Toward clinical applications of health status measures: sensitivity of scales to clinically important changes.
While the validity and reliability of many newer health status instruments have been reported, few data are available regarding the sensitivity of these instruments to clinically discernible changes in patient status. We studied this feature of the Sickness Impact Profile (SIP) in a group of patients with rheumatoid arthritis, comparing it with more traditional measures of functional status (the American Rheumatism Association (ARA) functional classification and a patient self-rating scale). Four different approaches were devised to measure "sensitivity to clinical change." These involved comparisons of functional score changes with clinical changes in patient status which were independently agreed upon by both clinician and patient, and also comparisons with several clinical disease severity indicators. When applied to groups of patients, the SIP and the patient self-rating scale were modestly superior to the ARA scale, but neither the SIP nor the self-rating scale was clearly superior to the other. For considering individual patients, all of the scales were relatively insensitive, and predictive accuracy for clinically estimated change was low. New strategies for assessing sensitivity to small changes should be developed and applied to health status and functional scales. Attention to this characteristic should allow refinement of existing scales and may enhance their clinical usefulness.


An improved test for detecting multiplicative homeostatic synaptic scaling.
Homeostatic scaling of synaptic strengths is essential for maintenance of network "gain", but also poses a risk of losing the distinctions among relative synaptic weights, which are possibly cellular correlates of memory storage. Multiplicative scaling of all synapses has been proposed as a mechanism that would preserve the relative weights among them, because they would all be proportionately adjusted. It is crucial for this hypothesis that all synapses be affected identically, but whether or not this actually occurs is difficult to determine directly. Mathematical tests for multiplicative synaptic scaling are presently carried out on distributions of miniature synaptic current amplitudes, but the accuracy of the test procedure has not been fully validated. We now show that the existence of an amplitude threshold for empirical detection of miniature synaptic currents limits the use of the most common method for detecting multiplicative changes. Our new method circumvents the problem by discarding the potentially distorting subthreshold values after computational scaling. This new method should be useful in assessing the underlying neurophysiological nature of a homeostatic synaptic scaling transformation, and therefore in evaluating its functional significance.


Longitudinal study of socio-economic differences in mortality among South Asian and West Indian migrants.
To examine socio-economic differences in mortality of South Asian and West Indian migrants. ONS Longitudinal Study, a 1% sample of the population of England and Wales. First generation South Asian and West Indian migrants present at the 1971 and 1981 censuses, aged under 65 years in 1981, were classified by occupational class, access to cars, housing, and a deprivation score. Relative risks of mortality, 1981-1997. There was considerable upward social mobility but most people remained socially stable, mainly relatively disadvantaged compared to all other study members. High levels of owner occupied housing among South Asians at both censuses were an exception. Socio-economic differentials in mortality were significant with car access in 1981 (relative risk 1.32, 95% confdence interval 1.03-1.70), and housing (rented vs owner occupied relative risk 1.50, 95% confidence interval 1.08-2.08) among South Asians and, among West Indians with car access in 1981 (relative risk 1.43, 95% confidence interval 1.05-1.94). Mortality was not significantly associated with socioeconomic circumstances in 1971. The disparity between those who remained advantaged or who moved up, and those who remained persistently disadvantaged, would have been a factor underlying the stronger differentials by later socio-economic circumstances. As these migrants age an increase in health inequalities could be expected.


[Evidence-based practices published in Brazil: identification and analysis of their types and methodological approches].
This is an integrative review of Brazilian studies on evidence-based practices (EBP) in health, published in ISI/JCR journals in the last 10 years. The aim was to identify the specialty areas that most accomplished these studies, their foci and methodological approaches. Based on inclusion criteria, 144 studies were selected. The results indicate that most EBP studies addressed childhood and adolescence, infectious diseases, psychiatrics/mental health and surgery. The predominant foci were prevention, treatment/rehabilitation, diagnosis and assessment. The most used methods were systematic review with or without meta-analysis, protocol review or synthesis of available evidence studies, and integrative review. A strong multiprofessional expansion of EBP is found in Brazil, contributing to the search for more selective practices by collecting, recognizing and critically analyzing the produced knowledge. The study also contributes to the analysis itself of ways to do research and new research possibilities.


Microsatellite marker set for genetic diversity assessment of primitive Chitala chitala (Hamilton, 1822) derived through SMRT sequencing technology.
In present study, single molecule-real time sequencing technology was used to obtain a validated set of microsatellite markers for application in population genetics of the primitive fish, Chitala chitala. Assembly of circular consensus sequencing reads resulted into 1164 sequences which contained 2005 repetitive motifs. A total of 100 sequences were used for primer designing and amplification yielded a set of 28 validated polymorphic markers. These loci were used to genotype n = 72 samples from three distant riverine populations of India, namely Son, Satluj and Brahmaputra, for determining intraspecific genetic variation. The microsatellite loci exhibited high level of polymorphism with PIC values ranging from 0.281 to 0.901. The genetic parameters revealed that mean heterozygosity ranged from 0.6802 to 0.6826 and the populations were found to be genetically diverse (Fst 0.03-0.06). This indicated the potential application of these microsatellite marker set that can used for stock characterization of C. chitala, in the wild. These newly developed loci were assayed for cross transferability in another notopterid fish, Notopterus notopterus.


Photoisomerisation of dibenzobarrelenes--a facile route to polycyclic synthons.
Triplet state mediated di-pi-methane rearrangements of dibenzobarrelenes give a variety of interesting synthons, formed as primary and secondary photoproducts. These synthons could find use for the synthesis of complex synthetic targets. This tutorial review highlights the photoisomerisation of some bridgehead substituted dibenzobarrelenes and the products derived from them. Selected examples of photoisomerisations proceeding through a tri-pi-methane pathway are also included.


Neurogenic abnormalities in Alzheimer's disease differ between stages of neurogenesis and are partly related to cholinergic pathology.
Neurogenesis occurs in the subventricular zone and the sub-granular layer of the hippocampus and is thought to take place in 5 stages, including proliferation, differentiation, migration, targeting, and integration phases, respectively. In Alzheimer's disease (AD) both increased and decreased neurogenesis has been reported and cholinergic activity is assumed to be involved in neurogenesis. The aim of this study was to systematically assess different phases of neurogenesis and their relation to AD and cholinergic pathology. We investigated post-mortem brain tissue from 20 AD patients and 21 non-demented controls that was neuropathologically characterized according to standardized criteria. Hippocampal sections were stained with antibodies against neurogenic markers Musashi-1, nestin, PSA-NCAM, doublecortin, and β-III-tubulin as well as ChAT (choline-acetyltransferase). Using image analysis immunoreactivity was assessed in the subventricular zone, the sub-granular layer, and the granule cell layer by determining the integrated optical density. In the sub-granular layer and the granule cell layer Musashi-1 and ChAT immunoreactivities were significantly lower in AD and decreased with increasing Braak stages. Conversely, immunorreactivities of both nestin and PSA-NCAM were significantly higher in AD and increased with increasing Braak stages while no changes were seen for doublecortin and β-III-tubulin, except for significantly higher doublecortin levels in the granule cell layer of AD cases. Of note, Musashi-1 immunoreactivity significantly correlated with ChAT immuonoreactivity across different Braak stages. In the subventricular zone only nestin immunoreactivity was significantly higher in AD and significantly increased with increasing Braak stages, while no significant differences were seen for all other markers. Our finding of a reduction of ChAT and Musashi-1 levels in AD is compatible with the assumption that cholinergic pathology per se has a detrimental influence on neurogenesis. We conclude that neurogenic abnormalities in AD differ between phases and areas of neurogenesis and stages of AD; while hippocampal stem cells (Musashi-1) decrease, proliferation (nestin) increases and differentiation/migration phase as well as axonal/dendritic targeting (doublecortin and β-III-tubulin) remains virtually unchanged. This suggests an attenuation of stem cells together with compensatory increased proliferation that, however, does not result in an increased number of migratory neuroblasts and differentiated neurons in AD.


Validity and reliability of the Neilands sexual stigma scale among Kenyan gay, bisexual, and other men who have sex with men.
We evaluated the validity and reliability of the Neilands sexual stigma scale administered to 871 gay, bisexual, and other men who have sex with men (GBMSM) at two research locations in Kenya. Using cross-validation, exploratory factor analysis (EFA) was performed on a randomly selected subset of participants and validated using confirmatory factor analysis (CFA) on the remaining participants. Associations of the initial and final stigma scale factors with depressive symptoms, alcohol use, and other substance use were examined for the entire dataset. EFA produced a two-factor scale of perceived and enacted stigma. The CFA model fit to the two-factor scale was improved after removing three cross-loaded items and adding correlated errors (chi-squared = 26.5, df 17, p = 0.07). Perceived stigma was associated with depressive symptoms (beta = 0.34, 95% CI 0.24, 0.45), alcohol use (beta = 0.14, 95% CI 0.03, 0.25) and other substance use (beta = 0.19, 95% CI 0.07, 0.31), while enacted stigma was associated with alcohol use (beta = 0.17, 95% CI 0.06, 0.27). Our findings suggest enacted and perceived sexual stigma are distinct yet closely related constructs among GBMSM in Kenya and are associated with poor mental health and substance use.


Early autism detection: are we ready for routine screening?
BACKGROUND. Autism is a serious neurodevelopmental disorder that has a reportedly rising prevalence rate. The American Academy of Pediatrics recommends that screening for autism be incorporated into routine practice. It is important to consider the pros and cons of conducting autism screening as part of routine practice and its implications on the community. We have explored this question in the context of screening from a scientific point of view. A literature search was conducted to assess the effectiveness of community screening programs for autism. Judged against critical questions about autism, screening programs failed to fulfill most criteria. Good screening tools and efficacious treatment are lacking, and there is no evidence yet that such a program would do more good than harm. On the basis of the available research, we believe that we do not have enough sound evidence to support the implementation of a routine population-based screening program for autism. Ongoing research in this field is certainly needed, including the development of excellent screening instruments and demonstrating with clinical trials that such programs work and do more good than harm.


