Effect of oviductal proteins on sperm functions and lipid peroxidation levels during cryopreservation in buffaloes.
A study was undertaken to find out the effect of addition of oviductal proteins on sperm functions and lipid peroxidation (LPO) levels in buffaloes. Oviductal flushings were collected from apparently healthy buffalo genital tracts (nonluteal and luteal stage of estrous cycle), centrifuged (3000 rpm; 30 min), filtered (0.2 microm) and frozen at -20 degrees C. The proteins in pooled nonluteal and luteal oviductal fluid were precipitated overnight using ammonium sulphate, centrifuged (10,000 rpm; 30 min) and dialyzed (>10 kDa). After protein estimation, aliquots of samples containing 10 mg proteins were lyophilized in cryovials and stored frozen at -20 degrees C. Six pooled good quality ejaculates collected by artificial vagina method from two Murrah buffalo bulls were utilized for the study. After fresh semen analysis, each pooled ejaculate was split into three parts and extended in Tris-Egg yolk-Citrate extender (20% egg yolk: 7% glycerol), so that final dilution yielded approximately 60 million sperm cells/ml and cryopreserved in 0.5 ml French straws (30 million sperm cells per straw) in LN2 (-196 degrees C). Before freezing, the nonluteal and luteal oviductal proteins (NLOP &LOP) were incorporated at the concentration of 1mg/ml of extended semen. The equilibrated and frozen thawed (37 degrees C for 30s) semen was evaluated for motility, viability and acrosomal integrity, bovine cervical mucus penetration test and hypo-osmotic sperm swelling test. Besides these tests, LPO level was assessed in sperm and seminal plasma in equilibrated and frozen thawed semen. Results revealed that addition of oviductal proteins to semen before freezing convey beneficial effect in terms of spermatozoan motility, viability and acrosomal integrity. Nonluteal oviductal proteins favored significantly (P < 0.05) higher sperm penetration distance in cervical mucus (23.00+/-1.15 mm) than the control group (15.00+/-3.46 mm) in frozen thawed semen. Similarly, swollen sperm percentage was also significantly (P < 0.05) higher in NLOP treated group than the LOP included and control groups. In frozen thawed spermatozoa, the LPO level was significantly (P < 0.05) lower in NLOP added group than the LOP added and control group. It was inferred that incorporation of oviductal proteins in extender before freezing reduced the lipid peroxidation levels in buffalo spermatozoa during cryopreservation and thereby improved the post-thaw semen quality.


Depletion of Olig2 in oligodendrocyte progenitor cells infected by Theiler's murine encephalomyelitis virus.
Theiler's murine encephalomyelitis virus (TMEV) infects the central nervous system of mice and causes a demyelinating disease that is a model for multiple sclerosis. During the chronic phase of the disease, TMEV persists in oligodendrocytes and macrophages. Lack of remyelination has been attributed to insufficient proliferation and differentiation of oligodendrocyte progenitor cells (OPCs), but the molecular mechanisms remain unknown. Here, we employed pluripotent stem cell technologies to generate pure populations of mouse OPCs to study the temporal and molecular effects of TMEV infection. Global transcriptome analysis of RNA sequencing data revealed that TMEV infection of OPCs caused significant up-regulation of 1926 genes, whereas 1853 genes were significantly down-regulated compared to uninfected cells. Pathway analysis revealed that TMEV disrupted many genes required for OPC growth and maturation. Down-regulation of Olig2, a transcription factor necessary for OPC proliferation, was confirmed by real-time PCR, immunofluorescence microscopy, and western blot analysis. Depletion of Olig2 was not found to be specific to viral strain and did not require expression of the leader (L) protein, which is a multifunctional protein important for persistence, modulation of gene expression, and cell death. These data suggest that direct infection of OPCs by TMEV may inhibit remyelination during the chronic phase of TMEV-induced demyelinating disease.


Development of a clinical assay for detection of GAA mutations and characterization of the GAA mutation spectrum in a Canadian cohort of individuals with glycogen storage disease, type II.
Glycogen storage disease, type II (GSDII; Pompe disease; acid maltase deficiency) is an autosomal recessive disease caused by mutations of the GAA gene that lead to deficient acid alpha-glucosidase enzyme activity and accumulation of lysosomal glycogen. Although measurement of acid alpha-glucosidase enzyme activity in fibroblasts remains the gold standard for the diagnosis of GSDII, analysis of the GAA gene allows confirmation of clinical or biochemical diagnoses and permits predictive and prenatal testing of individuals at risk of developing GSDII. We have developed a clinical molecular test for the detection of GAA mutations based on cycle sequencing of the complete coding region. GAA exons 2-20 are amplified in six independent PCR using intronic primers. The resulting products were purified and sequenced. Preliminary studies using this protocol were conducted with DNA from 21 GSDII-affected individuals from five centers across Canada. In total, 41 of 42 mutations were detected (96.7% detection rate). Mutations spanned intron 1 through exon 19 and included nine novel mutations. Haplotype analysis of recurrent mutations further suggested that three of these mutations are likely to have occurred independently at least twice. Additionally, we report the identification of the c.-32-13T>G GAA mutation in an individual with infantile variant GSDII, despite reports of this mutation being associated almost exclusively with late-onset forms of the disease. The development of a clinical molecular test provides an important tool for the management and counseling of families and individuals with GSDII, and has provided useful information about the GAA mutation spectrum in Canada.


Characterization of further association of the trimeric membrane protein porin by low-angle laser-light scattering photometry coupled with high-performance gel chromatography.
Porin (OmpF), a trimeric membrane protein, in an extract of the outer membrane of Escherichia coli gave a twin-peaked elution pattern on Sephacryl S-300HR gel chromatography in the presence of sodium dodecyl sulphate. The species eluting earlier and later were found to be the hexamer and trimer, respectively, from molecular mass determination by low-angle laser-light scattering photometry coupled with TSK-G3000SWXL gel chromatography. As the hexamer was dissociated into the trimer under the conditions of sodium dodecyl sulphate polyacrylamide gel electrophoresis, its presence had been overlooked. The addition of lipopolysaccharide, another component of the outer membrane, and subsequent dialysis induced association of the trimer, the product containing an appreciable amount of the hexamer.


Re-establishment of cerebral metabolism after carotid endarterectomy.
The purpose of this study was to evaluate the metabolic changes that occur in the human brain in patients with a symptomatic carotid artery stenosis. N-acetyl-aspartate (NAA), choline, creatine and lactate were measured both before, and 4 days after, carotid endarterectomy, by magnetic resonance spectroscopic imaging (MRSI). Eight controls and 16 patients were examined. MRI and MRSI studies were performed on a Philips Gyroscan S15 whole body system operating at 1.5 Tesla. 1H spectra were selected from regions in the centrum semi-ovale outside areas showing white matter hyper intensities on MRI. All patients showed a decrease of the NAA/choline and NAA/creatine ratio in the symptomatic hemisphere compared to the contralateral hemisphere and also compared to the controls. Lactate was present in some patients (5/16). After endarterectomy, the NAA/choline and NAA/creatine ratios increased significantly compared to the ratios preoperative. Lactate was absent or more than 50% reduced after the operation. MRSI showed metabolic changes in areas of the brain that did not show any abnormalities on MRI. There are marked changes in brain metabolism in the symptomatic hemisphere of patients with a severe carotid artery stenosis. These metabolic changes normalise four days after endarterectomy.


Combined intervention with pioglitazone and n-3 fatty acids in metformin-treated type 2 diabetic patients: improvement of lipid metabolism.
The marine n-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) exert numerous beneficial effects on health, but their potency to improve treatment of type 2 diabetic (T2D) patients remains poorly characterized. We aimed to evaluate the effect of a combination intervention using EPA + DHA and the insulin-sensitizing drug pioglitazone in overweight/obese T2D patients already treated with metformin. In a parallel-group, four-arm, randomized trial, 69 patients (66 % men) were assigned to 24-week-intervention using: (i) corn oil (5 g/day; Placebo), (ii) pioglitazone (15 mg/day; Pio), (iii) EPA + DHA concentrate (5 g/day, containing ~2.8 g EPA + DHA; Omega-3), or (iv) pioglitazone and EPA + DHA concentrate (Pio& Omega-3). Data from 60 patients were used for the final evaluation. At baseline and after intervention, various metabolic markers, adiponectin and cytokines were evaluated in serum using standard procedures, EPA + DHA content in serum phospholipids was evaluated using shotgun lipidomics and mass spectrometry, and hyperinsulinemic-euglycemic clamp and meal test were also performed. Indirect calorimetry was conducted after the intervention. Primary endpoints were changes from baseline in insulin sensitivity evaluated using hyperinsulinemic-euglycemic clamp and in serum triacylglycerol concentrations in fasting state. Secondary endpoints included changes in fasting glycemia and glycated hemoglobin (HbA1c), changes in postprandial glucose, free fatty acid and triacylglycerol concentrations, metabolic flexibility assessed by indirect calorimetry, and inflammatory markers. Omega-3 and Pio& Omega-3 increased EPA + DHA content in serum phospholipids. Pio and Pio& Omega-3 increased body weight and adiponectin levels. Both fasting glycemia and HbA1c were increased by Omega-3, but were unchanged by Pio& Omega-3. Insulin sensitivity was not affected by Omega-3, while it was improved by Pio& Omega-3. Fasting triacylglycerol concentrations and inflammatory markers were not significantly affected by any of the interventions. Lipid metabolism in the meal test and metabolic flexibility were additively improved by Pio& Omega-3. Besides preventing a modest negative effect of n-3 fatty acids on glycemic control, the combination of pioglitazone and EPA + DHA can be used to improve lipid metabolism in T2D patients on stable metformin therapy. EudraCT number 2009-011106-42.


Dynamics of EGFR mutations in plasma recapitulates the clinical response to EGFR-TKIs in NSCLC patients.
Genomic profiling using plasma cell-free DNA (cfDNA) represents a non-invasive alternative to tumor re-biopsy, which is challenging in clinical practice. The feasibility of dynamically monitoring epidermal growth factor receptor (EGFR) mutation status using serial plasma samples from non-small cell lung cancer (NSCLC) patients treated by tyrosine kinase inhibitors (TKIs) and its application in tracking clinical response and detection of resistance were investigated. Forty-five NSCLC patients with EGFR mutation-positive pre-TKI plasma and at least two post-TKI plasma collections were recruited to this study. EGFR mutations including L858R, exon 19 deletion (19-del) and T790M were analyzed using droplet digital PCR (ddPCR) in longitudinally collected plasma samples. We observed a significant reduction in plasma EGFR mutation abundance during the first two-month of TKI treatment. Acquiring of secondary T790M gatekeeper mutation or completed "loss" of EGFR mutations represented two major categories of resistance profiles. Moreover, we demonstrated that levels of plasma EGFR mutations highly correlated with changes of tumor diameter as determined by radiographic imaging, or development of new lesions. In a subset of patients, we further showed that reappearance of EGFR mutations could be detected in plasma up to 5 months ahead of progressive disease (PD), suggesting an early detection of drug resistance. Our findings suggest that genomic analysis using plasma cfDNA may offer an effective approach to monitor clinical response and emergence of resistance.


