Cotton Leafroll Dwarf Virus US Genomes Comprise Divergent Subpopulations and Harbor Extensive Variability.
Cotton leafroll dwarf virus (CLRDV) was first reported in the United States (US) in 2017 from cotton plants in Alabama (AL) and has become widespread in cotton-growing states of the southern US. To investigate the genomic variability among CLRDV isolates in the US, complete genomes of the virus were obtained from infected cotton plants displaying mild to severe symptoms from AL, Florida, and Texas. Eight CLRDV genomes were determined, ranging in size from 5865 to 5867 bp, and shared highest nucleotide identity with other CLRDV isolates in the US, at 95.9-98.7%. Open reading frame (ORF) 0, encoding the P0 silencing suppressor, was the most variable gene, sharing 88.5-99.6% and 81.2-89.3% amino acid similarity with CLRDV isolates reported in cotton growing states in the US and in Argentina and Brazil in South America, respectively. Based on Bayesian analysis, the complete CLRDV genomes from cotton in the US formed a monophyletic group comprising three relatively divergent sister clades, whereas CLRDV genotypes from South America clustered as closely related sister-groups, separate from US isolates, patterns reminiscent of phylogeographical structuring. The CLRDV isolates exhibited a complex pattern of recombination, with most breakpoints evident in ORFs 2 and 3, and ORF5. Despite extensive nucleotide diversity among all available CLRDV genomes, purifying selection (dN/dS < 1) was implicated as the primary selective force acting on viral protein evolution.


Effect of dimethyl sulfoxide on the function and structure of the intact and ischemic canine ileum.
The action of dimethyl sulfoxide (DMSO) was investigated employing an experimental model of intestinal perfusion in vivo of isolated loops of dog ileum before, during, and following 1-h ischemia. DMSO was administered either into the intestinal lumen or by a continuous injection via a branch of the artery supplying the experimental loop. In the intact intestine, intraluminal DMSO significantly decreased the net movement of water, electrolytes, and glucose without affecting either the active transport of phenylalanine and beta-methylglucoside or morphology. This inhibition was irreversible since, following DMSO removal, there was only partial recovery of water absorption, while the other parameters remained significantly low. Inhibition by DMSO was delayed when the substance was injected intraarterially (i.a.). Regardless of its route of administration, DMSO did not reduce the extent of ischemic injury in comparison with non-treated "Controls": during ischemia, all functional parameters were practically non-existent. Following the re-establishment of circulation, a net loss of water and electrolytes ensued, and active transport did not improve. In both instances, the structural alterations were those associated with ischemia of the experimental model employed: short, broad, club-shaped villi which had completely lost their epithelium, but fairly intact crypts. It can be concluded from the data presented that in the dog DMSO inhibits absorption in the intact ileum and exerts no protection against ischemic lesion.


Role of chemokine receptor CXCR2 expression in mammary tumor growth, angiogenesis and metastasis.
Chemokines and their receptors have long been known to regulate metastasis in various cancers. Previous studies have shown that CXCR2 expression is upregulated in malignant breast cancer tissues but not in benign ductal epithelial samples. The functional role of CXCR2 in the metastatic phenotype of breast cancer still remains unclear. We hypothesize that the chemokine receptor, CXCR2, mediates tumor cell invasion and migration and promotes metastasis in breast cancer. The objective of this study is to investigate the potential role of CXCR2 in the metastatic phenotype of mouse mammary tumor cells. We evaluated the functional role of CXCR2 in breast cancer by stably downregulating the expression of CXCR2 in metastatic mammary tumor cell lines Cl66 and 4T1, using short hairpin RNA (shRNA). The effects of CXCR2 downregulation on tumor growth, invasion and metastatic potential were analyzed in vitro and in vivo. We demonstrated knock down of CXCR2 in Cl66 and 4T1 cells (Cl66-shCXCR2 and 4T1-shCXCR2) cells by reverse transcriptase polymerase chain reaction (RT-PCR) at the transcriptional level and by immunohistochemistry at the protein level. We did not observe a significant difference in in vitro cell proliferation between vector control and CXCR2 knock-down Cl66 or 4T1 cells. Next, we examined the invasive potential of Cl66-shCXCR2 cells by in vitro Matrigel invasion assay. We observed a significantly lower number (52 ± 5) of Cl66-shCXCR2 cells invading through Matrigel compared to control cells (Cl66-control) (182 ± 3) (P < 0.05). We analyzed the in vivo metastatic potential of Cl66-shCXCR2 using a spontaneous metastasis model by orthotopically implanting cells into the mammary fat pad of female BALB/c mice. Animals were sacrificed 12 weeks post tumor implantation and tissue samples were analyzed for metastatic nodules. CXCR2 downregulation significantly inhibited tumor cell metastasis. All the mice (n = 10) implanted with control Cl66 cells spontaneously developed lung metastasis, whereas a significantly lower number of mice (40%) implanted with Cl66-shCXCR2 cells exhibited lung metastases. Together, these results suggest that CXCR2 may play a critical role in breast cancer invasion and metastasis.


Mitochondrial and nuclear DNA damage and repair in age-related macular degeneration.
Aging and oxidative stress seem to be the most important factors in the pathogenesis of age-related macular degeneration (AMD), a condition affecting many elderly people in the developed world. However, aging is associated with the accumulation of oxidative damage in many biomolecules, including DNA. Furthermore, mitochondria may be especially important in this process because the reactive oxygen species produced in their electron transport chain can damage cellular components. Therefore, the cellular response to DNA damage, expressed mainly through DNA repair, may play an important role in AMD etiology. In several studies the increase in mitochondrial DNA (mtDNA) damage and mutations, and the decrease in the efficacy of DNA repair have been correlated with the occurrence and the stage of AMD. It has also been shown that mitochondrial DNA accumulates more DNA lesions than nuclear DNA in AMD. However, the DNA damage response in mitochondria is executed by nucleus-encoded proteins, and thus mutagenesis in nuclear DNA (nDNA) may affect the ability to respond to mutagenesis in its mitochondrial counterpart. We reported that lymphocytes from AMD patients displayed a higher amount of total endogenous basal and oxidative DNA damage, exhibited a higher sensitivity to hydrogen peroxide and UV radiation, and repaired the lesions induced by these factors less effectively than did cells from control individuals. We postulate that poor efficacy of DNA repair (i.e., is impaired above average for a particular age) when combined with the enhanced sensitivity of retinal pigment epithelium cells to environmental stress factors, contributes to the pathogenesis of AMD. Collectively, these data suggest that the cellular response to both mitochondrial and nuclear DNA damage may play an important role in AMD pathogenesis.


The evolving private psychiatric inpatient market.
The private psychiatric hospital market has exhibited great volatility over time. From 1976 to 1992, the number of hospitals more than doubled, while in the decade following, the number of facilities dropped by half. Recently, however, the industry has begun to grow again. The evolution of this market reflects the response of a private industry with access to capital markets to changes in both the supply of substitutes and the demand for services. Most recently, the limited supply of facilities and expanded demand for psychiatric services have spurred renewed growth. The two leading firms today, Universal Health Services, Inc., which rode the market crest and downturn since the 1980s, and Psychiatric Solutions, Inc., a newer entrant, have employed different strategies to take advantage of these opportunities. The rapid responsiveness of the private psychiatric hospital market, as exemplified by these two firms, presents significant potential for shaping future mental health policy.


Effects of long-acting and short-acting beta-agonists on methacholine dose-response curves in asthmatics.
Regular use of short-acting beta-agonists may decrease control of asthma and increase airway responsiveness to bronchoconstrictor stimuli. The aim of this study was to determine the effects of regular treatment with the long-acting beta-agonist, salmeterol, on the methacholine dose-response curve (DRC) in mild-to-moderate asthmatics. Changes in methacholine airway responsiveness were measured in 14 stable adult asthmatics, randomized in a double-blind, three-way cross-over design to receive salmeterol 50 micrograms, salbutamol 200 micrograms or placebo, each twice daily for 4 days. Two baseline methacholine DRC, were performed, one without premedication and one following a single dose of 200 micrograms salbutamol. Following 4 days of regular treatment, methacholine DRC to plateau were carried out commencing 15 min after the final dose of trial medication. There were no significant differences in mean baseline forced expiratory volume in one second (FEV1) between treatments. Four days treatment with salmeterol and salbutamol shifted the DRC to the right, but salmeterol provided less protection than salbutamol. The point of inflection of the curve from baseline moved 1.9 and 3.2 doubling doses, respectively, compared to placebo (p < or = 0.001), and the provocative concentration of methacholine required to produce a 20% fall in FEV1 (PC20) increased 1.6 and 3.1 doubling doses, respectively (p < or = 0.001). The slope of the DRC was increased slightly by both beta-agonists compared to placebo (log slope 3.11, 3.06 and 2.77 for salmeterol, salbutamol and placebo, respectively). This effect of regular salmeterol on slope was more marked in subjects with lower baseline FEV1. Maximal response plateaus did not differ between the three treatments. These results suggest that regular use either of short- or long-acting beta-agonists could increase the risk of a more precipitous asthma episode associated with "breakthrough" bronchoconstrictor responses, particularly in those with more severe initial airflow obstruction, if subjects are exposed to a sufficiently potent stimulus.


Asthma episodes: stigma, children, and Hollywood films.
Asthma has been systematically stigmatized in Hollywood feature films, including films seen by children. Through content analysis of 66 movies containing one or more scenes showing asthma, and through informant interviews with a dozen U.S. children about representative scenes, the study explores how stigmatizing portrayals are interpreted, accepted, or resisted. Children suffering from asthma actively counterargued with incriminating excerpts, but in some respects their healthy friends were less critical. Overall, children viewed stigmatizing scenes in terms of the social interaction and the social ethics entailed. They did not scrutinize the characters for damaged selfhood, per se, but dwelled on the social processes out of which stigma is erected.


