Bone marrow-derived cells are sufficient and necessary targets to mediate glomerulonephritis and vasculitis induced by anti-myeloperoxidase antibodies.
Clinical and experimental evidence indicate that ANCA cause pauci-immune necrotizing and crescentic glomerulonephritis (NCGN) and systemic small vessel vasculitis in humans. One of the major target antigens for ANCA is myeloperoxidase (MPO). An animal model that closely resembles the human disease is induced by intravenous injection of anti-MPO IgG into mice. The likely primary pathogenic targets for the anti-MPO IgG are circulating neutrophils and monocytes, although other cells have been implicated, including endothelial cells and epithelial cells. Herein is reported a new model for anti-MPO-mediated glomerulonephritis and vasculitis that further documents the pathogenic potential of ANCA and demonstrates that bone marrow (BM)-derived cells are sufficient targets to cause anti-MPO disease in the absence of MPO in other cell type. MPO knockout (Mpo-/-) mice that were immunized with mouse MPO were exposed to irradiation and received a transplant of Mpo+/+ or Mpo-/- BM. Engraftment in mice with circulating anti-MPO resulted in development of pauci-immune NCGN in all mice and pulmonary capillaritis and splenic necrotizing arteritis in some. Anti-MPO IgG also was introduced intravenously into chimeric mice by transplantation of Mpo+/+ BM into irradiated Mpo-/- mice or Mpo-/- BM into irradiated Mpo+/+ mice. Chimeric Mpo-/- mice with circulating MPO-positive neutrophils developed NCGN, whereas chimeric Mpo+/+ mice with circulating MPO-negative neutrophils did not, thereby indicating that BM-derived cells are not only sufficient but also necessary for induction of anti-MPO disease. This novel animal model further documents ANCA IgG interactions with neutrophils as a cause of ANCA-associated glomerulonephritis and vasculitis.


Local circuitry of identified projection neurons in cat visual cortex brain slices.
The relationship between pyramidal cell morphology and efferent target was investigated in layer 6 of cat primary visual cortex (area 17). Layer 6 has 2 projections, one to the lateral geniculate nucleus (LGN) and another to the visual claustrum. The cells of origin of each projection were identified by retrograde transport of fluorescent latex microspheres. The labeled cells were visualized in brain slices prepared from area 17, using an epifluorescence compound microscope modified for intracellular recording. Individual retrogradely labeled cells were penetrated and intracellularly stained with Lucifer yellow to visualize the patterns of axons and dendrites associated with each projection. The neurons that give rise to the 2 projections had very different patterns of dendrites and local axonal collaterals, but the patterns within each group were highly stereotyped. The differences between their axonal collaterals were particularly dramatic. Claustrum projecting cells had fine, horizontally directed collaterals that arborized exclusively in layer 6 and lower layer 5. Most LGN projecting cells had virtually no horizontal arborization in layer 6. Instead, they sent widespread collaterals vertically, which arborized extensively in layer 4. The apical dendrites of the 2 groups also differed markedly. Claustrum projecting cells had apical dendrites reaching to layer 1, with branches in layer 5 only, while LGN projecting cells never had an apical dendrite reaching higher than layer 3, with side branches in layers 5 and 4. Therefore, each efferent target must receive inputs from neurons whose synaptic connections within area 17 are significantly different from those of neurons projecting to other targets. This further suggests that distinct visual response properties should be associated with each projection. In addition to the claustrum and LGN projecting cells, about 20% of layer 6 pyramidal neurons lacked an efferent axon. Morphologically, most resembled LGN projecting neurons, but a few had characteristics of claustrum projecting cells. These neurons may represent cells that either failed to make an efferent connection or cells that lost an efferent axon during development. Their frequency suggests that such intrinsic, presumably excitatory, neurons may play a significant role in cortical processing.


Oxygen and lactate concentrations measured in vivo in the intervertebral discs of patients with scoliosis and back pain.
Oxygen concentrations in intervertebral discs were measured in 10 patients during discography and in 13 patients with scoliosis and 11 patients with back pain during spinal surgery. Lactate concentration profiles were measured in 12 of these discs. The discs were graded for degeneration by magnetic resonance imaging and histology where possible. To determine if oxygen and lactate levels in human discs vary with degree of degeneration. Failure of nutrient transport is thought to lead to disc degeneration. The disc is avascular. Oxygen is used by the disc cells, and lactate is produced. Low oxygen and high lactate concentrations have been measured in the center of healthy animal discs. Oxygen concentrations were measured amprometrically. The sterilized gold-needle electrode was introduced into the disc during discography or after the disc was exposed surgically via an anterior approach. Concentration profiles of each disc took approximately 5 minutes to measure. Lactate concentrations were measured biochemically on the excised disc segment. Oxygen concentrations were highest at the disc surface and fell toward the center. Lactate concentrations showed the reverse profile. Oxygen levels were very variable, ranging from 5-150 mm Hg in the center of the nucleus. No correlation was seen with age, pathology, or degree of degeneration. Lactate concentrations ranged for the most part from 2 mmol/L to 6 mmol/L. Concentrations of metabolites depend on cellular activity and on transport of the metabolite between the blood supply and the cell. The correlation between degeneration and nutrition cannot be determined only from metabolite concentrations; measurements of metabolic activity and nutrient transport rates also are required.


Enhanced versus simply positive: A new condition-based regression analysis to disentangle effects of self-enhancement from effects of positivity of self-view.
Despite a large body of literature and ongoing refinements of analytical techniques, research on the consequences of self-enhancement (SE) is still vague about how to define SE effects, and empirical results are inconsistent. In this paper, we point out that part of this confusion is due to a lack of conceptual and methodological differentiation between effects of individual differences in how much people enhance themselves (SE) and in how positively they view themselves (positivity of self-view; PSV). We show that methods commonly used to analyze SE effects are biased because they cannot differentiate between the effects of PSV and the effects of SE. We provide a new condition-based regression analysis (CRA) that unequivocally identifies effects of SE by testing intuitive and mathematically derived conditions on the coefficients in a bivariate linear regression. Using data from 3 studies on intellectual SE (total N = 566), we then illustrate that the CRA provides novel results as compared with traditional methods. Results suggest that many previously identified SE effects are in fact effects of PSV alone. The new CRA approach thus provides a clear and unbiased understanding of the consequences of SE. It can be applied to all conceptualizations of SE and, more generally, to every context in which the effects of the discrepancy between 2 variables on a third variable are examined. (PsycINFO Database Record


The effectiveness and cost-effectiveness of shared care: protocol for a realist review.
Shared care (an enhanced information exchange over and above routine outpatient letters) is commonly used to improve care coordination and communication between a specialist and primary care services for people with long-term conditions. Evidence of the effectiveness and cost-effectiveness of shared care is mixed. Informed decision-making for targeting shared care requires a greater understanding of how it works, for whom it works, in what contexts and why. This protocol outlines how realist review methods can be used to synthesise evidence on shared care for long-term conditions.A further aim of the review is to explore economic evaluations of shared care. Economic evaluations are difficult to synthesise due to problems in accounting for contextual differences that impact on resource use and opportunity costs. Realist review methods have been suggested as a way to overcome some of these issues, so this review will also assess whether realist review methods are amenable to synthesising economic evidence. Database and web searching will be carried out in order to find relevant evidence to develop and test programme theories about how shared care works. The review will have two phases. Phase 1 will concentrate on the contextual conditions and mechanisms that influence how shared care works, in order to develop programme theories, which partially explain how it works. Phase 2 will focus on testing these programme theories. A Project Reference Group made up of health service professionals and people with actual experience of long-term conditions will be used to ground the study in real-life experience. Review findings will be disseminated through local and sub-national networks for integrated care and long-term conditions. This realist review will explore why and for whom shared care works, in order to support decision-makers working to improve the effectiveness of care for people outside hospital. The development of realist review methods to take into account cost and cost-effectiveness evidence is particularly innovative and challenging, and if successful will offer a new approach to synthesising economic evidence. This systematic review protocol is registered on the PROSPERO database (registration number: CRD42012002842).


In Vivo Retargeting of Poly(beta aminoester) (OM-PBAE) Nanoparticles is Influenced by Protein Corona.
One of the main bottlenecks in the translation of nanomedicines from research to clinics is the difficulty in designing nanoparticles actively vectorized to the target tissue, a key parameter to ensure efficacy and safety. In this group, a library of poly(beta aminoester) polymers is developed, and it is demonstrated that adding specific combinations of terminal oligopeptides (OM-PBAE), in vitro transfection is cell selective. The current study aims to actively direct the nanoparticles to the liver by the addition of a targeting molecule. To achieve this objective, retinol, successfully attached to OM-PBAE, is selected as hepatic targeting moiety. It is demonstrated that organ biodistribution is tailored, achieving the desired liver accumulation. Regarding cell type transfection, antigen presenting cells in the liver are those showing the highest transfection. Thanks to proteomics studies, organ but not cellular biodistribution can be explained by the formation of differential protein coronas. Therefore, organ biodistribution is governed by differential protein corona formed when retinol is present, while cellular biodistribution is controlled by the end oligopeptides type. In summary, this work is a proof of concept that demonstrates the versatility of these OM-PBAE nanoparticles, in terms of the modification of the biodistribution of OM-PBAE nanoparticles adding active targeting moieties.


Identifying Drivers of Parallel Evolution: A Regression Model Approach.
Parallel evolution, defined as identical changes arising in independent populations, is often attributed to similar selective pressures favoring the fixation of identical genetic changes. However, some level of parallel evolution is also expected if mutation rates are heterogeneous across regions of the genome. Theory suggests that mutation and selection can have equal impacts on patterns of parallel evolution; however, empirical studies have yet to jointly quantify the importance of these two processes. Here, we introduce several statistical models to examine the contributions of mutation and selection heterogeneity to shaping parallel evolutionary changes at the gene-level. Using this framework, we analyze published data from forty experimentally evolved Saccharomyces cerevisiae populations. We can partition the effects of a number of genomic variables into those affecting patterns of parallel evolution via effects on the rate of arising mutations, and those affecting the retention versus loss of the arising mutations (i.e., selection). Our results suggest that gene-to-gene heterogeneity in both mutation and selection, associated with gene length, recombination rate, and number of protein domains drive parallel evolution at both synonymous and nonsynonymous sites. While there are still a number of parallel changes that are not well described, we show that allowing for heterogeneous rates of mutation and selection can provide improved predictions of the prevalence and degree of parallel evolution.


