activity with respect to subsequently remembered emotional items correlated with the degree of left hippocampal pathology. Our data indicate a reciprocal dependence between amygdala and hippocampus during the encoding of emotional memories.


'Obsessed with goals': functions and mechanisms of teleological interpretation of actions in humans.
Humans show a strong and early inclination to interpret observed behaviours of others as goal-directed actions. We identify two main epistemic functions that this 'teleological obsession' serves: on-line prediction and social learning. We show how teleological action interpretations can serve these functions by drawing on two kinds of inference ('action-to-goal' or 'goal-to-action'), and argue that both types of teleological inference constitute inverse problems that can only be solved by further assumptions. We pinpoint the assumptions that the three currently proposed mechanisms of goal attribution (action-effect associations, simulation procedures, and teleological reasoning) imply, and contrast them with the functions they are supposed to fulfil. We argue that while action-effect associations and simulation procedures are generally well suited to serve on-line action monitoring and prediction, social learning of new means actions and artefact functions requires the inferential productivity of teleological reasoning.


Status Complexicus? The Emergence of Pediatric Complex Care.
Discourse about childhood chronic conditions has transitioned in the last decade from focusing primarily on broad groups of children with special health care needs to concentrating in large part on smaller groups of children with medical complexity (CMC). Although a variety of definitions have been applied, the term CMC has most commonly been defined as children and youth with serious chronic conditions, substantial functional limitations, increased health and other service needs, and increased health care costs. The increasing attention paid to CMC has occurred because these children are growing in impact, represent a disproportionate share of health system costs, and require policy and programmatic interventions that differ in many ways from broader groups of children with special health care needs. But will this change in focus lead to meaningful changes in outcomes for children with serious chronic diseases, or is the pediatric community simply adopting terminology with resonance in adult-focused health systems? In this article, we will explore the implications of the rapid emergence of pediatric complex care in child health services practice and research. As an emerging field, pediatric care systems should thoughtfully and rapidly develop evidence-based solutions to the new challenges of caring for CMC, including (1) clearer definitions of the target population, (2) a more appropriate incorporation of components of care that occur outside of hospitals, and (3) a more comprehensive outcomes measurement framework, including the recognition of potential limitations of cost containment as a target for improved care for CMC.


[Organophosphate-induced myotoxicity].
Organophosphates comprise a group of chemical compounds extensively used in farming as insecticides, which cause accidental poisoning in animals and men and are also used in suicide attempts. The toxicity of these compounds is due especially to the cardiac and respiratory impairment in consequence of autonomic nervous system disorders. However, it is known that some of these products induce a myopathy in experimental animals and humans. This myopathy is characterized by muscle cell degeneration, involving above all the respiratory muscles. Based on the fact that this involvement certainly enhances the respiratory impairment, this study offers an experimental method for routine evaluation of organophosphate myotoxicity, using a minimal and sufficient battery of stains and histochemical reactions, for muscle necrosis quantification. For this purpose, albino rats (Wistar) treated with the organophosphate paraoxon, were used both with and without antidotes (atropine or pralidoxime). Muscle fiber necrosis in the diaphragm of the rats treated with paraoxon or paraoxon and atropine, that affected about 15% of the fibers in some areas, was detected. In the group treated with paraoxon and pralidoxime, a minimal necrosis was seen, revealing a protective role of this later antidote during the development of myopathy.


Role of glutathione and dependent enzymes in anthracycline-resistant HL60/AR cells.
We studied the cellular enzymatic defenses against anthracycline-induced free radical damage in the HL60 human myelogenous leukemia cell line and in its anthracycline-resistant subline, HL60/AR. Intracellular glutathione (GSH) levels and gamma-glutamyl transpeptidase activity were lower in HL60/AR than in HL60 cells. Glutathione-S-transferase (GST) and glutathione peroxidase activities were similar in both cell lines. The intracellular distribution of GSH/GST was visualized by digitized video fluorescence microscopy, utilizing the fluorescent probe monochlorobimane fluorescence microscopy, utilizing the fluorescent probe monochlorobimane (MBCl), which is specifically conjugated to GSH by GST. In HL60 cells stained with the MBCl probe, a bright diffuse cytoplasmic and nuclear fluorescence pattern was observed, whereas in HL60/AR cells, the fluorescence was mostly localized to the Golgi apparatus with a lesser component of diffuse cytoplasmic and nuclear fluorescence. Pretreatment of HL60/AR cells with buthionine sulfoximine (BSO) partially reversed resistance to daunorubicin. This effect of BSO on resistance was associated not only with the abolition of localized MBCl fluorescence to the Golgi apparatus but also with increased intracellular accumulation and retention of daunorubicin. The results of our studies demonstrate that inhibition of GSH synthesis in HL60/AR cells results in significant sensitization to daunorubicin and suggest that changes in the intracellular distribution of GSH/GST and/or increased drug retention may be involved in mediating this effect.