Online monitoring of hepatic rat metabolism by coupling a liver biochip and a mass spectrometer.
A microfluidic liver biochip was coupled with a mass spectrometer to detect in real time the drug metabolism of hepatocytes. The hepatocytes were cultivated in the biochip for 35 h. The biochip was placed in a small-scale incubator in which the temperature and CO2 concentration were controlled. The biochip was connected serially to a mass spectrometer, a peristaltic pump and a culture medium reservoir. The injection in the mass spectrometer was performed every 10 min for 11 h. The metabolism of midazolam, phenacetin, omeprazole, dextromethorphan, repaglinide, rosuvastatin, tolbutamide and caffeine was investigated. We monitored the apparition of omeprazole sulfone, hydroxy omeprazole, repaglinide glucuronide, rosuvastatin lactone, dextrorphan, 1-hydroxy midazolam, 4-hydroxy midazolam, 1,4-hydroxy midazolam, paracetamol and 1,3-methylxanthine. Although these were observed, hydroxytolbutamide, 3-methoxymorphinan and midazolam glucuronide, hydroxy repaglinide were not detected. Based on a pharmacokinetic model, we calculated in vitro intrinsic clearances in which adsorption onto the perfusion circuit was taken into account. Then, using a liver organ model, we extrapolated the in vitro intrinsic clearances to the in vivo clearances. The estimated in vivo clearances were in agreement with the literature data on rats for midazolam, dextromethorphan, phenacetin, tolbutamide and caffeine. Rosuvastatin, omeprazole and repaglinide prediction underestimated the in vivo data.


The complex genetic epidemiology of prostate cancer.
Prostate cancer is the most frequent cancer among men in most developed countries, yet little is known about its causes. Older age, African ancestry and a positive family history of prostate cancer have long been recognized as important risk factors. The evidence that genetics probably plays a critical role is based on a variety of study designs, including case-control, cohort, twin and family-based, all of which are reviewed in detail. The search for prostate cancer susceptibility genes by linkage studies offered early hope that finding genes would be as 'easy' as finding genes for breast cancer and colon cancer susceptibilities. However, this hope has been dampened by the difficulty of replicating promising regions of linkage. This review provides updates on recent developments, and a broad view of the disparate findings from different linkage studies. Early linkage results have provided targeted candidate regions for prostate cancer susceptibility loci, including HPC1 on chromosome 1q23-25, PCAP on chromosome 1q42-43, CAPB on chromosome 1p36, linkage to chromosome 8p22-23, HPC2 on chromosome 17p, HPC20 on chromosome 20q13, and HPCX on chromosome Xq27-28. These linkage findings lead to refined mapping and mutation screening of several strong candidate genes, including ELAC2, RNASEL and MSR1. Up to now, a total of 10 genome-wide linkage scans for prostate cancer susceptibility have been completed, and are reviewed. Furthermore, recent findings that Gleason's grade, a measure of aggressiveness of prostate cancer, is linked to several genomic regions are reviewed. Finally, the roles of environmental and dietary risk factors, and common genetic polymorphisms of genes likely to play a role in common forms of prostate cancer, are briefly discussed within in the context of searching for genes that influence prostate cancer risk.


Seasonal dynamics of canine antibody response to Phlebotomus perniciosus saliva in an endemic area of Leishmania infantum.
Canine leishmaniosis (CanL) is an important zoonotic parasitic disease, endemic in the Mediterranean basin. In this region, transmission of Leishmania infantum, the etiological agent of CanL, is through the bite of phlebotomine sand flies. Therefore, monitoring host-vector contact represents an important epidemiological tool, and could be used to assess the effectiveness of vector-control programmes in endemic areas. Previous studies have shown that canine antibodies against the saliva of phlebotomine sand flies are specific markers of exposure to Leishmania vectors. However, this method needs to be further validated in natural heterogeneous dog populations living in CanL endemic areas. In this study, 176 dogs living in 12 different locations of an L. infantum endemic area in north-east Spain were followed for 14 months. Blood samples were taken at 5 pre-determined time points (February, August and October 2016; January and April 2017) to assess the canine humoral immune response to whole salivary gland homogenate (SGH) and to the single salivary 43 kDa yellow-related recombinant protein (rSP03B) of Phlebotomus perniciosus, a proven vector of L. infantum naturally present in this region. Simultaneously, in all dogs, L. infantum infection status was assessed by serology. The relationship between anti-SGH and anti-rSP03B antibodies with the sampling month, L. infantum infection and the location was tested by fitting multilevel linear regression models. The dynamics of canine anti-saliva IgG for both SGH and rSP03B followed the expected trends of P. perniciosus activity in the region. Statistically significant associations were detected for both salivary antigens between vector exposure and sampling month or dog seropositivity to L. infantum. The correlation between canine antibodies against SGH and rSP03B was moderate. Our results confirm the frequent presence of CanL vectors in the study area in Spain and support the applicability of SGH- and rSP03B-based ELISA tests to study canine exposure to P. perniciosus in L. infantum endemic areas.


Characteristics of ethylene biosynthesis-inducing xylanase movement in tobacco leaves.
(125)I-Labeled ethylene biosynthesis-inducing xylanase (EIX) was used to study the movement of this protein in tobacco (Nicotiana tabacum) tissues. A biologically active (125)I-labeled EIX was obtained using chloramine-T as the oxidizing agent. Labeled EIX was detected in the far most edges of the leaf 5 min after it was applied to the petiole of a detached leaf. EIX was distributed uniformly throughout the leaf, including the mesophyll area within 5 to 15 min, after which there was only little change in the distribution of radioactivity in the leaf. (125)I-Labeled EIX was extracted from treated leaves, and EIX translocation in the leaf was blocked by preincubation of labeled EIX with anti-EIX antibodies, indicating that the intact peptide moves in the leaf. Injection of anti-EIX antibodies into the intercellular spaces of the leaf mesophyll prevented induction of necrosis by EIX, suggesting the mesophyll as the site of EIX action. EIX was translocated both to upper and lower parts of the plant when applied to a whole plant through the petiole of a cut leaf. Radioactivity was found in all leaves and in the stem, although some leaves accumulated much more EIX than others; EIX was not found in the roots. There was no difference between the accumulation pattern of EIX in fresh and ethylene-treated leaves or between sensitive (Xanthi) and insensitive (Hicks) tobacco cultivars. These data support the hypothesis that intact EIX protein is translocated to the leaf mesophyll, where it directly elicits plant defense responses.


Effects of refractive turbulence on ground-based verification of coherent Doppler lidar performance.
The effects of refractive turbulence on ground-based verification of the far-field performance of coherent Doppler lidar are determined with numerical simulation and compared with the first-order terms of a theoretical expansion. The collimated small-beam far-field test has better performance than the focused-beam test. For typical ground-based conditions, higher-order terms of the theoretical expansion are required for convergence.


Expression and mutation analysis of TIG1 (tazarotene-induced gene 1) in human gastric cancer.
Tazarotene-induced gene 1 (TIG1) has been known to function as a cell adhesion molecule, which leads to better cell to cell contact and reduced proliferation. We investigated expression and mutation status of TIG1 in primary gastric tumors and cell lines to explore the candidacy of the gene as a tumor suppressor. A total of 172 gastric tissue specimes, including 80 primary adenocarcinomas, 12 benign tumors, and 80 adjacent normal mucosa, and 15 gastric cancer cell lines were used. TIG1 expression was analyzed by semiquantitative RT-PCR and immunoblot analysis. To screen for the presence of somatic mutations, RT-PCR-SSCP analysis was carried out. The effect of 5-aza-2'-deoxycytidine treatment was examined to elicit whether TIG1 reduction is associated with abnormal DNA hypermethylation. Compared to noncancerous tissues, a substantial reduction of TIG1 expression was observed in 73.3% (11115) cancer cell lines, and seven of these exhibited nearly undetectable levels of expression. Decreased expression of TIG1 was also found in 62 (77.5%) primary carcinoma tissues compared to adjacent noncancerous tissues, indicating a tumor-specific reduction of TIG1. Expression levels of TIG1 were significantly low in primary carcinomas and cancer cell lines compared to those of normal tissues. Moreover, loss or reduction of TIG1 was significantly high in advanced tumors compared to early tumors and more frequent in poorly differentiated tumors than well or moderately differentiated tumors. TIG1 expression was reactivated or its level was elevated following 5-aza-2'-deoxycytidine treatment, indicating that TIG1 expression is transcriptionally silenced in these cancer cells by abnormal DNA hypermethylation. These data indicate that TIG1 undergoes frequent epigenetic inactivation due to aberrant DNA hypermethylation in gastric cancers, and its altered expression is associated with the malignant progression of tumors.


A systematic review of intervention thresholds based on FRAX : A report prepared for the National Osteoporosis Guideline Group and the International Osteoporosis Foundation.
This systematic review identified assessment guidelines for osteoporosis that incorporate FRAX. The rationale for intervention thresholds is given in a minority of papers. Intervention thresholds (fixed or age-dependent) need to be country-specific. In most assessment guidelines, treatment for osteoporosis is recommended in individuals with prior fragility fractures, especially fractures at spine and hip. However, for those without prior fractures, the intervention thresholds can be derived using different methods. The aim of this report was to undertake a systematic review of the available information on the use of FRAX® in assessment guidelines, in particular the setting of thresholds and their validation. We identified 120 guidelines or academic papers that incorporated FRAX of which 38 provided no clear statement on how the fracture probabilities derived are to be used in decision-making in clinical practice. The remainder recommended a fixed intervention threshold (n = 58), most commonly as a component of more complex guidance (e.g. bone mineral density (BMD) thresholds) or an age-dependent threshold (n = 22). Two guidelines have adopted both age-dependent and fixed thresholds. Fixed probability thresholds have ranged from 4 to 20 % for a major fracture and 1.3-5 % for hip fracture. More than one half (39) of the 58 publications identified utilised a threshold probability of 20 % for a major osteoporotic fracture, many of which also mention a hip fracture probability of 3 % as an alternative intervention threshold. In nearly all instances, no rationale is provided other than that this was the threshold used by the National Osteoporosis Foundation of the USA. Where undertaken, fixed probability thresholds have been determined from tests of discrimination (Hong Kong), health economic assessment (USA, Switzerland), to match the prevalence of osteoporosis (China) or to align with pre-existing guidelines or reimbursement criteria (Japan, Poland). Age-dependent intervention thresholds, first developed by the National Osteoporosis Guideline Group (NOGG), are based on the rationale that if a woman with a prior fragility fracture is eligible for treatment, then, at any given age, a man or woman with the same fracture probability but in the absence of a previous fracture (i.e. at the 'fracture threshold') should also be eligible. Under current NOGG guidelines, based on age-dependent probability thresholds, inequalities in access to therapy arise especially at older ages (≥70 years) depending on the presence or absence of a prior fracture. An alternative threshold using a hybrid model reduces this disparity. The use of FRAX (fixed or age-dependent thresholds) as the gateway to assessment identifies individuals at high risk more effectively than the use of BMD. However, the setting of intervention thresholds needs to be country-specific.


