Exercise-induced improvement in endothelial dysfunction is not mediated by changes in CV risk factors: pooled analysis of diverse patient populations.
We have pooled data from a series of our exercise training studies undertaken in groups with a broad range of vascular (dys) function to the examine the hypothesis that exercise-induced improvements in the conduit and/or resistance vessel function are related to improvements in risk factors for cardiovascular (CV) disease. Endothelium-dependent and -independent conduit vessel function were assessed by using wall tracking of high-resolution ultrasound images of the brachial artery response to flow-mediated dilation (FMD) and glyceryl trinitrate. Resistance vessel function was assessed using intrabrachial administration of acetylcholine (ACh), sodium nitroprusside, and NG-monomethyl-l-arginine. Randomized cross-over studies of 8-wk exercise training were undertaken in untreated hypercholesterolemic (n = 11), treated hypercholesterolemic (n = 11), coronary artery disease (n = 10), chronic heart failure (n = 12), Type 2 diabetic (n = 15), and healthy control subjects (n = 16). Exercise training did not significantly alter plasma lipids, blood pressure, blood glucose, waist-to-hip ratio, or body mass index values, despite significant improvement in both FMD and ACh responses. There were no correlations between changes in any risk factor variables and indexes of either resistance or conduit vessel function. We conclude that, in these subjects with antecedent vascular dysfunction, the beneficial effects of relatively short-term exercise training on vascular function are not solely mediated by the effects of exercise on CV risk factors.


Two-dimensional thin-layer chromatography for simultaneous detection of bacterial beta-lactam acylases and beta-lactamases.
A rapid and specific procedure was developed for the simultaneous detection of bacterial acylases and beta-lactamases, using ampicillin and cephalexin as substrates. Bacterial suspensions from agar plates were incubated separately with each beta-lactam substrate for 1 h at 37 degrees C. The supernatant of the reaction mixture was dansylated, and the dansyl derivatives were separated by two-dimensional thin-layer chromatography on polyamide sheets. The end products resulting from acylase hydrolysis, including the intact beta-lactam nucleus, 6-aminopenicillanic acid or 7-aminodeacetoxycephalosporanic acid, and the acyl side chain acid, D-(-)-alpha-aminophenylacetic acid, and the end product resulting from beta-lactamase hydrolysis (D-phenylglycylpenicilloic acid or D-phenylglycyldeacetoxycephalosporoic acid) were separated from each unhydrolyzed substrate and amino acids by this procedure. The presence of the intact beta-lactam nucleus in the reaction mixture is the indication of acylase activity. This method is sensitive and reproducible and has been successfully applied to screening for acylase activity in a variety of bacteria. It may be pharmaceutically useful for identifying organisms capable of removing the acyl side chain from naturally occurring beta-lactam antibiotics such as penicillin G, penicillin V, and cephalosporin C for production of the beta-lactam nuclei which serve as the starting materials for semisynthetic beta-lactam antibiotics.


Formation of celecoxib nanoparticles from volatile microemulsions.
A new composition of a fully water-dilutable microemulsion system stabilized by natural surfactants is presented as a template for preparation of celecoxib nanoparticles. Nanoparticles are obtained as a dry powder upon rapid conversion of microemulsion droplets with dissolved celecoxib into nanoparticles, followed by evaporation of all the liquid in a spray dryer. The resultant powder is easily re-dispersible in water to form a clear, transparent dispersion. The celecoxib nanoparticles are amorphous and their average size in the dispersion is 17 nm, in agreement with cryo-TEM results and concentration measurements after filtration. As a result of the nanometric size and amorphous state, about 10-fold increase in dissolution of the powder was obtained, compared to that for particulate celecoxib in the presence of surfactants.


Feasibility, safety, and outcome of a challenging transseptal puncture facilitated by radiofrequency energy delivery: a prospective single-centre study.
Although it has been shown that a transseptal (TS) puncture in the electrophysiology laboratory is associated with a high success and a low complication rate, this procedure remains challenging particularly in difficult septum anatomies (aneurismal septum and thick septum) and during repeat TS catheterization. Radiofrequency (RF) electrocautery current delivery through the TS needle has been shown to facilitate the TS puncture. The aim of this study was to verify prospectively the feasibility, safety, and outcome of RF energy delivery associated with the standard TS technique in patients undergoing a challenging TS puncture. Over a 14-month period, 162 consecutive patients underwent left atrial (LA) arrhythmia ablation in our centre. Among them, we enrolled 18 patients who failed LA access after two TS puncture attempts. In these patients, an RF delivery through TS (RF-TS) needle approach was used to reach the LA. All 18 patients had a successful RF-TS at the first attempt. A transoesophageal echocardiography (TEE) guidance and fluoroscopy views were used in all patients. No acute complications were reported. There have been no clinical sequelae after 10 +/- 4 months of follow-up following the RF-TS approach. Challenging TS punctures were more frequent in repeat LA catheterization when compared with the first LA catheterization, respectively, in 35% (13 of 37) and 4% (5 of 125) of the patients. Radiofrequency electrocautery delivery associated with the standard TS approach is a safe and reproducible technique to reach the left atrium, using the TEE guidance. This technique is helpful during repeat TS catheterization and in the presence of anatomical atrial septum abnormalities.