Cotinine concentration in serum correlates with tobacco smoke-induced emphysema in mice.
Secondhand smoke (SHS) has been associated with a variety of adverse health outcomes in nonsmokers, including emphysema (a chronic obstructive pulmonary disease). One way to detect SHS exposure is to measure the concentration of cotinine, the primary metabolite of nicotine, in bodily fluids. We have developed a method for cotinine analysis by combining micellar electrokinetic chromatography with enrichment techniques. We employed the method to measure cotinine concentrations in serum samples of mice exposed to tobacco smoke for 12 or 24 weeks and found that it was 3.1-fold or 4.8-fold higher than those exposed to room air for the same period. Further, we investigated the morphological changes in lungs of mice and observed tobacco smoke induced emphysema. Our results indicate that the method can be used to measure cotinine and there is an association between the serum cotinine concentration and tobacco smoke-induced emphysema in mice.


Neural Mechanisms of Temporal Resolution of Attention.
The dynamic nature of the world requires that our visual representations are continuously updated. These representations are more precise if there is a narrow time window over which information is averaged. We assess the neural processes of visual updating by testing patients with lesions including inferior parietal cortex, control patients and healthy adults on a continuous visual monitoring task. In Experiment 1, observers kept track of the changing spatial period of a luminance grating and identified the final spatial period after the stimulus disappeared. Healthy older adults and neurological controls were able to perform better than simulated guesses, but only 3 of 11 patients with damage including parietal cortex were able to reach performance that differed from simulated guesses. The effects were unrelated to lesion size. Poor performance on this task is consistent with an inability to selectively attend to the final moment at which the stimulus was seen. To investigate the temporal limits of attention, we varied the rate of stimulus change in Experiment 2. Performance remained poor for some patients even with slow 2.5 Hz change rates. The performance of 4 patients with parietal damage displayed poor temporal precision, namely recovery of performance with slower rates of change.


Pandemic's Experience Questioning Capitalistic Dominance Comment on "Ensuring Global Health Equity in a Post-pandemic Economy".
Reflecting on the up-to-date global experience of the coronavirus disease 2019 (COVID-19) pandemic is of crucial importance in order to draw conclusions needed for the design of policies aiming the prevention of new epidemics and the effective protection, preparedness and response of any new emerging. Ongoing environmental destruction, excess mortality by COVID-19 and non-COVID diseases reflecting the dismantlement and commodification of both public health services and healthcare services, deep economic crisis, increasing and deepening social inequalities are the main characteristics raised by the pandemic. The causes of the causes of all these are the dominant rules of the capitalistic system, driven mainly by the unlimited greed for profit on the expenses of the majority of the society. The effectiveness of any proposed correction of this system is discussed and the need for another society responding to the needs of the population is argued.


Hereditary retinal degeneration in Drosophila melanogaster. A mutant defect associated with the phototransduction process.
Two genes in Drosophila, rdgA and rdgB, which when defective cause retinal degeneration, were discovered by Hotta and Benzer (Hotta, Y., and S. Benzer. 1970. Proc. Natl, Acad. Sci. U. S, A. 67:1156-1163). These mutants have photoreceptor cells that are histologically normal upon eclosion but subsequently degenerate. The defects in the rdgA and rdgB mutants were localized by the study of genetic mosaics to the photoreceptor cells. In rdgB mutants retinal degeneration is light induced. It can be prevented by rearing the flies in the dark or by blocking the receptor potential with a no-receptor-potential mutation, norpA. Vitamin A deprivation and genetic elimination of the lysosomal enzyme acid phosphatase alsoprotect the photoreceptors of rdgB flies against light-induced damage. The photopigment kinetics of dark-reared rdgB flies appear normal in vitro by spectrophotometric measurements, and in vivo by measurements of the M potential. In normal Drosophila, a 1-s exposure to intense 470-nm light produces a prolonged depolarizing afterpotential (PDA) which can last for several hours. In dark-reared rdgB mutants the PDA lasts less than 2 min;; it appears to initiate the degeneration process, since the photoreceptors become permanently unresponsive after a single such exposure. Another mutant was isolated which prevents degeneration in rdgB flies but which has a normal receptor potential. This suppressor of degeneration is an allele of norpA. It is proposed that the normal norpA gene codes for a product which, when activated, leads to the receptor potential, and which is inactivated by the product of the normal rdgB gene.