Analysis of Present Status for Surgery of Benign Prostatic Hyperplasia in Korea Using Nationwide Healthcare System Data.
There are no established statistical data available for the comparison of different surgical methods adopted for the resection of benign prostatic hyperplasia (BPH). This study investigates the present status related to BPH surgery in Korea for the past 8 years. National-level data from the National Health Insurance Service and National Statistical Office were analyzed in this study. From 2010 to 2017, the trends of surgeries for BPH were reviewed according to the procedure code including transurethral resection of the prostate (TURP), holmium laser enucleation of the prostate (HoLEP), or high-power potassium titanyl phosphate (KTP), and this trend also analyzed by age, geographic distribution, and hospital type. Over the past 8 years, there was not much change in the total number of BPH-related surgeries (range, 10,393- 11,072). Although there was not much alteration in the number of conventional TURP (from 6,801 in 2010 to 6,645 in 2017), the number of HoLEP has dramatically increased (from 278 in 2010 to 3,805 in 2017). The number of HoLEP surgeries after 2011 exceeded the number of surgeries using KTP, and the gap is anticipated to rise. The number of surgeries by age group was most common in the 70s and the total number of surgeries is decreasing in all age groups; for HoLEP, the trend is steadily increasing over the age of 60 years. Most of the BPH surgeries were performed in metropolitan areas, such as Seoul, Gyeonggido, and Busan, and in larger hospitals compared to smaller hospital settings. Through the data of the National Health Insurance Service, we could apprehend the present status of BPH-related surgery in Korea. Then, we could know about the trend according to several factors and we think these results will be valuable as academic references as well.


Radio frequency heating of metallic and semiconducting single-walled carbon nanotubes.
Here we report the effect of metallic (m-) and semiconducting (s-) properties of single-walled carbon nanotubes (SWCNTs) on the response of SWCNT films to radio frequency (RF) heating. We separated high-purity m- and s-SWCNTs from an initial SWCNTs mixture and prepared thin films using vacuum filtration method. The areal density of the films is 9.6 μg cm-2, and the DC conductivities are in the range of 7800-49 000 S m-1. We show rapid and non-contact Joule heating of films using a fringing-field RF applicator, and we observe maximum heating rates in the frequency range of 60-70 MHz. We determine that the more conductive m-SWCNT films reflect RF fields and heat at a maximum rate of 1.51 °C s-1 compared to maximum heating rate of 25.6 °C s-1 for s-SWCNT films. However, m-SWCNTs heat up faster than s-SWCNTs when dispersed in a dielectric medium. Our results confirm the non-monotonic relationship between RF heating rate and conductivity for CNT-based materials such that conductivity is required for heating but high values are correlated with reflections. Our findings also suggest that RF heating could be a possible metric for evaluating film purity because impurities in the films affect the conductivity and thus RF heating rate. We anticipate that RF heating may occur in SWCNT-based electronics and affect their performance.


Treatment of hepatocellular carcinoma associated with cirrhosis in the era of liver transplantation.
To review the treatment of cirrhotic patients with hepatocellular carcinoma in the era of liver transplantation and to determine the most appropriate approach to the treatment of patients at different stages of disease. A MEDLINE search of English-language articles published between 1981 and 1997 and the clinical experience of the Mount Sinai Liver Transplant Program. Selected studies were 1) original articles reporting results of resection and transplantation in the treatment of hepatocellular carcinoma in cirrhotic patients and 2) initial reports from major transplantation centers of multimethod therapies combining chemotherapy with transplantation. Study designs were assessed with careful attention to methods and aims. Relevant data on patient population, tumor stage distribution, treatment, survival, and rate of recurrent disease were extracted and analyzed. Options for the treatment of hepatocellular carcinoma in cirrhotic patients vary according to tumor stage and severity of underlying liver disease. Resection remains an important method primarily in eastern countries, where the screening of high-risk populations has been associated with early detection of small asymptomatic lesions. Long-term survival after resection, however, is low. In western countries, liver transplantation is becoming the treatment of choice in patients with advanced cirrhosis and small, unresectable lesions; resection is reserved for cirrhotic patients with small, peripheral lesions and preserved hepatic function. Minimally invasive procedures (such as percutaneous ethanol injection and transarterial chemoembolization) have been developed to treat unresectable tumors. Transarterial chemoembolization may also be effective in patients with advanced cirrhosis and unresectable lesions who are awaiting transplantation. The efficacy of liver transplantation for hepatocellular carcinoma has been proven mainly in patients with advanced cirrhosis and small lesions. Future studies may clarify the role of approaches combining neoadjuvant chemotherapy with transplantation for large (stage III) tumors.


Aldehyde dehydrogenase 2-mediated aldehyde metabolism promotes tumor immune evasion by regulating the NOD/VISTA axis.
Aldehyde dehydrogenase 2 (ALDH2) is a crucial enzyme involved in endogenous aldehyde detoxification and has been implicated in tumor progression. However, its role in tumor immune evasion remains unclear. Here, we analyzed the relationship between ALDH2 expression and antitumor immune features in multiple cancers. ALDH2 knockout tumor cells were then established using CRISPR/Cas9 system. In immunocompetent breast cancer EMT6 and melanoma B16-F10 mouse models, we investigated the impact of ALDH2 blockade on cytotoxic T lymphocyte function and tumor immune microenvironment by flow cytometry, mass cytometry, Luminex liquid suspension chip detection, and immunohistochemistry. Furthermore, RNA sequencing, flow cytometry, western blot, chromatin immunoprecipitation assay, and luciferase reporter assays were employed to explore the detailed mechanism of ALDH2 involved in tumor immune evasion. Lastly, the synergistic therapeutic efficacy of blocking ALDH2 by genetic depletion or its inhibitor disulfiram in combination with immune checkpoint blockade (ICB) was investigated in mouse models. In our study, we uncovered a positive correlation between the expression level of ALDH2 and T-cell dysfunction in multiple cancers. Furthermore, blocking ALDH2 significantly suppressed tumor growth by enhancing cytotoxic activity of CD8+ T cells and reshaping the immune landscape and cytokine milieu of tumors in vivo. Mechanistically, inhibiting ALDH2-mediated metabolism of aldehyde downregulated the expression of V-domain Ig suppressor of T-cell activation (VISTA) via inactivating the nucleotide oligomerization domain (NOD)/nuclear factor kappa-B (NF-κB) signaling pathway. As a result, the cytotoxic function of CD8+ T cells was revitalized. Importantly, ALDH2 blockade markedly reinforced the efficacy of ICB treatment. Our data delineate that ALDH2-mediated aldehyde metabolism drives tumor immune evasion by activating the NOD/NF-κB/VISTA axis. Targeting ALDH2 provides an effective combinatorial therapeutic strategy for immunotherapy.


SWATH mass spectrometry as a tool for quantitative profiling of the matrisome.
Proteomic analysis of extracellular matrix (ECM) and ECM-associated proteins, collectively known as the matrisome, is a challenging task due to the inherent complexity and insolubility of these proteins. Here we present sequential window acquisition of all theoretical fragment ion spectra mass spectrometry (SWATH MS) as a tool for the quantitative analysis of matrisomal proteins in both non-enriched and ECM enriched tissue without the need for prior fractionation. Utilising a spectral library containing 201 matrisomal proteins, we compared the performance and reproducibility of SWATH MS over conventional data-dependent analysis mass spectrometry (DDA MS) in unfractionated murine lung and liver. SWATH MS conferred a 15-20% increase in reproducible peptide identification across replicate experiments in both tissue types and identified 54% more matrisomal proteins in the liver versus DDA MS. We further use SWATH MS to evaluate the quantitative changes in matrisome content that accompanies ECM enrichment. Our data shows that ECM enrichment led to a systematic increase in core matrisomal proteins but resulted in significant losses in matrisome-associated proteins including the cathepsins and proteins of the S100 family. Our proof-of-principle study demonstrates the utility of SWATH MS as a versatile tool for in-depth characterisation of the matrisome in unfractionated and non-enriched tissues. SIGNIFICANCE: The matrisome is a complex network of extracellular matrix (ECM) and ECM-associated proteins that provides scaffolding function to tissues and plays important roles in the regulation of fundamental cellular processes. However, due to its inherent complexity and insolubility, proteomic studies of the matrisome typically require the application of enrichment workflows prior to MS analysis. Such enrichment strategies often lead to losses in soluble matrisome-associated components. In this study, we present sequential window acquisition of all theoretical fragment ion spectra mass spectrometry (SWATH MS) as a tool for the quantitative analysis of matrisomal proteins. We show that SWATH MS provides a more reproducible coverage of the matrisome compared to data-dependent analysis (DDA) MS. We also demonstrate that SWATH MS is capable of accurate quantification of matrisomal proteins without prior ECM enrichment and fractionation, which may simplify sample handling workflows and avoid losses in matrisome-associated proteins commonly linked to ECM enrichment.


Sensory and motor denervation influence epidermal thickness in rat foot glabrous skin.
Denervation in man often results in shiny, dry, thin skin. A previous study has shown that the epidermis of glabrous skin in the rat becomes approximately 40% thinner within 1 week following sciatic nerve transection, but which nerve fiber type or types influence epidermal thickness is unknown. In this study, we compared the effects on the epidermis of selective sensory, motor, and sympathetic denervation. Protein gene product 9.5 and calcitonin gene-related peptide immunocytochemical staining were used to determine the extent of denervation of epidermis, dermis, and sweat glands in the footpads. Epidermal thickness of the glabrous plantar skin of the foot was measured. To verify the specificity and reliability of each animal model, the relevant regions of the peripheral nervous system were examined by light or electron microscopy or both. Epidermal thickness decreased significantly following sciatic nerve transection (58% of control, P < 0.05) and dorsal root ganglionectomy (59%; P < 0.05). The thickness also decreased following lumbar ventral rhizotomy (61%; P < 0.01), destruction of lumbar spinal motor neurons (66%; P < 0.05), and botulinum toxin-induced paralysis of the tibialis anterior and gastrocnemius muscles (70%; P < 0.05). A slight decrease followed dorsal rhizotomy (84%; P < 0.01). In contrast, no significant alterations in epidermal thickness were detected following sham operation and sympathectomy. Epidermal thinning was paralleled by reductions in the amounts of transcripts for glyceraldehyde-3-phosphate dehydrogenase and beta-actin. These results suggest that selective loss of both sensory and motor fibers to the hind limb can contribute to reducing epidermal thickness in rat foot glabrous skin.