Facilitating factors in same-day discharge after pediatric laparoscopic appendectomy.
Appendicitis has been cited to be the most common abdominal disorder that requires acute care surgery in the pediatric population. Enhanced Recovery After Surgery protocols are multimodal perioperative care pathways designed to achieve early recovery after surgical procedures by maintaining preoperative organ function and reducing the profound stress response following surgery. Such pathways have been found to enhance quality of care for surgical patients as well as improve recovery and shorten hospital stays. The purpose of this study was to determine the key factors that facilitate same-day discharge (SDD) and early return to normal activities after laparoscopic appendectomies in children. This is a single-center retrospective chart review of pediatric patients (<18 y old) who underwent appendectomies for acute appendicitis from January 2015 to April 2017. The patient population was divided into two groups: those with SDD and those who were discharged one or more days after surgery. SDD was defined as discharge less than 24 h of surgical admission. Patient factors, including prehospital, preoperative, perioperative, and postoperative factors, were compared and analyzed between the two groups and statistically evaluated using Fisher's exact test for categorical data and student t-test for continuous variables. Two hundred forty eight patients were found under International Classification of Diseases-9 and International Classification of Diseases-10 codes for acute appendicitis. Of these, 63 were excluded due to perforated appendicitis, nonoperative management, interval appendectomies, or misdiagnosis. The remaining 185 had laparoscopic appendectomies; 59.5% (n = 110) were SDDs and 40.5% (n = 75) stayed more than one day. No significant difference was found for time between emergency room arrival and surgical admission (5.27 versus 5.4 h; P = 08.) but SDD patients had a significantly shorter time between surgical admission and operation (5.8 versus 11.4 h; P <0.001). SDD patients and non-SDD patients had no significant difference in rate of complications (0% versus 1.3%; P = 0.4). There was no significant difference in readmission rates between the two groups (2.7% versus 2.7%; P = 1). Total hospital charges, which indirectly reflect costs, were significantly less for SDD ($29,195 versus $33,703; P <0.001). Surgical leadership can be effective in facilitating SDD without increasing readmission rates or complications and helps reduce hospital costs, decreases chances of nosocomial infection, and increases patient and family satisfaction.


[Gene therapy of monogenic hereditary diseases. Duchenne myodystrophy].
The paper highlights the new trends in gene therapy research area and clinical trials. It should be noted that the majority of firms involved in development of the scientific approaches to gene therapy are concentrated in the United States. The investments of the given companies on development and research of new genetic constructs also delivery systems make hundred millions dollars. The greatest part (more than 80%) of gene therapy clinical trials projects are also connected with the US research departments; the majority of them is related to tumor therapy. The advantages and drawbacks of the main methods of nucleic acids delivery to the cells are considered; diseases that are attempted to be treated using gene therapy methods are listed. A special attention of the review is devoted to the modern stand in research on cell and Duchenne muscular dystrophy (DMD) gene therapy, also brief description of basic results achieved in the authors laboratory is given. Basic original results of transfection of mdx mice (DMD biological models) with dystrophin cDNA delivered by gene gun, cationic liposomes, synthetic microspheres, viral olygopeptides and lactoferrine are summarized.