Applying a New Framework for Public Health Systems Recovery following Emergencies and Disasters: The Example of Haiti following a Major Earthquake and Cholera Outbreak.
Emergencies can often directly impact health systems of an affected region or country, especially in resource-constrained areas. Health system recovery following an emergency is a complex and dynamic process. Health system recovery efforts have often been structured around the World Health Organization's health systems building blocks as demonstrated by the Post-Disaster Needs Assessment. Although this structure is valuable and well known, it can overlook the intricacies of public health systems. We retrospectively examine public health systems recovery, a subset of the larger health system, following the 2010 Haiti earthquake and cholera outbreak, through the lens of the 10 essential public health services. This framework illustrates the comprehensive nature of and helps categorize the activities necessary for a well-functioning public health system and can complement other assessments. Outlining the features of a public health system for recovery in structured manner can also help lay the foundation for sustainable long-term development leading to a more robust and resilient health system.


Optimizing the early phase development of new analgesics by human pain biomarkers.
Human pain biomarkers are based on standardized acute activation of pain pathways/mechanisms and quantitative assessment of the evoked responses. This approach can be applied to healthy volunteers, to pain patients, and before and after pharmacological interventions to help understanding and profile the mode of action (proof-of-concept) of new and existing analgesic compounds. Standardized stimuli of different modalities can be applied to different tissues (multimodal and multi-tissue) for profiling analgesic compounds with respect to modulation of pain transduction, transmission, specific mechanisms and processing. This approach substantiates which specific compounds may work in particular clinical pain conditions. Human pain biomarkers can be translational and may bridge animal findings in clinical pain conditions, which in turn can provide new possibilities for designing more successful clinical trials. Biomarker based proof-of-concept drug studies in either volunteers or selected patient populations provide inexpensive, fast and reliable mechanism-based information about dose-efficacy relationships. This is important information in the early drug development phase and for designing large expensive clinical trials.


Virtual qubits, virtual temperatures, and the foundations of thermodynamics.
We argue that thermal machines can be understood from the perspective of "virtual qubits" at "virtual temperatures": The relevant way to view the two heat baths which drive a thermal machine is as a composite system. Virtual qubits are two-level subsystems of this composite, and their virtual temperatures can take on any value, positive or negative. Thermal machines act upon an external system by placing it in thermal contact with a well-selected range of virtual qubits and temperatures. We demonstrate these claims by studying the smallest thermal machines. We show further that this perspective provides a powerful way to view thermodynamics, by analyzing a number of phenomena. This includes approaching Carnot efficiency (where we find that all machines do so essentially by becoming equivalent to the smallest thermal machines), entropy production in irreversible machines, and a way to view work in terms of negative temperature and population inversion. Moreover we introduce the idea of "genuine" thermal machines and are led to considering the concept of "strength" of work.


Use of a Smartphone App to Assess Neonatal Jaundice.
The assessment of jaundice in outpatient neonates is problematic. Visual assessment is inaccurate, and more exact methodologies are cumbersome and/or expensive. Our goal in this study was to assess the accuracy of a technology based on the analysis of digital images of newborns obtained using a smartphone application called BiliCam. Paired BiliCam images and total serum bilirubin (TSB) levels were obtained in a diverse sample of newborns (<7 days old) at 7 sites across the United States. By using specialized software, data on color values in the images ("features") were extracted. Machine learning and regression analysis techniques were used to identify features for inclusion in models to predict an estimated bilirubin level for each newborn. The correlation between estimated bilirubin levels and TSB levels was calculated. In addition, the sensitivity and specificity of the estimated bilirubin levels in identifying newborns with high TSB levels were calculated by using 2 recommended decision rules for jaundice screening. Estimated bilirubin levels were calculated and compared with TSB levels in a diverse sample of 530 newborns (20.8% African American, 26.3% Hispanic, and 21.2% Asian American). The overall correlation was 0.91, and correlations among white, African American, Hispanic, and Asian American newborns were 0.92, 0.90, 0.91, and 0.88, respectively. The sensitivities of BiliCam in identifying newborns with high TSB levels were 84.6% and 100%, respectively, by using 2 decision rules; specificities were 75.1% and 76.4%, respectively. BiliCam provided accurate estimates of TSB values, demonstrating that an inexpensive technology that uses commodity smartphones could be used to effectively screen newborns for jaundice.


Fouling of reverse osmosis and nanofiltration membranes by dairy industry effluents.
Fouling experiments of nanofiltration (NF) and reverse osmosis (RO) are reported for treatment of the effluent of chemical-biological treatment plant and the original effluent of dairy industry respectively. In the experiments, a thin film composite type of spiral wound was used and fitted with flowmeters and pressure sensors. The feed water was stored into a feed tank and passed a fine filter and was pumped to membrane. Brine and permeate were recirculated back to the feed tank. Membrane fouling was investigated with 16 and 30% water recovery of a single membrane at different pressures and flowrates for RO and NF membranes respectively. Fouling is evaluated with a relationship between relative flux (J/Jo) which is the ratio of the flux at any time during the fouling test to the initial flux and relative resistance (Rf/Rm) which is the ratio of fouling (cake) layer resistance to clean membrane resistance. Turbidity, conductivity, chemical oxygen demand (COD), total suspended solids (TSS) and total hardness were measured in the feed and permeate side of each membrane. The effluent total hardness concentrations of chemical-biological treatment plant were found greater than the influents. The results are presented in terms of the relative flux as a function of time related to hydrodynamic conditions and pollution characteristics of wastewater. The permeate water flux of RO membrane decreases more rapidly than NF membrane, the relative flux decreases with increasing the fouling layer resistance, Rf onto membrane surface. 50% the drop of permeate flux was observed for RO and NF membranes after 50 h and 80 h of operation, respectively. The fouling rate increases with an increase in the concentration of the wastewater constituents in the dairy industry. The relative flux decreased 10 and 20% with increasing chemical oxygen demand (COD) from 5,000 mgl-1 to 10,000 mgl-1 and from 45 mgl-1 to 450 mgl-1 for RO and NF membranes, respectively after 45 h of time. Fouling of membranes resulted in 100% increase of specific energy consumption as the relative permeate fluxes of NF and RO membranes decreased 30 and 40% respectively. The average of specific energy consumption was obtained at 6 and 10 kWhm-3; consequently, operational costs were estimated at U.S. $0.45 m-3 and U.S. $0.75 m-3 for NF and RO units respectively. Also, operational cost for chemical-biological treatment was found at U.S. $0.30 m-3.


Relationship between Silent Brain Infarction and Amount of Daily Coffee Consumption in Middle Age.
In aging societies such as that of Japan, it is important to characterize lifestyle-related factors that minimize the occurrence of silent brain infarction (SBI) among the middle aged population for preventing vascular dementia in older age. Little is known about the relationship between amount of coffee consumption and SBI. To assess the association between the amount of coffee consumption and SBI in middle age, we statistically analyzed magnetic resonance imaging findings and data from questionnaires of consecutive 242 healthy Japanese individuals whose ages were less than 65 years and who participated in a medical brain-screening program at Teikyo University Chiba Medical Center from June 2008 to June 2009. In comparison with noncoffee drinkers (reference group), coffee drinkers who took 3-4 cups/day and 5 or more cups/day had a statistically lower incidence of SBI (.22, .07-.64, .004 and .43, .19-.99, .043, respectively). Upward logistic regression analysis indicated that SBI was influenced by 3 factors: coffee intake of 3 or more cups/day (.43, .22~.84, .014), history of hypertension (4.2, 2.0~8.8, .0001), and unemployment (2.1, 1.0~4.4, .037). As for consecutive 62 participants whose ages were 65 years or older in the same period, logistic regression analysis did not indicate that drinking coffee affected SBI incidence. Our report demonstrated that SBI was observed less frequently in middle aged Japanese who consumed 3 cups or more of coffee per day. To avoid senile dementia and/or symptomatic infarction in older age, the middle aged individuals might have to drink more than 3 cups of coffee every day.


Rural and urban differences in metabolic profiles in a Cameroonian population.
The difference between modern lifestyle in urban areas and the traditional way of life in rural areas may affect the population's health in developing countries proportionally. In this study, we sought to describe and compare the metabolic (fasting blood sugar and lipid profile) profile in an urban and rural sample of a Cameroonian population, and study the association to anthropometric risk factors of obesity. 332 urban and 120 rural men and women originating from the Sanaga Maritime Department and living in the Littoral Region in Cameroon voluntarily participated in this study. In all participants, measurement of height, weight, waist circumference, hip circumference, blood pressure systolic (SBP) and blood pressure diastolic (DBP), resting heart rate (RHR), blood glucose and lipids was carried out using standard methods. Total body fat (BF%) was measured using bio-impedancemetry. Body mass index (BMI) and waist to hip ratio (WHR) were calculated. Low Density Lipoprotein-cholesterol (LDL-c) concentrations were calculated using the Friedwald formula. World Health Organization criteria were used to define high and low levels of blood pressure, metabolic and anthropometric factors. The highest blood pressure values were found in rural men. Concerning resting heart rate, only the youngest women's age group showed a significant difference between urban and rural areas (79 ± 14 bpm vs 88 ± 12 bpm, p = 0.04) respectively. As opposed to the general tendency in our population, blood glucose was higher in rural men and women compared to their urban counterparts in the older age group (6.00 ± 2.56 mmol/L vs 5.72 ± 2.72 mmol/L, p = 0.030; 5.77 ± 3.72 vs 5.08 ± 0.60, p = 0,887 respectively). Triglycerides (TG) were significantly higher in urban than rural men (1.23 ± 0.39 mmol/L vs 1.17 ± 0.64 mmol/L, p = 0.017). High Density Lipoprotein-cholesterol (HDL-c) levels were higher in rural compared to urban men (2.60 ± 0.10 35mmol/L vs 1.97 ± 1.14 mmol/L, p<0.001 respectively). However, total Cholesterol (TC) and LDL-c were significantly higher in urban than in rural men (p<0.001 and p = 0.005) and women (p<0.001 respectively. Diabetes' rate in this population was 6.6%. This rate was higher in the rural (8.3%) than in the urban area (6.0%). Age and RHR were significantly higher in diabetic women than in non-diabetics (p = 0.007; p = 0.032 respectively). In a multiple regression, age was an independent predictor of SBP, DBP and RHR in the entire population. Age predicted blood glucose in rural women only. BMI, WC and BF% were independent predictors of RHR in rural population, especially in men. WC and BF% predicted DBP in rural men only. Anthropometric parameters did not predict the lipid profile. Lipid profile was less atherogenic in rural than in urban area. The rural population was older than the urban one. Blood pressure and blood glucose were positively associated to age in men and women respectively; this could explain the higher prevalence of diabetes in rural than in urban area. The association of these metabolic variables to obesity indices is more frequent and important in urban than in rural area.