Aspergillus brain abscess in a patient with normal immunity--case report.
A 53-year-old male with normal immunity presented with Aspergillus brain abscess manifesting as frontal headache. T2-weighted magnetic resonance imaging revealed a hypointense lesion in the left frontal lobe extending into the right frontal lobe. The hypointense appearance on T2-weighted images appears to be characteristic of aspergillosis. Bifrontal craniotomy exposed an elastic-hard mass in the base of the left frontal lobe extending into the right frontal lobe, and into the left ethmoid sinus. The mass contained a cavity with white fluid. The abscess was removed almost totally. The histological diagnosis was Aspergillus abscess. Antibiotic treatment with amphotericin B and fluconazole was given for 2 months postoperatively. No recurrence was identified during 15-month follow-up.


Anti-JCV antibody prevalence in a French cohort of MS patients under natalizumab therapy.
To measure the prevalence of JCV-specific antibodies in a French cohort of MS patients treated with natalizumab and to identify risk factor(s) of JCV seropositivity. Progressive multifocal leukoencephalopathy (PML) risk may be stratified by anti-JCV antibody status, duration of natalizumab therapy (≥24 months) and prior exposure to immunosuppressive (IS) drugs. No data are available in France on the prevalence of anti-JCV antibodies and distribution of PML risk factors in patients treated with natalizumab. Sera of 361 patients under natalizumab therapy in two MS centers were analyzed using a previously validated ELISA test. We studied different characteristics: demographic, ethnic, radiological, clinical, prior use of immunomodulatory (IM) or IS drugs and natalizumab exposure duration. The JCV seropositivity rate was 51 % for the whole cohort. Mean natalizumab exposure duration was 27.27 months ± 15.57 (mean ± SD), and prior use of IS drugs was observed in 15.24 % of patients. Twenty-three patients (6.4 %) presented the three PML risk factors. By multivariate analysis, presence of anti-JCV antibodies was significantly linked to age, North African origin and natalizumab exposure duration. Anti-JCV antibody prevalence was similar to previously published data. Anti-JCV antibody status was linked to age. We also suggested that anti-JCV antibody status could be linked to natalizumab exposure duration and ethnic characteristics.


Clinical features and prognosis of primary biliary cirrhosis associated with systemic sclerosis.
To evaluate the prognosis of primary biliary cirrhosis (PBC) together with systemic sclerosis (SSc), as this is unknown. A PBC database of 580 patients identified 43 with PBC and SSc: two patients with PBC alone were matched to each PBC-SSc patient for serum bilirubin concentration at the initial visit. Forty (93%) patients had limited cutaneous SSc. At diagnosis of PBC, median values were: 49.7 years, bilirubin 17 micromol/l, and albumin 40.5 g/l. Liver diagnosis occurred a median 4.9 years after SSc in 24 (56%) patients. In matched patients, median values at diagnosis were: 53.2 years, bilirubin 12 micromol/l, and albumin 41 g/l. Median follow up was similar: 3.16 years (PBC-SSc) and 4.8 years (PBC alone). The risk of transplantation or death from diagnosis, adjusting for sex, age, log bilirubin, and alkaline phosphatase was significantly lower in PBC-SSc (hazard ratio 0.116, p=0.01) due to less transplantation (hazard ratio 0.068, p=0.006). The rate of bilirubin increase was less in PBC-SSc (p=0.04). Overall survival was similar (hazard ratio 1.11, p=0.948); there were nine deaths (21%) in PBC-SSc (six SSc related and two liver related) and nine (11%) in PBC alone (six liver related). Liver disease has a slower progression in PBC-SSc compared with matched patients with PBC alone.


Decision support for using mobile Rapid DNA analysis at the crime scene.
Mobile Rapid DNA technology is close to being incorporated into crime scene investigations, with the potential to identify a perpetrator within hours. However, the use of these techniques entails the risk of losing the sample and potential evidence, because the device not only consumes the inserted sample, it is also is less sensitive than traditional technologies used in forensic laboratories. Scene of Crime Officers (SoCOs) therefore will face a 'time/success rate trade-off' issue when making a decision to apply this technology. In this study we designed and experimentally tested a Decision Support System (DSS) for the use of Rapid DNA technologies based on Rational Decision Theory (RDT). In a vignette study, where SoCOs had to decide on the use of a Rapid DNA analysis device, participating SoCOs were assigned to either the control group (making decisions under standard conditions), the Success Rate (SR) group (making decisions with additional information on DNA success rates of traces), or the DSS group (making decisions supported by introduction to RDT, including information on DNA success rates of traces). This study provides positive evidence that a systematic approach for decision-making on using Rapid DNA analysis assists SoCOs in the decision to use the rapid device. The results demonstrated that participants using a DSS made different and more transparent decisions on the use of Rapid DNA analysis when different case characteristics were explicitly considered. In the DSS group the decision to apply Rapid DNA analysis was influenced by the factors "time pressure" and "trace characteristics" like DNA success rates. In the SR group, the decisions depended solely on the trace characteristics and in the control group the decisions did not show any systematic differences on crime type or trace characteristic. Guiding complex decisions on the use of Rapid DNA analyses with a DSS could be an important step towards the use of these devices at the crime scene.


Exposure and risk assessment of the Czech population to chlorinated pesticides and polychlorinated biphenyls using archived serum samples from the period 1970 to 1990.
The serum samples from the years 1970-1990 archived at the temperature of -20°C in the biobank primarily intended for serological survey performed in the CR since 1960 were pooled and analyzed for DDT, its metabolites, HCB, HCHs, and indicator PCB congeners using up-to-date GC/MS/MS methods to retrospectively assess health risks according to current health guidelines. Samples were pooled based on the decade of sampling, age, gender, and three geographical areas; in adults, one pooled samples consisted of ten and in children of twenty individual samples. Altogether 233 pooled samples were analyzed. For all organochlorine pesticides (OCPs), significant downward trends were observed in the period 1970-1990 (p<0.001). The levels of HCB exceeded the Biomonitoring Equivalent (BE) value. The hazard quotient (HQ) in Prague and Ostrava during the 1970s and 1980s was about 40 and in the 1990s it had dropped to about five. In Uherské Hradiště, the HQ in 1975 was one order of magnitude higher (about 170), and had decreased to approximately 12 by 1987. For both HCB and the DDT sum, the BE-related carcinogenic risk of actual concentrations in the past exceeded significantly the individually accepted cancer risk level of 10(-4). The levels of the main PCB congeners in the 1970s through 1990s revealed an upward time trend in all analyzed strata. The highest concentrations were found in the serum of residents from the hot-spot area Uherské Hradiště. Critical PCB sum concentration levels (700ng/g lipid for vulnerable population groups and 1800ng/g lipid for other population groups) were substantially exceeded with an increasing time trend. PCB sum had exceeded HBM II values of 7μg/L of serum since 1980 in all age strata. In conclusion, the body burden of the Czech general population relative to persistent organic pollutants (POPs) in the period 1970 through 1990 significantly exceeded currently existing health based limit values. The past exposure might adversely affect the health status of the Czech population.


Development of a high quantum yield dye for tumour imaging.
A fluorescent dye, FEB, with high fluorescence quantum yield for tumour imaging is reported. FEB dyes can be efficiently synthesized in three steps and then easily modified with either PEG or PEG-iRGD to yield FEB-2000 or FEB-2000-iRGD, respectively. Both modified dyes showed negligible toxicity and were thus able to be adopted for in vivo tumour imaging. PEG modification endowed the dye FEB-2000 with both long circulating times and good tumour targeting properties in a MDA-MB-231 xenograft model. Further conjugation with iRGD to generate FEB-2000-iRGD showed minimal targeting enhancement. These results provide a template for the efficient preparation of FEB dyes for use in tumour imaging, thus providing a foundation for future modifications.


Characterization of aromatic aminotransferases from Ephedra sinica Stapf.
Ephedra sinica Stapf (Ephedraceae) is a broom-like shrub cultivated in arid regions of China, Korea and Japan. This plant accumulates large amounts of the ephedrine alkaloids in its aerial tissues. These analogs of amphetamine mimic the actions of adrenaline and stimulate the sympathetic nervous system. While much is known about their pharmacological properties, the mechanisms by which they are synthesized remain largely unknown. A functional genomics platform was established to investigate their biosynthesis. Candidate enzymes were obtained from an expressed sequence tag collection based on similarity to characterized enzymes with similar functions. Two aromatic aminotransferases, EsAroAT1 and EsAroAT2, were characterized. The results of quantitative reverse transcription-polymerase chain reaction indicated that both genes are expressed in young stem tissue, where ephedrine alkaloids are synthesized, and in mature stem tissue. Nickel affinity-purified recombinant EsAroAT1 exhibited higher catalytic activity and was more homogeneous than EsAroAT2 as determined by size-exclusion chromatography. EsAroAT1 was highly active as a tyrosine aminotransferase with α-ketoglutarate followed by α-ketomethylthiobutyrate and very low activity with phenylpyruvate. In the reverse direction, catalytic efficiency was similar for the formation of all three aromatic amino acids using L-glutamate. Neither enzyme accepted putative intermediates in the ephedrine alkaloid biosynthetic pathway, S-phenylacetylcarbinol or 1-phenylpropane-1,2-dione, as substrates.


Enhancement of laser action in ZnO nanorods assisted by surface plasmon resonance of reduced graphene oxide nanoflakes.
We report the discovery of an enhancement of the random laser action in a nanocomposite comprising reduced graphene oxide nanoflakes and ZnO nanorods. We show that both emission intensity and lasing threshold exhibit an obvious improvement. Based on our theoretical calculations, the mechanism underlying the enhanced stimulated emission can be attributed to coupling between the optical transition and the surface plasmon resonance of the reduced graphene oxide nanoflakes, induced by the ZnO nanorod surface roughness. The approach we describe here will be very useful for the future development of high-efficiency optoelectronic devices and offers an alternative route for application of reduced graphene oxide.