Assessing eukaryotic biodiversity in the Florida Keys National Marine Sanctuary through environmental DNA metabarcoding.
Environmental DNA (eDNA) is the DNA suspended in the environment (e.g., water column), which includes cells, gametes, and other material derived from but not limited to shedding of tissue, scales, mucus, and fecal matter. Amplifying and sequencing marker genes (i.e., metabarcoding) from eDNA can reveal the wide range of taxa present in an ecosystem through analysis of a single water sample. Metabarcoding of eDNA provides higher resolution data than visual surveys, aiding in assessments of ecosystem health. This study conducted eDNA metabarcoding of two molecular markers (cytochrome c oxidase I (COI) and 18S ribosomal RNA (rRNA) genes) to survey eukaryotic diversity across multiple trophic levels in surface water samples collected at three sites along the coral reef tract within the Florida Keys National Marine Sanctuary (FKNMS) during four research cruises in 2015. The 18S rRNA gene sequences recovered 785 genera while the COI gene sequences recovered 115 genera, with only 33 genera shared between the two datasets, emphasizing the complementarity of these marker genes. Community composition for both genetic markers clustered by month of sample collection, suggesting that temporal variation has a larger effect on biodiversity than spatial variability in the FKNMS surface waters. Sequences from both marker genes were dominated by copepods, but each marker recovered distinct phytoplankton groups, with 18S rRNA gene sequences dominated by dinoflagellates and COI sequences dominated by coccolithophores. Although eDNA samples were collected from surface waters, many benthic species such as sponges, crustaceans, and corals were identified. These results show the utility of eDNA metabarcoding for cataloging biodiversity to establish an ecosystem baseline against which future samples can be compared in order to monitor community changes.


Recent advances of radiofrequency ablation for early hepatocellular carcinoma.
Hepatocellular carcinoma (HCC) is the third leading cause of death in the malignant neoplastic diseases in the world. Surgical operation is sometimes not indicated because of complicated liver cirrhosis and extrahepatic disorders. Radiofrequency ablation has been developed as a less invasive treatment for HCC since 1999, and long-term outcome has been shown. There are several complications which should be paid attention, and to improve the prognosis, combination treatment with transarterial chemoembolization should be discussed. Overall survival after between RFA and surgical resection should be compared prospectively. Establishment of staging system for treatment allocation of HCC and prevention of HCC recurrence is important issue to be examined.


Prevention of dementia by antihypertensive drugs: how AT1-receptor-blockers and dihydropyridines better prevent dementia in hypertensive patients than thiazides and ACE-inhibitors.
Our review of cohort studies and clinical trials evaluating antihypertensive drugs in the prevention of cognition decline and all dementia in patients with hypertension indicates that two antihypertensive drug classes have greater protective effects, independent of blood pressure decrease: dihydropyridine calcium-channel blockers as shown in the Syst-Eur trial and angiotensin-AT1 receptor blockers as found in the MOSES and ONTARGET trials. By contrast, diuretics and angiotensin-converting enzyme-inhibitors (ACEIs) prevent dementia only in patients with a stroke history, provided they are combined, and prevent stroke recurrence. A Japanese cohort study and a small trial in patients already suffering from Alzheimer's disease (AD) suggest, however, that the BBB-penetrating ACEI may slow down cognitive decline. Only cohort studies support the hypothesis that diuretics, (especially potassium-sparing diuretics), may decrease the risk of AD. beta-blockers worsen cognition decline, or are neutral, according to whether or not they cross the BBB. Centrally-acting sympatholytic agent have a negative impact on cognition as BBB-penetrating beta-blockers, probably by blunting the adrenergic pathways. The AD protective effect of DHP appears related to the blockade of neuronal calcium channels. The ambiguous effect of ACEI on cognitive decline and dementia prevention may be explained by the fact that brain ACE is not specific for angiotensin-I. Brain ACE also catabolizes cognition-enhancing brain peptides, amyloid peptides and converts toxic Abeta(42) into less toxic Abeta(40). Therefore, ACEIs may have short-term cognition-enhancing properties and may increase in the long term Abeta(42) brain burden and cognitive decline. The clinical relevance of this scenario, mainly observed in animals, cannot be excluded in man, since the ACE gene has been associated with AD via the human whole genome analysis. To support the hypothesized deleterious effect of ACEI on human AD, confirmation that the ACE gene polymorphism DD is associated with protection against AD is necessary, since this polymorphism increases ACE activity. Independently of their preventive impact on beta-amyloid degenerative neuropathological process by overexpressing insulin degrading enzyme which catabolyses amyloid, the angiotensin AT1-receptor-blockers may have greater cognition protective effects than ACEI (observed in the ONTARGET trial), as they share with ACEI cognition-enhancing effects directly linked with a common AT1-blunting effect. In addition, they increase angiotensin II and IV formation and therefore stimulate non-opposed AT2 and AT4 receptors, whose activation in cognitive processes is well established.


Role of omega-3 polyunsaturated fatty acids in preventing gastrointestinal cancers: current status and future perspectives.
Although inflammation is defensive and healing process that maintains organ homeostasis, unresolved inflammation can lead to diseases. Polyunsaturated fatty acids (PUFAs), especially n-6 PUFAs abundant in Western diet, are precursors of pro-inflammatory mediators, whereas n-3 PUFAs possess anti-inflammatory properties. Therefore, interest in the cancer-preventive effect of n-3 PUFAs is increasing. Areas covered: We have observed significant reductions of gastrointestinal tumorigenesis in the Fat-1 transgenic mouse as evidenced that the decrease in Helicobacter pylori-infected gastric tumorigenesis, colon, biliary, and pancreatic cancer was seen in Fat-1 mice producing n-3 PUFAs. However, despite many studies showing benefits, evidence-based medicine regarding molecular pathology, epidemiology, and clinical achievement of cancer prevention of n-3 PUFAs are still limited. Expert commentary: Primary deficiency of eicosapentaenoic acids and docosahexaenoic acids in Western diets can explain the risk of cancer development and the importance of n-3/n-6 PUFA ratio in reducing cancer risk. Alteration of cell membrane composition during carcinogenesis is particularly important, due to increased rate of lipid/cholesterol synthesis in cancerous tissues. Here, we discuss that direct incorporation of n-3 PUFAs in the cell membrane corrects abnormal cellular proliferation and decreases inflammation-associated carcinogenesis. This is exemplified by cancer-preventive effects of n-3 PUFAs as fat sources for gastrointestinal cancers.


Direct Comparison of the Enzymatic Characteristics and Superoxide Production of the Four Aldehyde Oxidase Enzymes Present in Mouse.
Aldehyde oxidases (AOXs) are molybdoflavoenzymes with an important role in the metabolism and detoxification of heterocyclic compounds and aliphatic as well as aromatic aldehydes. The enzymes use oxygen as the terminal electron acceptor and produce reduced oxygen species during turnover. Four different enzymes, mAOX1, mAOX3, mAOX4, and mAOX2, which are the products of distinct genes, are present in the mouse. A direct and simultaneous comparison of the enzymatic properties and characteristics of the four enzymes has never been performed. In this report, the four catalytically active mAOX enzymes were purified after heterologous expression in Escherichia coli The kinetic parameters of the four mouse AOX enzymes were determined and compared with the use of six predicted substrates of physiologic and toxicological interest, i.e., retinaldehyde, N1-methylnicotinamide, pyridoxal, vanillin, 4-(dimethylamino)cinnamaldehyde (p-DMAC), and salicylaldehyde. While retinaldehyde, vanillin, p-DMAC, and salycilaldehyde are efficient substrates for the four mouse AOX enzymes, N1-methylnicotinamide is not a substrate of mAOX1 or mAOX4, and pyridoxal is not metabolized by any of the purified enzymes. Overall, mAOX1, mAOX2, mAOX3, and mAOX4 are characterized by significantly different KM and kcat values for the active substrates. The four mouse AOXs are also characterized by quantitative differences in their ability to produce superoxide radicals. With respect to this last point, mAOX2 is the enzyme generating the largest rate of superoxide radicals of around 40% in relation to moles of substrate converted, and mAOX1, the homolog to the human enzyme, produces a rate of approximately 30% of superoxide radicals with the same substrate.


Dorsolateral prefrontal and orbitofrontal cortex interactions during self-control of cigarette craving.
Drug-related cues induce craving, which may perpetuate drug use or trigger relapse in addicted individuals. Craving is also under the influence of other factors in daily life, such as drug availability and self-control. Neuroimaging studies using drug cue paradigms have shown frontal lobe involvement in this contextual influence on cue reactivity, but have not clarified how and which frontal area accounts for this phenomenon. We explored frontal lobe contributions to cue-induced drug craving under different intertemporal drug availability conditions by combining transcranial magnetic stimulation and functional magnetic resonance imaging in smokers. We hypothesized that the dorsolateral prefrontal cortex (DLPFC) regulates craving during changes in intertemporal availability. Subjective craving was greater when cigarettes were immediately available, and this effect was eliminated by transiently inactivating the DLPFC with transcranial magnetic stimulation. Functional magnetic resonance imaging demonstrated that the signal most proportional to subjective craving was located in the medial orbitofrontal cortex across all contexts, whereas the DLPFC most strongly encoded intertemporal availability information. The craving-related signal in the medial orbitofrontal cortex was attenuated by inactivation of the DLPFC, particularly when cigarettes were immediately available. Inactivation of the DLPFC also reduced craving-related signals in the anterior cingulate and ventral striatum, areas implicated in transforming value signals into action. These findings indicate that DLPFC builds up value signals based on knowledge of drug availability, and support a model wherein aberrant circuitry linking dorsolateral prefrontal and orbitofrontal cortices may underlie addiction.


Identification of CYP2B6 sequence variants by use of multiplex PCR with allele-specific genotyping.
Cytochrome P450 2B6 (CYP2B6) has a role in the metabolism of many clinically important substances, but the variation within the CYP2B6 gene has not been fully characterized. The aim of the present study was to develop a reliable and robust assay for determining genotypic variants. We used a two-stage procedure. An initial multiplex PCR reaction amplified the relevant gene fragments in exonic and regulatory regions to ensure isolation of CYP2B6 from its similar pseudogene (CYP2B7). This product was then genotyped by allele-specific PCR. The assay detected the following published single-nucleotide polymorphisms: C64T (Arg22Cys), C78T, G216C, G516T (Gln172His), C777A (Ser259Arg), A785G (Lys262Arg), and C1459T (Arg487Cys), as well as additional loci found within the single-nucleotide polymorphism (SNP) databases: A1190G, C1268A, C1330T, A1382G, A1402T, and an A/T SNP in intron 2 (A12917T). This approach detected all common, previously reported alleles and identified a new allele (CYP2B6*4C) present in 2.2% of a Caucasian population. Genotypic frequencies obtained were consistent with previously published results. This method is simple, reliable, rapid, and amenable to automation and could facilitate the large-scale genotypic analysis of CYP2B6.