Diagnosis of malignant pleural mesothelioma from pleural fluid by Fourier transform-infrared spectroscopy coupled with chemometrics.
This study was conducted to differentiate malignant pleural mesothelioma (MPM) from lung cancer (LC) and benign pleural effusion (BPE) from pleural fluids using the diagnostic power of Fourier transform-infrared spectroscopy with attenuated total reflectance mode coupled with chemometrics. Infrared spectra of MPM (n = 24), LC (n = 20), and BPE (n = 25) were collected, and hierarchical cluster analysis (HCA) and principal component analysis (PCA) were applied to their spectra. HCA results indicated that MPM was differentiated from LC with 100% sensitivity and 100% specificity and from BPE, with 100% sensitivity and 88% specificity, which were also confirmed by PCA score plots. PCA loading plots indicated that these separations originated mainly from lipids, proteins, and nucleic acids-related spectral bands. There was significantly higher lipid, protein, nucleic acid, and glucose contents in the MPM and LC. However, the significant changes in triglyceride and cholesterol ester content, protein and nucleic acid structure, a lower membrane fluidity, and higher membrane order were only observed in the MPM. To check the classification success of some test samples/each group, soft independent modeling of class analogies was performed and 96.2% overall classification success was obtained. This approach can provide a rapid and inexpensive methodology for the efficient differentiation of MPM from other pleural effusions.


Effect of Light Quality and Vernalization on Late-Flowering Mutants of Arabidopsis thaliana.
We have analyzed the response to vernalization and light quality of six classes of late-flowering mutants (fb, fca, fe, fg, ft, and fy) previously isolated following mutagenesis of the early Landsberg race of Arabidopsis thaliana (L.) Heynh. When grown in continuous fluorescent illumination, four mutants (fca, fe, ft, and fy) and the Landsberg wild type exhibited a reduction in both flowering time and leaf number following 6 weeks of vernalization. A significant decrease in flowering time was also observed for all the mutants and the wild type when constant fluorescent illumination was supplemented with irradiation enriched in the red and far red regions of the spectrum. In the most extreme case, the late-flowering phenotype of the fca mutant was completely suppressed by vernalization, suggesting that this mutation has a direct effect on flowering. The fe and fy mutants also showed a more pronounced response than wild type to both vernalization and incandescent supplementation. The ft mutant showed a similar response to that of the wild type. The fb and fg mutants were substantially less sensitive to these treatments. These results are interpreted in the context of a multifactorial pathway for induction of flowering, in which the various mutations affect different steps of the pathway.


Repetitive treatment with diluted bee venom reduces neuropathic pain via potentiation of locus coeruleus noradrenergic neuronal activity and modulation of spinal NR1 phosphorylation in rats.
We previously demonstrated that a single injection of diluted bee venom (DBV) temporarily alleviates thermal hyperalgesia, but not mechanical allodynia, in neuropathic rats. The present study was designed to determine whether repetitive injection of DBV produces more potent analgesic effects on neuropathy-induced nociception and whether those effects are associated with increased neuronal activity in the locus coeruleus (LC) and with the suppression of spinal NMDA receptor NR1 subunit phosphorylation (pNR1). DBV (.25 mg/kg) was administered subcutaneously twice a day for 2 weeks beginning on day 15 post-chronic constrictive injury surgery. Pain responses were examined and potential changes in LC Fos expression and spinal pNR1 expression were determined. Repetitive DBV administration significantly reduced mechanical allodynia, as well as thermal hyperalgesia. The activity of LC noradrenergic neurons was increased and spinal pNR1 expression was significantly suppressed by repetitive DBV as compared with those of vehicle or single DBV injection. These suppressive effects of repetitive DBV on neuropathic pain and spinal pNR1 were prevented by intrathecal pretreatment of idazoxan, an alpha-2 adrenoceptor antagonist. These results indicate that repetitive DBV produces potent analgesic effects on neuropathic pain and this is associated with the activation of the LC noradrenergic system and with a reduction in spinal pNR1. The results of current study demonstrate that repetitive administration of DBV significantly suppresses neuropathic pain. Furthermore, this study provides mechanistic information that repetitive treatment of DBV can produce more potent analgesic effect than single DBV treatment, indicating a potential novel strategy for the management of chronic pain.


Designing genome-wide association studies: sample size, power, imputation, and the choice of genotyping chip.
Genome-wide association studies are revolutionizing the search for the genes underlying human complex diseases. The main decisions to be made at the design stage of these studies are the choice of the commercial genotyping chip to be used and the numbers of case and control samples to be genotyped. The most common method of comparing different chips is using a measure of coverage, but this fails to properly account for the effects of sample size, the genetic model of the disease, and linkage disequilibrium between SNPs. In this paper, we argue that the statistical power to detect a causative variant should be the major criterion in study design. Because of the complicated pattern of linkage disequilibrium (LD) in the human genome, power cannot be calculated analytically and must instead be assessed by simulation. We describe in detail a method of simulating case-control samples at a set of linked SNPs that replicates the patterns of LD in human populations, and we used it to assess power for a comprehensive set of available genotyping chips. Our results allow us to compare the performance of the chips to detect variants with different effect sizes and allele frequencies, look at how power changes with sample size in different populations or when using multi-marker tags and genotype imputation approaches, and how performance compares to a hypothetical chip that contains every SNP in HapMap. A main conclusion of this study is that marked differences in genome coverage may not translate into appreciable differences in power and that, when taking budgetary considerations into account, the most powerful design may not always correspond to the chip with the highest coverage. We also show that genotype imputation can be used to boost the power of many chips up to the level obtained from a hypothetical "complete" chip containing all the SNPs in HapMap. Our results have been encapsulated into an R software package that allows users to design future association studies and our methods provide a framework with which new chip sets can be evaluated.


Clinical application of pharmacokinetic analysis as a biomarker of solitary pulmonary nodules: dynamic contrast-enhanced MR imaging.
The purpose of this study is to evaluate perfusion indices and pharmacokinetic parameters in solitary pulmonary nodules (SPNs). Thirty patients of 34 enrolled with SPNs (15-30 mm) were evaluated in this study. T1 and T2-weighted structural images and 2D turbo FLASH perfusion images were acquired with shallow free breathing. B-spline nonrigid image registration and optimization by χ² test against pharmacokinetic model curve were performed on dynamic contrast-enhanced MRI. This allowed voxel-by-voxel calculation of k(ep) , the rate constant for tracer transport to and from plasma and the extravascular extracellular space. Mean transit time, time-to-peak, initial slope, and maximum enhancement (E(max) ) were calculated from time-intensity curves fitted to a gamma variate function. After blinded data analysis, correlation with tissue histology from surgical resection or biopsy samples was performed. Histologic evaluation revealed 25 malignant and five benign SPNs. All benign SPNs had k(ep) < 1.0 min⁻¹. Nineteen of 25 (76%) malignant SPNs showed k(ep) > 1.0 min⁻¹. Sensitivity to diagnose malignant SPNs at a cutoff of k(ep) = 1.0 min⁻¹ was 76%, specificity was 100%, positive predictive value was 100%, negative predictive value was 45%, and accuracy was 80%. Of all indices studied, k(ep) was the most significant in differentiating malignant from benign SPNs.


Deduced primary structure of vitellogenin in the giant freshwater prawn, Macrobrachium rosenbergii, and yolk processing during ovarian maturation.
A cDNA encoding vitellogenin (Vg) in the giant freshwater prawn, Macrobrachium rosenbergii, was cloned based on the cDNA sequence of vitellin (Vn) fragments A-N and B-42 determined previously, and its amino acid sequence deduced. The open reading frame (ORF) encoded 2,537 amino acid residues and its deduced amino acid sequence possessed three consensus cleavage sites, R-X-R-R, similar to those reported in Vgs of insects. The deduced primary structure of Vg in M. rosenbergii was seen to be similar to that of Penaeus japonicus, especially in the N-terminal region. It is therefore likely that Vgs in crustacean species including prawns and other related decapods exhibit a similar structural pattern. Based on the deduced primary structure of Vg and analysis of the various Vg and Vn subunits found in the hemolymph and ovary during ovarian maturation, we demonstrated the post-translational processing of Vg in M. rosenbergii. This is the first time that Vg processing has been clearly demonstrated in a crustacean species. Vg, after being synthesized in the hepatopancreas, is considered to be cleaved by a subtilisin-like endoprotease to form two subunits, A and proB, which are then released into the hemolymph. In the hemolymph, proB is possibly cleaved by a processing enzyme of unknown identity to give rise to subunits B and C/D. The three processed subunits A, B, and C/D are sequestered by the ovary to give rise to three yolk proteins, Macr-VnA, VnB, and VnC/D.


Characterizing poly(epsilon-caprolactone)-b-chitooligosaccharide-b-poly(ethylene glycol) (PCP) copolymer micelles for doxorubicin (DOX) delivery: effects of crosslinked of amine groups.
New amine-groups containing tri-block copolymers and micelles that consisting of poly(epsilon-caprolactone)-b-chitooligosaccharide-b-poly(ethylene glycol) (PCL-b-COS-b-PEG, PCP), were synthesized, characterized, and evaluated for delivering doxorubicin (DOX) with or without crosslinked amine groups by genipin. The characteristics of the PCP copolymers of Fourier-transform infrared spectrometry (FT-IR) verify the amine and ester groups of the COS and the PCL of the copolymers, respectively. 1H nuclear magnetic resonance (1H NMR) spectra verify the structures of the PCP copolymers consisting two PCL and PEG polymers reacted onto the COS block. In addition, gel permeation chromatography (GPC) determines the number average molecular weight of the tri-block copolymers (Mn) of approximately 11340 Da/mole. The PCP copolymers can self-assemble to form polymeric micelles at the critical micelle concentration (CMC) of 1.0 microM as determined by the UV-VIS absorption spectra. The mean diameter of the PCP micelles is 90 nm, as determined using a dynamic light-scattering (DLS) analyzer. Moreover, the zeta potentials of PCP micelles change from neutral to cationic state when pH of suspension mediums varied from 7.4 to 3.0. For evaluating delivery characteristics of hydrophobic DOX, it was loaded into PCP micelles with or without crosslinked by genipin. The burst release and release period of DOX for the crosslinked micelles are significantly reduced (P < 0.003, n = 3, for pH = 7.4) and sustained (e.g., 8 days), respectively, than those non-crosslinked ones (e.g., 4 days). In conclusion, new tri-block amine groups containing PCP copolymers are synthesized that can self-assemble as PCP micelles. After post-crosslinked amine groups of DOX loaded the micelles, they can effectively reduce the burst release and sustain the release of DOX at different pH dissolution mediums. Further applications of PCP copolymers and micelles for drug delivery can be explored in future.