Proficiency testing of allergen measurements in residential dust.
Analyses of household dust for allergens are more common with increased efforts to reduce and control asthma. Currently, no laboratory accreditation or quality assurance program specific to household dust allergen analyses exists. Moreover, there is an absence of peer-reviewed data on within-laboratory and between-laboratory variability that is achievable for these analyses. The purpose of this study was to characterize the levels of intralaboratory and interlaboratory variability in analyses of allergen concentrations in residential dust and to investigate the utility of quality control samples for monitoring laboratory performance. Aliquots from homogeneous batches of dust and dust extracts were provided to 8 commercial, academic, and municipal laboratories to be analyzed for as many as 6 allergens (Der p 1, Der f 1, Fel d 1, Can f 1, Bla g 1, Mus m 1) by using ELISA techniques. Coefficients of variation on the estimated geometric means from the analytical results ranged between 61% and 93%. In most cases, between-laboratory variability was the dominant component of total variability. In spite of this between-laboratory variability, reasonable agreement was observed between the means of allergen levels in the reference laboratory characterizations and the estimated geometric means from the model fitting of results across participating laboratories. The results from this study indicate that, in most cases, participating laboratories could measure the concentrations of allergens by using ELISA procedures with a level of accuracy and precision that may be acceptable in many situations.


Assignment of the gene coding for human beta-glucocerebrosidase to the region q21-q31 of chromosome 1 using monoclonal antibodies.
A series of man-Chinese hamster somatic cell hybrids with a variable content of human chromosomes was used to study the localization of the human gene coding for the lysosomal enzyme beta-glucocerebrosidase (EC 22.214.171.124). In lysates made from hybrid cells, the human enzyme was specifically recognized by a mouse monoclonal antibody raised against human placental beta-glucocerebrosidase. This monoclonal antibody did not cross-react with Chinese hamster beta-glucocerebrosidase. After reaction of the antibody with the enzyme, beta-glucocerebrosidase was precipitated by addition of Protein A-Sepharose beads, and was detected on the beads by its enzymatic activity. From the analysis of a series of man-Chinese hamster hybrids, among which were hybrids with specific segments of chromosome 1, we conclude that the gene coding for human beta-glucocerebrosidase is localized in the region q21-q31 of chromosome 1.


Engineering human MEK-1 for structural studies: A case study of combinatorial domain hunting.
Structural biology studies typically require large quantities of pure, soluble protein. Currently the most widely-used method for obtaining such protein involves the use of bioinformatics and experimental methods to design constructs of the target, which are cloned and expressed. Recently an alternative approach has emerged, which involves random fragmentation of the gene of interest and screening for well-expressing fragments. Here we describe the application of one such fragmentation method, combinatorial domain hunting (CDH), to a target which historically was difficult to express, human MEK-1. We show how CDH was used to identify a fragment which covers the kinase domain of MEK-1 and which expresses and crystallizes significantly better than designed expression constructs, and we report the crystal structure of this fragment which explains some of its superior properties. Gene fragmentation methods, such as CDH, thus hold great promise for tackling difficult-to-express target proteins.


Evaluating the impact of turbidity, precipitation, and land use on nutrient levels and atrazine concentrations in Illinois surface water as determined by citizen scientists.
The objective of this study was to evaluate the impact of turbidity, precipitation, land use, and five-week variation on nutrient levels and atrazine concentrations across Illinois state. To acquire the greatest number of samples in a cost and time-sensitive manner, data were collected by citizen scientists. Volunteers collected data regarding five water quality metrics: nitrites, nitrates, phosphates, atrazine, and turbidity once per week from April 19 until May 17, 2017. A subset (24 %) of volunteers also collected turbidity measurements. Data regarding precipitation was obtained from the Community Collaborative Rain, Hail and Snow Network (CoCoRaHS), a long-standing grassroots volunteer network of backyard weather observers. Three ordinal regression analyses were performed: one without a blocking effect, a second with week as a blocking effect, and a third with watershed as a blocking effect. In all cases, turbidity was significantly associated with elevated levels of nitrate (Pseudo R2-0.48 to 0.94) and phosphate (Pseudo R2-0.60 to 0.80), while precipitation was significantly associated with elevated levels of nitrate (Pseudo R2-0.25 to 0.35). While analyzing five-week variation, the nitrite and nitrate levels, but not phosphate or atrazine, tended to increase at each site. Further, nitrite and nitrate levels significantly varied between the four land uses - agricultural, urban, suburban, and park. When data were analyzed by the three most well-sampled watersheds, Kankakee, Des Plaines, and Chicago, it was identified that the nutrient levels in the Kankakee and Chicago watersheds were significantly elevated relative to the Des Plaines watershed. Finally, cluster analysis identified that clusters dominated by agricultural land, and to a lesser extent suburban land use, had the most elevated nutrient concentration and the greatest frequency of atrazine hits. Data collected by citizen scientists can provide insight into the geospatial variability of nutrients and agrichemicals and can do so across large geographies.


Predicting in vitro lung deposition behavior of combined dry powder inhaler via rheological properties.
Dry powder inhaler (DPI) for pulmonary delivery is currently the primary treatment for asthma and COPD (chronic obstructive pulmonary disease), an increasing number of combined DPIs (containing two or more drugs in one inhaler) have been developed to complement the effect of single DPIs. Based on our previous studies, the rheological properties can be a potential tool used to predict the in vitro lung deposition behavior of DPI formulations. However, it is unknown whether such a prediction model is suitable for combination systems. Therefore, this study aimed to verify the applicability of using powder rheological properties to predict in vitro drug deposition behavior in combined DPI formulations. Two drugs (fluticasone propionate and salmeterol xinafoate) and their combination of DPI formulations were prepared using fine lactose content (in the range of 1%-20%) as a variable. The physicochemical properties of the powder mixtures such as particle size and content uniformity were characterized. The rheological properties of the powder mixtures were measured by FT4 rheometer, the aerodynamic behavior of the DPI formulations was evaluated by a new generation impactor (NGI), and the effect of flowability and adhesion on the deposition of the fine particle fraction (FPF) was investigated by principal component analysis (PCA). The results showed that the combined DPI formulations with larger particle interaction forces have certain differences from the aerodynamic behavior of the single DPI formulations. The regularity of rheological properties affecting FPF revealed in single DPI is still applicable to combined DPI, the parameters basic flowability energy (BFE), representing flowability, and flow factor (ff), Cohesion representing adhesion, can be well linearly related to the FPF. The results of the principal component analysis showed that better flowability and suitable adhesion contributed to higher in vitro deposition of the drug in the formulation, and the contribution of adhesion (75.42%) was greater than that of flowability (24.58%). In conclusion, rheological properties is an effective tool for predicting the deposition behavior of DPI not only in single but also in combined DPIs.


Efficacy of micafungin in pediatric immunocompromised patients with invasive fungal infection.
Micafungin, an antifungal echinocandin, has been indicated for pediatric patients with invasive fungal infection (IFI) in Japan and Europe. Its efficacy in immunocompromised pediatric patients with IFI, however, has not been fully investigated. The safety and efficacy of micafungin as an antifungal therapy were analyzed in nine consecutive severe immunocompromised patients with IFI. Three patients with proven or probable Candida infections had complete response to micafungin therapy. Of the other six patients with proven, probable or possible Aspergillus infection, four had complete response and one had partial response to micafungin treatment. No severe adverse events were observed. In this small series, micafungin was effective for IFI caused by both Candida and Aspergillus species and no severe adverse events were observed in these immunocompromised patients.


Specific polyclonal and monoclonal antibody prevents paraquat accumulation into rat lung slices.
Sheep polyclonal and mouse monoclonal antibodies have been produced that bind to the bipyridyl herbicide, paraquat. The binding capacities and affinities of the various antibody solutions (serum, ascites, purified tissue culture supernatant) to paraquat were determined using a radioimmunoassay. All antibody solutions bound paraquat with high affinity (Ka = 10(9)-10(10) l/mol). The sheep polyclonal antisera, the mouse ascites fluid, and the purified culture supernatant had mean binding capacities of 8, 1 and 22 micrograms paraquat/ml respectively. All the antibody preparations were able to prevent the in vitro accumulation of paraquat into rat lung tissue. The amount of antibody to achieve this was dependent upon the binding capacity of the antibody solution, i.e. when the binding capacity of the antibody was equal to the amount of paraquat present in the incubation medium a total blockade of uptake was achieved. When antibody was added to lung tissue that had been accumulating paraquat for 1 hr, the inhibition of uptake was immediate and was complete for at least 2 hr. Both the radioimmunoassay and lung slice experiments indicate that an equivalent of 1 mg of IgG is required to bind 2.5 micrograms of paraquat ion. Preincubation of lung tissue with antibody did not affect the subsequent accumulation of paraquat, nor did it result in a detectable degree of intracellular neutralisation of paraquat as measured by paraquat's ability to stimulate the pentose phosphate pathway. The rate of efflux of paraquat from lung slices prepared from rats dosed intravenously with paraquat was not increased by the presence of antibody in the incubation medium. In conclusion, neutralising antibodies to paraquat have been produced. They bind to paraquat in solution with high affinity and render the paraquat unavailable for its in vitro accumulation into lung cells.


Interferon induction by the immunomodulating polyanion lambda carrageenan.
The lambda fraction of carrageenan induced murine interferon, which was characterized by sensitivity to trypsin and inhibition by pretreatment of target cells with actinomycin D.


Microglial metabolic disturbances and neuroinflammation in cerebral infarction.
Cerebral ischemia/reperfusion injury activates microglia, resident immune cells in the brain, and allows the infiltration of circulating immune cells into the ischemic lesions. Microglia play both exacerbating and protective roles in pathological processes and are thus often referred to as "double-edged swords." In ischemic brains, blood-borne macrophages play a role that is distinct from that of resident activated microglia. Recently, the metabolic alteration of immune cells in the pathogenesis of inflammatory disorders including cerebral infarction has become a critical target for investigation. We begin this review by describing the multifaceted functions of microglia in cerebral infarction. Next, we focus on the metabolic alterations that occur in microglia during pathological processes. We also discuss morphological changes that take place in the mitochondria, leading to functional disturbances, accompanied by alterations in microglial function. Moreover, we describe the involvement of the reactive oxygen species that are produced during aberrant metabolic activity. Finally, we discuss therapeutic strategies to ameliorate aggravative changes in metabolism.