GABAA receptor subtype-selective modulators. II. α5-selective inverse agonists for cognition enhancement.
Benzodiazepine site agonists (such as diazepam) are well-known to impair cognition. Since benzodiazepines exert their effects via modulation of α1-, α2-, α3- and α5-containing GABA(A) receptors, the cognition-impairing effects of diazepam must be associated with one or several of these subtypes. Of these different subtypes, α5-containing GABA(A) receptors represent an attractive option as the "cognition" subtype based upon the preferential localization of these receptors within the hippocampus and the well-established role of the hippocampus in learning and memory. As a result, it is hypothesized that an inverse agonist selective for the α5 subtype should enhance cognition. For example, L-655708, a partial inverse agonist with 50-100-fold higher affinity for the α5 relative to the α1, α2 and α3 subtypes of GABA(A) receptors, enhanced cognitive performance in rats. Unfortunately, however, pharmacokinetic properties of this compound prevented it being developed further. In order to try achieve binding selectivity in a series structurally distinct from the imidazobenzodiazepines, the group at Merck, Sharp & Dohme commenced studies within the triazolopyridazine series. Although a degree of binding selectivity could be achieved (a maximum of 22-125-fold for α5 versus α1, α2 or α3) this approach was dropped in favour of a strategy to identify compounds with either a combination of selective affinity and selective efficacy or purely selective efficacy. With respect to the former, screening of the Merck chemical collection identified a novel, moderately α5-binding selective thiophene series and further optimization of this series produced MRK-536, which demonstrated a modest α5 binding selectivity (~10-fold) as well as α5-efficacy selectivity. However, the structure-activity relationship within this and the analogous tetralone series proved unpredictable and these series were not pursued further. The success of the selective efficacy approach on the α2/α3-selective agonist project led a similar paradigm being adopted for the α5 project. The starting point for this strategy was the triazolopyridazine 3 which, like MRK-536, possessed a degree of both α5 binding- and efficacy-selectivity. By changing the core from a triazolopyridazine to a triazolophthalazine structure, α5 binding selectivity was lost but with subsequent optimization, compounds with the desired profile (low or antagonist efficacy at the α1, α2 and α3 subtypes and marked inverse agonism at α5-containing receptors) could be achieved, allowing the clinical candidate α5IA as well as the structurally-related pharmacological tool compound α5IA-II to be identified. By appending features of the prototypic α2/α3-selective triazolopyridazine L-838417 (t-butyl and 1,2,4 triazole) along with the isoxazole of α5IA to a pyrazolotriazine core, an additional clinical candidate, MRK-016, was identified. Finally, a degree of α5 efficacy selectivity was achieved the pyridazine series but metabolic instability within this chemotype limited its further optimization. Overall, these studies demonstrate the feasibility of adopting a selective efficacy approach in the identification of α5 selective GABA(A) receptor inverse agonists.


The complete nucleotide sequence of tobacco necrosis virus strain D.
The complete sequence of the RNA genome of tobacco necrosis virus strain D (TNV-D) consisting of 3759 nucleotides has been determined. The positive strand contains five open reading frames (ORFs). The 5'-proximal ORF encodes a 22K protein terminating with an amber codon which may be read through to produce a 82K protein (p82). Two small centrally located ORFs each encode two out-of-frame 7K proteins (p7a and p7b). The 3'-proximal ORF encodes the 29K coat protein (CP), the N terminus of which has been sequenced directly. The genomic organization of TNV-D is very similar to that of TNV-A but differs in the placement of the p7a ORF, which does not overlap the p82 ORF in TNV-D, and in the absence of an ORF downstream of the CP gene in TNV-D. The p82 ORF shows extensive sequence similarity with the putative polymerases of the carmovirus group. This ORF is also as closely related to the corresponding ORF of TNV-A as it is to the corresponding ORF of the tombusvirus cucumber necrosis virus. The amino acid sequence of the TNV-D CP gene is similar to both the TNV-A and southern bean mosaic virus CP genes. Of the two p7 ORFs, p7a exhibits amino acid sequence similarity with corresponding proteins from TNV-A, melon necrotic spot virus, carnation mottle virus, turnip crinkle virus and maize chlorotic mottle virus, whereas the p7b ORF appears to be unique to TNV-A and TNV-D. Only the 3'-terminal three nucleotides of TNV-D genomic RNA are identical to the 3'-terminal nucleotides of the TNV satellite virus.


Identification of the current path for a conductive molecular wire on a tripodal platform.
We present the chemical synthesis as well as charge transport measurements and calculations for a new tripodal platform based on a rigid 9,9'-spirobifluorene equipped with a phenylene-ethynylene wire. The transport experiments are performed with the help of the low-temperature mechanically controlled break junction technique with gold electrodes. By combining experimental and theoretical investigations of elastic and inelastic charge transport, we show that the current proceeds through the designated molecular wire and identify a binding geometry that is compatible with the experimental observations. The conductive molecular wire on the platform features a well-defined and relatively high conductance of the order of 10(-3)G0 despite the length of the current path of more than 1.7 nm, demonstrating that this platform is suitable to incorporate functional units like molecular switches or sensors.


Role of antibiotics and fungal microbiota in driving pulmonary allergic responses.
Over the past four decades, there has been a significant increase in allergy and asthma in westernized countries, which correlates with alterations in fecal microbiota (microflora) and widespread use of antibiotics (the "hygiene hypothesis"). Antibiotics also lead to overgrowth of the yeast Candida albicans, which can secrete potent prostaglandin-like immune response modulators. We have developed a mouse model of antibiotic-induced microbiota disruption that includes stable increases in gastrointestinal (GI) enteric bacteria and GI Candida levels with no introduction of microbes into the lungs. Mice are treated for 5 days with cefoperazone in the drinking water, followed by a single oral gavage of C. albicans. This results in alterations of GI bacterial populations and increased yeast numbers in the GI microbiota for at least 2 to 3 weeks and can drive the development of a CD4 T-cell-mediated allergic airway response to subsequent mold spore (Aspergillus fumigatus) exposure in immunocompetent mice without previous systemic antigen priming. The allergic response in the lungs is characterized by increased levels of eosinophils, mast cells, interleukin-5 (IL-5), IL-13, gamma interferon, immunoglobulin E, and mucus-secreting cells. In the absence of antibiotics, mice exposed to Aspergillus spores do not develop an allergic response in the airways. This study provides the first experimental evidence to support a role for antibiotics and fungal microbiota in promoting the development of allergic airway disease. In addition, these studies also highlight the concept that events in distal mucosal sites such as the GI tract can play an important role in regulating immune responses in the lungs.


Initiation of human astrocytoma by clonal evolution of cells with progressive loss of p53 functions in a patient with a 283H TP53 germ-line mutation: evidence for a precursor lesion.
Little is known about the genetic and molecular events leading to the early stages of human astrocytoma formation. To examine this issue, we analyzed the significance of sequential accumulation of two somatic point mutations (R267W and E258D) in the TP53 gene during the initiation of astrocytoma in a patient born with a single germ-line p53 point mutation (R283H). We adapted a p53 transcriptional assay in yeast to establish the temporal occurrence and allelic distribution of the p53 mutations present in the patient and characterized these mutations through functional assays and structural modeling. Our results show that the first somatic mutation occurred at codon 267 on the p53 allele harboring the germ-line mutation R283H, whereas the second somatic mutation occurred in the remaining wild-type (wt) allele at codon 258. These two mutations induced the formation of tumor cells with the genotype p53(267W+283H/258D), which comprised 70% of the cells in the primary WHO grade II astrocytoma. Another 8% of cells within the tumor had the partially mutated genotype p53(267W+283H/WT) and represented the remnants of a clinically undetectable intermediate stage of astrocytic neoplastic transformation. The remaining 22% of cells had the constitutive p53(283H/WT) genotype and likely consisted of nontumor cells. Functional analysis of the p53 alleles present in the patient's tumor indicated that the germ-line p53(R283H) could transactivate the CDKN1A((p21, WAF1, cip1, SDI1)) but not the BAX gene and retained the ability to induce growth arrest of human glioblastoma cells. The p53(R267W+R283H) and p53(E258D) were incapable of transactivating either promoter or inducing growth arrest. Modeling of p53 interaction with DNA suggests that R283H mutation may weaken the sequence-specific interaction of p53 lysine 120 with the BAX gene but not the CDKN1A p53-responsive elements. Taken together, these results have characterized, for the first time, the genetic events defining a clinically undetectable precursor lesion leading to a grade II astrocytoma. They also suggest that astrocytoma initiation in this patient resulted from monoclonal evolution driven by a sequential loss of proapoptotic and growth arrest functions of p53.


The PTK7 and ROR2 Protein Receptors Interact in the Vertebrate WNT/Planar Cell Polarity (PCP) Pathway.
The non-canonical WNT/planar cell polarity (WNT/PCP) pathway plays important roles in morphogenetic processes in vertebrates. Among WNT/PCP components, protein tyrosine kinase 7 (PTK7) is a tyrosine kinase receptor with poorly defined functions lacking catalytic activity. Here we show that PTK7 associates with receptor tyrosine kinase-like orphan receptor 2 (ROR2) to form a heterodimeric complex in mammalian cells. We demonstrate that PTK7 and ROR2 physically and functionally interact with the non-canonical WNT5A ligand, leading to JNK activation and cell movements. In the Xenopus embryo, Ptk7 functionally interacts with Ror2 to regulate protocadherin papc expression and morphogenesis. Furthermore, we show that Ptk7 is required for papc activation induced by Wnt5a. Interestingly, we find that Wnt5a stimulates the release of the tagged Ptk7 intracellular domain, which can translocate into the nucleus and activate papc expression. This study reveals novel molecular mechanisms of action of PTK7 in non-canonical WNT/PCP signaling that may promote cell and tissue movements.