Comparison of computed tomography and magnetic resonance imaging in the evaluation of facet tropism and facet arthrosis in degenerative cervical spondylolisthesis.
The aim of this study was to determine the reliability of magnetic resonance imaging (MRI) in the assessment of facet tropism and facet arthrosis of spondylolisthesis levels in degenerative cervical spondylolisthesis as compared to computed tomography (CT). The discrepancies in the interpretation of CT and MRI data in the evaluation of facet tropism and arthrosis have given rise to questions regarding the reliability of comparisons of the two techniques. Using a 4-point scale, 3 blinded readers independently graded the severity of facet tropism and facet arthrosis of 79 cervical facet joints on axial T2-weighted and sagittal T1 and T2-weighted turbo spin echo images as well as the corresponding axial CT scans. All results were subjected to the kappa coefficient statistic for strength of agreement. In the assessment of the severity of facet arthrosis, intermethod agreement (weighted κ) between CT scanning with a moderate inter-rater reliability (range κ = 0.43-0.57) and MRI with fair inter-rater reliability (range κ = 0.23-0.38) was 0.76 and 0.43 for the severity of facet tropism and facet arthrosis, respectively. Intra-rater reliability for the severity of facet arthrosis was moderate to substantial for CT and was moderate for MRI scans. Intra-rater reliability for the severity of facet tropism was substantial to very good for CT and substantial for MRI scans. MRI can reliably determine the presence or degree of facet tropism but not facet arthrosis. Therefore, for a comprehensive assessment of cervical facet joint degeneration, both a CT and an MRI scan should be performed.


HSP90AB1 is a host factor that promotes Porcine deltacoronavirus replication.
Porcine deltacoronavirus (PDCoV) is an emerging enteropathogenic coronavirus. It causes mortality in neonatal piglets and is of growing concern because of its broad host range, including humans. To date, the mechanism of PDCoV infection remains poorly understood. Here, based on a genome-wide CRISPR screen of PDCoV-infected cells, we found that HSP90AB1 promotes PDCoV infection. Knockdown or knockout of HSP90AB1 in LLC-PK cells resulted in a significantly suppressed PDCoV infection. Infected cells treated with HSP90 inhibitors 17-AAG and VER-82576 also showed a significantly suppressed PDCoV infection, although KW-2478, which does not affect the ATPase activity of HSP90AB1, had no effect on PDCoV infection. We found that HSP90AB1 interacts with the N, NS7, and NSP10 proteins of PDCoV. We further evaluated the interaction between N and HSP90AB1 and found that the C-tail domain of the N protein is the HSP90AB1-interacting domain. Further studies showed that HSP90AB1 protects N protein from degradation via the proteasome pathway. In summary, our results reveal a key role for HSP90AB1 in the mechanism of PDCoV infection and contribute to provide new host targets for PDCoV antiviral research.


Coping style and depression influence the healing of diabetic foot ulcers: observational and mechanistic evidence.
Experimental evidence suggests that the healing of diabetic foot ulcers is affected by psychosocial factors such as distress. We examined this proposal in a prospective study, in which we considered the role of psychological distress and coping style in the healing of diabetic foot ulcers over a 24 week period. We also explored the role of salivary cortisol and matrix metalloproteinases (MMPs) as potential mechanisms. For this prospective observational study we recruited 93 (68 men; mean age 60 years) patients with neuropathic or neuroischaemic diabetic foot ulcers from specialist podiatry clinics in secondary care. Clinical and demographic determinants of healing, psychological distress, coping, salivary cortisol and both MMP2 and MMP9 were assessed at baseline. Ulcers were assessed at baseline and at 6, 12 and 24 weeks post-baseline. The primary outcome was ulcer status at 24 weeks, i.e. healed vs not healed. After controlling for clinical and demographic determinants of healing, ulcer healing at 24 weeks was predicted by confrontation coping, but not by depression or anxiety. Patients with unhealed ulcers exhibited greater confrontation coping (model including depression: OR 0.809, 95% CI 0.704-0.929, p = 0.003; model including anxiety: OR 0.810, 95% CI 0.704-0.930, p = 0.003). However, change in ulcer size over the observation period was associated with depression only (p = 0.04, d = 0.31). Healed ulcers by 24 weeks were also associated with lower evening cortisol, higher precursor MMP2 and a greater cortisol awakening response. Confrontation coping and depression predict ulcer healing. Our preliminary enquiry into biological mechanisms suggests that cortisol and precursor MMP2 may underlie these relationships.


Distribution of genetic markers of fecal pollution on a freshwater sandy shoreline in proximity to wastewater effluent.
Water, sand, and sediment from a Lake Superior harbor site continuously receiving wastewater effluent was sampled monthly for June to October 2010 and from May to September 2011. Understanding the dynamics of genetic markers of fecal bacteria in these matrices is essential to accurately characterizing health risks. Genetic markers for enterococci, total Bacteroides, and human-associated Bacteroides were measured in site-water, sand, and sediment and in final effluent by quantitative PCR. The similarity between the quantity of molecular markers in the water column and effluent indicated that the abundance of genetic markers in the water column was likely controlled by effluent inputs. Effluent turbidity was positively correlated (p ≤ 0.05) with AllBac and HF183 in final effluent and AllBac in the water column. In sand and sediment, Entero1 and AllBac were most abundant in the upper 1-3 cm depths, whereas HF183 was most abundant in the upper 1 cm of sand and at 7 cm in sediment. The AllBac and Entero1 markers were 1- and 2-orders of magnitude more abundant in sand and sediment relative to the water column per unit mass. These results indicate that sand and sediment may act as reservoirs for genetic markers of fecal pollution at some freshwater sites.