Differential induction of PD-1/PD-L1 in Neuroimmune cells by drug of abuse.
Interaction of programmed cell death protein 1 (PD-1) and programmed death-ligand 1 (PD-L1) plays a critical role in regulating the delicate balance between protective immunity and tolerance. Human neuroimmune cells express very low or undetectable levels of PD-1/PD-L1 in normal physiological condition.We seek to examine if exposure of these cells to drug of abuse such as methamphetamine (METH) alters the profile of PD-1/PD-L1 levels, thereby dampens the innate immune response of the host cells. Thus, we assessed the changes in the levels of PD-1/PD-L1 in primary human macrophages, brain endothelial cells (hBECs), astrocytes, microglia, and neurons after exposure to METH. We observed that stimulation of these neuroimmune cells by METH responded differentially to PD-1/PD-L1 expression. Stimulation of macrophage culture with 50 μM of METH exhibited immediate gradual upregulation of PD-L1, while increase in PD-1 took 2-4 hours later than PD-L1. The response of hBECs to PD-1/PD-L1 induction occurred at 24 hours, while increase of PD-1/PD-L1 levels in neurons and microglia was immediate following METH exposure. We found that astrocytes expressed moderate levels of endogenous PD-1/PD-L1, which was diminished by METH exposure. Our findings show a differential expression of PD-1/PD-L1 in neuroimmune cells in response to METH stimulation, suggesting that PD-1/PD-L1 interplay in these cell types could orchestrate the intercellular interactive communication for neuronal death or protection in the brain environment.


Search for scalar bottom quarks from gluino decays in collisions at.
We searched for scalar bottom quarks 156 pb(-1) of pp collisions at radicalS = 1.96 recorded by the Collider Detector at Fermilab II experiment at the Tevatron. Scalar bottom quarks can be produced from gluino decays in -parity conserving models of supersymmetry when the mass of the gluino exceeds that of the scalar bottom quark. Then, a scalar bottom quark can decay into a bottom quark and a neutralino. To search for this scenario, we investigated events with large missing transverse energy and at least three jets, two or more of which were identified as containing a secondary vertex from the hadronization of quarks. We found four candidate events, where 2.6 +/- 0.7 are expected from standard model processes, and placed 95% confidence level lower limits on gluino and scalar bottom quark masses of up to 280 and 240 GeV/c(2), respectively.


Estimation of TCA cycle flux, aminotransferase flux, and anaplerosis in heart: validation with syntactic model.
Weiss et al. (Circ. Res. 70: 392-408, 1992) proposed a model of the citric acid cycle (CAC) in myocytes and a system of 17 differential equations that can be used to describe the changes over time in enrichment of carbons C-2 and C-4 of glutamate under conditions of metabolic steady state. They also proposed an empirical measure (KT) of flux through the CAC, which has been shown to be correlated to O2 consumption in rat hearts perfused with acetate or a mixture of glucose and acetate. We report a new method for estimation of the absolute rate of the flux through the CAC in heart (vTCA), without the numerical solution of differential equations. Unlike KT, our estimate is equal to the rate of flux catalyzed by the alpha-ketoglutarate dehydrogenase complex (vTCA), not merely correlated with it. We also estimate the rate of flux catalyzed by aspartate aminotransferase (vTA) and by NADP(+)-dependent malic enzyme (an anaplerotic reaction). The formula for vTCA during administration of [2-13C]acetate is as follows: vTCA = M[(C-2ssLC-4)/[C-4ss(LC-4-LC-2)]], where C-2ss and C-4ss represent steady-state fractional enrichment, LC-2 and LC-4 represent dominant rate constants of C-2 and C-4 of glutamate, respectively, and M is the sum of concentrations of aspartate, glutamate, and intermediates of the CAC. The assumptions underlying our formula are as follows: 1) metabolic steady state is maintained, 2) exchange of molecules between cytosolic and mitochondrial compartments is rapid, 3) 13C enters pools of the CAC only from acetyl CoA via citrate synthase, 4) [citrate]/[glutamate] < 1 + (vTCA/vTA), and 5) (m-[glutamate])/M < C-2ss/C-4ss.


Suck, swallow and breathing coordination in infants with infantile colic.
There appears to be a perception amongst parents and in popular literature that infantile colic is caused by feeding difficulties. Limited support for this perception is found in scientific literature. Whilst there is scientific evidence that suck, swallow and breathing are key components of successful feeding, these components and the coordination thereof in infants with colic have not been extensively researched. The objective of the study was to explore the suck, swallow and breathing coordination in infants with infantile colic and compare it with infants without the condition. An assessment protocol for suck, swallow and breathing coordination was compiled from literature. This protocol was performed on a research group of 50 infants, independently diagnosed with infantile colic, and a control group of 28 infants without the condition. All participants were from two rural towns in the North–West province, South Africa, selected with a snowball selection method and strict selection criteria. The study followed a static comparison group design. A significant difference in the key components of feeding and the presence of colic in participants of four age categories were found. The correlation between postural control and the presence of infantile colic were sustained in participants from 2–19 weeks old. Suck, swallow and breathing were found to be significantly associated with infantile colic. The findings should be investigated further. It appears that speech-language therapists may play an expanding role in infantile colic.


Overall survival time prediction for high-grade glioma patients based on large-scale brain functional networks.
High-grade glioma (HGG) is a lethal cancer with poor outcome. Accurate preoperative overall survival (OS) time prediction for HGG patients is crucial for treatment planning. Traditional presurgical and noninvasive OS prediction studies have used radiomics features at the local lesion area based on the magnetic resonance images (MRI). However, the highly complex lesion MRI appearance may have large individual variability, which could impede accurate individualized OS prediction. In this paper, we propose a novel concept, namely brain connectomics-based OS prediction. It is based on presurgical resting-state functional MRI (rs-fMRI) and the non-local, large-scale brain functional networks where the global and systemic prognostic features rather than the local lesion appearance are used to predict OS. We propose that the connectomics features could capture tumor-induced network-level alterations that are associated with prognosis. We construct both low-order (by means of sparse representation with regional rs-fMRI signals) and high-order functional connectivity (FC) networks (characterizing more complex multi-regional relationship by synchronized dynamics FC time courses). Then, we conduct a graph-theoretic analysis on both networks for a jointly, machine-learning-based individualized OS prediction. Based on a preliminary dataset (N = 34 with bad OS, mean OS, ~400 days; N = 34 with good OS, mean OS, ~1030 days), we achieve a promising OS prediction accuracy (86.8%) on separating the individuals with bad OS from those with good OS. However, if using only conventionally derived descriptive features (e.g., age and tumor characteristics), the accuracy is low (63.2%). Our study highlights the importance of the rs-fMRI and brain functional connectomics for treatment planning.


Top-down expectation effects of food labels on motivation.
Labels on food packages inform our beliefs, shaping our expectations of food properties, such as its expected taste and healthiness. These beliefs can influence the processing of caloric rewards beyond objective sensory properties and have the potential to impact decision making. However, no studies, within or beyond the food domain, have assessed how written information, such as food labels, affect implicit motivation to obtain rewards, even though choices in daily life might be strongly driven by implicit motivational biases. We investigated how written information affects implicit motivation to obtain caloric rewards in healthy young adults. We used food labels (high- and low-calorie), associated with an identical fruit-flavored sugar-sweetened beverage, to study motivation for caloric rewards during fMRI. In a joystick task, hungry participants (N = 31) were instructed to make fast approach or avoid movements to earn the cued beverages. Behaviorally, we found a general approach bias, which was stronger for the beverage that was most preferred during a subsequent choice test, i.e., the one labeled as low-calorie. This behavioral effect was accompanied by increased BOLD signal in the sensorimotor cortex during the response phase of the task for the preferred, low-calorie beverage compared with the non-preferred, high-calorie beverage. During the anticipation phase, the non-preferred, high-calorie beverage label elicited stronger fMRI signal in the right ventral anterior insula, a region associated with aversion and taste intensity, than the preferred, low-calorie label. Together, these data suggest that high-calorie labeling can increase avoidance of beverages and reduce neural activity in brain regions associated with motor control. In conclusion, we show effects of food labeling on fMRI responses during anticipation and subsequent motivated action and on behavior, in the absence of objective taste differences, demonstrating the influence of written information on implicit biases. These findings contribute to our understanding of implicit biases in real-life eating behavior.


SPECT imaging of inflammatory response in ischemic-reperfused rat hearts using a 99mTc-labeled dual-domain cytokine ligand.
Soluble tumor necrosis factor (TNF) receptor-2 (TNFR2) and interleukin-1 receptor antagonist (IL-1ra) were fused to the Fc portion of IgG1 using recombinant DNA technology. The resulting dual-domain cytokine ligand, TNFR2-Fc-IL-1ra, specifically binds to TNF and to the type I IL-1 receptor (IL-1RI). This study was designed to characterize the kinetic profile of (99m)Tc-labeled TNFR2-Fc-IL-1ra (TFI) for imaging inflammatory response in an ischemic-reperfused (IR) rat heart model. The IR model was created by ligating the left coronary artery for 45 min, followed by 2-h reperfusion. Cardiac SPECT images of TFI in the IR model (n = 6) were dynamically acquired for 3 h. Correlative data of myocardial TFI distribution versus microsphere-determined tissue blood flow were acquired in 3 extra IR hearts. Inflammation targeting affinity of TFI was compared with 2 individual cytokine radioligands, (99m)Tc-IL-1ra-Fc (IF) and (99m)Tc-TNFR2-Fc (TF) (n = 6 each group). Myocardial cytokine expression was evaluated by immunochemical assay. Increased TFI uptake was found in the ischemic area and correlated with the severity of ischemia. At 3 h after injection, the ratio of hot-spot accumulation in the ischemic area to a remote viable zone was 5.39 ± 1.11 for TFI, which was greater than that for IF (3.28 ± 0.81) and TF (3.29 ± 0.75) (P < 0.05). The in vivo uptake profiles of TFI, TF, and IF were consistent with ex vivo radioactive measurements and correlated with upregulated IL-1 and TNF expression. The dual-domain TFI is promising for noninvasive detection of inflammatory reactions in IR myocardium because of its more potent affinity to the inflammatory sites compared with TF and IF.