Regulation of neuronal high-voltage activated Ca(V)2 Ca(2+) channels by the small GTPase RhoA.
High-Voltage-Activated (HVA) Ca(2+) channels are known regulators of synapse formation and transmission and play fundamental roles in neuronal pathophysiology. Small GTPases of Rho and RGK families, via their action on both cytoskeleton and Ca(2+) channels are key molecules for these processes. While the effects of RGK GTPases on neuronal HVA Ca(2+) channels have been widely studied, the effects of RhoA on the HVA channels remains however elusive. Using heterologous expression in Xenopus laevis oocytes, we show that RhoA activity reduces Ba(2+) currents through CaV2.1, CaV2.2 and CaV2.3 Ca(2+) channels independently of CaVβ subunit. This inhibition occurs independently of RGKs activity and without modification of biophysical properties and global level of expression of the channel subunit. Instead, we observed a marked decrease in the number of active channels at the plasma membrane. Pharmacological and expression studies suggest that channel expression at the plasma membrane is impaired via a ROCK-sensitive pathway. Expression of constitutively active RhoA in primary culture of spinal motoneurons also drastically reduced HVA Ca(2+) current amplitude. Altogether our data revealed that HVA Ca(2+) channels regulation by RhoA might govern synaptic transmission during development and potentially contribute to pathophysiological processes when axon regeneration and growth cone kinetics are impaired.


Oestrogens and progestogens for preventing and treating postnatal depression.
Postnatal depression, with a prevalence of at least 10%, is probably the most common complication of the puerperium. A deficiency or imbalance of sex hormones has repeatedly been suggested as a cause. The objective of this review was to evaluate the role of oestrogens and progestogens in the prevention and treatment of postnatal depression. The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. All trials were considered in which pregnant or postpartum women (up to 18 months) were randomised to receive postpartum oestrogen or progestogen or placebo for the treatment or prevention of postnatal depression. Two published randomised placebo controlled trials were identified for inclusion in the analyses for this review. One study was excluded. Depot norethisterone enanthate given within 48 hours of delivery and lasting 8-12 weeks was associated with significantly higher postpartum depression scores than placebo. Oestrogen therapy in severely depressed women was associated with a greater improvement in depression scores than placebo. There is no place for synthetic progestogens in the prevention of treatment of postnatal depression. Long-acting norethisterone enanthate is associated with an increased risk of postnatal depression. It and other long-acting progestogen contraceptives should be used with caution in the postnatal period, especially in women with a history of depression. The role of progesterone in the prevention and treatment of postnatal depression has yet to be evaluated in a randomised placebo-controlled trial. Oestrogen therapy may be of modest value at a late stage of severe postnatal depression. Its role in the prevention of recurrent postnatal depression has not been evaluated. Further research on its value is unlikely for ethical reasons.


Neurochemical compartmentalization within the pigeon basal ganglia.
The goals of this study were to use multiple informative markers to define and characterize the neurochemically distinct compartments of the pigeon basal ganglia, especially striatum and accumbens. To this end, we used antibodies against 12 different neuropeptides, calcium-binding proteins or neurotransmitter-related enzymes that are enriched in the basal ganglia. Our results clarify boundaries between previously described basal ganglia subdivisions in birds, and reveal considerable novel heterogeneity within these previously described subdivisions. Sixteen regions were identified that each displayed a unique neurochemical organization. Four compartments were identified within the dorsal striatal region. The neurochemical characteristics support previous comparisons to part of the central extended amygdala, somatomotor striatum, and associational striatum of mammals, respectively. The medialmost part of the medial striatum, however, has several unique features, including prominent pallidal-like woolly fibers and thus may be a region unique to birds. Four neurochemically distinct regions were identified within the pigeon ventral striatum: the accumbens, paratubercular striatum, ventrocaudal striatum, and the ventral area of the lateral part of the medial striatum that is located adjacent to these regions. The pigeon accumbens is neurochemically similar to the mammalian rostral accumbens. The pigeon paratubercular and ventrocaudal striatal regions are similar to the mammalian accumbens shell. The ventral portions of the medial and lateral parts of the medial striatum, which are located adjacent to accumbens shell-like areas, have neurochemical characteristics as well as previously reported limbic connections that are comparable to the accumbens core. Comparisons to neurochemically identified compartments in reptiles, mammals, and amphibians indicate that, although most of the basic compartments of the basal ganglia were highly conserved during tetrapod evolution, uniquely avian compartments may exist as well.


Malaria surveillance--United States, 2009.
Malaria in humans is caused by intraerythrocytic protozoa of the genus Plasmodium. These parasites are transmitted by the bite of an infective female Anopheles mosquito. The majority of malaria infections in the United States occur among persons who have traveled to areas with ongoing malaria transmission. In the United States, cases can occur through exposure to infected blood products, congenital transmission, or local mosquitoborne transmission. Malaria surveillance is conducted to identify episodes of local transmission and to guide prevention recommendations for travelers. This report summarizes cases in persons with onset of illness in 2009 and summarizes trends during previous years. Malaria cases diagnosed by blood film, polymerase chain reaction or rapid diagnostic tests are mandated to be reported to local and state health departments by health-care providers or laboratory staff. Case investigations are conducted by local and state health departments, and reports are transmitted to CDC through the National Malaria Surveillance System (NMSS), National Notifiable Diseases Surveillance System (NNDSS), or direct CDC consults. Data from these reporting systems serve as the basis for this report. CDC received reports of 1,484 cases of malaria, including two transfusion-related cases, three possible congenital cases, one transplant case and four fatal cases, with an onset of symptoms in 2009 among persons in the United States. This number represents an increase of 14% from the 1,298 cases reported for 2008. Plasmodium falciparum, P. vivax, P. malariae, and P. ovale were identified in 46%, 11%, 2%, and 2% of cases, respectively. Thirteen patients were infected by two or more species. The infecting species was unreported or undetermined in 38% of cases. Among the 1,484 cases 1,478 were classified as imported. Among the 103 U.S. civilians for whom information on chemoprophylaxis use and travel area was known, only 34 (33%) reported that they had followed and adhered to a chemoprophylactic drug regimen recommended by CDC for the area to which they had traveled. Nineteen cases were reported in pregnant women, among whom none adhered to chemoprophylaxis. Almost 22% of the cases among pregnant women were treated with an inappropriate treatment drug regimen, of which 39% were among cases with either a P. vivax or P. ovale infection where primaquine was not taken. Among all the reasons for travel, travelers visiting friends and relatives (VFR) and missionaries were the groups with the lowest proportion of chemoprophylexis use. A notable increase in the number of malaria cases was reported from 2008 to 2009; however, the number of cases in 2009 is consistent with the average number of cases reported during the preceding 4 years. In the majority of reported cases, U.S. civilians who acquired infection abroad had not adhered to a chemoprophylaxis regimen that was appropriate for the country in which they acquired malaria. Furthermore, treatment of malaria, while appropriate for the majority of cases, was insufficient for a large number of P. vivax and P. ovale infections, putting patients at risk for relapsing malaria. Decreasing the number of malaria cases in subsequent years will require conveying the importance of adhering to appropriate preventive measures for malaria specifically targeting travelers visiting friends and relatives, missionary, and pregnant populations. Clinicians require education on the need to encourage use of malaria prophylaxis and need further information on the appropriate diagnostic and treatment guidelines for malaria. Malaria prevention recommendations are available online (http://www.cdc.gov/malaria/travelers/ or http://wwwnc.cdc.gov/travel/yellowbook/2010/chapter-2/malaria.aspx#990). Malaria infections can be fatal if not diagnosed and treated promptly with antimalarial medications appropriate for the individual patient's age and medical history, the likely site of malaria acquisition, and previous use of antimalarial chemoprophylaxis. Clinicians should consult the CDC Guidelines for Treatment and contact the CDC's Malaria Hotline for case management advisement when needed. Malaria treatment recommendations can be obtained online (http://www.cdc.gov/malaria/diagnosis_treatment) or by calling the Malaria Hotline (770-488-7788).


Personalized prediction of early childhood asthma persistence: A machine learning approach.
Early childhood asthma diagnosis is common; however, many children diagnosed before age 5 experience symptom resolution and it remains difficult to identify individuals whose symptoms will persist. Our objective was to develop machine learning models to identify which individuals diagnosed with asthma before age 5 continue to experience asthma-related visits. We curated a retrospective dataset for 9,934 children derived from electronic health record (EHR) data. We trained five machine learning models to differentiate individuals without subsequent asthma-related visits (transient diagnosis) from those with asthma-related visits between ages 5 and 10 (persistent diagnosis) given clinical information up to age 5 years. Based on average NPV-Specificity area (ANSA), all models performed significantly better than random chance, with XGBoost obtaining the best performance (0.43 mean ANSA). Feature importance analysis indicated age of last asthma diagnosis under 5 years, total number of asthma related visits, self-identified black race, allergic rhinitis, and eczema as important features. Although our models appear to perform well, a lack of prior models utilizing a large number of features to predict individual persistence makes direct comparison infeasible. However, feature importance analysis indicates our models are consistent with prior research indicating diagnosis age and prior health service utilization as important predictors of persistent asthma. We therefore find that machine learning models can predict which individuals will experience persistent asthma with good performance and may be useful to guide clinician and parental decisions regarding asthma counselling in early childhood.


Alexithymia, not autism, predicts poor recognition of emotional facial expressions.
Despite considerable research into whether face perception is impaired in autistic individuals, clear answers have proved elusive. In the present study, we sought to determine whether co-occurring alexithymia (characterized by difficulties interpreting emotional states) may be responsible for face-perception deficits previously attributed to autism. Two experiments were conducted using psychophysical procedures to determine the relative contributions of alexithymia and autism to identity and expression recognition. Experiment 1 showed that alexithymia correlates strongly with the precision of expression attributions, whereas autism severity was unrelated to expression-recognition ability. Experiment 2 confirmed that alexithymia is not associated with impaired ability to detect expression variation; instead, results suggested that alexithymia is associated with difficulties interpreting intact sensory descriptions. Neither alexithymia nor autism was associated with biased or imprecise identity attributions. These findings accord with the hypothesis that the emotional symptoms of autism are in fact due to co-occurring alexithymia and that existing diagnostic criteria may need to be revised.