Ultra broadband microwave frequency combs generated by an optical pulse-injected semiconductor laser.
We have demonstrated and characterized the generation of ultra broadband microwave frequency combs with an optical pulse-injected semiconductor laser. Through optical pulse injection, the microwave frequency combs generated in the slave laser (SL) have bandwidths greater than 20 GHz within a +/-5 dB amplitude variation, which is almost 3-fold of the 7 GHz relaxation oscillation frequency of the laser used. The line spacing of the comb is tunable from 990 MHz to 2.6 GHz, determined by the repetition frequency of the injection optical pulses produced by the master laser (ML) with optoelectronic feedback. At an offset frequency of 200 kHz, a single sideband (SSB) phase noise of -60 dBc/kHz (-90 dBc/Hz estimated) in the 1(st) harmonic is measured while a noise suppression relative to the injected regular pulsing state of the ML of more than 25 dB in the 17(th) harmonic is achieved. A pulsewidth of 29 ps and a ms timing jitter of 18.7 ps are obtained in the time domain for the microwave frequency comb generated. Further stabilization is realized by modulating the ML at the fundamental frequency of the injected regular pulsing state. The feasibility of utilizing the generated microwave frequency comb in frequency conversion and signal broadcasting is also explored. The conversion gain of each channel increases linearly as the signal power increases with a ratio of about 0.81 dB/dBm.


The role of serious games in the iManageCancer project.
Within the iManageCancer project, two serious games were developed, one for adults and one for children and adolescents. The adult's game was developed by the University of Bedfordshire (UK), the kid's game by Promotion Software GmbH (Germany). The aim was to support adult and young cancer patients with serious games to manage the impact of the disease on their psychological status, such as negative emotions, anxiety or depression, and motivate them to stay positive and to participate in social life [Patterson and Garwick (1994) Ann Behav Med16 131-142; Brennan et al. (2002) J Consul Clin Psychol70 1075-1085; Robinson et al. (2007) J Pediatr Psychol32 400-410]. Early demonstrators showed how important it is that players understand the meaning behind the game design. Improvements in the design of the games addressed that issue. Also, technical and use case issues were found that had a significant impact on the outcome. Both games interact with the iManageCancer platform, record game results and make them available for research. Two pilots are on the way and another evaluation cycle will follow.


Methylation Levels of CADM1, MAL, and MIR124-2 in Cervical Scrapes for Triage of HIV-Infected, High-Risk HPV-Positive Women in Kenya.
To evaluate the value of cervical cell methylation markers in screening HIV-infected women also positive for high-risk human papillomavirus (hrHPV). Cross-sectional and prospective. Two hundred forty-eight HIV-infected hrHPV-positive women enrolled in a cervical cancer screening study in Nairobi, Kenya, had colposcopy-directed biopsy and histological diagnoses. Exfoliated cervical cells were used to measure methylation levels of the CADM1, MAL, and MIR124-2 genes using quantitative methylation-specific polymerase chain reaction. Methylation levels were summarized as cycle threshold (Ct) ratios compared with the β-actin gene. Median Ct ratios were compared across histological diagnoses, with 95% confidence intervals calculated by bootstrapping. Methylation levels at 6 months were assessed in 128 women who remained hrHPV positive. All 3 methylation markers showed significantly (P < 0.001) raised median Ct ratios in women with cervical intraepithelial neoplasia (CIN) grade 3 compared with women with a normal cervix. When markers were combined into a single test, the area under the receiver operating characteristic curve for prediction of CIN2 or worse (CIN2+) was 0.80. When the test was calibrated to have similar specificity, sensitivity of the combined tri-marker test for CIN2+ was comparable with cytology [atypical squamous cells of undetermined significance or worse] (89% and 95%, respectively) and superior to visual inspection with acetic acid (85% vs 70%) and HPV16/18 genotyping (65% vs 40%). Among women with no CIN2+ at baseline and persistent hrHPV at 6-month follow-up, MAL-m1 and MIR124-2 Ct ratios increased significantly. Methylation markers in combination with HPV testing may offer a full molecular screening strategy to the many HIV-infected women who are also hrHPV positive.


Rapid decrement in the effects of the Ponzo display dissociates action and perception.
It has been demonstrated that pictorial illusions have a smaller influence on grasping than they do on perceptual judgments. Yet to date this work has not considered the reduced influence of an illusion as it is measured repeatedly. Here we studied this decrement in the context of a Ponzo illusion to further characterize the dissociation between vision for perception and for action. Participants first manually estimated the lengths of single targets in a Ponzo display with their thumb and index finger, then actually grasped these targets in another series of trials, and then manually estimated the target lengths again in a final set of trials. The results showed that although the perceptual estimates and grasp apertures were equally sensitive to real differences in target length on the initial trials, only the perceptual estimates remained biased by the illusion over repeated measurements. In contrast, the illusion's effect on the grasps decreased rapidly, vanishing entirely after only a few trials. Interestingly, a closer examination of the grasp data revealed that this initial effect was driven largely by undersizing the grip aperture for the display configuration in which the target was positioned between the diverging background lines (i.e., when the targets appeared to be shorter than they really were). This asymmetry between grasping apparently shorter and longer targets suggests that the sensorimotor system may initially treat the edges of the configuration as obstacles to be avoided. This finding highlights the sensorimotor system's ability to rapidly update motor programs through error feedback, manifesting as an immunity to the effects of illusion displays even after only a few trials.


Specialist Insect Herbivore and Light Availability Do Not Interact in the Evolution of an Invasive Plant.
Release from specialist insect herbivores may allow invasive plants to evolve traits associated with decreased resistance and increased competitive ability. Given that there may be genetic trade-off between resistance and tolerance, invasive plants could also become more tolerant to herbivores. Although it is widely acknowledged that light availability affects tolerance to herbivores, little information is available for whether the effect of light availability on tolerance differ between the introduced and native populations. We conducted a common garden experiment in the introduced range of Alternanthera philoxeroides using ten invasive US and ten native Argentinean populations at two levels of light availability and in the presence or absence of a specialist stem-boring insect Agasicles hygrophila. Plant biomass (total and storage root biomass), two allocation traits (root/shoot ratio and branch intensity, branches biomass/main stem biomass) and two functional traits (specific stem length and specific leaf area), which are potentially associated with herbivore resistance and light capture, were measured. Overall, we found that A. philoxeroides from introduced ranges had comparable biomass and tolerance to specialist herbivores, lower branch intensity, lower specific stem length and specific leaf area. Moreover, introduced populations displayed higher shade tolerance of storage root biomass and lower plastic response to shading in specific stem length. Finally, light availability had no significant effect on evolution of tolerance to specialist herbivores of A. philoxeroides. Our results suggest that post-introduction evolution might have occurred in A. philoxeroides. While light availability did not influence the evolution of tolerance to specialist herbivores, increased shade tolerance and release from specialist insects might have contributed to the successful invasion of A. philoxeroides.


Current challenges in clinical development of "targeted therapies": the case of acute myeloid leukemia.
A fundamental difficulty in testing "targeted therapies" in acute myeloid leukemia (AML) is the limitations of preclinical models in capturing inter- and intrapatient genomic heterogeneity. Clinical trials typically focus on single agents despite the routine emergence of resistant subclones and experience in blast-phase chronic myeloid leukemia and acute promyelocytic leukemia arguing against this strategy. Inclusion of only relapsed-refractory, or unfit newly diagnosed, patients risks falsely negative results. There is uncertainty as to whether eligibility should require demonstration of the putative target and regarding therapeutic end points. Although use of in vivo preclinical models employing primary leukemic cells is first choice, newer preclinical models including "organoids" and combinations of pharmacologic and genetic approaches may better align models with human AML. We advocate earlier inclusion of combinations ± chemotherapy and of newly diagnosed patients into clinical trials. When a drug plausibly targets a pathway uniquely related to a specific genetic aberration, eligibility should begin with this subset, including patients with other malignancies, with subsequent extension to other patients. In other cases, a more open-minded approach to initial eligibility would facilitate quicker identification of responsive subsets. Complete remission without minimal residual disease seems a particularly useful short-term end point. Genotypic and phenotypic studies should be prespecified and performed routinely to distinguish responders from nonresponders.


Improved blood pressure, nitric oxide and asymmetric dimethylarginine are independent after bariatric surgery.
Obesity is associated with hypertension, but the exact mechanism is not fully understood. Bariatric surgery significantly decreases weight and blood pressure (BP). Low plasma nitric oxide (NO) and raised asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NO, concentrations are associated with both obesity and hypertension. Correlations between the changes in these parameters were studied after bariatric surgery. Weight, BP, plasma ADMA and NO were measured in 29 obese patients (24 female, 5 male) before and six weeks after bariatric surgery. Patients were 39.2 ± 1.2 (mean ± SEM) years old and weighed 126 ± 3 kg. Six weeks after the surgery, patients had lost 10 ± 0.7 kg (P < 0.0001) and mean arterial pressure (MAP) decreased by 11 ± 1.0 mmHg (P < 0.0001). The plasma ADMA concentration decreased by 24 ± 2% from 5 ± 0.4 to 4.0 ± 0.3 μmol/L (P < 0.0001). The plasma total nitrite concentration increased by 15 ± 1% from 51.4 ± 2.6 to 60 ± 3 μmol/L (P < 0.0001). The correlation between the decrease of ADMA and increase of NO subsequent to weight loss was significant (P < 0.0001). However, MAP was not correlated to the changes in ADMA or NO. After bariatric surgery, beneficial changes in BP, NO and ADMA occur, but our findings suggest that these BP changes are independent of changes in the NO-ADMA axis. Other causes for the changes in BP should therefore be considered.


Phylogenetic analysis of betanodavirus isolates from Australian finfish.
In Australia, disease caused by betanodavirus has been reported in an increasing number of cultured finfish since the first report of mortalities in 1990. Partial coat protein gene sequences from the T2 or T4 regions of 8 betanodaviruses from barramundi Lates calcarifer, sleepy cod Oxyeleotris lineolata, striped trumpeter Latris lineata, barramundi cod Cromileptes altivelis, Australian bass Macquaria novemaculata and gold-spotted rockcod Epinephelus coioides from several Australian states were determined. Analysis of the 606 bp nucleotide sequences of the T2 region of 4 isolates demonstrated the close relationship with isolates from the red-spotted grouper nervous necrosis virus (RGNNV) genotype and the Cluster Ia subtype. Comparison of a smaller 289 bp sequence from the T4 region identified 2 distinct groupings of the Australian isolates within the RGNNV genotype. Isolates from barramundi from the Northern Territory, barramundi, sleepy cod, barramundi cod and gold-spotted rockcod from Queensland, and striped trumpeter from Tasmania shared a 96.2 to 99.7% nucleotide identity with each other. These isolates were most similar to the RGNNV genotype Cluster Ia. Isolates from Australian bass from New South Wales and from barramundi from South Australia shared a 98.6% sequence identity with each other. However, these isolates only shared an 85.8 to 87.9% identity with the other Australian isolates and representative RGNNV isolates. The closest nucleotide identity to sequences reported in the literature for the New South Wales and South Australian isolates was to an Australian barramundi isolate (Ba94Aus) from 1994. These 2 Australian isolates formed a new subtype within the RGNNV genotype, which is designated as Cluster Ic.