Lighting Deviation Correction for Integrating-Sphere Multispectral Imaging Systems.
In an integrating sphere multispectral imaging system, measurement inconsistency can arise when acquiring the spectral reflectances of samples. This is because the lighting condition can be changed by the measured samples, due to the multiple light reflections inside the integrating sphere. Besides, owing to non-uniform light transmission of the lens and narrow-band filters, the measured reflectance is spatially dependent. To deal with these problems, we propose a correction method that consists of two stages. The first stage employs a white board to correct non-uniformity and a small white patch to correct lighting deviation, both under the assumption of ideal Lambertian reflection. The second stage uses a polynomial regression model to further remove the lighting inconsistency when measuring non-Lambertian samples. The method is evaluated on image data acquired in a real multispectral imaging system. Experimental results illustrate that our method eliminates the measurement inconsistency considerably. This consequently improves the spectral and colorimetric accuracy in color measurement, which is crucial to practical applications.


Dimensionality in rhythmic bimanual coordination.
Newell and Vaillancourt (2001) hypothesized that the dimensionality of motor behavior is a function of the level of task performance and the task dynamic. The present study examined high (in-phase), moderate (antiphase) and low (45°, 90°, and 135° relative phase) levels of task performance in bimanual coordination. Estimates of dimensionality were calculated for the component (effector movements), coupling of components (coupling of effectors), and task output (the produced relative phase) levels of analysis. The in-phase coordination mode had lower Approximate Entropy within, and lower Cross-Approximate Entropy between, effector movements than all other modes. The in-phase mode had higher relative phase Approximate Entropy than all other modes. These findings indicate lower effector and coupling dimensionality, and higher relative phase dimensionality, in the in-phase mode. These results support the hypothesis that at the levels of analysis with limit-cycle dynamics high levels of task performance are characterized by lower dimensionality than lower levels of performance. The results also support the hypothesis that high task performance of the fixed-point task goal of maintaining a constant relative phase is characterized by higher dimensionality than low level performance. Together, these findings support and generalize the Newell and Vaillancourt hypothesis to the component, coupling, and task output levels of analysis.


The adaptability of H9N2 avian influenza A virus to humans: A comparative docking simulation study.
Influenza A virus, the H9N2 subtype, is an avian influenza virus that has long been circulating in the worldwide poultry industry and is occasionally found to be transmissible to humans. Evidence from genomic analysis suggests that H9N2 provides the genes for the H5N1 and H7N9 subtypes, which have been found to infect mammals and pose a threat to human health. However, due to the lack of a structural model of the interaction between H9N2 and host cells, the mechanism of the extensive adaptability and strong transformation capacity of H9N2 is not fully understood. In this paper, we collected 40 representative H9N2 virus samples reported recently, mainly in China and neighboring countries, and investigated the interactions between H9N2 hemagglutinin and the mammalian receptor, the polysaccharide α-2,6-linked lactoseries tetrasaccharide c, at the atomic level using docking simulation tools. We categorized the mutations of studied H9N2 hemagglutinin according to their effects on ligand-binding interactions and the phylogenetic analysis. The calculations indicated that all the studied H9N2 viruses can establish a tight binding with LSTc although the mutations caused a variety of perturbations to the local conformation of the binding pocket. Our calculations suggested that a marginal equilibrium is established between the conservative ligand-receptor interaction and the conformational dynamics of the binding pocket, and it might be this equilibrium that allows the virus to accommodate mutations to adapt to a variety of environments. Our results provided a way to understand the adaptive mechanisms of H9N2 viruses, which may help predict its propensity to spread in mammals.