Entrustable Professional Activities (EPAs) for Global Health.
As global health education and training shift toward competency-based approaches, academic institutions and organizations must define appropriate assessment strategies for use across health professions. The authors aim to develop entrustable professional activities (EPAs) for global health to apply across academic and workplace settings. In 2019, the authors invited 55 global health experts from medicine, nursing, pharmacy, and public health to participate in a multiround, online Delphi process; 30 (55%) agreed. Experts averaged 17 years of global health experience, and 12 (40%) were from low- to middle-income countries. In round one, participants listed essential global health activities. The authors used in vivo coding for round one responses to develop initial EPA statements. In subsequent rounds, participants used 5-point Likert-type scales to evaluate EPA statements for importance and relevance to global health across health professions. The authors elevated statements that were rated 4 (important/relevant to most) or 5 (very important/relevant to all) by a minimum of 70% of participants (decided a priori) to the final round, during which participants evaluated whether each statement represented an observable unit of work that could be assigned to a trainee. Descriptive statistics were used for quantitative data analysis. The authors used participant comments to categorize EPA statements into role domains. Twenty-two EPA statements reached at least 70% consensus. The authors categorized these into 5 role domains: partnership developer, capacity builder, data analyzer, equity advocate, and health promoter. Statements in the equity advocate and partnership developer domains had the highest agreement for importance and relevance. Several statements achieved 100% agreement as a unit of work but achieved lower levels of agreement regarding their observability. EPAs for global health may be useful to academic institutions and other organizations to guide the assessment of trainees within education and training programs across health professions.


Drug safety data mining with a tree-based scan statistic.
In post-marketing drug safety surveillance, data mining can potentially detect rare but serious adverse events. Assessing an entire collection of drug-event pairs is traditionally performed on a predefined level of granularity. It is unknown a priori whether a drug causes a very specific or a set of related adverse events, such as mitral valve disorders, all valve disorders, or different types of heart disease. This methodological paper evaluates the tree-based scan statistic data mining method to enhance drug safety surveillance. We use a three-million-member electronic health records database from the HMO Research Network. Using the tree-based scan statistic, we assess the safety of selected antifungal and diabetes drugs, simultaneously evaluating overlapping diagnosis groups at different granularity levels, adjusting for multiple testing. Expected and observed adverse event counts were adjusted for age, sex, and health plan, producing a log likelihood ratio test statistic. Out of 732 evaluated disease groupings, 24 were statistically significant, divided among 10 non-overlapping disease categories. Five of the 10 signals are known adverse effects, four are likely due to confounding by indication, while one may warrant further investigation. The tree-based scan statistic can be successfully applied as a data mining tool in drug safety surveillance using observational data. The total number of statistical signals was modest and does not imply a causal relationship. Rather, data mining results should be used to generate candidate drug-event pairs for rigorous epidemiological studies to evaluate the individual and comparative safety profiles of drugs.


Genomics, Signaling, and Treatment of Waldenström Macroglobulinemia.
Next-generation sequencing has revealed recurring somatic mutations in Waldenström macroglobulinemia (WM). Commonly recurring mutations include MYD88 (95% to 97%), CXCR4 (30% to 40%), ARID1A (17%), and CD79B (8% to 15%). Diagnostic discrimination of WM from overlapping B-cell malignancies is aided by MYD88 mutation status. Transcription is affected by MYD88 and CXCR4 mutations and includes overexpression of genes involved in VDJ recombination, CXCR4 pathway signaling, and BCL2 family members. Among patients with MYD88 mutations, those with CXCR4 mutations show transcriptional silencing of tumor suppressors associated with acquisition of mutated MYD88. Deletions involving chromosome 6q are common and include genes that modulate nuclear factor-κB, BCL2, BTK, apoptosis, differentiation, and ARID1B. Non-chromosome 6q genes are also frequently deleted and include LYN, a regulator of B-cell receptor signaling. MYD88 and CXCR4 mutations affect WM disease presentation and treatment outcome. Patients with wild-type MYD88 show lower bone marrow disease burden and serum immunoglobulin M levels but show an increased risk of death. Patients with CXCR4 mutations have higher bone marrow disease burden, and those with nonsense CXCR4 mutations have higher serum immunoglobulin M levels and incidence of symptomatic hyperviscosity. Mutated MYD88 triggers BTK, IRAK1/IRAK4, and HCK growth and survival signaling, whereas CXCR4 mutations promote AKT and extracellular regulated kinase-1/2 signaling and drug resistance in the presence of its ligand CXCL12. Ibrutinib is active in patients with WM and is affected by MYD88 and CXCR4 mutation status. Patients with mutated MYD88 and wild-type CXCR4 mutation status exhibit best responses to ibrutinib. Lower response rates and delayed responses to ibrutinib are associated with mutated CXCR4 in patients with WM. MYD88 and CXCR4 mutation status may be helpful in treatment selection for symptomatic patients. Novel therapeutic approaches under investigation include therapeutics targeting MYD88, CXCR4, and BCL2 signaling.


Elliptical mirror based imaging with aperture angle greater than π/2.
Elliptical mirror based imaging systems permit aperture angles greater than π/2 to be realized. It is therefore possible to collect part or all of both the forward and backward diffractive fields emitted from single molecules. In this paper we derive rigorous formulae for the image intensity when the single molecule is modeled as a dipole emitter. It is found in theory that the point spread function can be 2.44 times narrower at full-width-half-maximum in the axial direction when using an elliptical mirror with the maximum aperture angle of 2π/3 as compared with a parabolic mirror system with the aperture angle of π/2 whereas the side lobe level is increased by only 0.21% when the dipole is oriented along Z axis.


Genetic and Real-World Clinical Data, Combined with Empirical Validation, Nominate Jak-Stat Signaling as a Target for Alzheimer's Disease Therapeutic Development.
As genome-wide association studies (GWAS) have grown in size, the number of genetic variants that have been associated per disease has correspondingly increased. Despite this increase in the number of single-nucleotide polymorphisms (SNPs) identified per disease, their biological interpretation has in many cases remained elusive. To address this, we have combined GWAS results with orthogonal sources of evidence, namely the current knowledge of molecular pathways; real-world clinical data from six million patients; RNA expression across tissues from Alzheimer's disease (AD) patients, and purpose-built rodent models for experimental validation. In more detail, first we show that when examined at a pathway level, analysis of all GWAS studies groups AD in a cluster with disorders of immunity and inflammation. Using clinical data, we show that the degree of comorbidity of these diseases with AD correlates with the strength of their genetic association with molecular participants in the Janus kinases/signal transducer and activator of transcription (JAK-STAT) pathway. Using four independent RNA expression datasets we then find evidence for the altered regulation of JAK-STAT pathway genes in AD. Finally, we use both in vitro and in vivo rodent models to demonstrate that Aβ induces gene expression of the key drivers of this pathway, providing experimental evidence to validate these data-driven observations. These results therefore nominate JAK-STAT anomalies as a prominent aetiopathological event in AD and hence a potential target for therapeutic development, and moreover demonstrate a de novo multi-modal approach to derive information from rapidly increasing genomic datasets.


Stereoselectivity of the TDxADx/FLx Amphetamine/Methamphetamine II amphetamine/methamphetamine immunoassay--response of urine specimens following nasal inhaler use.
The TDxADx/FLx Amphetamine/Methamphetamine II fluorescence polarization immunoassay (Abbott Diagnostic) for the detection of amphetamine and methamphetamine in urine was evaluated for stereoselectivity and response to specimens collected following as recommended and double the recommended dose use of Vicks Nasal Inhaler. The assay is calibrated with d-amphetamine, at cutoff concentration of 300 ng/mL for a positive response. Cross-reactivity studies demonstrated a positive result with 1000 ng/mL l-amphetamine, 300 ng/mL d-methamphetamine, and approximately 2000 ng/mL desoxyephedrine. A good correlation was observed between urines obtained following ingestion of d-amphetamine simultaneously analyzed by TDxADx/FLx Amphetamine/Methamphetamine II and gas chromatography/mass spectrometry: r2 = 0.954, N = 100. However, urine obtained following ingestion of racemic methamphetamine showed no correlation between TDxADx/FLx Amphetamine/Methamphetamine II and gas chromatography/mass spectrometry results: r2 = 0.420, N = 28. Urines collected following as recommended use for five consecutive days of Vicks Nasal Inhaler containing l-desoxyephedrine did not yield positive TDxADx/FLx Amphetamine/Methamphetamine II results. Only two urines from a subject using twice the recommended inhaler dose yielded positive TDxADx/FLx Amphetamine/Methamphetamine II results. These urines contained 1560 and 1530 ng/mL desoxyephedrine when analyzed by gas chromatography/mass spectrometry. Greater than recommended use of Vicks Nasal Inhaler may yield false positive TDxADx/FLx Amphetamine/Methamphetamine II results for amphetamine use when calibrated at 300 ng/mL d-amphetamine. If calibrated at 1000 ng/mL, d-amphetamine, the TDxADx/FLx Amphetamine/Methamphetamine II is unlikely to yield false positive results for amphetamine, even following excessive inhaler use.


Attitudes of neuroscience nurses toward patients with conversion symptoms.
Patients with conversion symptoms on neurology wards often report negative interactions with nursing staff. The authors therefore systematically studied, by means of an anonymous questionnaire, the attitudes of neuroscience nurses (N=68) toward these patients. Negative attitudes were common: 16% disagreed that conversion symptoms were "real;" 46% thought the patients were "manipulative;" and 34% disagreed that neurology was "an appropriate place" for these patients. Levels of self-perceived knowledge were low. These negative attitudes toward patients are likely to impair effective management. Illness beliefs of patients with conversion symptoms need to be understood in the context of the beliefs of the health professionals looking after them.


Multiple enhancer regions govern the transcription of CCN2 during embryonic development.
CCN2 is a critical matricellular protein that is expressed in several cells with major implications in physiology and different pathologies. However, the transcriptional regulation of this gene remains obscure. We used the Encyclopaedia of DNA Elements browser (ENCODE) to visualise the region spanning from 300 kb upstream to the CCN2 start site in silico in order to identify enhancer regions that regulate transcription of this gene. Selection was based on three criteria associated with enhancer regions: 1) H3K4me1 and H3K27ac histone modifications, 2) DNase I hypersensitivity of chromatin and 3) inter-species conservation. Reporter constructs were created with sequences spanning each of the regions of interest placed upstream of an Hsp68 silent proximal promoter sequence in order to drive the expression of β-galactosidase transgene. Each of these constructs was subsequently used to create transgenic mice in which reporter gene production was assessed at the E15.5 developmental stage. Four functional enhancers were identified, with each driving distinct, tissue-specific patterns of transgene expression. An enhancer located -100 kb from the CCN2 transcription start site facilitated expression within vascular tissue. An enhancer -135 kb upstream of CCN2 drove expression within the articular chondrocytes of synovial joints. The other two enhancers, located at -198 kb and -229 kb, mediated transgene expression within dermal fibroblasts, however the most prevalent activity was found within hypertrophic chondrocytes and periosteal tissue, respectively. These findings suggest that the global expression of CCN2 during development results from the activity of several tissue-specific enhancer regions in addition to proximal regulatory elements that have previously been demonstrated to drive transcription of the gene during development.