Prediction of mortality in very premature infants: a systematic review of prediction models.
Being born very preterm is associated with elevated risk for neonatal mortality. The aim of this review is to give an overview of prediction models for mortality in very premature infants, assess their quality, identify important predictor variables, and provide recommendations for development of future models. Studies were included which reported the predictive performance of a model for mortality in a very preterm or very low birth weight population, and classified as development, validation, or impact studies. For each development study, we recorded the population, variables, aim, predictive performance of the model, and the number of times each model had been validated. Reporting quality criteria and minimum methodological criteria were established and assessed for development studies. We identified 41 development studies and 18 validation studies. In addition to gestational age and birth weight, eight variables frequently predicted survival: being of average size for gestational age, female gender, non-white ethnicity, absence of serious congenital malformations, use of antenatal steroids, higher 5-minute Apgar score, normal temperature on admission, and better respiratory status. Twelve studies met our methodological criteria, three of which have been externally validated. Low reporting scores were seen in reporting of performance measures, internal and external validation, and handling of missing data. Multivariate models can predict mortality better than birth weight or gestational age alone in very preterm infants. There are validated prediction models for classification and case-mix adjustment. Additional research is needed in validation and impact studies of existing models, and in prediction of mortality in the clinically important subgroup of infants where age and weight alone give only an equivocal prognosis.


Towards improved butanol production through targeted genetic modification of Clostridium pasteurianum.
Declining fossil fuel reserves, coupled with environmental concerns over their continued extraction and exploitation have led to strenuous efforts to identify renewable routes to energy and fuels. One attractive option is to convert glycerol, a by-product of the biodiesel industry, into n-butanol, an industrially important chemical and potential liquid transportation fuel, using Clostridium pasteurianum. Under certain growth conditions this Clostridium species has been shown to predominantly produce n-butanol, together with ethanol and 1,3-propanediol, when grown on glycerol. Further increases in the yields of n-butanol produced by C. pasteurianum could be accomplished through rational metabolic engineering of the strain. Accordingly, in the current report we have developed and exemplified a robust tool kit for the metabolic engineering of C. pasteurianum and used the system to make the first reported in-frame deletion mutants of pivotal genes involved in solvent production, namely hydA (hydrogenase), rex (Redox response regulator) and dhaBCE (glycerol dehydratase). We were, for the first time in C. pasteurianum, able to eliminate 1,3-propanediol synthesis and demonstrate its production was essential for growth on glycerol as a carbon source. Inactivation of both rex and hydA resulted in increased n-butanol titres, representing the first steps towards improving the utilisation of C. pasteurianum as a chassis for the industrial production of this important chemical.


Hyperkalemia caused by rapid red cell transfusion and the potassium absorption filter.
We report a case of transient hyperkalemia during hysterectomy after cesarean section, due to preoperatively undiagnosed placenta accreta that caused unforeseen massive hemorrhage and required rapid red cell transfusion. Hyperkalemia-induced by rapid red cell transfusion is a well-known severe complication of transfusion; however, in patients with sudden massive hemorrhage, rapid red cell transfusion is necessary to save their life. In such cases, it is extremely important to monitor serum potassium levels. For an emergency situation, a system should be developed to ensure sufficient preparation for immediate transfusion and laboratory tests. Furthermore, sufficient stock of preparations to treat hyperkalemia, such as calcium preparations, diuretics, glucose, and insulin is required. Moreover, a transfusion filter that absorbs potassium has been developed and is now available for clinical use in Japan. The filter is easy to use and beneficial, and should be prepared when it is available.


Quantitative analyses of cross-sectional shape of the distal radius in three species of macaques.
I conducted quantitative analyses of the cross-sectional shape of the distal radial shaft in three species of macaques, which differ in locomotor behavior: semi-terrestrial Japanese macaques ( Macaca fuscata), arboreal long tailed macaques ( M. fascicularis), and relatively terrestrial rhesus macaques ( M. mulatta). I took CT scans of the distal radial shafts of a total of 180 specimens at the level of the inferior radio-ulnar articulation. From each CT image, the periosteal outline of the radius was traced automatically by a digital imaging technique. I determined five points (landmarks) on the outline by developing a standardized morphometric technique. Bone surface lengths were measured by using these landmarks and their soft tissue correlates were investigated. The results of this study were as follows: (1) Semi-terrestrial M. fuscata has features that are approximately intermediate between those of the other two species. M. fuscata has a relatively small groove for M. abductor pollicis longus and a large groove for Mm. extensor carpi radialis longus et brevis. These characters resemble those of M. fascicularis. On the other hand, the ulnar notch of M. fuscata is relatively large, a character which is similar to that of M. mulatta. Moreover, compared to the other two macaques, the surface of the flexor muscles of M. fuscata is intermediate in size. (2) The more terrestrial M. mulatta has a relatively large groove for M. abductor pollicis longus and a small groove for Mm. extensor carpi radialis longus et brevis. Moreover, M. mulatta has a relatively large ulnar notch and a small surface for the flexor muscles. (3) The arboreal M. fascicularis has similar features to those of M. fuscata for the first and second relative size index. However, in the ulnar notch, M. fascicularis has a peculiar character and the surface for the flexor muscles is relatively large compared to those of the other two species. These results can be interpreted in terms of positional habits and presumed functional demands. A form-functional study by Lemelin and Schmitt also corroborates the interpretations of the present study. Thus, the distal region of the forearm strongly reflects muscular development and joint resultant force, and is an important region for investigating locomotor adaptations in primates. The present study reveals the possibility of using this type of morphometric analysis for reconstructing the positional behavior of fossil primates.


Initial prognostic factors and lymphoblast-erythrocyte rosette formation in 109 children with acute lymphoblastic leukemia.
Bone marrow lymphoblasts from 109 children admitted with untreated acute lymphoblastic leukemia (ALL) were tested for spontaneous rosette formation with sheep erythrocytes. Twenty-six children (24%) had lymphoblasts that formed rosettes (E+). Of 13 initial clinical characteristics, 8 were significantly associated with E+ lymphoblasts: mediastinal enlargement (86% of patients E+), leukocyte counts over 100 X 10(9)/liter (65% E+), nodes greater than 2 cm in any diameter (65% E+), age over 5 yr (46% E+), hemoglobin over 8 g/dl (44% E+), hepatomegaly greater than 5 cm (38% E+), boys (35% E+), and lymph node enlargement outside of the cervical area (28% E+). Spleen size, initial platelet counts, and periodic acid-Schiff scores did not distinguish E+ from E- patients. Since few patients were black and few presented with central nervous system leukemia, the association of these two characteristics with E+ blasts could not be determined. A hierarchical classification scheme and a linear logistic regression model were used to define the patterns of characteristics associated with E+ lymphoblasts. The initial clinical characteristics and the poorer course of E+ patients suggest that ALL comprises at least two biologically and clinically distinct types. The E+ ALL may result from a leukemic transformation of a non-Hodgkin lymphoma.


Recombinant human MDM2 oncoprotein shows sequence composition selectivity for binding to both RNA and DNA.
MDM2 is a 90 kDa nucleo-phosphoprotein that binds p53 and other proteins contributing to its oncogenic properties. Its structure includes an amino proximal p53 binding site, a central acidic domain and a carboxy region which incorporates Zinc and Ring Finger domains suggestive of nucleic acid binding or transcription factor function. It has previously been reported that a bacculovirus expressed MDM2 protein binds RNA in a sequence-specific manner through the Ring Finger domain, however, its ability to bind DNA has yet to be examined. We report here that a bacterially expressed human MDM2 protein binds both DNA as well as the previously defined RNA consensus sequence. DNA binding appears selective and involves the carboxy-terminal domain of the molecule. RNA binding is inhibited by an MDM2 specific antibody, which recognises an epitope within the carboxy region of the protein. Selection cloning and sequence analysis of MDM2 DNA binding sequences, unlike RNA binding sequences, revealed no obvious DNA binding consensus sequence, but preferential binding to oligopurine:pyrimidine-rich stretches. Our results suggest that the observed preferential DNA binding may occur through the Zinc Finger or in a charge-charge interaction through the Ring Finger, thereby implying potentially different mechanisms for DNA and RNA MDM2 binding.


Occurrence of Turnip Scab Caused by Phytotoxin-Producing Streptomyces spp.
Turnips (Brassica rapa L var. rapifera) purchased from a New Brunswick supermarket in the winter of 2003 contained discernable corky scab-like lesions on their surfaces. The turnips were surface-disinfested with a warm-water rinse, and microorganisms from excised lesions were isolated with methodologies developed for the specific isolation of Streptomyces spp. (1). Nine isolates morphologically characteristic of Streptomyces scabies strains (2) were subsequently examined for pathogenicity on potato mini-tubers and production of the scab phytotoxin, thaxtomin A (1). Four of the nine isolates proved pathogenic on potato mini-tubers. When grown on an oatmeal broth medium, only the pathogenic isolates generated thin-layer chromatographic detectable quantities of the requisite phytotoxin, thaxtomin A (1). Previous greenhouse studies have demonstrated the ability of pathogenic Streptomyces spp. isolated from scab lesions on potatoes to infect turnips (2). To our knowledge, this is the first investigation to confirm the presence of thaxtomin A producing isolates on field-grown turnips. References: (1) R. R. King et al. Am. Potato J. 68:675, 1991. (2) R. Loria et al. Plant Dis. 81:836, 1997.