[Characteristics of soil microbial populations in depressions between karst hills under different land use patterns].
Based on the investigation and analysis of six soil microbial indices, eight soil conventional nutrient indices, six soil mineral nutrient indices, and 15 vegetation indices in the farmland, grassland, scrub, forest plantation, secondary forest, and primary forest in the depressions between karst hills, this paper analyzed the main soil microbial populations, soil microbial biomass carbon (MBC), nitrogen (MBN), and phosphorus (MBP) and their fractal characteristics, and the relationships of the soil microbes with vegetation, soil nutrients, and soil mineral components under different land use patterns. The soil microbial populations differed in their quantity and composition under different land use patterns. Primary forest and farmland had the highest quantity of soil microbial populations, while forest plantation had the lowest one. The three forests had a higher proportion of soil bacteria, the farmland, grassland, and scrub had a higher proportion of actinomycetes, and all the six land use patterns had a low proportion of soil fungi. Under the six land use patterns, the soil MBC, MBN, and MBP were all high, with the maximum in primary forest. There was a good fractal relationship between the soil MBC and microbial populations, but no fractal relationships between the soil MBN and MBP and the microbial populations. Significant relationships were observed between the soil microbes and the vegetation, soil nutrients, and soil mineral components, and the soil MBC had the closest relationships with the Shannon index of tree layer and the soil total nitrogen, Fe2O3, and CaO contents.


Storage Medium Affects the Surface Porosity of Dental Cements.
Calcium silicate-based cements physical properties is influenced by environmental changes. Here, we intended to evaluate the effect of storage medium on surface porosity of root Mineral Trioxide Aggregate (MTA) and Biodentine cement. A total of 40 polyethylene tubes were selected and divided into two groups: Group A (MTA) and Group B (Biodentine). Each group was subdivided into two subgroups (n=10). In subgroups A1 and B1, tubes were transferred to Distilled Water (DW), while samples of subgroup A2 and B2 were transferred to Synthetic Tissue Fluid (STF) as storage medium and samples were stored for three days. All specimens were then placed in a desiccator for 24 hours and then subject to surface porosity evaluation by Scanning Electron Microscopy (SEM) at ×500, ×1000, ×2000 and ×5000 magnifications. The number and the surface porosities were determined by Image J analysis. Data were analyzed by ANOVA at level of significance of p<0.05. The lowest surface porosity was observed in MTA samples stored in STF and the highest was in Biodentine samples stored in DW. Significant differences were noted between groups and subgroups of each group (p< 0.05). MTA samples stored in DW and STF showed significantly lower surface porosities compared to Biodentine samples (p < 0.05). Storage medium can drastically affect the surface porosity of tested calcium silicate-based cements. However, MTA showed lower surface porosity compared to Biodentine cement, which can result in lower microleakage in applied area.


Widening area-based socioeconomic inequalities in cancer mortality in Germany between 2003 and 2019.
Cancer mortality has declined in recent decades, but-due to a lack of national individual-level data-it remains unclear whether this applies equally to all socioeconomic groups in Germany. Using an area-based approach, this study investigated socioeconomic inequalities in cancer mortality and their secular trends on a German nationwide scale for the first time. Official cause-of-death data from 2003 to 2019 were linked to the district-level German Index of Socioeconomic Deprivation. Age-standardised mortality rates for all cancers combined and the most common site-specific cancers were calculated according to the level of regional socioeconomic deprivation. To quantify the extent of area-based socioeconomic inequalities in cancer mortality, absolute (SII) and relative (RII) indices of inequality were estimated using multilevel Poisson models. On average, cancer mortality was 50% (women) and 80% (men) higher in Germany's most deprived than least deprived districts (absolute difference: 84 deaths per 100,000 in women and 185 deaths per 100,000 in men). As declines in cancer mortality were larger in less deprived districts, the socioeconomic gap in cancer mortality widened over time. This trend was observed for various common cancers. Exceptions were cancers of the lung in women and of the pancreas in both sexes, for which mortality rates increased over time, especially in highly deprived districts. Our study provides first evidence on increasing socioeconomic inequalities in cancer mortality on a nationwide scale for Germany. Area-based linkage allows to examine socioeconomic inequalities in cancer mortality across Germany and identify regions with high needs for cancer prevention and control.