A randomized controlled trial of acupuncture added to usual treatment for fibromyalgia.
To evaluate the effectiveness of acupuncture for fibromyalgia. Fifty-eight women with fibromyalgia were allocated randomly to receive either acupuncture together with tricyclic antidepressants and exercise (n=34), or tricyclic antidepressants and exercise only (n=24). Patients rated their pain on a visual analogue scale. A blinded assessor evaluated both the mean pressure pain threshold value over all 18 fibromyalgia points and quality of life using SF-36. At the end of 20 sessions, patients who received acupuncture were significantly better than the control group in all measures of pain and in 5 of the SF-36 subscales. After 6 months, the acupuncture group was significantly better than the control group in numbers of tender points, mean pressure pain threshold at the 18 tender points and 3 subscales of SF-36. After one year, the acupuncture group showed significance in one subscale of the SF-36; at 2 years there were no significant differences in any outcome measures. Addition of acupuncture to usual treatments for fibromyalgia may be beneficial for pain and quality of life for 3 months after the end of treatment. Future research is needed to evaluate the specific effects of acupuncture for fibromyalgia.


Nickel-Catalyzed Asymmetric Synthesis of α-Arylbenzamides.
A nickel-catalyzed asymmetric reductive hydroarylation of vinyl amides to produce enantioenriched α-arylbenzamides is reported. The use of a chiral bisimidazoline (BIm) ligand, in combination with diethoxymethylsilane and aryl halides, enables the regioselective introduction of aryl groups to the internal position of the olefin, forging a new stereogenic center α to the N atom. The use of neutral reagents and mild reaction conditions provides simple access to pharmacologically relevant motifs present in anticancer, SARS-CoV PLpro inhibitors, and KCNQ channel openers.


The role of IL-10 in inflammatory bowel disease: "of mice and men".
Inflammatory bowel disease (IBD) is a generic term typically used to describe a group of idiopathic inflammatory intestinal conditions in humans that are generally divided into Crohn's disease and ulcerative colitis. Although the etiology of these diseases remains unknown, a number of rodent models of IBD have recently been identified, all sharing the concept that the development of chronic intestinal inflammation occurs as a consequence of alterations in the immune system that lead to a failure of normal immunoregulation in the intestine. On the basis of these models, it has been hypothesized that the development of IBD in humans may be related to a dysregulated immune response to normal flora in the gut. Immunodeficient scid mice injected with CD4+ CD45RB(high) T cells and mice deficient in interleukin (IL)-10 (IL-10-/-) are among the rodent models of IBD. In both models, there is inflammation and evidence of a Th1-like response in the large intestine, characterized by CD4+ T-cell and macrophage infiltrates, and elevated levels of interferon-gamma. Because IL-10 is an immunomodulatory cytokine that is capable of controlling Th1-like responses, the role of IL-10 was investigated in these models. IL-10 was shown to be important in regulating the development of intestinal inflammation in both models. These results provided key data that supported initiation of clinical trials evaluating the efficacy of IL-10 in patients with IBD.


Surgery in Greece.
The expenditure on health care reached 7.8% of the gross domestic product in Greece in 1991. Almost 9000 specialized surgeons in 11 recognized specialties provide services to more than 10 million people living in Greece and 2 million visitors annually. Medical studies comprise a 6-year course of 5000 to 5500 teaching hours in 7 medical schools and are free. Organizational reform of the health care system in vital and long overdue to move the system into the 21st century. The quality of surgical services provided is high by international standards; cardiac surgery, organ transplantation, surgical oncology, and anesthesiology are some areas of excellence. In addition, Greece has a good record in medical technology and instrumentation. Research activities cover a range of subjects, and the number of clinical medicine articles from Greece that appear in peer-reviewed journals compares favorably with other larger countries. Reforms in the postgraduate training system and ongoing education are necessary.


Gene-specific requirement for P-TEFb activity and RNA polymerase II phosphorylation within the p53 transcriptional program.
Activation of the p53 pathway mediates cellular responses to diverse forms of stress. Here we report that the p53 target gene p21(CIP1) is regulated by stress at post-initiation steps through conversion of paused RNA polymerase II (RNAP II) into an elongating form. High-resolution chromatin immunoprecipitation assays (ChIP) demonstrate that p53-dependent activation of p21(CIP1) transcription after DNA damage occurs concomitantly with changes in RNAP II phosphorylation status and recruitment of the elongation factors DSIF (DRB Sensitivity-Inducing Factor), P-TEFb (Positive Transcription Elongation Factor b), TFIIH, TFIIF, and FACT (Facilitates Chromatin Transcription) to distinct regions of the p21(CIP1) locus. Paradoxically, pharmacological inhibition of P-TEFb leads to global inhibition of mRNA synthesis but activation of the p53 pathway through p53 accumulation, expression of specific p53 target genes, and p53-dependent apoptosis. ChIP analyses of p21(CIP1) activation in the absence of functional P-TEFb reveals the existence of two distinct kinases that phosphorylate Ser5 of the RNAP II C-terminal domain (CTD). Importantly, CTD phosphorylation at Ser2 is not required for p21(CIP1) transcription, mRNA cleavage, or polyadenylation. Furthermore, recruitment of FACT requires CTD kinases, yet FACT is dispensable for p21(CIP1) expression. Thus, select genes within the p53 pathway bypass the requirement for P-TEFb and RNAP II phosphorylation to trigger a cellular response to inhibition of global mRNA synthesis.


Single-cell isoform RNA sequencing characterizes isoforms in thousands of cerebellar cells.
Full-length RNA sequencing (RNA-Seq) has been applied to bulk tissue, cell lines and sorted cells to characterize transcriptomes, but applying this technology to single cells has proven to be difficult, with less than ten single-cell transcriptomes having been analyzed thus far. Although single splicing events have been described for ≤200 single cells with statistical confidence, full-length mRNA analyses for hundreds of cells have not been reported. Single-cell short-read 3' sequencing enables the identification of cellular subtypes, but full-length mRNA isoforms for these cell types cannot be profiled. We developed a method that starts with bulk tissue and identifies single-cell types and their full-length RNA isoforms without fluorescence-activated cell sorting. Using single-cell isoform RNA-Seq (ScISOr-Seq), we identified RNA isoforms in neurons, astrocytes, microglia, and cell subtypes such as Purkinje and Granule cells, and cell-type-specific combination patterns of distant splice sites. We used ScISOr-Seq to improve genome annotation in mouse Gencode version 10 by determining the cell-type-specific expression of 18,173 known and 16,872 novel isoforms.


DNA polymerase template switching at specific sites on the phi29 genome causes the in vivo accumulation of subgenomic phi29 DNA molecules.
The accumulation of subgenomic phage phi29 DNA molecules with specific sizes was observed after prolonged infection times with delayed lysis phage mutants. Whereas the majority of the molecules had a size of 4 kb, additional DNA species were observed with sizes of 8.2, 6.5, 2.3, 2 and 1 kb. Most of the molecules were shown to originate from the right end of the linear Bacillus subtilis phage phi29 genome. The nature of the 4, 2.3, 2 and 1 kb molecules was studied. The 2 kb molecules were shown to be single-stranded self-complementary strands forming hairpin structures. The other molecules consisted of palindromic linear double-stranded DNA molecules. Most probably, the subgenomic DNA molecules were formed when the moving phage replication fork from the right origin encountered a block that induces the DNA polymerase to switch template. Once formed, the subgenomic molecules are then amplified in vivo. Determination of the centres of symmetry of the 4 and 1 kb molecules revealed that both contained the almost 16 bp perfect dyad symmetry element (DSE): 5'-TGTTtCAC-GTGg-AACA-3' being a likely candidate for a protein binding site. Database analysis showed that this sequence occurs four times in the phi29 genome. In addition, the almost identical sequence 5'-TgGTTTCAC-GTGGAAtCA-3' was found once. These five DSEs are all located in the right half of the phi29 genome, and the same sequences are also present in the linear DNA of related B. subtilis phages. Most interestingly, this sequence is also found in the spoOJ gene of the B. subtilis chromosome. Recently, it has been shown that the SpoOJ protein is associated in vivo with the same DSE. As the same subgenomic phi29 DNA molecules accumulate after infection of B. subtilis spoOJ deletion strains, it is likely that, in addition to and/or independently of SpoOJ, other protein(s) bind to DSE.


Recent development of two-photon fluorescent probes for bioimaging.
Fluorescent probes are essential tools for studying biological systems. The last decade has witnessed particular interest in the development of two-photon excitable probes, owing to their advantageous features in tissue imaging compared to the corresponding one-photon probes. This review summarizes various types of two-photon probes that have been applied to bioimaging, categorized by the principles in the probe design and the target analytes, which would provide a basis for the future development of novel two-photon probes for tackling important biological issues.


Race/ethnicity, genetic ancestry, and breast cancer-related lymphedema in the Pathways Study.
Breast cancer-related lymphedema (BCRL) is a serious chronic condition after breast cancer (BC) surgery and treatment. It is unclear if BCRL risk varies by race/ethnicity. In a multiethnic prospective cohort study of 2953 BC patients, we examined the association of self-reported BCRL status with self-reported race/ethnicity and estimated genetic ancestry. Hazard ratios (HR) and 95 % confidence intervals (CI) were calculated by multivariable Cox proportional hazards models, with follow-up starting 6 months post-BC diagnosis. Estimates were further stratified by body mass index (BMI). By 48 months of follow-up, 342 (11.6 %) women reported having BCRL. Younger age at BC diagnosis, higher BMI at baseline, and lower physical activity were associated with greater BCRL risk. African American (AA) women had a 2-fold increased risk of BCRL compared with White women (HR = 2.04; 95 % CI 1.35-3.08). African genetic ancestry was also associated with an increased risk (HR = 2.50; 95 % CI 1.43, 4.36). Both risks were attenuated but remained elevated after adjusting for known risk factors and became more pronounced when restricted to the nonobese women (adjusted HR = 2.31 for AA and HR = 3.70 for African ancestry, both p < 0.05). There was also evidence of increased BCRL risk with Hispanic ethnicity in the nonobese women. Nonobese AA women had a higher risk of BCRL than White women, which cannot be fully explained by known risk factors. This is the first large-scale, prospective study demonstrating differences in BCRL risk according to race/ethnicity as assessed by both self-report and genetic ancestry data, with a potential ancestry-obesity interaction.