Dual-chamber pacing in the cardiomyopathy of primary amyloidosis.
Cardiac function in a 75-year-old woman with primary amyloidosis and a restrictive cardiomyopathy deteriorated markedly after the occurrence of high-grade atrioventricular block. Atrial synchronous cardiac pacing, which was unreported previously in this setting, produced a marked improvement in cardiac performance.


Liver disease in pregnancy.
Changes in the liver biochemical profile are normal in pregnancy. However, up to 3% to 5% of all pregnancies are complicated by liver dysfunction. It is important that liver disease during pregnancy is recognized because early diagnosis may improve maternal and fetal outcomes, with resultant decreased morbidity and mortality. Liver diseases that occur in pregnancy can be divided into 3 different groups: liver diseases that are unique to pregnancy, liver diseases that are not unique to pregnancy but can be revealed or exacerbated by pregnancy, and liver diseases that are unrelated to but occur coincidentally during pregnancy.


Closure mechanism of the anal canal in women: assessed by three-dimensional ultrasound imaging.
To describe the functional correlates of anal canal anatomy using 3 dimensional ultrasound imaging. Ten nulliparous women were studied by using a 10-cm bag of 20-mm diameter. The bag was placed along the anal canal and inflated with 20 to 45 ml water, in 5-ml increments. At each volume, a three-dimensional ultrasound volume of the anal canal was obtained while the subjects were at rest and squeeze. The ultrasound images were analyzed to determine the relationship between the bag cross-sectional area and bag pressure. At low distension volumes, the bag is shaped like an "hourglass." The flared ends of the funnels correspond with the proximal and distal margins of the puborectalis muscle and external anal sphincter respectively. With increasing bag volumes, the length of completely closed segment of anal canal decreased. The last anal segment to open at rest was the one surrounded by all three structures. Anal contraction resulted in reduction of the anal canal cross-sectional area; the least compliant part of the anal canal was the one surrounded by external anal sphincter. The internal anal sphincter, external anal sphincter, and puborectalis muscle are all involved in the anal canal closure function. During contraction, the external anal sphincter is the strongest component of anal canal closure mechanism.


The Plasmodium liver-stage parasitophorous vacuole: A front-line of communication between parasite and host.
The intracellular development and differentiation of the Plasmodium parasite in the host liver is a prerequisite for the actual onset of malaria disease pathology. Since liver-stage infection is clinically silent and can be completely eliminated by sterilizing immune responses, it is a promising target for urgently needed innovative antimalarial drugs and/or vaccines. Discovered more than 65 years ago, these stages remain poorly understood regarding their molecular repertoire and interaction with their host cells in comparison to the pathogenic erythrocytic stages. The differentiating and replicative intrahepatic parasite resides in a membranous compartment called the parasitophorous vacuole, separating it from the host-cell cytoplasm. Here we outline seminal work that contributed to our present understanding of the fundamental dynamic cellular processes of the intrahepatic malarial parasite with both specific host-cell factors and compartments.


Effects of motion correction, sampling rate and parametric modelling in dynamic contrast enhanced MRI of the temporomandibular joint in children affected with juvenile idiopathic arthritis.
The temporomandibular joint (TMJ) is typically involved in 45-87% of children with Juvenile Idiopathic Arthritis (JIA). Accurate diagnosis of JIA is difficult as various clinical tests, including MRI, disagree. The purpose of this study is to optimize the methodological aspects of Dynamic Contrast Enhanced (DCE) MRI of the TMJ in children. In this cross-sectional study, including data from 73 JIA affected children, aged 6-15 years, effects of motion correction, sampling rate and parametric modelling on DCE-MRI data is investigated. Consensus among three radiologists determined the regions of interest. Quantitative perfusion parameters were estimated using four perfusion models; the Adiabatic Approximation to Tissue Homogeneity (AATH), Distributed Capillary Adiabatic Tissue Homogeneity (DCATH), Gamma Capillary Transit Time (GCTT) and Two Compartment Exchange (2CXM) models. Effects of motion correction were evaluated by a sum of least squares between corrected raw data and the GCTT model. The effect of systematically down sampling the raw data was tested. The sum of least squares was computed across all pharmacokinetic models. Relative difference perfusion parameters between the left and right TMJ were used for an unsupervised k-means based stratification of the data based on a principal component analysis, as well as for a supervised random forest classification. Diagnostic sensitivity and specificity were computed relative to structural image scorings. Paired sample t-tests, as well as ANOVA tests, were used (significant threshold: p < 0.05) with Tukeys post hoc test. High-level elastic motion correction provides the best least square fit to the GCTT model (percental improvement: 72-84%). A 4 s sampling rate captures more of the potentially disease relevant signal variations. The various parametric models all leave comparable residues (relative standard deviation: 3.4%). In further evaluation of DCE-MRI as a potential diagnostic tool for JIA a high-level elastic motion correction scheme should be adopted, with a sampling rate of at least 4 s. Results suggest that DCE-MRI data can be a valuable part in JIA diagnostics in the TMJ.


Rapid and dynamic processing of face pareidolia in the human brain.
The human brain is specialized for face processing, yet we sometimes perceive illusory faces in objects. It is unknown whether these natural errors of face detection originate from a rapid process based on visual features or from a slower, cognitive re-interpretation. Here we use a multifaceted approach to understand both the spatial distribution and temporal dynamics of illusory face representation in the brain by combining functional magnetic resonance imaging and magnetoencephalography neuroimaging data with model-based analysis. We find that the representation of illusory faces is confined to occipital-temporal face-selective visual cortex. The temporal dynamics reveal a striking evolution in how illusory faces are represented relative to human faces and matched objects. Illusory faces are initially represented more similarly to real faces than matched objects are, but within ~250 ms, the representation transforms, and they become equivalent to ordinary objects. This is consistent with the initial recruitment of a broadly-tuned face detection mechanism which privileges sensitivity over selectivity.


Shear-Thinning Viscous Materials for Subconjunctival Injection of Microparticles.
While drug-loaded microparticles (MPs) can serve as drug reservoirs for sustained drug release and therapeutic effects, needle clogging by MPs poses a challenge for ocular drug delivery via injection. Two polymers commonly used in ophthalmic procedures-hyaluronic acid (HA) and methylcellulose (MC)-have been tested for their applicability for ocular injections. HA and MC were physically blended with sunitinib malate (SUN)-loaded PLGA MPs for subconjunctival (SCT) injection into rat eyes. The HA and MC viscous solutions facilitated injection through fine-gauged needles due to their shear-thinning properties as shown by rheological characterizations. The diffusion barrier presented by HA and MC reduced burst drug release and extended overall release from MPs. The significant level of MP retention in the conjunctiva tissue post-operation confirmed the minimal leakage of MPs following injection. The safety of HA and MC for ocular applications was demonstrated histologically.


De novo characterization of Panax japonicus C. A. Mey transcriptome and genes related to triterpenoid saponin biosynthesis.
Panax japonicus C.A.Mey, the traditional medicinal herb in the Araliaceae family, has been used as a tonic, anti-inflammatory and haemostatic agent in China for more than thousand years. Its clinical effects are mainly due to the presence of triterpenoid saponins. However, little is known at the genetic level about how saponins are biosynthesized in this plant. We have therefore performed the de novo transcriptome assembly and high throughput RNA-seq analysis for P. japonicus. 66,403 unigenes were assembled from 19.6 Gbp raw data, and 34,639 unigenes were annotated. After annotation, 29 unigenes involved in putative backbone biosynthesis of triterpenoid saponin were selected. Additionally, 34 Cytochrome P450 and 18 UDP-glycosyltransferase unigenes were predicted based on the annotation, which were related to the saponin backbone modification. The expression level of related key genes were further verified by qPCR analysis. The results of this study provide the most comprehensive expressed sequence resources for P. japonicus, which will enlarge the available P. japonicus gene pool and facilitate further genome-wide research and analyses in this species.


Resistance to cerebral ischemia in developing gerbils.
Two-, three-, four-, five-, and twelve-week-old gerbils were subjected to various periods of bilateral carotid occlusion (BCO). Rectal and cranial temperatures were maintained at 37 degrees C during BCO, and only rectal temperature was monitored for 30 min of reperfusion. Seven days after ischemia, animals were perfusion-fixed and the neuronal densities in the hippocampal CA1 subfields were counted. The extent of cerebral ischemia during BCO was evaluated with [14C]iodoantipyrine autoradiography. The rectal temperature spontaneously fell to 33-34 degrees C during reperfusion in 2-week-old gerbils, although animals over 3 weeks old presented postischemic hyperthermia. Two-week-old animals therefore were divided into three experimental groups: In one group (2-week-old group I) rectal temperature was not regulated during 30 min of reperfusion, while in the other two groups (2-week-old groups II and III) rectal temperature was regulated at 37 and 38 degrees C, respectively, during reperfusion. Five-minute BCO produced almost complete destruction of the CA1 neurons in 12-week-old animals. In contrast, most CA1 neurons survived 30 min of BCO in 2-week-old group I and 15 min of BCO in 2-week-old groups II and III. [14C]Iodoantipyrine autoradiography revealed that BCO produced severe forebrain ischemia in 2-week-old gerbils as well as in 12-week-old gerbils. These findings indicate that developing gerbils have a greater tolerance to cerebral ischemia and that such ischemic tolerance is not due to a collateral network between the vertebrobasilar and the carotid circulations previously reported to develop more abundantly in developing gerbils.


Continuous intravenous lidocaine in the treatment of paralytic ileus due to severe spinal cord injury.
Paralytic ileus is a major concern in the acute phase of spinal cord injury. Classical treatment with neostigmine is often ineffective. Continuous intravenous (i.v.) lidocaine infusion has been previously proposed intra and post-operatively in order to decrease the duration of post-operative ileus after abdominal surgery. We report the cases of seven patients suffering from complete paralytic spinal cord injury-related ileus with colectasy resistant to neostigmine, who were treated by i.v. lidocaine infusion.