[Sequelae of imported tropical diseases in Germany].
20-50% of all travellers to tropical and subtropical countries experience health problems during or after travel. Mainly respiratory tract infections or gastrointestinal disorders are predominant. As specific disorders imported from the tropics traveller's diarrhoea is prevailing, however amebic and helminthic infections, hepatitis A, malaria, sexually transmitted diseases as well skin disorders are rather common. Classical tropical diseases such as cholera, sleeping sickness or trachoma play only a very minor role as imported infections. The majority of health impairments during or after travel are uncomplicated or self limiting. However, falciparum malaria, viral hepatitides, typhoid fever, tropical viral infections and infections of the CNS can take a malicious course. Early diagnosis and treatment generally can provide complete cure without sequels. Sequels are most commonly seen following hepatitis B and C as well as HIV infection but also as a result of CNS infections (e.g. encephalitis) and of imported tuberculosis. For medical expert opinion it is essential that sequels were present already during the acute phase of illness. The socio-economical impact of infections imported from the tropics is considerable due to the high morbidity figures. Preventive measures before and after a stay in tropical countries could markedly reduce the health risks involved.


Effects of temperature on identified central neurons that control jumping in the grasshopper.
Grasshoppers, like many poikilotherms, are generally more active at warmer body temperatures. In particular, they jump more frequently when warm. To determine the neuronal basis of this increase in jumping activity, we investigated the effects of temperature on the properties of identified central neurons known to be involved in the control of the jump; these included the fast extensor tibiae (FETi) motoneuron and the C, G, and M interneurons. Heating did not result in a reduction in the current or voltage threshold for action potentials; in most cases, there was an increase in the current threshold with heating. At higher temperatures, the frequency-current relations of interneurons and motoneurons had steeper slopes. With strong current pulses, increasing the temperature resulted in an increase in the initial peak firing frequencies of central neurons and usually also in their steady state firing frequencies. A second temperature effect favoring increased CNS activity in warm grasshoppers was increased afferent input from the periphery. In a broad variety of sensory receptors, there was a dramatic increase in their sensitivity to sensory stimuli at both threshold and suprathreshold intensities. Various identified central neurons differed in the way in which some of their properties were influenced by temperature. The C and G interneurons showed a striking similarity in the unusual way in which their repetitive firing properties were influenced by heating. Since these neurons are sibling progeny of a single neuroblast, this shared physiological property is correlated with their developmental history.


Lipofuscin granules in human photoreceptor cells.
Twenty-four human retinas were structurally examined in order to study the degradative pathway in inner segment turnover. Lipofuscin granules were found in myoids of photoreceptor inner segments. Cone lipofuscin granules exhibited autofluorescence, in ultraviolet light. Both the cone lipofuscin granules (-1.6 micron) and the rod ones (-0.6 micron) were membrane-limited inclusions comprising different contents. Various vacuoles related to autophagy were found in the myoids of rods and cones. Acid phosphatase activity was demonstrated in lipofuscin granules of the rods and cones, as well as adjacent various vacuoles. A survey of the cone lipofuscin granules in the semithin Epon sections revealed that more than one-half of the eyes from humans over 30 years of age contained cone lipofuscin granules, whereas eyes from those under 30 years of age did not. These results strongly suggest that lipofuscin granules represent an accumulation of residual bodies of autophagy in the photoreceptor inner segments.


Effects of metal nanoparticles on the lateral line system and behaviour in early life stages of zebrafish (Danio rerio).
The unique physicochemistry and potential toxicity of manufactured nanoparticles (NPs) requires innovative approaches for the assessment of toxicity to aquatic organisms. Here, the toxicity of Cu-NPs, Ag-NPs and TiO2-NPs on the lateral line system of free-swimming zebrafish embryos was investigated and compared to appropriate metal salts or bulk material controls. Fish were exposed for 4h at 96-h post-fertilization. Metal salt (CuSO4 and AgNO3) controls reduced the number of functional lateral line neuromasts (LLN) to <5% of unexposed controls, but no effect on LLN was observed for TiO2-NPs or Ag-NPs. Exposure to Cu-NPs caused only a 15% reduction in LLN. Performance of positive rheotaxis was reduced by Cu-NPs, Ag-NPs, and the metal salt controls. The data show that some metal NPs can affect LLN and fish behaviour (rheotaxis) important for survival, and that effects were different from those of comparable metal ion controls. Capsule: We demonstrate that behaviour is a particularly sensitive indicator of metal NP exposure in fish and highlight the interaction between behaviour and external tissue surfaces.


Assessing Rotation-Invariant Feature Classification for Automated Wildebeest Population Counts.
Accurate and on-demand animal population counts are the holy grail for wildlife conservation organizations throughout the world because they enable fast and responsive adaptive management policies. While the collection of image data from camera traps, satellites, and manned or unmanned aircraft has advanced significantly, the detection and identification of animals within images remains a major bottleneck since counting is primarily conducted by dedicated enumerators or citizen scientists. Recent developments in the field of computer vision suggest a potential resolution to this issue through the use of rotation-invariant object descriptors combined with machine learning algorithms. Here we implement an algorithm to detect and count wildebeest from aerial images collected in the Serengeti National Park in 2009 as part of the biennial wildebeest count. We find that the per image error rates are greater than, but comparable to, two separate human counts. For the total count, the algorithm is more accurate than both manual counts, suggesting that human counters have a tendency to systematically over or under count images. While the accuracy of the algorithm is not yet at an acceptable level for fully automatic counts, our results show this method is a promising avenue for further research and we highlight specific areas where future research should focus in order to develop fast and accurate enumeration of aerial count data. If combined with a bespoke image collection protocol, this approach may yield a fully automated wildebeest count in the near future.


The E3 ubiquitin ligase MARCH2 regulates ERGIC3-dependent trafficking of secretory proteins.
The E3 ubiquitin ligase membrane-associated ring-CH-type finger 2 (MARCH2) is known to be involved in intracellular vesicular trafficking, but its role in the early secretory pathway between the endoplasmic reticulum (ER) and Golgi compartments is largely unknown. Human ER-Golgi intermediate compartment protein 2 (ERGIC2) and ERGIC3 are orthologs of Erv41 and Erv46 in yeast, proteins that form a heteromeric complex, cycle between the ER and Golgi, and function as cargo receptors in both anterograde and retrograde protein trafficking. Here, we report that MARCH2 directs ubiquitination and subsequent degradation of ERGIC3 and that MARCH2 depletion increases endogenous ERGIC3 levels. We provide evidence that the lysine residues at positions 6 and 8 of ERGIC3 are the major sites of MARCH2-mediated ubiquitination. Of note, MARCH2 did not significantly decrease the levels of an ERGIC3 variant with lysine-to-arginine substitutions at residues 6 and 8. We also show that ERGIC3 binds to itself or to ERGIC2, whereas ERGIC2 is unable to interact with itself. Our results indicate that α1-antitrypsin and haptoglobin are likely to be cargo proteins of ERGIC3. We further observed that α1-antitrypsin and haptoglobin specifically bind to ERGIC3 and that ERGIC3 depletion decreases their secretion. Moreover, MARCH2 reduced secretion of α1-antitrypsin and haptoglobin, and coexpression of the ubiquitination-resistant ERGIC3 variant largely restored their secretion, suggesting that MARCH2-mediated ERGIC3 ubiquitination is the major cause of the decrease in trafficking of ERGIC3-binding secretory proteins. Our findings provide detailed insights into the regulation of the early secretory pathway by MARCH2 and into ERGIC3 function.


Distribution of pontomesencephalic cholinergic neurons projecting to substantia nigra differs significantly from those projecting to ventral tegmental area.
Locations of pontomesencephalic cholinergic projection neurons from the laterodorsal tegmental (LDTg) and pedunculopontine tegmental (PPTg) nuclei to midbrain dopaminergic nuclei were mapped. Stereotaxic microinjections of Fluoro-Gold- or rhodamine-labeled microspheres were made either to substantia nigra (SN) or ventral tegmental area (VTA) in rat. Choline acetyltransferase was visualized immunohistochemically. Labeled cells were digitally mapped at multiple levels of the nuclei using an interactive computer/microscope system. SN-projecting neurons were distributed predominantly ipsilaterally in distinct regions of the PPTg: either at its rostral pole or caudally in an area ventromedial to the superior cerebellar peduncle. Few SN-projecting neurons were found in LDTg. VTA-projecting neurons were distributed bilaterally throughout the cholinergic group, primarily in the densest regions of the LDTg and caudal PPTg. Neurons were not strictly segregated into these patterns. Scattered cells belonging to either projection could be found throughout the cholinergic group on either side. Hierarchical log-linear analysis showed these differences in topographic distribution to be statistically significant. Subtraction of cell density images demonstrated well delineated regions of the cholinergic group where the projections were predominately either to SN or VTA. These data indicate a high degree of internal organization within the pontomesencephalic cholinergic group based on the location of efferent projections to SN or VTA. These findings support the concept that this cholinergic group is functionally organized in a manner which selectively innervates motor (SN) and limbic (VTA) dopaminergic nuclei.