Clinical outcomes according to primary treatment in gastric cancer patients with peritoneal seeding.
Peritoneal seeding of gastric cancer is known to have a poor prognosis. With the diagnosis of peritoneal seeding, there is no effective treatment modality. Gastrectomy with chemotherapy or primary chemotherapy is basically one of major options for this condition. This study was conducted to compare the clinical outcomes of these treatments and to identify the better way to improve the prognosis of patients with peritoneal seeding. Between 2001 and 2007, gastric cancer patients with peritoneal seeding by preoperative or intraoperative diagnosis were reviewed retrospectively. The enrolled patients were divided as primary gastrectomy and primary chemotherapy group. Clinicopathologic characteristics and clinical outcomes of groups were analyzed and compared. Fifty-four patients were enrolled. 21 patients belonged to the group of primary gastrectomy and 33 patients were to the primary chemotherapy group. Among 33 patients of the primary chemotherapy group, 17 patients were received only chemotherapy and 16 patients were received gastrectomy due to the good responses of primary chemotherapy. The 3 years survival rates were 14% in primary gastrectomy group, 55% in patients who received gastrectomy after primary chemotherapy, and 0% in patients with primary chemotherapy only. Although this study had many limitations, some valuable information was produced. In terms of survival benefits for the gastric cancer patients with peritoneal seeding, primary gastrectomy and additional gastrectomy after primary chemotherapy revealed the better clinical outcomes. But, prospective randomized clinical study and multi-center study should be performed to decide proper treatment for gastric cancer patients with peritoneal seeding [corrected].


Prophylactic cholecystectomy is not indicated following renal transplantation.
The appropriate management of gallstones in patients undergoing renal transplantation is controversial. Screening for gallstones and subsequent prophylactic cholecystectomy has been recommended by some authors for kidney transplant candidates. Our program does not practice routine pretransplant screening for gallstones, and we reviewed our data to determine the outcome of our management approach. We reviewed the records of the 1,364 currently followed patients who have undergone kidney transplant at our institution since 1985 in order to evaluate the morbidity and mortality of biliary disease in the post-transplant period. We attempted to contact all patients by telephone or mail survey for the presence of biliary tract disease or operations. Six hundred and sixty-two patients were fully evaluated. Fifty-two (7.85%) required cholecystectomy for stone disease. Seven patients underwent incidental cholecystectomy during other operations, 2 patients developed acalculus cholecystitis, and 14 patients with asymptomatic cholelithiasis are being followed up. Surgical indications included 38 biliary colic, 9 acute cholcystitis, 3 gallstone pancreatitis, and 2 patients who were asymptomatic. Fifty-two patients underwent 30 laparoscopic cholecystectomies, 20 open cholecystectomies, and 2 conversions. Surgery occurred from 7 days to 9.6 years following transplantation. Overall, the median hospital stay (no postoperative stay) was 4 days (range 1 to 57). Patients undergoing laparoscopy had a median stay of 2 days compared with 7 days for those undergoing an open procedure. Complications were seen in 6 patients (11.5%) with no morbidity and no graft loss. The 1-, 2-, and 5-year graft survival was 98%, 96%, and 85%, respectively, in patients undergoing cholecystectomy. Transplant patients are not at an increased risk for developing biliary tract disease compared with nontransplant patients. Gallstone disease does not have a negative impact on graft survival. Treatment of gallstones has a low risk and does not represent an increased risk of complications in patients following renal transplantation.


Palladium-mediated 11C-carbonylations using aryl halides and cyanamide.
A robust and high-yielding radiochemical synthesis of 11C-N-cyanobenzamides using a palladium-mediated aminocarbonylation with 11C-CO, aryl halides and cyanamide is described. The bidentate ligand 1,1'-bis(diphenylphosphino)ferrocene provided 11C-N-cyanobenzamides from aryl-iodides, bromides, triflates and even chlorides in 28-79% radiochemical yield after semi-preparative HPLC. To further highlight the utility of this method, novel 11C-N-cyanobenzamide analogs of flufenamic acid, meflanamic acid, dazoxiben and tamibarotene were synthesized in 34-71% radiochemical yields.


Deletion of CTLA-4 on regulatory T cells during adulthood leads to resistance to autoimmunity.
Cytotoxic T lymphocyte antigen-4 (CTLA-4) is an essential negative regulator of T cell responses. Germline Ctla4 deficiency is lethal, making investigation of the function of CTLA-4 on mature T cells challenging. To elucidate the function of CTLA-4 on mature T cells, we have conditionally ablated Ctla4 in adult mice. We show that, in contrast to germline knockout mice, deletion of Ctla4 during adulthood does not precipitate systemic autoimmunity, but surprisingly confers protection from experimental autoimmune encephalomyelitis (EAE) and does not lead to increased resistance to MC38 tumors. Deletion of Ctla4 during adulthood was accompanied by activation and expansion of both conventional CD4(+)Foxp3(-) (T conv) and regulatory Foxp3(+) (T reg cells) T cell subsets; however, deletion of CTLA-4 on T reg cells was necessary and sufficient for protection from EAE. CTLA-4 deleted T reg cells remained functionally suppressive. Deletion of Ctla4 on T reg cells alone or on all adult T cells led to major changes in the Ctla4 sufficient T conv cell compartment, including up-regulation of immunoinhibitory molecules IL-10, LAG-3 and PD-1, thereby providing a compensatory immunosuppressive mechanism. Collectively, our findings point to a profound role for CTLA-4 on T reg cells in limiting their peripheral expansion and activation, thereby regulating the phenotype and function of T conv cells.


Differential role of CCR2 in the dynamics of microglia and perivascular macrophages during prion disease.
The expansion of the microglial population is one of the hallmarks of numerous brain disorders. The addition of circulating progenitors to the pool of brain macrophages can contribute to the progression of brain disease and needs to be precisely defined to better understand the evolution of the glial and inflammatory reactions in the brain. We have analyzed the degree of infiltration/recruitment of circulating monocytes to the microglial pool, in a prion disease model of chronic neurodegeneration. Our results indicate a minimal/absent level of CCR2-dependent recruitment of circulating monocytes, local proliferation of microglia is the main driving force maintaining the amplification of the population. A deficiency in CCR2, and thus the absence of recruitment of circulating monocytes, does not impact microglial dynamics, the inflammatory profile or the temporal behavioral course of prion disease. However, the lack of CCR2 has unexpected effects including the failure to recruit perivascular macrophages in diseased but not healthy CNS and a small reduction in microglia proliferation. These data define the composition of the CNS-resident macrophage populations in prion disease and will help to understand the dynamics of the CNS innate immune response during chronic neurodegeneration.


Murine muscle-specific enolase: cDNA cloning, sequence, and developmental expression.
In vertebrates, the glycolytic enzyme enolase (EC 22.214.171.124) is present as homodimers and heterodimers formed from three distinct subunits of identical molecular weight, alpha, beta, and gamma. We report the cloning and sequencing of a cDNA encoding the beta subunit of murine muscle-specific enolase. The corresponding amino acid sequence shows greater than 80% homology with the beta subunit from chicken obtained by protein sequencing and with alpha and gamma subunits from rat and mouse deduced from cloned cDNAs. In contrast, there is no homology between the 3' untranslated regions of mouse alpha, beta, and gamma enolase mRNAs, which also differ greatly in length. The short 3' untranslated region of beta enolase mRNA accounts for its distinct length, 1600 bases. It is known that a progressive transition from alpha alpha to beta beta enolase occurs in developing skeletal muscle. We show that this transition mainly results from a differential regulation of alpha and beta mRNA levels. Analysis of myogenic cell lines shows that beta enolase gene is expressed at the myoblast stage. Moreover, transfection of premyogenic C3H10T1/2 cells with MyoD1 cDNA shows that the initial expression of beta transcripts occurs during the very first steps of the myogenic pathway, suggesting that it could be a marker event of myogenic lineage determination.


Slitrk5 deficiency impairs corticostriatal circuitry and leads to obsessive-compulsive-like behaviors in mice.
Obsessive-compulsive disorder (OCD) is a common psychiatric disorder defined by the presence of obsessive thoughts and repetitive compulsive actions, and it often encompasses anxiety and depressive symptoms. Recently, the corticostriatal circuitry has been implicated in the pathogenesis of OCD. However, the etiology, pathophysiology and molecular basis of OCD remain unknown. Several studies indicate that the pathogenesis of OCD has a genetic component. Here we demonstrate that loss of a neuron-specific transmembrane protein, SLIT and NTRK-like protein-5 (Slitrk5), leads to OCD-like behaviors in mice, which manifests as excessive self-grooming and increased anxiety-like behaviors, and is alleviated by the selective serotonin reuptake inhibitor fluoxetine. Slitrk5(-/-) mice show selective overactivation of the orbitofrontal cortex, abnormalities in striatal anatomy and cell morphology and alterations in glutamate receptor composition, which contribute to deficient corticostriatal neurotransmission. Thus, our studies identify Slitrk5 as an essential molecule at corticostriatal synapses and provide a new mouse model of OCD-like behaviors.


Enhanced Biologically Inspired Model for Object Recognition.
The biologically inspired model (BIM) proposed by Serre presents a promising solution to object categorization. It emulates the process of object recognition in primates' visual cortex by constructing a set of scale- and position-tolerant features whose properties are similar to those of the cells along the ventral stream of visual cortex. However, BIM has potential to be further improved in two aspects: mismatch by dense input and randomly feature selection due to the feedforward framework. To solve or alleviate these limitations, we develop an enhanced BIM (EBIM) in terms of the following two aspects: 1) removing uninformative inputs by imposing sparsity constraints, 2) apply a feedback loop to middle level feature selection. Each aspect is motivated by relevant psychophysical research findings. To show the effectiveness of the EBIM, we apply it to object categorization and conduct empirical studies on four computer vision data sets. Experimental results demonstrate that the EBIM outperforms the BIM and is comparable to state-of-the-art approaches in terms of accuracy. Moreover, the new system is about 20 times faster than the BIM.


Gate-controlled conductance enhancement from quantum Hall channels along graphene p-n junctions.
The formation of quantum Hall channels inside the bulk of graphene is studied using various contact and gate geometries. p-n junctions are created along the longitudinal direction of samples, and enhanced conductance is observed in the case of bipolar doping due to the new conducting channels formed in the bulk, whose position, propagating direction and, in one geometry, coupling to electrodes are determined by the gate-controlled filling factor across the device. This effect could be exploited to probe the behavior and interaction of quantum Hall channels protected against uncontrolled scattering at the edges.