Assessment of the ecological quality status of soft-bottoms in Reunion Island (tropical Southwest Indian Ocean) using AZTI marine biotic indices.
The ability of the two synthetic marine biotic indices, AMBI and M-AMBI, to account for changes in the ecological quality of coastal soft bottoms of Reunion Island according to disturbances was assessed from macrobenthic samples collected in five sectors between 1994 and 2004. Samples were collected under non-perturbed conditions and at two sites subjected to heavy organic enrichment. Both indices are based on a classification of macrofauna into ecological groups (EG), and their transfer to tropical waters required some adaptations. These indices proved efficient in detecting a degradation of habitat quality. Their use resulted in the classification of all sites sampled between 1996 and 1998 as "good" or "high". M-AMBI nevertheless tended to result in the attribution of a slightly worse ecological quality status than AMBI. Together with an update of the EG species list for the Indian Ocean area, our results support the extension of both indices for the assessment of tropical soft bottoms.


Acute waterborne and chronic sediment toxicity of silver and titanium dioxide nanomaterials towards the oligochaete, Lumbriculus variegatus.
The use of silver (Ag) and titanium dioxide (TiO2) nanomaterials (NMs) in industrial processes and consumer products has experienced considerable growth since the late 20th century. Throughout their lifecycle, both Ag NM and TiO2NM are released into the environment, with benthic systems anticipated to be the final sink. Their potential toxicity towards benthic species is therefore of major concern. This study investigated the toxicity of silver (Ag; NM-300 K) and titanium dioxide (TiO2; NM-104) NMs to the freshwater oligochaete, Lumbriculus variegatus in acute (0-96-h) waterborne and chronic (28-d) sediment studies. Toxicity was investigated via assessment of mortality, behaviour, and antioxidant enzyme activity. The 96-h LC50 for Ag NMs in water was 0.51 mg/l (95% CI, 0.45-0.56), with L. variegatus displaying inhibited predation-avoidance behaviour compared to controls (6.66 ± 10%) successful response at 24-h), as well as significant increases (p < 0.05) in catalase (CAT) activity at sub-lethal concentrations at 24-h. Behavioural improvement and the return of antioxidant enzymes to control levels was observed after 48 and 72-h. AgNO3 exposure proved more toxic than Ag NM (96-h LC50 = 0.034 mg/l, 95% CI, 0.031-0.037) but resulted in no changes to antioxidant enzymes following sub-lethal exposure. Furthermore, Ag dissolution from Ag NM (~2-4%) could not account for the full extent of toxicity observed, suggesting a nano-specific effect. Increased environmental relevance via the inclusion of Suwannee River Humic Acid (SRHA, 5 mg/l) alleviated sub-lethal Ag NM toxicity despite a comparable 96-h LC50 (0.54 mg/l, 95% CI, 0.51-0.57). Significant effects of Ag NMs in formulated sediments (mortality, biomass) were only recorded according to OECD 225 at the highest test concentration (1333 mg/kg) for Ag NM indicating a potential attenuating effect of sediments towards toxicity. No toxicity was observed for TiO2 NM in aquatic or sediment exposures up to concentrations of 2000 mg/l and 1333 mg/kg, respectively.


Acculturation and sociocultural influences on dietary intake and health status among Puerto Rican adults in Massachusetts.
Previous studies have shown negative consequences of acculturation on lifestyle factors, health status, and dietary intake of Hispanic immigrants in the US. Despite prevalent type 2 diabetes and low socioeconomic status (SES) among Puerto Rican adults living on the US mainland, little is known about acculturation in this group. We investigated associations among acculturation, lifestyle characteristics, health status, and carbohydrate nutrition in Puerto Rican adults. A secondary objective was to investigate possible confounding and/or effect modification on these associations by SES. Cross-sectional data from the Boston Puerto Rican Health Study, which included 1219 Puerto Ricans in the Boston area, aged 45-75 years. Characteristics were compared using ANCOVA, linear trend and Pearson's chi-square tests across quartiles of acculturation. Tests for interaction by poverty status were conducted. Proportional contributions of foods to intake of total carbohydrate and fiber were assessed using SAS RANK. Levels of acculturation were low, despite young age at first arrival to the US mainland (25.4 ± 12.3 y) and long length of stay (34.2 ± 12.2 y). Greater English language use was associated with higher SES, alcohol consumption, physical activity, better perceived health, and less central obesity. Acculturation was associated with lower legume fiber and greater cereal fiber intake. Among those above the poverty threshold, acculturation was associated with lower dietary glycemic index and starch intake, and greater fruit and non-starchy vegetable intake. In contrast to studies with Mexican Americans, the association of acculturation with dietary quality in these Puerto Rican adults was mixed, but tended toward better carbohydrate quality. Dietary recommendations should include maintenance of traditional, healthful dietary practices including consumption of legumes, but also reduction in refined grains, and greater inclusion of fruit, non-starchy vegetables, and whole grains. Interventions to improve access to better quality carbohydrate sources are necessary for this group disproportionately affected by diabetes.


Cost-Effective H2 O2 -Regeneration of Powdered Activated Carbon by Isolated Fe Sites.
The reuse of powdered activated carbon (PAC) vitally determines the economics and security of the PAC-based adsorption process, while state-of-the-art PAC regeneration technologies are usually unsatisfactory. Here, it is demonstrated that isolated Fe sites anchored on commercial PAC enable fast H2 O2 activation to produce Fe-based reactive oxygen species for highly efficient PAC regeneration at room temperature. Taking rhodamine B as a representative pollutant, PAC decorated with isolated Fe sites realize H2 O2 based regeneration with negligible adsorption capacity degradation for 10 cycles. Moreover, in terms of the PAC loss rate, this technology is greatly superior to traditional Fenton-based regeneration technology. Further operando experiments and theoretical calculations reveal that the high regeneration performance can be attributed to the isolated HOFeO motifs, which activate H2 O2 via a nonradical reaction pathway. These findings provide a very promising strategy toward reducing the cost of H2 O2 -based PAC regeneration technology.


Gender and mortality in sepsis: do sex hormones impact the outcome?
Comparative assessment of the mortality rates of two septic patients' ages and/or gender subgroups, admitted to the intensive care unit of a university hospital. From December 2005 to April 2008, from a total of 628 patients, 133 were admitted to the intensive care unit with sepsis and included into two age subgroups: (G1) 14 - 40 years old and (G2) more than 50 years old. Patients aged between 41 and 50 years old (n = 8) were excluded. Demographic data, prognostic indicators (APACHE II score, organ dysfunction and circulatory shock) and outcome (mortality) were analyzed. Of the G1 patients (n = 44), 27 were female (61.4%), and in G2 (n = 81), 40 were female (49.4%). For both groups, mean APACHE II scores, multi-organ dysfunction and progression to circulatory shock rates were not significantly different between female and male patients. For G1, overall mortality rate was lower in female than in male patients (P = 0.04), while for G2, the opposite trend was observed. In this sample, reproductive age female patients younger than 40 years old showed lower mortality rates compared with age-matched male patients; for patients older than 50 years old, male patients had lower mortality rates than female patients.


Adenylate cyclase activity during growth and maturation of keratinocytes: comparison of two methods of study.
The adenylate cyclase activity of primary cultures of guinea-pig ear keratinocytes was studied at different times by challenge with various agonists after labelling of ATP by exposure of the cells to [3H]-adenine. The [3H]-cAMP (cyclic adenosine-3',5-monophosphate) formed was assayed by scintillation counting. In other experiments, cells were exposed to the equivalent amount of unlabelled adenine, challenged and cAMP was then assayed by radioimmunoassay. These absolute amounts were compared to [3H]-cAMP levels expressed as percentage conversion of [3H]-ATP. Comparison of the two methods suggested that the use of the prelabelling method would result in underestimation of cAMP formed. It was found that the response of the enzyme to different agonists changed with the length of time of the cells in culture.


Molybdenum-phosphorus triple bond stabilization by ancillary alkoxide ligation: synthesis and structure of a terminal phosphide tris-1-methylcyclohexanoxide complex.
Using alcoholysis, we converted terminal phosphide PMo(N[i-Pr]Ar)3 into a new, monomeric terminal phosphide PMo(OR)3, where R = 1-methylcyclohexyl or 1-adamantyl. Dimerization of the PMo unit was observed upon alcoholysis with 2,6-dimethylphenol, and the dimer [PMo(N[i-Pr]Ar)(O-2,6-C6H3Me2)2]2 was isolated and characterized by X-ray crystallography.


The positive correlation of TIPRL with LC3 and CD133 contributes to cancer aggressiveness: potential biomarkers for early liver cancer.
Studies have reported dysregulation of TIPRL, LC3 and CD133 in liver cancer tissues. However, their respective relationships to liver cancer and roles as biomarkers for prognosis and diagnosis of liver cancer have never been studied. Here we report that the level of TIPRL is significantly correlated with levels of LC3 (Spearman r = 0.9) and CD133 (r = 0.7) in liver cancer tissues. We observed significant upregulations of TIPRL, LC3 and CD133 in hepatocellular carcinomas (HCCs) compared with adjacent normal tissues. Importantly, TIPRL, tested among additional variables, showed a significant impact on the prognosis of HCC patients. TIPRL knockdown significantly reduced expressions of LC3, CD133, stemness-related genes, as well as viability and stemness of liver cancer cell-lines, which were promoted by ectopic TIPRL expression. Either alone or as a combination, TIPRL, LC3 and CD133 showed significant values of area under the curve (AUC) and sensitivity/specificity in early liver cancer tissues. Furthermore, the statistical association and the diagnostic efficacies of TIPRL, LC3 and CD133 in HCC tissues were confirmed in a different IHC cohort. This data demonstrates that the complex involvement of TIPRL/LC3/CD133 in liver cancer aggressiveness can together or individually serve as potential biomarkers for the early detection of liver cancer.


Multiplex Reverse-Transcription Loop-Mediated Isothermal Amplification Coupled with Cascade Invasive Reaction and Nanoparticle Hybridization for Subtyping of Influenza A Virus.
Considering the fatal human victims and economic loss caused by influenza virus infection every year, methodologies for rapid and on-site detection of influenza viruses are urgently needed. LAMP is the most commonly used nucleic acid isothermal amplification technology suitable for on-site use. However, for multiplex LAMP, differentiation of the amplicons derived from multiple targets is still challengeable currently. Here we developed a multiplex RT-LAMP assay for simultaneous amplification of three prominent subtypes of influenza viruses (A/H5, A/H7 and 2009A/H1). The amplicons were further identified by cascade invasive reaction and nanoparticle hybridization in separate target-specific detection tubes (referred to as mRT-LAMP-IRNH). The analytic sensitivities of the assay are 10 copies of RNA for all the three HA subtypes, and the specificity reached 100%. Clinical specimen analysis showed this assay had a combined sensitivity and specificity of 98.1% and 100%, respectively. Overall, the mRT-LAMP-IRNH assay can be used as a cost-saving method that utilizes a simple instrument to detect A/H5, A/H7, and 2009A/H1 influenza viruses, especially in resource-limited settings.