Neurocognitive deficits in HIV-infected women and victims of childhood trauma.
The study investigated the behavioral and brain effects of childhood trauma and human immunodeficiency virus (HIV) infection, both separately and in combination, and assessed potential interactions in women who were dually affected. Eighty-three HIV-positive and 47 matched HIV-negative South African women underwent neuromedical, neuropsychiatric, and neurocognitive assessments. Univariate tests of significance assessed if either HIV infection or childhood trauma, or the combination, had a significant effect on neurocognitive performance. The majority of women were Black (96%) and had an average age of 30 years. An analysis of covariance revealed significant HIV effects for the Hopkins Verbal Learning Test (HVLT) learning and delay trials (p < 0.01) and the Halstead Category Test (HCT) (p < 0.05). A significant trauma effect was seen on the HVLT delay trial (p < 0.05). The results provide evidence for neurocognitive dysfunction in memory and executive functions in HIV-infected women and memory disturbances in trauma exposed women.


Mechanisms of cervical spine injury in rugby union: a systematic review of the literature.
Severe cervical spine injury in rugby union (rugby) exerts a major impact on the individual who sustains the injury and on the broader society. Since the late 1970s, authors of rugby case report studies have postulated that the underlying mechanism of cervical spine injury is hyperflexion of the neck. However, this is in conflict with findings from more recent experimental studies. These have shown that it is more likely that the majority of cervical spine injuries occur due to buckling of the cervical spinal column. To investigate the primary mechanism of cervical spine injury in rugby. A comprehensive and systematic review of the literature was undertaken. Six key factors were identified and subsequently used to investigate the two principally postulated mechanisms of cervical spine injury: hyperflexion and buckling. Facet dislocations, in particular bilateral facet dislocations, were identified as the most common types of cervical spine injury in rugby. Trauma occurred most often at lower cervical spinal levels, notably the C4/5 and C5/6 motion segments. Experimental studies demonstrate that bilateral facet dislocations occurring at the lower cervical spinal levels are primarily produced via buckling. Our analysis of key factors for cervical spine injury in rugby shows that it is unlikely that the majority of injuries occur after hyperflexion of the neck. It appears more likely that they are the result of buckling of the cervical spinal column.


Analysis of the association between the incidence of two spatially aggregated foliar diseases of strawberry.
ABSTRACT Association of the incidence of leaf blight (caused by Phomopsis obscurans) and leaf spot of strawberry (caused by Mycosphaerella fragariae) was assessed at multiple scales in perennial plantings at several commercial farms over 3 years (1996 to 1998). For each field, the presence or absence of each disease was recorded from n = 15 leaflets in each of N approximately 70 evenly spaced sampling units, and the proportion of leaflets with blight, spot, and total disease (blight or spot) was determined. Individual diseases and total disease incidence were all well described by the beta-binomial distribution but not by the binomial distribution, indicating overdispersion of disease. The Jaccard similarity index was used to measure disease co-occurrence at the leaflet, sampling-unit, and field scales. Standard errors of this index for the lower two scales were obtained using the jackknife (resampling) procedure, and data randomizations were used to determine the expected Jaccard index for an independent arrangement of the two diseases, conditioned on the incidence and spatial heterogeneity of the observed disease data. Results based on these statistics showed that only 4 of 52 data sets at the leaflet level and no data sets at the sampling-unit level had Jaccard index values significantly different from that expected under an independent rearrangement of the two diseases. Rank correlation and cross-correlation statistics were calculated to determine the degree of covariation in incidence between the two diseases. Additionally, covariation between diseases was tested using a new procedure in the Spatial Analysis by Distance IndicEs (SADIE) class of tests. Covariation was detected in 21% of the data sets using rank correlation methods and in 15% of the data sets using the SADIE-based approach. The discrepancy between these two methods may be due to the rank correlation procedure not taking into account the effects of spatial pattern of disease incidence. There was no relationship between mean disease incidence per field of spot and blight or between degree of heterogeneity of the two diseases (as measured by theta of the beta-binomial distribution), demonstrating lack of covariation at the field scale. Incidence of leaflets with either disease (total disease incidence) could be well predicted using a linear combination of the estimated probabilities of leaf blight and leaf spot incidence based on independence of the two diseases. Heterogeneity of total disease incidence, measured with the estimated theta parameter of the beta-binomial distribution, could also be well predicted using a linear combination of the weighted theta values for leaf blight and leaf spot, with weights proportional to incidence of the individual diseases.