A novel platform for peptide-mediated affinity capture and LC-MS/MS identification of host receptors involved in Plasmodium invasion.
Successful Plasmodium falciparum invasion of red blood cells includes the orderly execution of highly specific receptor-ligand molecular interactions between the parasite's proteins and the red blood cell membrane proteins. There is a growing need for elucidating receptor-ligand pairings, which will help in understanding the parasite's biology and provide the fundamental basis for developing prophylactic or therapeutic alternatives leading to mitigating or eliminating this type of malaria. We have thus used Plasmodium falciparum RH5 - derived peptides and ghost red blood cell proteins in synthetic peptide affinity capture assays to identify important host receptors used by Plasmodium spp. in the invasion of red blood cells. LC-MS/MS analysis confirmed the extensively described interaction between PfRH5 and the basigin receptor on the red blood cell membrane. As shown here, tagged synthetic peptides displaying high binding ability to erythrocytes can be used to identify receptors present in protein extracts from ghost red blood cells via affinity capture and LC-MS/MS. SIGNIFICANCE: The article describes a novel approach for identifying red blood cell receptors based on the ability of synthetic peptides having high red blood cell binding capacity to capture Plasmodium spp. receptors on proteins extracted from ghost red blood cells. Specifically, novel methods to identify Plasmodium falciparum reticulocyte binding protein homolog 5 PfRH5 and basigin interaction using a combination of affinity capture and LC-MS/MS assays is described. Identification of these host RBC receptors interacting with malarial parasite proteins is of utmost importance in studying the disease's pathogenesis and will provide crucial information in understanding the parasite's biology. In addition, data from these studies can be used to identify potential therapeutic target(s) to mitigate or eliminate this debilitating disease.


Iron deficiency anemia at admission for labor and delivery is associated with an increased risk for Cesarean section and adverse maternal and neonatal outcomes.
Maternal iron deficiency anemia (IDA) impacts placenta and fetus. We evaluated effects of IDA at admission for delivery on cesarean rates, and adverse maternal and neonatal outcomes. Medical records from Jerusalem (2005-2012) identified women with a live-birth singleton fetus in cephalic presentation of any gestational age and excluded planned cesarean, chronic/gestational diseases identified with anemia. Study population was divided into anemic and non-anemic women using WHO criteria. cesarean rate, and adverse outcomes (maternal: packed cells transfusion, early post-partum hemorrhage, preterm delivery; and neonatal: 5' Apgar < 7, Neonatal Intensive Care Unit [NICU] admission, extreme birthweights). Continuous variable analysis and multivariate backward step-wise logistic regression models were prepared with Odds Ratios (OR) and 95% confidence intervals (CI). In all, 96,066 deliveries were registered, of which 75,660 (78.8%) were included. IDA was present in 7,977 women (10.5%). Anemia at birth was significantly associated with cesarean section (OR 1.30; 95%CI, 1.13-1.49, p < 0.001), packed cells transfusion (OR 5.48; 95%CI, 4.57-6.58, p < 0.001), preterm delivery (OR 1.54; 95%CI, 1.36-1.76, p < 0.001), macrosomia (OR 1.23; 95%CI, 1.12-1.35, p < 0.001), Large for Gestational Age (OR 1.29; 95%CI, 1.20-1.39, p < 0.001), Apgar 5' < 7 (OR 2.21; 95%CI, 1.84-2.64, p < 0.001), and NICU admission (OR 1.28; 95%CI, 1.04-1.57, p = 0.018). Iron deficiency anemia at delivery is associated with an increased risk for cesarean section and adverse maternal and neonatal outcomes in otherwise healthy women. Monitoring/correction of hemoglobin concentrations even in late pregnancy may prevent these adverse events.


Assessing the impact of TRAP laws on abortion and women's health in the USA: a systematic review.
Targeted Regulation of Abortion Providers (TRAP) laws impose extensive and sometimes costly requirements on abortion providers and facilities, potentially leading to barriers to care. Understanding the impact of these laws is important given their prevalence in the USA, but no review to date has summarised the available evidence. We conducted a systematic review of literature on TRAP laws and their impact on abortion trends and women's health. We searched MEDLINE, PubMed and EconLit for original, quantitative studies where the exposure was at least one TRAP policy and the outcome was abortion and/or any women's physical or mental health outcome. Six articles met our inclusion criteria. The most common outcome was population-level abortion trends; studies also assessed the effect of TRAP laws on gestational age at presentation and measures of self-perceived burden. While certain TRAP laws (eg, admitting privilege requirements) appeared to have an effect on abortion outcomes, the impact of other laws - or combinations of laws - was unclear, due in part to heterogeneity between studies with respect to study design, geography, and exposure definition. TRAP laws may have an impact on the experience of obtaining an abortion in the USA. However, our review revealed a paucity of empirical research on their population and individual-level impact, as well as some disagreement about the effect of different TRAP laws on subsequent abortion outcomes. Future research should prioritise the specific TRAP laws that may have a uniquely strong effect on state-level abortion rates and other outcomes.


Dynamic changes in genomic and social structures in third millennium BCE central Europe.
Europe's prehistory oversaw dynamic and complex interactions of diverse societies, hitherto unexplored at detailed regional scales. Studying 271 human genomes dated ~4900 to 1600 BCE from the European heartland, Bohemia, we reveal unprecedented genetic changes and social processes. Major migrations preceded the arrival of "steppe" ancestry, and at ~2800 BCE, three genetically and culturally differentiated groups coexisted. Corded Ware appeared by 2900 BCE, were initially genetically diverse, did not derive all steppe ancestry from known Yamnaya, and assimilated females of diverse backgrounds. Both Corded Ware and Bell Beaker groups underwent dynamic changes, involving sharp reductions and complete replacements of Y-chromosomal diversity at ~2600 and ~2400 BCE, respectively, the latter accompanied by increased Neolithic-like ancestry. The Bronze Age saw new social organization emerge amid a ≥40% population turnover.


A profile of academic physicians in British Columbia.
To determine the extent of involvement of British Columbia's physician community in the operation of the province's only medical school, the authors sent questionnaires to all physicians who had any affiliation with the University of British Columbia (UBC). About 20 percent of the province's physicians were involved in some capacity with the UBC Faculty of Medicine, which accepts about 120 students into the first year annually. Most faculty held "clinical" appointments, meaning that they pursued largely non-academic careers. Full-time academic appointees worked more than 20 percent more hours annually than did their "clinical" counterparts, and average hours for men exceeded those for women. As many as two-thirds of the full-time faculty were also engaged in sufficient clinical practice activity to be classified as full-time practicing physicians by a definition adopted by a provincial Joint Medical Manpower Committee.


Sacral Reconstruction with a 3D-Printed Implant after Hemisacrectomy in a Patient with Sacral Osteosarcoma: 1-Year Follow-Up Result.
Pelvic reconstruction after sacral resection is challenging in terms of anatomical complexity, excessive loadbearing, and wide defects. Nevertheless, the technological development of 3D-printed implants enables us to overcome these difficulties. Here, we present a case of sacral osteosarcoma surgically treated with hemisacrectomy and sacral reconstruction using a 3D-printed implant. The implant was printed as a customized titanium prosthesis from a 3D real-sized reconstruction of a patient's CT images. It consisted mostly of a porous mesh and incorporated a dense strut. After 3-months of neoadjuvant chemotherapy, the patient underwent hemisacretomy with preservation of contralateral sacral nerves. The implant was anatomically installed on the defect and fixed with a screw-rod system up to the level of L3. Postoperative pain was significantly low and the patient recovered sufficiently to walk as early as 2 weeks postoperatively. The patient showed left-side foot drop only, without loss of sphincter function. In 1-year follow-up CT, excellent bony fusion was noticed. To our knowledge, this is the first report of a case of hemisacral reconstruction using a custom-made 3D-printed implant. We believe that this technique can be applied to spinal reconstructions after a partial or complete spondylectomy in a wide variety of spinal diseases.


Developing a Health Equity and Criminal Justice Concentration for a Master of Public Health (MPH) Program: Results From a Needs Assessment Among Community Partners and Potential Employers.
The United States has experienced a 4-fold increase in jail and prison populations over the last 40 years, disproportionately burdening African American and Hispanic/Latinx communities. Mass incarceration threatens the health of individuals, families, and communities, and requires a public health response. The Master of Public Health (MPH) Program at Touro University California (TUC) trains students to become skillful, socially-conscious public health professionals. We are developing a concentration focused on the public health impacts of incarceration. Along with the core public health curriculum, students of this new "Health Equity and Criminal Justice (HECJ)" concentration will receive training in criminal justice, reentry, reintegration, recidivism, restorative justice, structural racism, and social and community impacts of incarceration. Our study gauges interest in an HECJ concentration in our local community, including potential employers. We surveyed a cross-section of community partners including public health departments, other governmental agencies, California correctional facilities, county jails, community groups, health clinics, and hospitals. A majority (89%) of respondents consider mass incarceration a public health problem and 86% believe specialized training would make graduates employable by criminal justice related organizations. The HECJ track will fill a gap in the field and train a future generation of public health professionals to address the epidemic of mass incarceration.


Corneal Astigmatism Measurements Comparison among Ray-Tracing Aberrometry, Partial Coherence Interferometry, and Scheimpflug Imaging System.
To investigate interdevice agreement among corneal topography/ray-tracing aberrometry (iTrace), partial coherence interferometry (IOLMaster), and Scheimpflug imaging (Pentacam) for the measurement of corneal astigmatism. The analysis included 90 eyes of 90 subjects without ocular disease. The main outcome measures were corneal cylinder power and axis of astigmatism. All corneal astigmatism measurements were converted to two perpendicular components by using vector analysis. Interdevice agreement was assessed using Bland-Altman analysis, paired sample t-test, and one-way analysis of variance. No significant interdevice difference existed in the astigmatism magnitude, cardinal component, and oblique component (all P > 0.05). On comparing iTrace wavefront and simulated keratometry (SimK) astigmatism, significant differences were observed in the astigmatism magnitude and oblique component (both P < 0.01), but not in the cardinal component (P=0.687). On comparing Pentacam pupil 3 mm and corneal vertex 3 mm axial astigmatism, significant difference was observed in the astigmatism magnitude (P < 0.001), but not in the cardinal and oblique components (both P > 0.05). The iTrace, IOLMaster, and Pentacam devices could be used interchangeably for corneal astigmatism measurement. However, the measurement difference in iTrace wavefront and SimK astigmatism and Pentacam pupil 3 mm and vertex 3 mm axial astigmatism should be considered in clinic practice.