[Peer network for the therapeutic education of patients with type 2 diabetes in Mali].
Despite numerous health challenges, African healthcare systems are faced with limited financial and human resources. In this context, the epidemiological transition witnessing an explosion in the prevalence of non-communicable diseases, particularly diabetes, there are tremendous challenges to be met. Given the lack of human resources in these health systems, therapeutic education relies on networks of trained peer educator patients who also provide significant psychosocial support.


Subacute oral toxicity investigation of nanoparticulate and ionic silver in rats.
Subacute toxicity of 14 nm nanoparticulate silver (Ag-NP) stabilised with polyvinylpyrrolidone and ionic silver in the form of silver acetate (Ag-acetate) was investigated in four-week-old Wistar rats. Animals received orally by gavage the following: vehicle control (10 ♀, 6 ♂); Ag-NP at doses: 2.25 (8 ♀), 4.5 (8 ♀) or 9 mg/kg bw/day (10 ♀, 6 ♂); or Ag-acetate 9 mg silver/kg bw/day (8 ♀) for 28 days. Clinical, haematolological and biochemical parameters, organ weights, macro- and microscopic pathological changes were investigated. Caecal bacterial phyla and their silver resistance genes were quantified. For the Ag-NP groups, no toxicological effects were recorded. For Ag-acetate, lower body weight gain (day 4-7, 11-14, 14-16, P < 0.05; overall, day 1-28, P < 0.01), increased plasma alkaline phosphatase (P < 0.05), decreased plasma urea (P < 0.05) and lower absolute (P < 0.01) and relative (P < 0.05) thymus weight were recorded. In conclusion, these findings indicate toxicity of 9 mg/kg bw/day ionic silver but not of an equimolar Ag-NP dose. This is in accordance with previously reported data showing that oral Ag-acetate, in comparison with an equimolar dose of Ag-NP, resulted in higher silver plasma and organ concentrations.


Saturation of the tilt aftereffect.
The tilt aftereffect increases as a logarithmic function of adapting time, reaches saturation after approx 1 hr and decays on a symmetric, logarithmic time-course. This is similar to the time-course of contrast threshold elevation, suggesting that threshold and suprathreshold aftereffects are based on similar type of adaptation processes.


Augmenting physician examiner scoring in objective structured clinical examinations: including the standardized patient perspective.
In Canada, high stakes objective structured clinical examinations (OSCEs) administered by the Medical Council of Canada have relied exclusively on physician examiners (PEs) for scoring. Prior research has looked at using SPs to replace PEs. This paper reports on two studies that implement and evaluate a standardized patient (SP) scoring tool to augment PE scoring. The unique aspect of this study is that it explores the benefits of combining SP and PE scores. SP focus groups developed rating scales for four dimensions they labelled: Listening, Communication, Empathy/Rapport, and Global Impression. In Study I, 43 SPs from one site of a national PE-scored OSCE rated 60 examinees with the initial SP rating scales. In Study II, 137 SPs used slightly revised rating scales with optional narrative comments to score 275 examinees at two sites. Examinees were blinded to SP scoring and SP ratings did not count. Separate PE and SP scoring was examined using descriptive statistics and correlations. Combinations of SP and PE scoring were assessed using pass-rates, reliability, and decision consistency and accuracy indices. In Study II, SP and PE comments were examined. SPs showed greater variability in their scoring, and rated examinees lower than PEs on common elements, resulting in slightly lower pass rates when combined. There was a moderate tendency for both SPs and PEs to make negative comments for the same examinee but for different reasons. We argue that SPs and PE assess performance from different perspectives, and that combining scores from both augments overall reliability of scores and pass/fail decisions. There is potential to provide examinees with feedback comments from each group.


Consanguinity and advanced maternal age as risk factors for reproductive losses in Alexandria, Egypt.
Consanguinity has been a long-standing social habit among Egyptians. Estimates of consanguinity ratios in different parts of Egypt ranged from 29 to 50%. This study aimed at delineating the role of consanguinity and advanced maternal age on reproductive losses in Alexandria, Egypt. A case-control study, on 730 couples with history of reproductive losses and 2,081 normal couples, was done during the period October, 1998 until August, 2000. Of the 730 couples with reproductive losses, consanguinity frequency was 68.8% with 56.2% first cousins. Prenatal loss and infant deaths were highly encountered among consanguineous marriages (p < 0.0001). In a multivariate logistic regression analysis, consanguinity between couples increased the relative risk of repeated abortion (OR: 3.95; 95% CI: 3.04-5.14), stillbirths (OR: 10.6; 95% CI: 6.7-17.0), neonatal death (OR: 17.2; 95% CI: 10.8-27.3), post-neonatal death (OR: 14.5; 95% CI: 10.6-19.9) and total reproductive losses (OR: 8.3; 95% CI: 6.9-10.1). A positive association was found between advanced maternal age and repeated abortion (OR: 3.19; 95% CI: 2.04-4.97) as well as total reproductive losses (OR: 2.37; 95% CI: 1.74-3.2). This study, the largest-reported case-control study on reproductive losses in Alexandria, strongly suggests that consanguinity may play a major role in the high rates of prenatal and infant mortality while advanced maternal age has a significant role in the causation of repeated abortion, and they must be taken into account for genetic counseling in Egypt. Because of the possibility of controlling, the study gives clear indications for prevention.


Posterior hepatodiaphragmatic interposition of the colon: ultrasonographic and computed tomographic appearance.
High posterior hepatodiaphragmatic interposition of the colon is an unusual normal variant that may mimic posterior hepatic lesions, posterior retroperitoneal masses, or a disrupted right diaphragm on sonography. The correct diagnosis of this entity may be suggested on plain abdominal films, and is made easily with computed tomography.


Protection mechanism of Tween 80 during freeze-thawing of a model protein, LDH.
The purpose of the study was to investigate the protective mechanism of a non-ionic surfactant, Tween 80, at freeze-thawing with controlled temperature history of a model protein, lactate dehydrogenase (LDH). The system was examined by differential scanning calorimetry (DSC) and infrared spectroscopy (IR). LDH activity assays were performed spectrophotometrically. In all samples, independent of temperature history and addition of surfactant, all water was crystallized to polycrystalline ice at temperatures below -20 degrees C. The size and perfection of the ice crystals could be varied by a range of cooling rates giving different degrees of undercooling. At Tween concentrations below the cmc at crystallization, lower concentrations were required at low cooling rates compared to higher cooling rates to protect LDH. Concentrations above cmc of Tween reduced the protection at a cooling rate of 5 degrees C min(-1) and at quenching in N(2)(l). The amount of Tween needed for complete protection correlated to the surface area of the ice crystals at a certain temperature history. Tween 80 protects LDH from denaturation at freeze-thawing by hindering its destructive interaction with the ice crystals. The protective effect might be obtained when Tween molecules compete with the protein for sites on the ice surface. The optimum concentration of Tween needed for complete protection is dependent on the temperature history.


Multicenter study of bitolterol and isoproterenol nebulizer solutions in nonsteroid-using patients.
Bitolterol mesylate, 1.0 mg, or isoproterenol hydrochloride, 1.5 mg, was administered three times daily for 3 months in a double-blind, multicenter study via closed-port, intermittent-flow, compressor-driven nebulizer system (CPIF) to 182 nonsteroid-using patients with asthma. Mean baseline FEV1 was approximately 60% of predicted normal for both groups. Pulmonary function tests and vital signs were measured before and for up to 8 hours after treatments on test days 1, 30, 60, and 90. Mean maximum increases in FEV1 were 51%, 54%, 52%, and 55% for bitolterol versus 48%, 46%, 50%, and 43% for isoproterenol on these monthly test days. The mean FEV1 response remained greater than 15% over zero time (baseline) for greater than or equal to 8 hours after medication with bitolterol on each of four monthly pulmonary function test days and 2 1/2 to 5 hours for isoproterenol. Median durations of bronchodilator activity for bitolterol were 7.3, 6.5, 6.5, and 6.0 hours versus 4.0, 1.7, 3.7, and 1.9 hours for isoproterenol on the monthly test days. On these test days, 37% to 49% of the patients treated with bitolterol had a duration of action of at least 8 hours compared with 16% to 29% after isoproterenol treatment. The onset of activity was within 5 minutes for both drugs. Bitolterol provided superior bronchodilator activity with fewer adverse effects compared with isoproterenol, and there was no evidence for drug tolerance during this 3-month study.


A Novel Foam Contrast Agent Suitable for Fluoroscopic Interventional Procedure: Comparative Study of Physical Properties and Experimental Intervention in Animal Model.
In fluoroscopic contrast study for interventional procedure, liquid contrast agent may be diluted in body fluid, losing its contrast effect. We developed a novel contrast agent of "foam state" to maintain contrast effect for enough time and performed a comparative study of physical properties and its usefulness in experimental intervention in animal model. The mean size of microbubble of foam contrast was 13.8 ± 3.6 µm. The viscosity was 201.0 ± 0.624 cP (centipoise) and the specific gravity was 0.616. The foam decayed slowly and it had 97.5 minutes of half-life. In terms of the sustainability in a slow flow environment, foam contrast washed out much more slowly than a conventional contrast. In experimental colonic stent placement, foam contrast revealed significantly better results than conventional contrast in procedure time, total amount of contrast usage, and the number of injections (p < 0.05). Our foam contrast has high viscosity and low specific gravity and maintains foam state for a sufficient time. Foam contrast with these properties was useful in experimental intervention in animal model. We anticipate that foam contrast may be applied to various kinds of interventional procedures.


Quantitative determination of potential genotoxic impurity 3-aminopyridine in linagliptin active pharmaceutical ingredient using HILIC-UV.
This study aimed to develop an analytical method to determine the quantity of the impurity 3-aminopyridine (3AP). 3-Aminopyridine is a reactive reagent in the synthesis of linagliptin. The method was sensitive at level of 30.0 ppm of 3AP relative to linagliptin. The analysis was carried out using hydrophilic interaction liquid chromatography. The analytical column was Tracer Extrasil Silica (150 × 4.0 mm, 3 μm). A mobile phase of water-acetonitrile (10:90, v/v) containing 10.0 mM ammonium acetate was prepared and adjusted to pH 6.0. A UV detector was used to detect the amount of 3AP at a wavelength of 298 nm. Validation of the method was performed as per the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use in terms of detection limit, quantitation limit, linearity, accuracy, precision, specificity and robustness. The calibration curve was linear (r2 = 0.999) for 3AP concentration in the range of 30.0-450.0 ppm. This method showed a good sensitivity with a detection limit and a quantitation limit of 7.5 and 25.0 ppm, respectively.