Continuously tunable terahertz metamaterial employing magnetically actuated cantilevers.
Terahertz metamaterial structures that employ flexing microelectromechanical cantilevers for tuning the resonance frequency of an electric split-ring resonator are presented. The tuning cantilevers are coated with a magnetic thin-film and are actuated by an external magnetic field. The use of cantilevers enables continuous tuning of the resonance frequency over a large frequency range. The use of an externally applied magnetic field for actuation simplifies the metamaterial structure and its use for sensor or filter applications. A structure for minimizing the actuating field is derived. The dependence of the tunable bandwidth on frequency is discussed.


Chitosan-modified PLGA nanoparticles with versatile surface for improved drug delivery.
Shortage of functional groups on surface of poly(lactide-co-glycolide) (PLGA)-based drug delivery carriers always hampers its wide applications such as passive targeting and conjugation with targeting molecules. In this research, PLGA nanoparticles were modified with chitosan through physical adsorption and chemical binding methods. The surface charges were regulated by altering pH value in chitosan solutions. After the introduction of chitosan, zeta potential of the PLGA nanoparticle surface changed from negative charge to positive one, making the drug carriers more affinity to cancer cells. Functional groups were compared between PLGA nanoparticles and chitosan-modified PLGA nanoparticles. Amine groups were exhibited on PLGA nanoparticle surface after the chitosan modification as confirmed by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. The modified nanoparticles showed an initial burst release followed by a moderate and sustained release profile. Higher percentage of drugs from cumulative release can be achieved in the same prolonged time range. Therefore, PLGA nanoparticles modified by chitosan showed versatility of surface and a possible improvement in the efficacy of current PLGA-based drug delivery system.


Extracellular matrix protein CCN1 regulates cardiomyocyte apoptosis in mice with stress-induced cardiac injury.
Expression of extracellular matrix protein CCN1 is induced in end-stage ischaemic cardiomyopathy in humans, and after cardiac ischaemia and reperfusion in experimental animal models. Despite its well-documented angiogenic activities, CCN1 increases the cytotoxicities of the tumour necrosis factor family cytokines, which promotes apoptosis in fibroblasts. We aimed to determine the physiological function of CCN1 in an injured heart. To assess the function of CCN1 in vivo, knock-in mice carrying the apoptosis-defective mutant allele Ccn1-dm were tested in an isoproterenol (ISO)-induced myocardial injury model (100 mg/kg/day of sc injected ISO for 5 days). Compared with wild-type mice, Ccn1(dm/dm) mice were remarkably resistant to ISO-induced cardiac injury; they showed no post-treatment cardiomyocyte apoptosis or myocardial tissue damage. ISO cardiotoxicity was dependent on Fas ligand (FasL) and its downstream signalling. Using primary cultures of cardiomyocytes isolated from rats, we demonstrated that CCN1 sensitized FasL-mediated apoptosis by engaging its cell-surface receptor integrin α6β1 and up-regulating intracellular reactive oxygen species (ROS), which activated mitogen-activated protein kinase p38, and increased cell-surface Fas expression. CCN1 is a critical pathophysiological regulator that mediates cardiomyocyte apoptosis during work-overload-induced cardiac injury. CCN1 increases cellular susceptibility to Fas-induced apoptosis by increasing ROS and cell-surface Fas expression.


Colocalization of inflammatory response with B7-h1 expression in human melanocytic lesions supports an adaptive resistance mechanism of immune escape.
Although many human cancers such as melanoma express tumor antigens recognized by T cells, host immune responses often fail to control tumor growth for as yet unexplained reasons. Here, we found a strong association between melanocyte expression of B7-H1 (PD-L1), an immune-inhibitory molecule, and the presence of tumor-infiltrating lymphocytes (TILs) in human melanocytic lesions: 98% of B7-H1(+) tumors were associated with TILs compared with only 28% of B7-H1(-) tumors. Indeed, B7-H1(+) melanocytes were almost always localized immediately adjacent to TILs. B7-H1/TIL colocalization was identified not only in melanomas but also in inflamed benign nevi, indicating that B7-H1 expression may represent a host response to tissue inflammation. Interferon-γ, a primary inducer of B7-H1 expression, was detected at the interface of B7-H1(+) tumors and TILs, whereas none was found in B7-H1(-) tumors. Therefore, TILs may actually trigger their own inhibition by secreting cytokines that drive tumor B7-H1 expression. Consistent with this hypothesis, overall survival of patients with B7-H1(+) metastatic melanoma was significantly prolonged compared with that of patients with B7-H1(-) metastatic melanoma. Therefore, induction of the B7-H1/PD-1 pathway may represent an adaptive immune resistance mechanism exerted by tumor cells in response to endogenous antitumor activity and may explain how melanomas escape immune destruction despite endogenous antitumor immune responses. These observations suggest that therapies that block this pathway may benefit patients with B7-H1(+) tumors.


Buffer-free integrative nanofluidic device for real-time continuous flow bioassays by ion concentration polarization.
To perform precision medicine in real-time, a sensor capable of continuously monitoring target biomolecules secreted from a patient under dynamic situations is essential. In this study, a novel portable device combining an aptamer probe and a nanofluidic component was developed, enabling the buffer-free continuous monitoring of small molecules in biological fluids. This integration is synergistic: the aptamer sensor is used to bind target biomolecules, triggering a fluorescence signal change, while the nanofluidic component is applied to achieve ion concentration polarization and convert serum into a clean buffer for aptamer signal regeneration. To demonstrate the system's versatility, we measured various adenosine triphosphate concentrations in human serum for hours with high sensitivity and specificity at minute temporal resolution. Our results demonstrate that this integrative device can be applied for the continuous measurement of target biomolecules and online signal regeneration in patient samples without the use of bulky clean buffer solutions for dynamic real-time healthcare.


Evidence for STAT4 as a common autoimmune gene: rs7574865 is associated with colonic Crohn's disease and early disease onset.
Recent studies demonstrated an association of STAT4 variants with systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), indicating that multiple autoimmune diseases share common susceptibility genes. We therefore investigated the influence of STAT4 variants on the susceptibility and phenotype of inflammatory bowel diseases (IBD) in a large patient and control cohort. Genomic DNA from 2704 individuals of Caucasian origin including 857 patients with Crohn's disease (CD), 464 patients with ulcerative colitis (UC), and 1383 healthy, unrelated controls was analyzed for seven SNPs in the STAT4 gene (rs11889341, rs7574865, rs7568275, rs8179673, rs10181656, rs7582694, rs10174238). In addition, a detailed genotype-phenotype analysis was performed. Our analysis revealed an association of the STAT4 SNP rs7574865 with overall decreased susceptibility to CD (p = 0.047, OR 0.86 [95% CI 0.74-0.99]). However, compared to CD patients carrying the wild type genotype, the STAT4 SNP rs7574865 was significantly associated with early CD onset (p = 0.021) and colonic CD (p = 0.008; OR = 4.60, 95% CI 1.63-12.96). For two other STAT4 variants, there was a trend towards protection against CD susceptibility (rs7568275, p = 0.058, OR 0.86 [95% CI 0.74-1.00]; rs10174238, p = 0.057, OR 0.86 [95% CI 0.75-1.00]). In contrast, we did not observe any association with UC susceptibility. Evidence for weak gene-gene interaction of STAT4 with the IL23R SNP rs11209026 was lost after Bonferroni correction. Our results identified the STAT4 SNP rs7574865 as a disease-modifying gene variant in colonic CD. However, in contrast to SLE and RA, the effect of rs7574865 on CD susceptibility is only weak.


Comparative Profiling of Ubiquitin Proteasome System Interplay with Influenza A Virus PB2 Polymerase Protein Recapitulating Virus Evolution in Humans.
The optimized exploitation of cell resources is one cornerstone of a successful infection. Differential mapping of host-pathogen protein-protein interactions (PPIs) on the basis of comparative interactomics of multiple strains is an effective strategy to highlight correlations between host proteome hijacking and biological or pathogenic traits. Here, we developed an interactomic pipeline to deliver high-confidence comparative maps of PPIs between a given pathogen and the human ubiquitin proteasome system (UPS). This subarray of the human proteome represents a range of essential cellular functions and promiscuous targets for many viruses. The screening pipeline was applied to the influenza A virus (IAV) PB2 polymerase proteins of five strains representing different levels of virulence in humans. An extensive PB2-UPS interplay has been detected that recapitulates the evolution of IAVs in humans. Functional validation with several IAV strains, including the seasonal H1N1pdm09 and H3N2 viruses, confirmed the biological relevance of most identified UPS factors and revealed strain-independent and strain-specific effects of UPS factor invalidation on IAV infection. This strategy is applicable to proteins from any other virus or pathogen, providing a valuable resource with which to explore the UPS-pathogen interplay and its relationship with pathogenicity. IMPORTANCE Influenza A viruses (IAVs) are responsible for mild-to-severe seasonal respiratory illness of public health concern worldwide, and the risk of avian strain outbreaks in humans is a constant threat. Elucidating the requisites of IAV adaptation to humans is thus of prime importance. In this study, we explored how PB2 replication proteins of IAV strains with different levels of virulence in humans hijack a major protein modification pathway of the human host cell, the ubiquitin proteasome system (UPS). We found that the PB2 protein engages in an extended interplay with the UPS that evolved along with the virus's adaptation to humans. This suggests that UPS hijacking underlies the efficient infection of humans and can be used as an indicator for evaluation of the potential of avian IAVs to infect humans. Several UPS factors were found to be necessary for infection with circulating IAV strains, pointing to potential targets for therapeutic approaches.


A policy to control the spread of HIV infection.
Prevention of transmission of HIV infection is the most important public health concern of the AIDS epidemic. To date, unfortunately, we have failed to contain the epidemic. The increasingly rapid spread of HIV into the IV drug-abusing population and subsequent heterosexual transmission represent a further failure of the public health system. Current organization of the public health programs, especially the lack of independence and adequate financial and personnel support, is an extremely serious problem. More funding may not be the answer, unless there is better organization. Identification of infected individuals and a vigorous education program must be implemented. HIV antibody-positive individuals should be followed carefully in order to evaluate the risk factors for AIDS and efficacy of specific interventions.