Study of the biologic effects of lapatinib, a reversible inhibitor of ErbB1 and ErbB2 tyrosine kinases, on tumor growth and survival pathways in patients with advanced malignancies.
This was a pilot study to assess the biologic effects of lapatinib on various tumor growth/survival pathways in patients with advanced ErbB1 and/or ErbB2-overexpressing solid malignancies. Heavily pretreated patients with metastatic cancers overexpressing ErbB2 and/or expressing ErbB1 were randomly assigned to one of five dose cohorts of lapatinib (GW572016) administered orally once daily continuously. The biologic effects of lapatinib on tumor growth and survival pathways were assessed in tumor biopsies obtained before and after 21 days of therapy. Clinical response was determined at 8 weeks. Sequential tumor biopsies from 33 patients were examined. Partial responses occurred in four patients with breast cancer, and disease stabilization occurred in 11 others with various malignancies. Responders exhibited variable levels of inhibition of p-ErbB1, p-ErbB2, p-Erk1/2, p-Akt, cyclin D1, and transforming growth factor alpha. Even some nonresponders demonstrated varying degrees of biomarker inhibition. Increased tumor cell apoptosis (TUNEL) occurred in patients with evidence of tumor regression but not in nonresponders (progressive disease). Clinical response was associated with a pretreatment TUNEL score > 0 and increased pretreatment expression of ErbB2, p-ErbB2, Erk1/2, p-Erk1/2, insulin-like growth factor receptor-1, p70 S6 kinase, and transforming growth factor alpha compared with nonresponders. Lapatinib exhibited preliminary evidence of biologic and clinical activity in ErbB1 and/or ErbB2-overexpressing tumors. However, the limited sample size of this study and the variability of the biologic endpoints suggest that further work is needed to prioritize biomarkers for disease-directed studies, and underscores the need for improved trial design strategies in early clinical studies of targeted agents.


Gender scripts and age at marriage in India.
Research on marriage in developing countries has been somewhat narrow in scope because of both conceptual and data limitations. While the feminist literature recognizes marriage as a key institutional site for the production and reproduction of gender hierarchies, little is known about the processes through which this relationship operates. This article uses data from the newly collected India Human Development Survey 2005 for 27,365 ever-married women aged 25-49 to explore ways in which different dimensions of gender in Indian society shape the decisions regarding age at marriage. We explore the impact of three dimensions of gender: (1) economic factors, such as availability of wage employment, dowry expectations, and wedding expenses; (2) indicators offamilial empowerment, such as women s role in household decision making and access to and control over resources; and (3) markers of gender performance, such as observance of purdah and male-female separation in the household. Results from hierarchical linear models confirm the importance of markers of gender performance but fail to demonstrate a large role for economic factors and familial empowerment.


Modulation of Na+-H+ exchange isoforms NHE1 and NHE3 by insulin-like growth factor-1 in isolated bovine articular chondrocytes.
Incubation with serum modulates the transporters that regulate intracellular pH (pH(i)) in articular chondrocytes, upregulating acid extrusion by Na(+)-H(+) exchange (NHE). There is stimulation of NHE1, together with induction of NHE3 activity. These isoforms exhibit differential responses to components of mechanical load experienced by chondrocytes during joint loading. The identity of the component(s) of serum responsible is unknown. A possibility, however, is insulin-like growth factor-1 (IGF-1), present in normal cartilage and found at enhanced levels in osteoarthritic tissue. In the present study, the effects of IGF-1 on pH(i) regulation have been characterized using fluorescence measurements of bovine articular chondrocytes, and the sensitivity of pH(i) regulation to hyperosmotic shock and raised hydrostatic pressure determined. For cells isolated in the absence of IGF-1, pH(i) recovery following acidification was predominantly mediated by NHE1. Recovery was enhanced when cells were incubated for 18 h with 20 ng mL(-1) IGF; this effect represented increased acid extrusion by NHE1, supplemented by NHE3 activity. NHE3 activity was not detected in IGF-1-treated cells that had been incubated with the protein synthesis inhibitor cycloheximide, although NHE1 activity was unaffected. In the absence of IGF-1, suspension in hyperosmotic solutions or raised hydrostatic pressure enhanced pH(i) recovery of acidified cells. This response was missing in cells incubated with IGF-1. Unresponsiveness to hyperosmotic shock represented inhibition of NHE3 activity, and was prevented using the protein kinase A inhibitor KT5720. For raised hydrostatic pressure, a decrease in NHE1 activity was responsible, and was prevented by the protein kinase C inhibitor chelerythrine.


Chronic active B-cell-receptor signalling in diffuse large B-cell lymphoma.
A role for B-cell-receptor (BCR) signalling in lymphomagenesis has been inferred by studying immunoglobulin genes in human lymphomas and by engineering mouse models, but genetic and functional evidence for its oncogenic role in human lymphomas is needed. Here we describe a form of 'chronic active' BCR signalling that is required for cell survival in the activated B-cell-like (ABC) subtype of diffuse large B-cell lymphoma (DLBCL). The signalling adaptor CARD11 is required for constitutive NF-kappaB pathway activity and survival in ABC DLBCL. Roughly 10% of ABC DLBCLs have mutant CARD11 isoforms that activate NF-kappaB, but the mechanism that engages wild-type CARD11 in other ABC DLBCLs was unknown. An RNA interference genetic screen revealed that a BCR signalling component, Bruton's tyrosine kinase, is essential for the survival of ABC DLBCLs with wild-type CARD11. In addition, knockdown of proximal BCR subunits (IgM, Ig-kappa, CD79A and CD79B) killed ABC DLBCLs with wild-type CARD11 but not other lymphomas. The BCRs in these ABC DLBCLs formed prominent clusters in the plasma membrane with low diffusion, similarly to BCRs in antigen-stimulated normal B cells. Somatic mutations affecting the immunoreceptor tyrosine-based activation motif (ITAM) signalling modules of CD79B and CD79A were detected frequently in ABC DLBCL biopsy samples but rarely in other DLBCLs and never in Burkitt's lymphoma or mucosa-associated lymphoid tissue lymphoma. In 18% of ABC DLBCLs, one functionally critical residue of CD79B, the first ITAM tyrosine, was mutated. These mutations increased surface BCR expression and attenuated Lyn kinase, a feedback inhibitor of BCR signalling. These findings establish chronic active BCR signalling as a new pathogenetic mechanism in ABC DLBCL, suggesting several therapeutic strategies.


PAGE4 is a cytoplasmic protein that is expressed in normal prostate and in prostate cancers.
PAGE4 is an X chromosome-linked cancer-testis antigen that was identified by expressed sequence tags database mining and a functional genomic approach. PAGE4 is preferentially expressed in normal male and female reproductive tissues and also in a variety of cancers including prostate. In the present study, we have used in situ hybridization to show that PAGE4 mRNA is expressed only in the epithelial cells of normal and prostate-cancer specimens. Analysis of the protein product encoded by the PAGE4 mRNA reveals that it encodes a Mr 16,000 protein and is detected in tissue extracts from both normal prostate and prostate cancer. Cell fractionation analysis of PAGE4 protein indicates that PAGE4 is localized in the cytoplasm of the cell. Furthermore, cDNA microarray analysis indicates that the expression of lipoprotein lipase, a gene frequently deleted in prostate cancer, is down-regulated in a cell line that expresses PAGE4.


Finger-powered agglutination lab chip with CMOS image sensing for rapid point-of-care diagnosis applications.
Agglutination is an antigen-antibody reaction with visible expression of aggregation of the antigens and their corresponding antibodies. Applications extend to the identification of acute bacterial infection, hemagglutination, such as blood grouping, and diagnostic immunology. Our finger-powered agglutination lab chip with external CMOS image sensing was developed to support a platform for inexpensive, rapid point-of-care (POC) testing applications related to agglutination effects. In this paper, blood grouping (ABO and Rh grouping) was utilized to demonstrate the function of our finger-powered agglutination lab chip with CMOS image sensing. Blood antibodies were preloaded into the antibody reaction chamber in the lab chip. The blood sample was pushed through the antibody reaction chamber using finger-powered pressure actuation to initiate a hemagglutination reaction to identify the blood type at the on-chip detection area using our homemade CMOS image sensing mini-system. Finger-powered actuation without the need for external electrical pumping is excellent for low-cost POC applications, but the pumping liquid volume per finger push is hard to control. In our finger-powered agglutination lab chip with CMOS image sensing, we minimized the effects of different finger push depths and achieved robust performance for the test results with different push depths. The driving sample volume per finger push is about 0.79 mm3. For different chips and different pushes, the driven sample volume per finger push was observed to vary in the range of 0.64 to 1.18 mm3. The red blood cells were separated from the plasma on-chip after the whole blood sample was finger pumped and before the red blood cells reached the antibody chamber via an embedded plasma-separation membrane. Our homemade CMOS image mini-system robustly read and identified the agglutination results on our agglutination lab chip.


Probing acid sites in solid catalysts with pyridine UV-Vis spectroscopy.
Solid acids hold widespread applications in the field of heterogeneous catalysis. In this work, we present pyridine UV-Vis spectroscopy as a novel and promising tool to study the acidic properties of such solid acids. It was found that upon interaction with acid sites, the electronic properties of pyridine change significantly. Monitoring of consecutive adsorption and desorption of pyridine revealed absorption bands in the UV-Vis region characteristic for (a) pyridinium ions formed on Brønsted acid sites, (b) pyridine coordinated to Lewis acid sites, (c) pyridine hydrogen-bonded to surface hydroxyl groups, and (d) physisorbed pyridine. The classical pyridine FT-IR method probes the presence of different Brønsted and Lewis acid sites as well, but lacks sensitivity towards the differentiation between surface hydroxyl groups. In contrast, the pyridine UV-Vis spectroscopy method proves especially useful for the identification and distinction of different surface hydroxyl groups, since the band position in the UV-Vis spectrum strongly depends on the chemical environment of the hydroxyl group. Moreover, utilizing a slow desorption procedure under N2 flow, it was possible to study the differences in acidic strength of the hydroxyl groups. This method and related measurement protocols were developed for the study of acidic properties within solid acids with different silica/alumina ratios, but are, in our opinion, more generally applicable to any solid acid.


Cooperativity of adaptive and innate immunity: implications for cancer therapy.
The dichotomy of immunology into innate and adaptive immunity has created conceptual barriers in appreciating the intrinsic two-way interaction between immune cells. An emerging body of evidence in various models of immune rejection, including cancer, indicates an indispensable regulation of innate effector functions by adaptive immune cells. This bidirectional cooperativity in innate and adaptive immune functions has broad implications for immune responses in general and for regulating the tumor-associated inflammation that overrides the protective antitumor immunity. Mechanistic understanding of this two-way immune cross-talk could provide insights into novel strategies for designing better immunotherapy approaches against cancer and other diseases that normally defy immune control.