Pre-notification letter type and response rate to a postal survey among women who have recently given birth.
Surveys are commonly used in health research to assess patient satisfaction with hospital care. Achieving an adequate response rate, in the face of declining trends over time, threatens the quality and reliability of survey results. This paper evaluates a strategy to increase the response rate in a postal satisfaction survey with women who had recently given birth. A sample of 2048 Australian women who had recently given birth at seven maternity units in New South Wales were invited to participate in a postal survey about their recent experiences with maternity care. The study design included a randomised controlled trial that tested two types of pre-notification letter (with or without the option of opting out of the survey). The study also explored the acceptability of a request for consent to link survey data with existing routinely collected health data (omitting the latter data items from the survey reduced survey length and participant burden). This consent was requested of all women. The survey had an overall response rate of 46% (913 completed surveys returned, total sample 1989). Women receiving the pre-notification letter with the option of opting out of the survey were more likely to actively decline to participate than women receiving the letter without this option, although the overall numbers of women declining were small (27 versus 12). Letter type was not significantly associated with the return of a completed survey. Among women who completed the survey, 97% gave consent to link their survey data with existing health data. The two types of pre-notification letters used in our study did not influence the survey response rate. However, seeking consent for record linkage was highly acceptable to women who completed the survey, and represents an important strategy to add to the arsenal for designing and implementing effective surveys. In addition to aspects of survey design, future research should explore how to more effectively influence personal constructs that contribute to the decision to participate in surveys.


Regulation of mRNA translation by protein folding in the endoplasmic reticulum.
The process of protein secretion is intimately linked to the rate and potential of proper folding and assembly of secretory proteins. The efficiency of protein folding is communicated to the cytoplasm via several signal transduction pathways. This regulates the rate of polypeptide chain synthesis and induction of genes encoding functions that reduce protein-folding load on the endoplasmic reticulum (ER). This review summarizes recent insights into the mechanisms that couple protein translation with protein folding in the ER.


Use of glomerular filtration rate estimating equations for drug dosing in HIV-positive patients.
Current HIV treatment guidelines recommend using the Cockcroft-Gault equation for drug dosing adjustments. The use of newer glomerular filtration rate (GFR) estimating equations for drug dosing and the appropriateness of physician antiretroviral dosing based on estimated kidney function have not been studied in an HIV-positive population. We evaluated concordance between measured and estimated GFR for the assignment of kidney function categories designated by the US Food and Drug Administration (FDA) Guidance for industry for pharmacokinetic studies, and appropriateness of physician antiretroviral drug dosing for level of kidney function in 200 HIV-positive patients on stable antiretroviral therapy. Estimated kidney function was determined using the Chronic Kidney Disease-Epidemiology collaboration (CKD-EPI), Modification of Diet in Renal Disease (MDRD) Study and Cockcroft-Gault equations. For assignment of FDA-designated kidney function categories, concordance rates between measured and estimated GFR using the CKD-EPI, MDRD Study and Cockcroft-Gault equations were 79%, 71% and 77%, respectively. This pattern was consistent across most subgroups. When actual prescribed dosages were compared with recommended dosages based on the level of estimated kidney function, 3-19% of study participants were prescribed higher than recommended dosages. The largest discordance between prescribed and recommended dosages was observed for the Cockcroft-Gault equation. The CKD-EPI equation has the highest concordance with measured GFR for the assignment of FDA-designated kidney function categories. Its use may lead to lower dosing-related errors in HIV-infected US adults on stable antiretroviral therapy. More education is required with respect to dose adjustment for level of kidney function.


Developmental and reproductive toxicity testing of vaccines.
The majority of new preventative and therapeutic vaccines are now assessed for developmental toxicity according to guidelines issued by the FDA in 2006. Despite the absence of confirmed effects in humans, vaccines are frequently suspected of having adverse side-effects on the development of children. Such suspicions are perhaps unavoidable considering the extremely widespread use of vaccines. The preclinical developmental toxicology studies are designed to assess possible influences of each component of the vaccine formulation-and the induced antibodies-on the development of the conceptus, neonate and suckling organism. Immune modulation by a vaccine or an adjuvant could, for instance, affect the outcome of pregnancy by interfering with the natural shift in immune balance of the mother during gestation. Maternal immunoglobulins are transferred from the mother to the offspring in order to confer passive immunity during early life. This maternal antibody transport is prenatal in humans and monkeys, but tends to be delayed until after birth in other species. Therefore, a suitable model species needs to be chosen for preclinical studies in order to ensure exposure of the foetus to the induced maternal antibodies following vaccination. Rabbits are the best laboratory model for prenatal immunoglobulin transfer, but rodents are more practical for the necessary postnatal investigations. Non-human primates are the only appropriate models for the testing of vaccines that are not immunogenic in lower species. It is advisable to test new adjuvants separately according to the ICH S5(R2) guidelines. Preclinical paediatric investigations are not currently required for vaccines, even though most vaccines are given to children. Other areas of regulatory concern include developmental immunotoxicity and effects on the preimplantation embryo. Because of the limitations of the available animal models for developmental toxicity testing, pharmacovigilance is essential.


Immunotheranostic Polymersomes Modularly Assembled from Tetrablock and Diblock Copolymers with Oxidation-Responsive Fluorescence.
Intracellular delivery is a key step for many applications in medicine and for investigations into cellular function. This is particularly true for immunotherapy, which often requires controlled delivery of antigen and adjuvants to the cytoplasm of immune cells. Due to the complex responses generated by the stimulation of diverse immune cell populations, it is critical to monitor which cells are targeted during treatment. To address this issue, we have engineered an immunotheranostic polymersome delivery system that fluorescently marks immune cells following intracellular delivery. N-(3-bromopropyl)phthalimide end-capped poly(ethylene glycol)-bl-poly(propylene sulfide) (PEG-PPS-PI) was synthesized by anionic ring opening polymerization and linked with PEG-PPS-NH2 via a perylene bisimide (PBI) bridge to form a tetrablock copolymer (PEG-PPS-PBI-PPS-PEG). Block copolymers were assembled into polymersomes by thin film hydration in phosphate buffered saline and characterized by dynamic light scattering, cryogenic electron microscopy and fluorescence spectroscopy. Polymersomes were injected subcutaneously into the backs of mice, and draining lymph nodes were extracted for flow cytometric analysis of cellular uptake and disassembly. Modular self-assembly of tetrablock / diblock copolymers in aqueous solutions induced π-π stacking of the PBI linker that both red-shifted and quenched the PBI fluorescence. Reactive oxygen species within the endosomes of phagocytic immune cell populations oxidized the PPS blocks, which disassembled the polymersomes for dequenching and shifting of the PBI fluorescence from 640 nm to 550 nm emission. Lymph node resident macrophages and dendritic cells were found to increase in 550 nm emission over the course of 3 days by flow cytometry. Immunotheranostic polymersomes present a versatile platform to probe the contributions of specific cell populations during the elicitation of controlled immune responses. Flanking PBI with two oxidation-sensitive hydrophobic PPS blocks enhanced π stacking and introduced a mechanism for disrupting π-π interactions to shift PBI fluorescence in response to oxidative conditions. Shifts from red (640 nm) to green (550 nm) fluorescence occurred in the presence of physiologically relevant concentrations of reactive oxygen species and could be observed within phagocytic cells both in vitro and in vivo.


Treatment of a malignant esophageal perforation with a prototype conical Wallstent.
A 60-year-old man with a malignant esophageal perforation could not be treated by conventional covered metallic stents because the upper esophagus was dilated. The perforation was eventually closed by deployment of a prototype, conical covered Wallstent.


Adaptive response of the murine collecting duct to alkali loading.
Fine-tuning of salt and acid-base homeostasis is achieved in the renal collecting duct through the action of intercalated and principal cells. Their activity is tightly regulated adapting to changes in systemic acid-base, fluid, or electrolyte status. The relative number of acid or bicarbonate secretory intercalated cells changes in response to acid or alkali loading. Several factors that may induce collecting duct plasticity in response to acid loading have been identified including cell proliferation, Growth Differentiation Factor 15 (Gdf15), hensin (DMBT1), and SDF1 (or CXCL12). Also, the transcription factors Foxi1 and CP2L1, or the Notch2-Jag1 signaling pathway, may play a role. However, little is known about the mechanisms mediating the adaptive response of the collecting duct to alkali loading. Here, we examined in mouse kidney the response of these factors to alkali loading. Mice were left untreated or received NaHCO3 or NaCl over 7 days. Cell proliferation in vivo was monitored by Ki67 labeling or BrdU incorporation and expression of cell markers, and regulatory factors were examined. Foxi1 and GDF15 were upregulated and CP2L1 downregulated during alkali loading. Ki67 staining and BrdU incorporation were frequent in AQP2-positive cells in the NaCl and NaHCO3 groups, but no evidence was found for increased Ki67 or BrdU staining in bicarbonate-secretory cells consistent with a model that AQP2 positive precursor cells may differentiate into intercalated cells. Thus, alkali loading alters the cellular profile of the collecting duct, which may involve cell proliferation and changes in the network of molecules determining the plasticity of the collecting duct.


An in-depth study of accidents involving collisions with utility poles.
This paper summarizes the objectives, methodology, findings, and recommendations of a large-scale study of automobile crashes involving utility poles. In an 8-month survey of accidents, information on site characteristics and accident severity was obtained for a sample of 879 crashes. Randomly-selected samples of 795 sites and 627 vehicles provided control information. An accident-predictor model which identifies accident risk on the basis of site measurements has been derived. The model reveals a range of relative risks in the population of exposed poles of the order of 1000:1, and enables the identification of the relatively small proportion of poles which account for the majority of accidents. The model was used in conjunction with estimates of the costs of accidents to show that a number of remedial treatments are warranted. The hazardous effects of low tyre tread depths and improper tyre inflation pressures are also demonstrated.