The potential impact of density dependent fecundity on the use of the faecal egg count reduction test for detecting drug resistance in human hookworms.
Current efforts to control human soil-transmitted helminth (STH) infections involve the periodic mass treatment of people, particularly children, in all endemic areas, using benzimidazole and imidothiazole drugs. Given the fact that high levels of resistance have developed to these same drugs in roundworms of livestock, there is a need to monitor drug efficacy in human STHs. The faecal egg count reduction test (FECRT), in which faecal egg output is measured pre- and post-drug treatment, is presently under examination by WHO as a means of detecting the emergence of resistance. We have examined the potential impact of density dependent fecundity on FECRT data. Recent evidence with the canine hookworm indicates that the density dependent egg production phenomenon shows dynamic properties in response to drug treatment. This will impact on measurements of drug efficacy, and hence drug resistance. It is likely that the female worms that survive a FECRT drug treatment in some human cases will respond to the relaxation of density dependent constraints on egg production by increasing their egg output significantly compared to their pre-treatment levels. These cases will therefore underestimate drug efficacy in the FECRT. The degree of underestimation will depend on the ability of the worms within particular hosts to increase their egg output, which will in turn depend on the extent to which their egg output is constrained prior to the drug treatment. As worms within different human cases will likely be present at quite different densities prior to a proposed FECRT, there is potential for the effects of this phenomenon on drug efficacy measurements to vary considerably within any group of potential FECRT candidates. Measurement of relative drug efficacy may be improved by attempting to ensure a consistent degree of underestimation in groups of people involved in separate FECRTs. This may be partly achieved by omission of cases with the heaviest infections from a FECRT, as these cases may have the greatest potential to increase their egg output upon removal of density dependent constraints. The potential impact of worm reproductive biology on the utility of the FECRT as a resistance detection tool highlights the need to develop new drug resistance monitoring methods which examine either direct drug effects on isolated worms with in vitro phenotypic assays, or changes in worm genotypes.


Diagnosis of Helicobacter pylori: invasive methods.
The methods which require endoscopy for the examination of the gastric mucosa are called invasive (direct) methods. Several tests can be performed on the gastric mucosa obtained by endoscopy: Rapid urease test, histology, smear (cytology), culture and polymerase chain reaction. A combination of at least two tests with high sensitivity and specificity is recommended for clinical trials, and, when possible, for clinical management of patients. In clinical practice the invasive methods should be carried out only in those patients to be treated with eradication regimes. All invasive methods rely on gastric biopsy samples. Therefore, the specimens should be taken under optimal conditions. Several factors guide the site and the number of biopsy specimens for identification of Helicobacter pylori infection. In pre-treatment diagnosis, the biopsies should be taken from the distal antrum (2 cm from the pylorus). The number required depends on the diagnostic method used: in Helicobacter pylori eradicated patients the same number of biopsies for histology and culture should be taken from the antrum and corpus. The sensitivity and specificity of the Rapid urease test varies from 80-90% the results are known very quickly and the cost is very low. The maximal accuracy of histology is obtained with: an optimal specimen processing, an adequate staining and an experienced observer. This method of biopsy processing facilitates the identification of Helicobacter pylori which is commonly located on the superficial and foveolar epithelium. Sensitivity depends on the observer's experience and the extent of biopsy sampling. In general, the histological method has a sensitivity and specificity of 90-95%. In patients treated by proton pump inhibitors, antibiotics or bismuth salts two-four weeks prior to biopsy, the bacteria may be restricted to the corpus or fundus. Culture is strictly indicated in patients after failure of two or more eradication regimens to test for susceptibility and resistence. Specificity is 100% and sensitivity depends on the experience and interest of the microbiologist. The indication of polymerase chain reaction is only for research, specially, in molecular epidemiology and for fingerprinting.


Combining Metabolic Pulse Labeling and Quantitative Proteomics to Monitor Protein Synthesis Upon Viral Infection.
Viruses like influenza A virus (IAV) hijack host cells in order to replicate. To actively and abundantly synthesize viral proteins, they reprogram the cellular transcriptional and translational landscape. Here, we present a proteomic approach that allows us to quantify the differences in host and viral protein synthesis comparatively for different strains of IAV. The method is based on combining quantitative proteomics using stable isotope labelling by amino acids in cell culture (SILAC) and bioorthogonal labeling with methionine analogs. This methodology accurately quantifies synthesis of host and viral proteins with high temporal resolution and faithfully detects global changes in cellular translation capacity. It thus provides unique insights into the dynamics of protein synthesis as the infection progresses.


[Therapy of back pain: what is evidence-based?].
Back pain has to be classified by localisation, duration, and existence or absence of neurological deficits. Therapy mainly depends on the severity of neurological symptoms. The time period between the onset of symptoms and efficient therapy should be short in order to prevent the development of chronic pain. Therefore a sufficient, consistent and evidence-based conservative treatment should be initiated early on, with the only exception of special emergency situations as described further down in this article. Patients should not be immobilized, a better outcome is proven in patients who immediately continued practising their daily activities. First line medication consists of non-steroidal-anti-inflammatory-drugs (NSAID). If necessary, opioids and muscle-relaxants may be applied additionally. Surgical procedures are only indicated in a small number of patients. A sufficient data base with regard to the benefit of acupuncture, orthesis, and chiropractic medicine is lacking. Local injections should only be done in very special cases. Concerning chronic back pain a multimodal, multi-disciplinary pain treatment concept with physical training and behavioural therapy should be applied in order to teach the patient to avoid a passive and maladjusted attitude. Antidepressant drugs may also be indicated in chronic back pain.