Multisite transcranial Random Noise Stimulation (tRNS) modulates the distress network activity and oscillatory powers in subjects with chronic tinnitus.
Tinnitus is a common disorder in which auditory and non-auditory areas are involved as a network. Therefore, the multisite protocol of brain stimulation (tRNS) seems effective in relieving tinnitus symptoms. This study aimed to verify the modulatory effects of multisite tRNS on the tinnitus network. Thirty-two tinnitus-suffering patients received two consecutive sessions of tRNS (SHAM+ REAL) applied over the T3, T4 (the auditory cortex group) and F4, FP1 followed by T3, T4 (the multisite group). A 3-min resting-state EEG was recorded before and immediately after each session. After the multisite-tRNS real session, there was a significantly increased power in the alpha-1 band at the auditory and prefrontal cortex accompanied by decreased power in the delta and beta-2 bands in the prefrontal cortex. Standardized low-resolution brain electromagnetic tomography (sLORETA) showed a significant decrease in beta-2 activity in the prefrontal cortex, anterior cingulate cortex, and the parahippocampus and decreased alpha connectivity between the right prefrontal cortex and the left auditory cortex. No significant effects were noticed for the sham session. This study showed that tRNS has modulatory effects on the electrical activity of the brain and that targeting prefrontal and auditory areas in sequence could modulate the distress network and multiple hubs in the tinnitus network.


Positive selection of tyrosine loss in metazoan evolution.
John Nash showed that within a complex system, individuals are best off if they make the best decision that they can, taking into account the decisions of the other individuals. Here, we investigate whether similar principles influence the evolution of signaling networks in multicellular animals. Specifically, by analyzing a set of metazoan species we observed a striking negative correlation of genomically encoded tyrosine content with biological complexity (as measured by the number of cell types in each organism). We discuss how this observed tyrosine loss correlates with the expansion of tyrosine kinases in the evolution of the metazoan lineage and how it may relate to the optimization of signaling systems in multicellular animals. We propose that this phenomenon illustrates genome-wide adaptive evolution to accommodate beneficial genetic perturbation.


SIN1/MIP1 maintains rictor-mTOR complex integrity and regulates Akt phosphorylation and substrate specificity.
Mammalian target of rapamycin (mTOR) controls cell growth and proliferation via the raptor-mTOR (TORC1) and rictor-mTOR (TORC2) protein complexes. Recent biochemical studies suggested that TORC2 is the elusive PDK2 for Akt/PKB Ser473 phosphorylation in the hydrophobic motif. Phosphorylation at Ser473, along with Thr308 of its activation loop, is deemed necessary for Akt function, although the regulatory mechanisms and physiological importance of each phosphorylation site remain to be fully understood. Here, we report that SIN1/MIP1 is an essential TORC2/PDK2 subunit. Genetic ablation of sin1 abolished Akt-Ser473 phosphorylation and disrupted rictor-mTOR interaction but maintained Thr308 phosphorylation. Surprisingly, defective Ser473 phosphorylation affected only a subset of Akt targets in vivo, including FoxO1/3a, while other Akt targets, TSC2 and GSK3, and the TORC1 effectors, S6K and 4E-BP1, were unaffected. Our findings reveal that the SIN1-rictor-mTOR function in Akt-Ser473 phosphorylation is required for TORC2 function in cell survival but is dispensable for TORC1 function.