Fabrication of ion doped WO3 photocatalysts through bulk and surface doping.
Na(+) doped WO3 nanowire photocatalysts were prepared by using post-treatment (surface doping) and in situ (bulk doping) doping methods. Photocatalytic degradation of Methyl Blue was tested under visible light irradiation, the results showed that 1wt.% Na(+) bulk-doped WO3 performed better, with higher photoactivity than surface-doped WO3. Photoelectrochemical characterization revealed the differences in the photocatalytic process for surface doping and bulk doping. Uniform bulk doping could generate more electron-hole pairs, while minimizing the chance of electron-hole recombination. Some bulk properties such as the bandgap, Fermi level and band position could also be adjusted by bulk doping, but not by surface doping.


Studies of introital colonization in women with recurrent urinary infections. II. A comparison of growth in normal vaginal fluid of common versus uncommon serogroups of Escherichia coli.
Vaginal fluid collected from volunteers who denied a history of urinary infections was bactericidal to Escherichia coli at pH 4.0. When the pH was increased to 6.5 with sodium hydroxide, the same vaginal fluids supported bacterial growth. If vaginal fluid was adjusted at pH increments between 4.0 and 5.0, and inoculated with common and uncommon serogroups of Escherichia coli, the uncommon O-groups that never cause urinary infections were significantly (p less than 0.005) more inhibited by the lower vaginal pH than were the common O-groups of Escherichia coli.


Neonatal pharyngoesophageal perforation mimicking esophageal atresia: clues to diagnosis.
Perforation of the newborn child's esophagus or pharynx can mimic esophageal atresia clinically. Fourteen cases of newborn hypopharyngeal or esophageal perforation are presented. In nine cases, esophageal atresia was the initial diagnosis entertained; seven of these on the first day of life. In four others, the diagnosis of a perforated pharynx or esophagus was evident on plain x-ray. Six patients had esophagograms. Two newborns underwent thoracotomies for repair of their initially diagnosed esophageal atresia. Subsequently, esophageal perforation was diagnosed intraoperatively on each and suture repair was undertaken. The 12 remaining neonates were treated nonoperatively. There were four deaths, all in the nonoperative group and three weighing less than 1,000 g. None of the deaths were directly attributable to the perforations. In retrospect, each child presented with one or more early clues to the diagnosis of pharyngoesophageal perforation, which included a history of difficult tracheal or nasogastric intubation, blood in the nasogastric tube, length and course of nasogastric tube inserted, and subtle chest x-ray changes. In these cases, nonoperative treatment is usually successful.


[Not Available].
Based on representative analyses of Austrian cereals, a distinct shift in the spectrum of Fusarium toxins and Fusarium species has been observed since the middle of the eighties.Although in the case of maizeF subglutinans - apart fromF graminearum - proved to be the most frequent and constant contaminant over the entire range of test series, there has been a shift in the spectrum of species which is not to be explained simply by seasonal variations or by the varying degree of occurrence of the European corn borer, which in Austria is considered to be the main vector for infections involving fusaria of the Liseola section. Compared to the results from earlier vegetation periods, the nineties brought a significant increase in the number of infections withF proliferatum, a fumonisin-producer. In all likelihood, this shift in the spectrum of species is due to the changed climatic conditions now prevailing in Austria - milder and more humid winters vs. drier and warmer summers - which favour the progress ofF proliferatum.The principal toxin-forming fungus on cereals in Austria isF graminearum. On maize, its respective populations are exclusively those which produce 15-acetyl-DON as a precursor to DON (deoxynivalenol). Whilst in the 1980s,F graminearum isolates from wheat yielded both 15-acetyl-DON and 3-acetyl-DON types, only 15-acetyl-DON populations could be detected in the last few years. One possible explanation for this phenomenon is the continual intensification of maize-wheat crop rotations. In the light of the above observations, the frequently used argument whereby EuropeanF graminearum isolates produce mainly 3-acetyl-DON and American strains prevalently 15-acetyl-DON will have to be reviewed.


Is antenatal bladder neck mobility a risk factor for postpartum stress incontinence?
To assess the contribution of constitutional factors, as demonstrated by antenatal bladder neck mobility, in the development of postpartum urinary stress incontinence. A prospective investigational study. General district hospital. One hundred and three primigravid women with no pre-existing urinary incontinence or neurological disorder. Antenatal and postnatal measurement of bladder neck mobility using perineal ultrasound. Urinary stress incontinence at 10-14 weeks postpartum. Women with postpartum urinary stress incontinence have significantly greater antenatal bladder neck mobility than those women continent postpartum. There were no significant differences in any labour or delivery variables, including mode of delivery, between the postpartum continent and incontinent women. There is evidence for a constitutional risk factor (eg, defective pelvic floor connective tissue in the development of postpartum stress incontinence).


Cervical spine trauma - Evaluating the diagnostic power of CT, MRI, X-Ray and LODOX.
Traumatic cervical spine (c-spine) injuries account for 10% of all spinal injuries. The c-spine is prone to injury by blunt acceleration/deceleration traumas. The Canadian C-Spine rule and NEXUS criteria guide clinical decision-making but lack consensus on imaging modality when necessary. This study aims to evaluate the sensitivity and specificity of CT, MRI, X-Ray, and, for the first time, LODOX-Statscan in identifying c-spine injuries in patients with blunt trauma and neck pain. We conducted a retrospective monocenter cohort study using patient data from the emergency department at Inselspital, Bern, Switzerland's largest level one trauma center. We identified patients presenting with trauma and neck pain during the recruitment period from 01.01.2012 to 31.12.2017. We included all patients that required a radiographic c-spine evaluation according to the NEXUS criteria. Certified spine surgeons reviewed each case, analyzed patient demographics, injury classification, trauma mechanism, and emergency management. The retrospective full case review was established as gold standard to decide whether the c-spine was injured. Sensitivity and specificity were calculated for CT, MRI, LODOX, and X-Ray imaging methods. We identified 4996 patients, of which 2321 met the inclusion criteria. 91.3% (n = 2120) patients received a CT scan, 8.9% (n = 206) a MRI, 9.3% (n = 215) an X-ray, and 21.5% (n = 498) a LODOX scan. By retrospective case review, 186 participants were classified as injured. The sensitivity of CT was 88.6% (specificity 99%), and 89.8% (specificity 99.2%) with orthopedic surgeon consultation. MRI had a sensitivity of 88.5% (specificity of 96.9%); highlighting 14 cases correctly diagnosed as injured by MRI and misdiagnosed by CT. Projection radiography (36.4% sensitivity, 95.1% specificity) and LODOX (5.3% sensitivity, 100% specificity) were unsuitable for ruling out spinal injury. While CT offers high sensitivity for detecting traumatic c-spine injury, MRI holds clinical significance in revealing injuries not recognized by CT in symptomatic patients. LODOX and projection radiography are insufficient for accurately ruling out c-spine injury. For patients with neurological symptoms, we recommend extended MRI use when CT scans are negative.


Reading room electives: say goodbye to the "radi-holiday".
Reading room electives are among the most challenging curricula to develop for medical student educators. Students tend to have few responsibilities and poorly defined learning objectives. The authors review the components of a successful reading room elective. The important components include a thorough orientation, written goals and expectations, attendance and feedback systems, objective mid-elective and end-of-elective evaluations, and an end-of-elective debriefing session. Many educational activities, tools, and assessments are available and should be considered to supplement the curriculum. Radiology electives must break free of the "radi-holiday" stereotype and adopt increased responsibilities and expectations, similar to rigorous clinical subinternships.


TIM3 comes of age as an inhibitory receptor.
T cell immunoglobulin and mucin domain-containing protein 3 (TIM3), a member of the TIM family, was originally identified as a receptor expressed on interferon-γ-producing CD4+ and CD8+ T cells. Initial data indicated that TIM3 functioned as a 'co-inhibitory' or 'checkpoint' receptor, but due to the lack of a definable inhibitory signalling motif, it was also suggested that TIM3 might act as a co-stimulatory receptor. Recent studies have shown that TIM3 is part of a module that contains multiple co-inhibitory receptors (checkpoint receptors), which are co-expressed and co-regulated on dysfunctional or 'exhausted' T cells in chronic viral infections and cancer. Furthermore, co-blockade of TIM3 and programmed cell death 1 (PD1) can result in tumour regression in preclinical models and can improve anticancer T cell responses in patients with advanced cancers. Here, we highlight the developments in understanding TIM3 biology, including novel ligand identification and the discovery of loss-of-function mutations associated with human disease. In addition, we summarize emerging data from human clinical trials showing that TIM3 indeed acts as a 'checkpoint' receptor and that inhibition of TIM3 enhances the antitumour effect of PD1 blockade.


Ductal Carcinoma in Situ: Molecular Changes Accompanying Disease Progression.
Ductal carcinoma in situ (DCIS) is a non-obligate precursor of invasive ductal carcinoma (IDC), whereby if left untreated, approximately 12% of patients develop invasive disease. The current standard of care is surgical removal of the lesion, to prevent potential progression, and radiotherapy to reduce risk of recurrence. There is substantial overtreatment of DCIS patients, considering not all DCIS lesions progress to invasive disease. Hence, there is a critical imperative to better predict which DCIS lesions are destined for poor outcome and which are not, allowing for tailored treatment. Active surveillance is currently being trialed as an alternative management practice, but this approach relies on accurately identifying cases that are at low risk of progression to invasive disease. Two DCIS-specific genomic profiling assays that attempt to distinguish low and high-risk patients have emerged, but imperfections in risk stratification coupled with a high price tag warrant the continued search for more robust and accessible prognostic biomarkers. This search has largely turned researchers toward the tumor microenvironment. Recent evidence suggests that a spectrum of cell types within the DCIS microenvironment are genetically and phenotypically altered compared to normal tissue and play critical roles in disease progression. Uncovering the molecular mechanisms contributing to DCIS progression has provided optimism for the search for well-validated prognostic biomarkers that can accurately predict the risk for a patient developing IDC. The discovery of such markers would modernize DCIS management and allow tailored treatment plans. This review will summarize the current literature regarding DCIS diagnosis, treatment, and pathology.