Plasma folate, vitamin B6, vitamin B12, and homocysteine and pancreatic cancer risk in four large cohorts.
Folate deficiency induces DNA breaks and may alter cellular capacity for mutation and epigenetic methylation. Few studies have examined the influence of one-carbon nutrients on pancreatic cancer risk, although recent studies suggest a potential protective effect for one-carbon nutrients from food sources, but not from supplements. We conducted a prospective nested case-control study to examine plasma concentrations of folate, vitamin B6 [whose main circulating form is pyridoxal-5'-phosphate (PLP)], vitamin B12, and homocysteine in relationship to pancreatic cancer, using four large prospective cohorts. Multivariable adjusted odds ratios (OR) and 95% confidence intervals (95% CI) were calculated using conditional logistic regression. All statistical tests were two sided. Among 208 cases and 623 controls, we observed no association between folate, PLP, vitamin B12, or homocysteine and pancreatic cancer risk. Comparing the highest to lowest quartiles of plasma concentration, the ORs were 1.20 (95% CI, 0.76-1.91) for folate, 0.80 (95% CI, 0.51-1.25) for B6, 0.91 (95% CI, 0.57-1.46) for B12, and 1.43 (95% CI, 0.90-2.28) for homocysteine. In analyses restricted to nonusers of multivitamins, we observe a modest inverse trend between folate, PLP, and B12 and pancreatic cancer risk. In contrast, no such inverse associations were observed among study subjects who reported multivitamin supplement use. Among all participants, plasma levels of folate, B6, B12, and homocysteine were not associated with a significant reduction in the risk of pancreatic cancer. Among participants who obtain these factors exclusively through dietary sources, there may be an inverse relation between circulating folate, B6, and B12 and risk.


Starvation resistance is associated with developmentally specified changes in sleep, feeding and metabolic rate.
Food shortage represents a primary challenge to survival, and animals have adapted diverse developmental, physiological and behavioral strategies to survive when food becomes unavailable. Starvation resistance is strongly influenced by ecological and evolutionary history, yet the genetic basis for the evolution of starvation resistance remains poorly understood. The fruit fly Drosophila melanogaster provides a powerful model for leveraging experimental evolution to investigate traits associated with starvation resistance. While control populations only live a few days without food, selection for starvation resistance results in populations that can survive weeks. We have previously shown that selection for starvation resistance results in increased sleep and reduced feeding in adult flies. Here, we investigate the ontogeny of starvation resistance-associated behavioral and metabolic phenotypes in these experimentally selected flies. We found that selection for starvation resistance resulted in delayed development and a reduction in metabolic rate in larvae that persisted into adulthood, suggesting that these traits may allow for the accumulation of energy stores and an increase in body size within these selected populations. In addition, we found that larval sleep was largely unaffected by starvation selection and that feeding increased during the late larval stages, suggesting that experimental evolution for starvation resistance produces developmentally specified changes in behavioral regulation. Together, these findings reveal a critical role for development in the evolution of starvation resistance and indicate that selection can selectively influence behavior during defined developmental time points.


Enzymatic kinetic parameters for polyfluorinated alkyl phosphate hydrolysis by alkaline phosphatase.
The hydrolysis kinetics of three polyfluorinated alkyl phosphate monoesters (monoPAPs), differing in fluorinated chain length, were measured using bovine intestinal alkaline phosphatase to catalyze the reaction. Kinetic values were also measured for analogous hydrogenated phosphate monoesters to elucidate the effects of the fluorinated chain on the rate of enzymatic hydrolysis. Michaelis constants (K(m)) were obtained by a competition kinetics technique in the presence of p-nitrophenyl phosphate (PNPP) using UV-vis spectroscopy. Compared with K(m) (PNPP), Michaelis constants for monoPAPs ranged from 0.9 to 2.1 compared with hydrogenated phosphates, which ranged from 4.0 to 13.0. Apparent bimolecular rate constants (k(cat)/K(m)) were determined by monitoring rates of product alcohol formation at low substrate concentrations using gas chromatography-mass spectrometry. The experimental values for k(cat)/K(m) averaged as 1.1 × 10(7) M(-1) s(-1) for monoPAPs compared with 3.8 × 10(5) M(-1) s(-1) for hexyl phosphate. This suggests that the electron-withdrawing nature of the fluorinated chain enhanced the alcohol leaving group ability. The results were used in a simple model to suggest that monoPAPs in a typical mammalian digestive tract would hydrolyze in approximately 100 s, supporting a previous study that showed its absence after a dosing study in rats.


Educational programs to enhance medical expertise in tropical diseases: the Gorgas Course experience 1996-2001.
In 1995, the American Society of Tropical Medicine and Hygiene (ASTMH) adopted defined criteria for accreditation of clinical training programs in tropical diseases. The first data on the development, enrollment, and outcomes of such a program are presented. A nine-week Diploma course, the Gorgas Course in Clinical Tropical Medicine, given on-site in the tropics (Lima, Cusco, and Iquitos, Peru) has trained 157 individuals from 38 countries from 1996 through 2001. The average age of participants was 38.3 with 11.3 years since graduation. Graduates were 44.5% primary care physicians, 22.2% infectious diseases specialists, 12.7% emergency medicine specialists, 13.5% other specialists, and 7.1% nurses. Residents and fellows accounted for 32.8% and full-time academic faculty for 11.0%. Approximately half of all eligible Gorgas graduates have taken the ASTMH certification examination. In response to the enrollees' profiles and needs, adult learning theory has been extensively used in course design. Stable professional relationships between multiple educational partners are required for an endeavor of this scope.


CXCL12/CXCR4 axis promotes mesenchymal stem cell mobilization to burn wounds and contributes to wound repair.
Bone marrow-derived mesenchymal stem cells (BM-MSCs) play a crucial role in tissue repair. Their role in thermal burn wound regeneration and the relevant mechanism, however, is rarely studied. BM-MSCs from green fluorescent protein transgenic male mice were transfused to irradiated recipient female C57BL/6 mice. Twenty-one days later, the female mice were inflicted with burn wounds. The size of the burned area was measured by an in vivo fluorescence imaging system, and BM-MSC chemotaxis and epithelialization were estimated by fluorescence in situ hybridization and immunofluorescence technology. The expression of CXCL12 and CXCR4 in the wound margin was detected by enzyme-linked immunosorbent assay and immunohistochemistry. The importance of CXCL12/CXCR4 signaling in BM-MSC chemotaxis was further estimated by blocking CXCR4 in vivo and in vitro. In vivo imaging results showed that BM-MSCs migrated to the injured margins. Fluorescence in situ hybridization and immunofluorescence technology revealed that Y chromosome-positive cells derived from green fluorescent protein transgenic mice were detected to be colocalized with keratin protein. Enzyme-linked immunosorbent assay revealed increased levels of CXCL12 and CXCR4 protein in the wound sites of BM-MSC-treated chimeric mice after burn. Immunohistochemistry also disclosed that CXCL12 levels were elevated at postburn day 7 compared with day 0. Furthermore, pretreatment of the BM-MSCs with the CXCR4 antagonist AMD3100 significantly inhibited the mobilization of BM-MSCs in vitro and in vivo, which attenuated wound closure. BM-MSC migration to the burned margins promotes the epithelialization of the wound, and mobilization of BM-MSCs is mediated by CXCL12/CXCR4 signaling.


The accuracy of physicians' perceptions of patients' suffering: findings from two teaching hospitals.
How accurately physicians perceive patient suffering remains unclear. The authors sought to quantitatively compare physicians' estimates of their patients' suffering with the patients' ratings of their own suffering, using a paired survey. Six major domains of suffering (DOSs) were derived from narrative descriptions of suffering by physicians and patients in a preliminary multicenter pilot study. From September 2005 through July 2006 at two teaching hospitals in Washington, DC, and Bethesda, Maryland, before a clinical encounter between a patient and physician, patients rated the impact of each of these DOSs on their overall suffering. After the same encounter, physicians rated the same DOSs according to their perception of suffering experienced by that patient. Patient responses were compared with physician responses using the Wilcoxon signed ranks and Spearman correlation tests. Two hundred twenty-seven adult patients and their treating physicians completed the survey. Cooperation rates among patients and physicians were 94% and 97%, respectively. For two of the six DOSs (pain and physically nonpainful symptoms), there was no significant difference between the physicians' estimates of suffering and the patients' ratings of the DOS. For the remaining four DOSs (communication, emotional factors, loss, and systems factors), there was significant disagreement between the physicians' estimates and the actual suffering of the patients (P < .01). When all six DOSs were combined to ascertain how well perceptions of overall suffering correlated, significant discordance was also observed between physicians' perceptions and patients' descriptions (P < .001). This study suggests that the physicians who participated might need more training in the recognition of patient suffering. Because these physicians were trained in medical schools across the country, the deficiencies noted here may not be limited to only the physicians in this study. More studies of physicians' ability to detect and manage suffering are needed, especially at nonteaching hospitals.


A Survey on Mental Health Status of Adult Population Aged 15 and above in the Province of North Khorasan, Iran.
This research aims to determine the mental health status of population aged 15 and over in the province of North Khorasan in 2015. The statistical population of this cross-sectional field survey consisted of residents of urban and rural areas of North Khorasan in Iran. Through systematic random cluster sampling, 1200 individuals were selected from the residents of urban and rural areas of of Bojnourd, Sfaraien and Shirvan. The 28-item version of the General Health Questionnaire was applied as the screening tool. The data were analyzed using SPSS, version 18.0 for windows. This study showed that using the traditional scoring method, 22.2% of the subjects (28% of females and 16.4% of males) were suspected of mental disorders. The prevalence of suspected psychiatric disorders in urban areas (23.9%) was higher than the prevalence of these disorders in rural areas (18.3%). The prevalence of suspected anxiety and the somatization of symptoms was higher than the prevalence of social dysfunction and depression, and the prevalence of these components was higher in women than men. The findings of this study also showed that the prevalence of suspected mental disorders increased significantly with age. The prevalence of suspected cases of these disorders was higher among women, the age group of 65 and older, people living in urban areas, divorced and widowed, illiterate, and retired people compared to the other groups. The results of this study show that more than one fifth of the sample were suspected of mental disorders. Therefore, health authorities and administrators need to take the principle measures to ensure and maintain the mental health of individuals as well as the evaluation and treatment of patients with mental disorders.


Comparison of Co-housing and Littermate Methods for Microbiota Standardization in Mouse Models.
The intestinal microbiota is a fundamental factor that broadly influences physiology. Thus, studies using transgenic animals should be designed to limit the confounding effects of microbiota variation between strains. Here, we report the impact on intestinal microbiota of co-housed versus F2-generation littermates, two commonly used techniques to standardize microbiota in animal models. Our results establish that while fecal microbiota is partially normalized by extended co-housing, mucosal communities associated with the proximal colon and terminal ileum remain stable and distinct. In contrast, strain inter-crossing to generate F2 littermates allows robust microbiota standardization in fecal, colon, and ileum sampling locations. Using reciprocal inter-crosses of P1 parents, we identify dissymmetry in F2 community structures caused by maternal transmission, in particular of the Verrucomicrobiaceae. Thus, F2 littermate animals from a unidirectional P1 cross should be used as a standard method to minimize the influence of the microbiota in genotype-phenotype studies.