Vertebral Implantation of NELL-1 Enhances Bone Formation in an Osteoporotic Sheep Model.
Vertebral compression fractures related to osteoporosis greatly afflict the aging population. One of the most commonly used therapy today is balloon kyphoplasty. However, this treatment is far from ideal and is associated with significant side effects. NELL-1, an osteoinductive factor that possesses both pro-osteogenic and anti-osteoclastic properties, is a promising candidate for an alternative to current treatment modalities. This study utilizes the pro-osteogenic properties of recombinant human NELL-1 (rhNELL-1) in lumbar spine vertebral defect model in osteoporotic sheep. Osteoporosis was induced through ovariectomy, dietary depletion of calcium and vitamin D, and steroid administration. After osteoporotic induction, lumbar vertebral body defect creation was performed. Sheep were randomly implanted with the control vehicle, comprised of hyaluronic acid (HA) with hydroxyapatite-coated β-tricalcium phosphate (β-TCP), or the treatment material of rhNELL-1 protein lyophilized onto β-TCP mixed with HA. Analysis of lumbar spine defect healing was performed by radiographic, histologic, and computer-simulated biomechanical testing. rhNELL-1 treatment significantly increased lumbar spine bone formation, as determined by bone mineral density, % bone volume, and mean cortical width as assessed by micro-computed tomography. Histological analysis revealed a significant increase in bone area and osteoblast number and decrease in osteoclast number around the implant site. Computer-simulated biomechanical analysis of trabecular bone demonstrated that rhNELL-1-treatment resulted in a significantly more stress-resistant composition. Our findings suggest rhNELL-1-based vertebral implantation successfully improved cortical and cancellous bone regeneration in the lumbar spine of osteoporotic sheep. rhNELL-1-based bone graft substitutes represent a potential new local therapy.


Perceived risk and condom use among adolescents in sub-Saharan Africa: a latent class analysis.
While studies have examined factors associated with condom use behaviors, few have assessed risk perception and condom use among SSA adolescents. This work sought to fill this gap. Data for this study are derived from the National Survey of Adolescents (2004-2005), a survey of 12-19 year olds in Burkina Faso, Ghana, Malawi, and Uganda. Latent class analysis was conducted. Two risk profiles emerged in the data. Percent of adolescents in the high perceived risk sub-groups ranged from 49.6% - 90.8% and moderate risk from 29.1-83.8%. Those in the high perceived risk class were 2.26 to 5.10 times more likely to report condom use. The findings provide additional information about the factors that influence condom use among adolescents in these four countries. Because an individual's development is influenced by his/her settings and systems, expanded views of behavior must be included in prevention efforts.


Reciprocal congenic lines for a major stroke QTL on rat chromosome 1.
We previously identified a quantitative trait locus (QTL) for stroke proneness between the kallikrein (Klk) and Mt1pa markers on rat chromosome 1. To gain functional insights, we constructed congenic strains by introgressing either the whole or selected chromosomal segments from the stroke-prone (SHRsp) onto the stroke-resistant (SHRsr) spontaneously hypertensive rat genome and vice versa. The phenotype was the latency to develop stroke under a Japanese high-salt, low-potassium diet for 3 mo [known as Japanese diet (JD)]. Blood pressure (BP) was measured by tail cuff throughout the experiment. Urinary protein excretion was monitored in all lines under JD. The SHRsp-derived lines carrying the SHRsr allele, and particularly the D1Rat134-Mt1pa chromosomal segment, had a significant delay of stroke occurrence and improved survival compared with SHRsp (P < 0.001). On the other hand, a significant occurrence of stroke events (20%) was detected in the reciprocal lines by the end of the 3-mo treatment with JD (P = 0.003). The stroke phenotype was also associated with increased proteinuria. Our results underscore the functional importance of the Chr 1 stroke QTL. Furthermore, they underscore the utility of stroke/congenic lines in dissecting the genetics of stroke.