Nutrition educator adoption and implementation of an experiential foods curriculum.
Describe changes in Nutrition Educator (NE) and Extension Agent (EA) motivation, self-efficacy, and behavioral capability over time after experiential food tasting curriculum training. Identify promoters of curriculum adoption, implementation, and future use. Mixed methods design including surveys, lesson implementation reports, and interviews. New Mexico limited-resource schools. Convenience sample of New Mexico Extension NE (n = 42) and their EA supervisors (n = 21). Three-hour curriculum training employing Social Cognitive Theory and Diffusion of Innovations. Perceived change in motivation, self-efficacy, and behavioral capability from post-training through 8-month post-training; promoters and challenges to curriculum adoption, implementation, and future use. Repeated-measures ANOVA analyzed perceived behavior change over time. Significance was set at P ≤ .05. Qualitative responses were categorized by theme. Gains in NE motivation, self-efficacy, and behavioral capability were sustained at 8 months post-training. High adoption/implementation rates (79%) were attributed to strong implementation expectations, observational learning, experiential training elements, and perceived curriculum compatibility. Environmental factors including time constraints, personnel turnover, and scheduling conflicts proved challenging. Maximizing curriculum simplicity and compatibility and incorporating behavioral capability, observational learning, and expectations into training support adoption and use. Adaptations and techniques to problem-solve challenges should be provided to new curricula implementers.


When things become what you call them.
Unprecise nomenclature may have unprecedented effects over time as is suggested in this article on the arguments surrounding donor insemination.


End-stage renal disease secondary to renal malignancy: Epidemiologic trends and survival outcomes.
Loss of renal parenchyma after surgery may contribute to chronic kidney disease; however, the long-term consequences of chronic kidney disease may differ by cause. We analyzed the outcomes of patients with end-stage renal disease (ESRD) based on various medical and surgical causes. In the United States Renal Data System from the period 1983 to 2007, patients with renal tumors, traumatic surgical loss, diabetes, or other known causes were identified. The annual incidence, prevalence, and influence of age, race, sex, and primary cause on survival were evaluated. Of 1.3 million patients, 6,812 (0.49%) had renal malignancy-related ESRD (RM-ESRD). An increased over time was noted in the standardized incidence rates of patients with RM-ESRD (R2 = 0.973, P<0.0001). Patients with RM-ESRD had a worse median survival (1.9 vs. 3.4 y, P<0.0001), whereas those with ESRD related to nonmalignant surgical loss had improved survival (3.8 y) compared to diabetic ESRD (P<0.0001). The 5-year cancer-specific mortality was higher for RM-ESRD (30.9% vs. 5.5%, P<0.0001) compared to ESRD from other known causes; however, the non-cancer-specific mortality was improved compared to patients with ESRD with diabetic causes (P<0.0001). Limitations include retrospective analysis and lack of specific clinical data, such as cancer grade. The incidence of RM-ESRD is increasing, possibly owing to the increased rate of renal cell carcinoma treatment. Although overall survival for RM-ESRD was worse than either that of nonmalignant surgical loss or other known causes, non-cancer-specific mortality was decreased compared to diabetic causes, likely due to systemic effects by cause of ESRD.


Elevated expression of an endogenous retroviral long terminal repeat in a mouse colon tumor.
We have previously identified a number of cloned poly(A)+ RNA sequences in a cDNA library, each of whose expression is markedly altered in a dimethylhydrazine-induced mouse colon tumor as compared to the normal colon. Transcripts which hybridize to one of the cloned cDNAs, pMCT-1, have now been shown to be elevated in expression in three mouse leukemia cell lines as well, and induction of differentiation in one of these lines (DS-19) is associated with a substantial decrease in expression of these transcripts. By hybridization and sequence analysis, pMCT-1 has been found to contain a region homologous to a long terminal repeat of an intracisternal A particle sequence. Intracisternal A sequences are endogenous, retroviral genomes repeated several thousandfold and dispersed in the mouse genome. The data show that the transcripts which are homologous to this long terminal repeat are highly heterogeneous, suggesting that many intracisternal type A particle transcriptional units may be expressed in the colon tumor and leukemia cell lines.


Determination of ceftazidime and cefepime in plasma and dialysate-ultrafiltrate from patients undergoing continuous veno-venous hemodiafiltration by HPLC.
We have developed and validated a new, rapid and reproducible HPLC method for the determination of cefepime and ceftazidime in plasma and dialysate-ultrafiltrate samples obtained from intensive care unit (ICU) patients undergoing continuous veno-venous hemodiafiltration (CVVHDF). The method for plasma samples involved protein precipitation with acetonitrile, followed by washing with dichloromethane to remove apolar lipophilic compounds. Dialysate-ultrafiltrate samples did not require any preparation. Separation was performed on a muBondapak C18 (30 cm x 3.9 mm x 10 microm) with UV detection. The mobile phase contained acetate buffer: ACN and was delivered at 2 ml/min. The coefficients of determination of the calibration curves were always > or = 0.998 and R.S.D.% of the response factors <10%. The intra and inter-assay precision and accuracy of the quality controls (QC) and limit of quantification (LOQ) were satisfactory in all cases. Plasma and dialysate-ultrafiltrate samples were stable at -20 and -80 degrees C for 2 months and also after three freeze/thaw cycles. Dialysate-ultrafiltrate samples were stable in the chromatographic rack for 24h at room temperature, but we recommend storing processed plasma samples at 4 degrees C until the analysis. The described method has proved to be useful to give accurate measurements of ceftazidime and cefepime in samples obtained from patients undergoing CVVHDF.


Analysis of the genetic structure and diversity of upland cotton groups in different planting areas based on SNP markers.
Genetic diversity, kinship and population genetic structure analyses of Gossypium hirsutum germplasm can provide a better understanding of the origin and evolution of G. hirsutum biodiversity. In this study, adopt 273 elite upland cotton cultivar accessions collected from around the world, and especially from China to get 1,313,331 SNP molecular markers, it were used to construct a phylogenetic tree of each sample using MEGAX, to perform population structure analysis by ADMIXTURE software and principal component analysis (PCA) by EIGENSOFT software, and to estimate relatedness using SPAGeDi. ADMIXTURE software divided the experimental cotton population into 16 subgroups, and the Gossypium hirsutum samples could be roughly clustered according to source place, but there were some overlapping characteristics among samples. The experimental cotton population was divided into six groups according to source to calculate the genetic diversity index (H), and the obtained value (0.306) was close to that for germplasm collected by others in China. Cluster 4 had a relatively high genetic diversity level (0.390). The degrees of genetic differentiation within the experimental cotton population groups were low (the population differentiation indexes ranged from 0.02368 to 0.10664). The genetic distance among cotton accessions varied from 0.000332651 to 0.562664014, with an average of 0.25240429. The results of this study may provide a basis for mining elite alleles and using them for subsequent association analysis.


[Natural regeneration of Larix principis-rupprechtii plantations with different densities and its influencing factors].
In this study, we analyzed the natural regeneration of Larix principis-rupprechtii pure plantations with stand densities of 128, 240, 320, 400, 480, 560, 640 and 720 trees·hm-2 in the Guandi Mountains and its influencing factors. The results showed that the regeneration index first increased and then decreased with the increases of stand density, and that the regeneration performance of stand with medium density (400-560 trees·hm-2) was significantly better than other stands. Light conditions, herbaceous plants and litter of the understory had a dual effect on the regeneration of L. principis-rupprechtii. Excessive light, herbaceous plant cover or over-thick litter was not instrumental to the regeneration. Soil organic matter promoted stand regeneration by providing soft soil texture, adequate water content, low phosphorus but high nitrogen. The effects of the examined factors on the regeneration index were as follows: soil water content (0.798) > total nitrogen (0.621) > litter thickness (-0.597) > soil porosity (0.504) > soil organic matter (0.493) > total phosphorus (-0.404) > transmitted total light (-0.274) > herbaceous plants cover (-0.021). In the plantation management, stand density could be controlled at about 480 trees·hm-2 by thinning or replanting, while litter could be cleared properly to improve soil condition and to promote natural regeneration of L. principis-rupprechtii.


Evolving connectionist systems for knowledge discovery from gene expression data of cancer tissue.
Microarray techniques have made it possible to observe the expression of thousands of genes simultaneously. They have recently been applied to study gene expression patterns in tissue samples. This may lead to highly desirable improvements in the diagnosis and treatment of human diseases. Statistical and machine learning methods have recently been used to classify cancer tissue based on gene expression data. Although some of these methods have achieved a high degree of accuracy, they generally lack transparency in their classification process. This, however, is crucial for the application in the medical field. In order to overcome this obstacle, we used knowledge-based neurocomputing (KBN), since KBN seeks to gain knowledge that is comprehensible to humans. In particular, we applied evolving fuzzy neural networks (EFuNNs) to classify cancer tissue, which is illustrated on the case studies of leukaemia and colon cancer. EFuNNs belong to the evolving connectionist system paradigm (ECOS) that has been recently introduced. They are well suited for adaptive learning and knowledge discovery. Fuzzy logic rules can be extracted from the trained networks and offer knowledge about the classification process in an easily accessible form. These rules point to genes that are strongly associated with specific types of cancer and may be used for the development of new tests and treatment discoveries.


Neonatal morbidity and mortality after elective cesarean delivery.
This article explores the effects of elective cesarean delivery (ECD) at term on neonatal morbidity and mortality. Available data have limitations, and do not provide conclusive evidence regarding the safety of planned ECD versus planned vaginal delivery. Some data suggest an association between ECD and increased neonatal respiratory morbidity and lacerations, and possibly decreased central and peripheral nervous system injury. Potentially increased risks of neonatal mortality with ECD at term may be counterbalanced by risks for fetal demise in ongoing pregnancies. Patients and physicians considering ECD should review competing risks and benefits; further research is needed to inform these discussions.


Subcutaneous panniculitis-like T-cell lymphoma with vacuolar interface dermatitis resembling lupus erythematosus panniculitis.
Lupus erythematosus (LE) panniculitis (LEP) is a form of chronic cutaneous LE most often characterized by erythematous subcutaneous nodules. The histopathology of LEP may be distinctive, allowing a diagnosis even in the absence of any other LE features. Lymphocytic infiltration of the panniculus is termed lymphocytic lobular panniculitis. This entity has been reported in both LEP and subcutaneous panniculitis-like T-cell lymphoma. We describe a 67-year-old woman who presented with multiple dermal and subcutaneous nodules on her legs and hips. Biopsy specimens of roughly 15 lesions were interpreted as being diagnostic of LEP, however, with each subsequent biopsy specimen more atypical cells were observed. The patient responded only to moderate doses of oral prednisone, and failed intralesional triamcinolone acetonide, oral immunosuppressive agents, oral antimalarial agents, and oral thalidomide. Three years after the onset of her disease, ulcerated plaques and nodules developed. At this time a T-cell receptor gene rearrangement was present and subcutaneous panniculitis-like T-cell lymphoma was diagnosed. Despite multiple doses of chemotherapy she died approximately 1 year after diagnosis. Patients with apparent LEP that have atypical lymphocytes in their biopsy specimens should be followed up closely for the development of T-cell malignancy.