Opportunities and limitations of natural killer cells as adoptive therapy for malignant disease.
Although natural killer (NK) cells can be readily generated for adoptive therapy with current techniques, their optimal application to treat malignant diseases requires an appreciation of the dynamic balance between signals that either synergize with or antagonize each other. Individuals display wide differences in NK function that determine their therapeutic efficacy. The ability of NK cells to kill target cells or produce cytokines depends on the balance between signals from activating and inhibitory cell-surface receptors. The selection of NK cells with a predominant activating profile is critical for delivering successful anti-tumor activity. This can be achieved through selection of killer immunoglobulin-like receptor-mismatched NK donors and by use of blocking molecules against inhibitory pathways. Optimum NK cytotoxicity may require licensing or priming with tumor cells. Recent discoveries in the molecular and cellular biology of NK cells inform in the design of new strategies, including adjuvant therapies, to maximize the cytotoxic potential of NK cells for adoptive transfer to treat human malignancies.


Comparative transcriptome analysis of Gossypium hirsutum L. in response to sap sucking insects: aphid and whitefly.
Cotton (Gossypium hirsutum L.) is a major fiber crop that is grown worldwide; it faces extensive damage from sap-sucking insects, including aphids and whiteflies. Genome-wide transcriptome analysis was performed to understand the molecular details of interaction between Gossypium hirsutum L. and sap-sucking pests, namely Aphis gossypii (Aphid) and Bemisia tabacci (Whiteflies). Roche's GS-Titanium was used to sequence transcriptomes of cotton infested with aphids and whiteflies for 2 h and 24 h. A total of 100935 contigs were produced with an average length of 529 bp after an assembly in all five selected conditions. The Blastn of the non-redundant (nr) cotton EST database resulted in the identification of 580 novel contigs in the cotton plant. It should be noted that in spite of minimal physical damage caused by the sap-sucking insects, they can change the gene expression of plants in 2 h of infestation; further change in gene expression due to whiteflies is quicker than due to aphids. The impact of the whitefly 24 h after infestation was more or less similar to that of the aphid 2 h after infestation. Aphids and whiteflies affect many genes that are regulated by various phytohormones and in response to microbial infection, indicating the involvement of complex crosstalk between these pathways. The KOBAS analysis of differentially regulated transcripts in response to aphids and whiteflies indicated that both the insects induce the metabolism of amino acids biosynthesis specially in case of whiteflies infestation at later phase. Further we also observed that expression of transcript related to photosynthesis specially carbon fixation were significantly influenced by infestation of Aphids and Whiteflies. A comparison of different transcriptomes leads to the identification of differentially and temporally regulated transcripts in response to infestation by aphids and whiteflies. Most of these differentially expressed contigs were related to genes involved in biotic, abiotic stresses and enzymatic activities related to hydrolases, transferases, and kinases. The expression of some marker genes such as the overexpressors of cationic peroxidase 3, lipoxygenase I, TGA2, and non-specific lipase, which are involved in phytohormonal-mediated plant resistance development, was suppressed after infestation by aphids and whiteflies, indicating that insects suppressed plant resistance in order to facilitate their infestation. We also concluded that cotton shares several pathways such as phagosomes, RNA transport, and amino acid metabolism with Arabidopsis in response to the infestation by aphids and whiteflies.


Optimization of neural network architecture using genetic programming improves detection and modeling of gene-gene interactions in studies of human diseases.
Appropriate definition of neural network architecture prior to data analysis is crucial for successful data mining. This can be challenging when the underlying model of the data is unknown. The goal of this study was to determine whether optimizing neural network architecture using genetic programming as a machine learning strategy would improve the ability of neural networks to model and detect nonlinear interactions among genes in studies of common human diseases. Using simulated data, we show that a genetic programming optimized neural network approach is able to model gene-gene interactions as well as a traditional back propagation neural network. Furthermore, the genetic programming optimized neural network is better than the traditional back propagation neural network approach in terms of predictive ability and power to detect gene-gene interactions when non-functional polymorphisms are present. This study suggests that a machine learning strategy for optimizing neural network architecture may be preferable to traditional trial-and-error approaches for the identification and characterization of gene-gene interactions in common, complex human diseases.


Pre-End-Stage Renal Disease Hemoglobin Variability Predicts Post-End-Stage Renal Disease Mortality in Patients Transitioning to Dialysis.
Hemoglobin variability (Hb-var) has been associated with increased mortality both in non-dialysis dependent chronic kidney disease (NDD-CKD) and end-stage renal disease (ESRD) patients. However, the impact of Hb-var in advanced NDD-CKD on outcomes after dialysis initiation remains unknown. Among 11,872 US veterans with advanced NDD-CKD transitioning to dialysis between October 2007 through September 2011, we assessed Hb-var calculated from the residual SD of at least 3 Hb values during the last 6 months before dialysis initiation (prelude period) using within-subject linear regression models, and stratified into quartiles. Outcomes included post-transition all-cause, cardiovascular, and infection-related mortality, assessed in Cox proportional hazards models and adjusted for demographics, comorbidities, length of hospitalization, medications, estimated glomerular filtration rate (eGFR), type of vascular access, Hb parameters (baseline Hb [i.e., intercept] and change in Hb [i.e., slope]), and number of Hb measurements. Higher prelude Hb-var was associated with use of iron and antiplatelet agents, tunneled dialysis catheter use, higher levels of baseline Hb, change in Hb, eGFR, and serum ferritin. After multivariable adjustment, higher prelude Hb-var was associated with higher post-ESRD all-cause and infection-related mortality, but not cardiovascular mortality (adjusted hazard ratios [95% CI] for the highest [vs. lowest] quartile of Hb-var, 1.10 [1.02-1.19], 1.28 [0.93-1.75], and 0.93 [0.79-1.10], respectively). High pre-ESRD Hb-var is associated with higher mortality, particularly from infectious causes rather than cardiovascular causes. Further research is required to clarify the underlying mechanisms and true causal nature of the observed association.


Information given to women invited for cervical screening: results of two postal surveys seven years apart.
In South Staffordshire, England, we compared women's views on information provided to them at different stages of the cervical screening program in 1994 with that provided in 2001. An age-stratified random sample of women aged 20-64 years who had a cervical smear taken between January and March 1994 (3856) or between January and March 2001 (4057) were sent postal questionnaires in June 1994 and July 2001, respectively. Response rates in 1994 (3124/3856, 81%) and 2001 (3288/4057, 81%) were similar. Compared to 1994, the proportion of women who thought the invitation letter was clear to read in 2001 increased (70% vs 98%, P < 0.0001); however, letters were thought to be less reassuring in 2001 compared to 1994 (P < 0.0001). In both study periods, 66% of women reported that the procedure was explained to them before the smear was taken. A greater proportion of women received their results by letter in 2001 compared to 1994 (57% vs 41%, P < 0.0001); however, 49% of women waited >4 weeks to receive their results in 2001 compared to 26% in 1994 (P < 0.0001). Bivariate analysis suggests that responses were age related, with older women (> or =45 years) experiencing poorer information provision. The issues highlighted by this study deserve further investigation in other areas.


The limits of counting accuracy in distributed neural representations.
Learning about a causal or statistical association depends on comparing frequencies of joint occurrence with frequencies expected from separate occurrences, and to do this, events must somehow be counted. Physiological mechanisms can easily generate the necessary measures if there is a direct, one-to-one relationship between significant events and neural activity, but if the events are represented across cell populations in a distributed manner, the counting of one event will be interfered with by the occurrence of others. Although the mean interference can be allowed for, there is inevitably an increase in the variance of frequency estimates that results in the need for extra data to achieve reliable learning. This lowering of statistical efficiency (Fisher, 1925) is calculated as the ratio of the minimum to actual variance of the estimates. We define two neural models, based on presynaptic and Hebbian synaptic modification, and explore the effects of sparse coding and the relative frequencies of events on the efficiency of frequency estimates. High counting efficiency must be a desirable feature of biological representations, but the results show that the number of events that can be counted simultaneously with 50% efficiency is fewer than the number of cells or 0.1-0.25 of the number of synapses (on the two models) - many fewer than can be unambiguously represented. Direct representations would lead to greater counting efficiency, but distributed representations have the versatility of detecting and counting many unforeseen or rare events. Efficient counting of rare but important events requires that they engage more active cells than common or unimportant ones. The results suggest reasons that representations in the cerebral cortex appear to use extravagant numbers of cells and modular organization, and they emphasize the importance of neuronal trigger features and the phenomena of habituation and attention.


Exploring patients' and carers' views about the clinical use of ketamine to inform policy and practical decisions: mixed-methods study.
Only one-third of patients with major depressive disorder achieve remission. One new and promising treatment, ketamine, may prove challenging to implement because of its abuse potential. Although clinicians' views have been sought, we need patients' views before large scale roll-out is considered. To explore patients' and carers' views to inform policy and practical decisions about the clinical use of ketamine. We carried out a mixed-methods study using data from 44 participants in 21 focus groups in three sessions and an online survey with patients, carers and advocates during a consultation day. Focus groups explored participant's views about ketamine as a form of treatment and the best way for ketamine to be prescribed and monitored. The qualitative data were analysed by two patient-researchers using an exploratory framework analysis and was supplemented by a survey. The ten themes generated were monitoring, information, effect on daily life, side-effects, recreational use, effectiveness, appropriate support, cost, stigma and therapy. Participants wanted better evidence on the safety of ketamine after long-term use and felt that monitoring was required. Collecting this information would provide evidence for ketamine's safe use and administration. There were, however, concerns about the misuse of this information. Practical issues of access were important: repeated travelling to clinics and a lack of sufficiently informed medical staff were key barriers. Clinicians have some similar and some different views to those of patients, carers and advocates, which need to be considered in any future roll-out of ketamine. R.M. has had UK National Institute for Health Research grant funding to study ketamine, is participating in trials of esketamine, runs a clinic that provides ketamine treatment, and has consulted for Johnson & Johnson and Eleusis.