Predicting Patterns of Intimate Partner Violence Perpetration From Late Adolescence to Young Adulthood.
Saint-Eloi Cadely et al. found longitudinal patterns for the perpetration of both psychological and physical intimate partner violence (IPV), including actively and minimally aggressive patterns. The current study builds on these findings by examining four theory-derived variables (interparental aggression, social-information processing [SIP] biases, relationship insecurities [preoccupied and fearful], and discontinuity in relationship partner over time) as predictors of membership within these patterns, using multinomial logistic regression. The analysis sample consisted of 484 participants who were romantically involved at least once during the eight waves of data collection from the ages of 18 to 25. In predicting psychological IPV, more SIP biases, higher levels of a preoccupied insecurity, and less discontinuity in relationship partners over time differentiated the actively aggressive patterns from the minimally aggressive pattern. In addition, two actively aggressive patterns of psychological IPV differed in terms of SIP biases and discontinuity in romantic partners. Specifically, more SIP biases and less discontinuity in romantic partnerships distinguished the extensively aggressive pattern from the pattern that mainly consisted of minor types of aggression. In predicting physical IPV, the aggressive pattern differed from the nonaggressive pattern in terms of more interparental aggression, more SIP biases, and more relationship insecurities. The findings that developmental patterns of IPV can be predicted by social and psychological factors may aid both developmental theory and practice.


Xenopus helveticus, an endangered species?
This review traces the history of Xenopus research in Switzerland, its worldwide beginnings and British chapters having been summarised previously (Gurdon and Hopwood, 2000). As in other countries, Xenopus was initially used in the pharmaceutical industry at Basel for pregnancy testing. Developmental biologists became interested in this peculiar amphibian because it may be induced to ovulate all year round. Swiss Xenopus research is reviewed over 50 years, from the introduction of Xenopus by Rudolf Weber to the University of Bern, the return from Great Britain of two Swiss expatriates, Michail Fischberg and Max Birnstiel through the numerous pupils of the founder labs to the independently arisen Xenopus research units in the country. Besides developmental biology, Swiss Xenopus research engaged in immunology, genetics and cell biology, the latter focusing mainly on the oocyte. It set highlights in molecular biology by isolating some of the first eukaryotic genes and analysing their transcriptional regulation and post-transcriptional modifications through 'surrogate genetic' approaches in the oocyte system. An important line of research applied this system to study nuclear trafficking. Presently, functional testing mainly serves to characterise the function of proteins produced from expression vectors injected into the oocyte. A main accent of developmental studies was, from the early beginnings and still today, set on molecular characterisation of gene function in the embryo.


Viewing angle enhancement for two- and three-dimensional holographic displays with random superresolution phase masks.
Holographic displays employing binary phase modulation have been demonstrated to be attractive on the grounds of efficiency and miniaturization, and they offer a plausible approach to two-dimensional (2D) and three-dimensional (3D) image projection and display. A novel algorithm--one-step phase retrieval--and corresponding hardware architecture have recently been proposed, providing the performance required for real-time holographic display. However, since viewing angle varies inversely with pixel size, very small display pixels are required to achieve a wide field of view. This is particularly problematic for 3D displays, as the requirement for a large display with small pixels has hitherto necessitated an unachievably large electrical bandwidth. We present a novel approach, utilizing fixed random pixelated quaternary phase masks of greater resolution than the displayed hologram, to dramatically increase the viewing angle for 2D and 3D holographic displays without incurring a bandwidth penalty or significantly degrading image quality. Furthermore, an algorithm is presented to generate holograms accounting for the presence of such a phase mask, so that only one mask is required.


Rapid detection of Clostridium difficile toxins and laboratory diagnosis of Clostridium difficile infections.
Clostridium difficile is an anaerobic, spore-forming and Gram-positive bacillus. It is the major cause of antibiotic-associated diarrhea prevailing in hospital settings. The morbidity and mortality of C. difficile infection (CDI) has increased significantly due to the emergence of hypervirulent strains. Because of the poor clinical different between CDI and other causes of hospital-acquired diarrhea, laboratory test for C. difficile is an important intervention for diagnosis of CDI. Laboratory tests for CDI can broadly detect either the organisms or its toxins. Currently, several laboratory tests are used for diagnosis of CDI, including toxigenic culture, glutamate dehydrogenase detection, nucleic acid amplification testing, cell cytotoxicity assay, and enzyme immunoassay towards toxin A and/or B. This review focuses on the rapid testing of C. difficile toxins and currently available methods for diagnosis of CDI, giving an overview of the role that the toxins rapid detecting plays in clinical diagnosis of CDI.


Eliminating mother-to-child HIV transmission in South Africa.
The World Health Organization has produced clear guidelines for the prevention of mother-to-child transmission (PMTCT) of the human immunodeficiency virus (HIV). However, ensuring that all PMTCT programme components are implemented to a high quality in all facilities presents challenges. Although South Africa initiated its PMTCT programme in 2002, later than most other countries, political support has increased since 2008. Operational research has received more attention and objective data have been used more effectively. In 2010, around 30% of all pregnant women in South Africa were HIV-positive and half of all deaths in children younger than 5 years were associated with the virus. Between 2008 and 2011, the estimated proportion of HIV-exposed infants younger than 2 months who underwent routine polymerase chain reaction (PCR) tests to detect early HIV transmission increased from 36.6% to 70.4%. The estimated HIV transmission rate decreased from 9.6% to 2.8%. Population-based surveys in 2010 and 2011 reported transmission rates of 3.5% and 2.7%, respectively. CRITICAL ACTIONS FOR IMPROVING PROGRAMME OUTCOMES INCLUDED: ensuring rapid implementation of changes in PMTCT policy at the field level through training and guideline dissemination; ensuring good coordination with technical partners, such as international health agencies and international and local nongovernmental organizations; and making use of data and indicators on all aspects of the PMTCT programme. Enabling health-care staff at primary care facilities to initiate antiretroviral therapy and expanding laboratory services for measuring CD4+ T-cell counts and for PCR testing were also helpful.


Hif1 is a component of yeast histone acetyltransferase B, a complex mainly localized in the nucleus.
Hat1 is the catalytic subunit of the only type B histone acetyltransferase known (HAT-B). The enzyme specifically acetylates lysine 12, and to a lesser extent lysine 5, of free, non-chromatin-bound histone H4. The complex is usually isolated with cytosolic fractions and is thought to be involved in chromatin assembly. The Saccharomyces cerevisiae HAT-B complex also contains Hat2, a protein stimulating Hat1 catalytic activity. We have now identified by two-hybrid experiments Hif1 as both a Hat1- and a histone H4-interacting protein. These interactions were dependent on HAT2, indicating a mediating role for Hat2. Biochemical fractionation and co-immunoprecipitation assays demonstrated that Hif1 is a component of a yeast heterotrimeric HAT-B complex, in which Hat2 bridges Hat1 and Hif1 proteins. In contrast to Hat2, this novel subunit does not appear to regulate Hat1 enzymatic activity. Nevertheless, similarly to Hat1, Hif1 influences telomeric silencing. In a localization analysis by immunofluorescence microscopy on yeast strains expressing tagged versions of Hat1, Hat2, and Hif1, we have found that all three HAT-B proteins are mainly localized in the nucleus. Thus, we propose that the distinction between A- and B-type enzymes should henceforth be based on their capacity to acetylate histones bound to nucleosomes and not on their location within the cell. Finally, by Western blotting assays, we have not detected differences in the in vivo acetylation of H4 lysine 12 (acK12H4) between wild-type and hat1Delta, hat2Delta, or hif1Delta mutant strains, suggesting that the level of HAT-B-dependent acK12H4 may be very low under normal growth conditions.


Pigmentiphaga daeguensis sp. nov., isolated from wastewater of a dye works, and emended description of the genus Pigmentiphaga.
A Gram-negative, non-spore-forming, rod-shaped Pigmentiphaga-like bacterial strain, K110(T), was isolated from wastewater collected from a dye works in Korea and was subjected to a polyphasic taxonomic analysis. Strain K110(T) grew optimally at pH 7.0--8.0 and 37 degrees C in the presence of 0.5 % (w/v) NaCl. It contained Q-8 as the predominant ubiquinone and C(16 : 0), cyclo C(17 : 0) and cyclo C(19 : 0)omega8c as the major fatty acids. The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine and two unidentified aminolipids. The DNA G+C content was 67.4 mol%. In a neighbour-joining phylogenetic tree constructed on the basis of 16S rRNA gene sequences, strain K110(T) joined Pigmentiphaga kullae, the sole species of the genus, at a bootstrap confidence level of 100 %. Strain K110(T) exhibited a 16S rRNA gene sequence similarity of 99.4 % with respect to the type strain of P. kullae. Although strain K110(T) was found to be similar to P. kullae in terms of phenotypic properties, it differed in terms of motility, polar lipids, DNA-DNA relatedness and repetitive extragenic palindromic PCR genomic fingerprinting patterns. On the basis of phenotypic, phylogenetic and genetic data, strain K110(T) represents a novel species of the genus Pigmentiphaga, for which the name Pigmentiphaga daeguensis sp. nov. is proposed. The type strain is K110(T) (=KCTC 12838(T)=JCM 14330(T)).


Effects of diet-age and streptomycin on virulence of Autographa californica M nucleopolyhedrovirus against the tobacco budworm.
Addition of the antibiotic streptomycin to two artificial diets routinely used in bioassays of neonate lar vae of Heliothis virescens (tobacco budworm) infected with Autographa californica M nucleopolyhedrovirus (AcMNPV) increased lethal times of the virus. After storage of diets for 3 weeks at 4 degrees C, lethal times of infected larvae were significantly slower compared to those for larvae bioassayed using diets stored for 2 weeks or less. The effect of diet-age on rate of mortality was not the result of a change in total protein content or pH of the diet, but was apparently the result of some other alteration in the quality of the diet (e.g. microbial spoilage, palatibility, and/or nutritional value unrelated to total protein). Although we did not determine why lethal times were slower in response to streptomycin concentration or diet-age, we did find that slower lethal times were correlated with slower relative growth rates (RGR) of infected larvae. In addition, RGR of infected larvae decreased as a function of increasing streptomycin concentration, diet-age, and the interaction of the two factors. These results demonstrate that it is difficult to obtain consistent and comparable bioassay results if antibiotic composition and diet-age are not controlled. We suggest a standardized diet or highly standardized procedures for a given diet be developed that permits comparison of bioassays among and within laboratories.