Modeling auditory-nerve responses for high sound pressure levels in the normal and impaired auditory periphery.
This paper presents a computational model to simulate normal and impaired auditory-nerve (AN) fiber responses in cats. The model responses match physiological data over a wider dynamic range than previous auditory models. This is achieved by providing two modes of basilar membrane excitation to the inner hair cell (IHC) rather than one. The two modes are generated by two parallel filters, component 1 (C1) and component 2 (C2), and the outputs are subsequently transduced by two separate functions. The responses are then added and passed through the IHC low-pass filter followed by the IHC-AN synapse model and discharge generator. The C1 filter is a narrow-band, chirp filter with the gain and bandwidth controlled by a nonlinear feed-forward control path. This filter is responsible for low and moderate level responses. A linear, static, and broadly tuned C2 filter followed by a nonlinear, inverted and nonrectifying C2 transduction function is critical for producing transition region and high-level effects. Consistent with Kiang's two-factor cancellation hypothesis, the interaction between the two paths produces effects such as the C1/C2 transition and peak splitting in the period histogram. The model responses are consistent with a wide range of physiological data from both normal and impaired ears for stimuli presented at levels spanning the dynamic range of hearing.


Outcomes of small for gestational age infants born at <27 weeks' gestation.
To determine whether small for gestational age (SGA) infants born at <27 weeks gestational age (GA) are at increased risk for mortality, morbidity, and growth and neurodevelopmental impairment at 18-22 months corrected age. This was a retrospective cohort study from National Institute of Child Health and Human Development Neonatal Research Network's Generic Database and Follow-Up Studies. Infants born at <27 weeks GA between January 2006 and July 2008 were included. SGA was defined as birth weight <10th percentile for GA based on Olsen growth curves. Infants with birth weight ≥ 10th percentile for GA were classified as non-SGA. Maternal and infant characteristics, neonatal outcomes, and neurodevelopmental data were compared in SGA and non-SGA infants. Neurodevelopmental impairment was defined as any of the following: cognitive score <70 on the Bayley Scales of Infant Development III, moderate or severe cerebral palsy, bilateral hearing loss (with and without amplification), or blindness (bilateral vision <20/200). Logistic regression analysis was applied to evaluate the associations between SGA status and death or neurodevelopmental impairment. The SGA group comprised 385 infants; the non-SGA group, 2586 infants. Compared with mothers of non-SGA infants, mothers of SGA infants were more likely to have a high school education, prenatal care, cesarean delivery, pregnancy-induced hypertension, and antenatal corticosteroid exposure. Compared with non-SGA infants, SGA infants had higher mortality and were more likely to have postnatal growth failure, prolonged mechanical ventilation, and postnatal steroid use. SGA status was associated with increased risk of death or neurodevelopmental impairment (OR, 3.91; 95% CI, 2.91-5.25; P < .001). SGA status in infants born at <27 weeks GA is associated with an increased likelihood of postnatal steroid use, mortality, growth failure, and neurodevelopmental impairment at 18-22 months corrected age.


Rituximab in paediatric onset multiple sclerosis: a case series.
Paediatric onset multiple sclerosis (POMS) is characterized by high inflammatory activity. No disease modifying treatment has been approved for POMS. The objective of this report was to report the use of rituximab, a B cell depleting monoclonal anti-CD20-antibody, in POMS. This is a retrospective case series at four specialized MS centres in Sweden. Participants were identified through the Swedish MS-registry and our own patient stocks. Data were collected through medical charts review. We identified 14 POMS patients treated with i.v. rituximab 500-1000 mg every 6th to 12th months. Median age at disease onset was 14.7 years, median age at rituximab treatment initiation was 16.5 years, and median treatment duration was 23.6 months. No relapses were reported, and the EDSS scores remained stable or decreased in 13 of 14 cases during rituximab treatment. Beyond 6 months from initiating rituximab treatment, only one new lesion was detected on MRI. No serious AEs were reported. The drug survival was 86%. Our data indicate that rituximab treatment is safe, effective and well tolerated in children with MS. Nine POMS cases treated with rituximab have previously been published. They had higher disease activity pre-rituximab, but similar safety and efficacy outcomes after treatment. An RCT of rituximab in POMS is warranted.