North Atlantic forcing of moisture delivery to Europe throughout the Holocene.
Century-to-millennial scale fluctuations in precipitation and temperature are an established feature of European Holocene climates. Changes in moisture delivery are driven by complex interactions between ocean moisture sources and atmospheric circulation modes, making it difficult to resolve the drivers behind millennial scale variability in European precipitation. Here, we present two overlapping decadal resolution speleothem oxygen isotope (δ(18)O) records from a cave on the Atlantic coastline of northern Iberia, covering the period 12.1-0 ka. Speleothem δ(18)O reveals nine quasi-cyclical events of relatively wet-to-dry climatic conditions during the Holocene. Dynamic Harmonic Regression modelling indicates that changes in precipitation occurred with a ~1500 year frequency during the late Holocene and at a shorter length during the early Holocene. The timing of these cycles coincides with changes in North Atlantic Ocean conditions, indicating a connectivity between ocean conditions and Holocene moisture delivery. Early Holocene climate is potentially dominated by freshwater outburst events, whilst ~1500 year cycles in the late Holocene are more likely driven by changes internal to the ocean system. This is the first continental record of its type that clearly demonstrates millennial scale connectivity between the pulse of the ocean and precipitation over Europe through the entirety of the Holocene.


Application and underlying mechanism of acupuncture for the nerve repair after peripheral nerve injury: remodeling of nerve system.
Peripheral nerve injury (PNI) is a structural event with harmful consequences worldwide. Due to the limited intrinsic regenerative capacity of the peripheral nerve in adults, neural restoration after PNI is difficult. Neurological remodeling has a crucial effect on the repair of the form and function during the regeneration of the peripheral nerve after the peripheral nerve is injured. Several studies have demonstrated that acupuncture is effective for PNI-induced neurologic deficits, and the potential mechanisms responsible for its effects involve the nervous system remodeling in the process of nerve repair. Moreover, acupuncture promotes neural regeneration and axon sprouting by activating related neurotrophins retrograde transport, such as nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF), N-cadherin, and MicroRNAs. Peripheral nerve injury enhances the perceptual response of the central nervous system to pain, causing central sensitization and accelerating neuronal cell apoptosis. Together with this, the remodeling of synaptic transmission function would worsen pain discomfort. Neuroimaging studies have shown remodeling changes in both gray and white matter after peripheral nerve injury. Acupuncture not only reverses the poor remodeling of the nervous system but also stimulates the release of neurotrophic substances such as nerve growth factors in the nervous system to ameliorate pain and promote the regeneration and repair of nerve fibers. In conclusion, the neurological remodeling at the peripheral and central levels in the process of acupuncture treatment accelerates nerve regeneration and repair. These findings provide novel insights enabling the clinical application of acupuncture in the treatment of PNI.


Working with glaucoma patients - prospects for "shared care".
To investigate the prospects for "shared care" by evaluating to what extent ophthalmic nurses are involved in ophthalmic care decision tasks in the management of glaucoma patients, how well prepared the nurses feel they are for these tasks and their level of satisfaction in those tasks they actually are performing. A questionnaire was sent to the ophthalmic nurses at 13 eye clinics in Sweden. The clinics were selected to represent different parts of Sweden as well as university clinics and smaller rural clinics. A difference was found between clinics regarding the number of tasks independently performed by the ophthalmic nurses and a significant correlation was found between level of participation and satisfaction (p=0.008). The nurses felt prepared for (61/62) and satisfied (58/62) when performing the glaucoma management tasks, and a high and very significant correlation (r=0.96, p= 0.0001) was found between the mean preparedness and satisfaction scores for each of the 14 tasks. The findings in the study show that the ophthalmic nurses feel prepared for their tasks and regard their involvement in glaucoma care as subjectively satisfactory. These observations support the idea of "shared care" in glaucoma management.


Hydrothermal aging affects the three-dimensional fit and fatigue lifetime of zirconia abutments.
Evaluate the effect of aging using two different methods on the three-dimensional fit of zirconia abutments at the implant-abutment connection and estimate the probability of survival of anterior crowns supported by straight and 17-degree angled abutments. Two different zirconia abutment designs, straight and 17-degree angled abutments (n = 63/group), were evaluated in the current study. The abutments were randomly allocated into three experimental groups according to laboratory aging condition (134°C, 2.2 bar, 20 h): (i) control, (ii) autoclave aging, and (iii) hydrothermal reactor aging. Crystalline content was determined by X-Ray diffraction (XRD) and Raman spectroscopy, and microstructure was analyzed using field-emission gun scanning electron microscope (FEG-SEM). Implant-abutment volume misfit was determined in the straight abutments by micro-computed tomography using the silicone replica technique. For fatigue testing, abutments were torqued to the implants and connected to standardized maxillary incisor zirconia crowns. The assemblies were subjected to step-stress accelerated life testing (SSALT) in water until fracture or suspension. The use level probability Weibull curves and probability of survival for a mission of 50,000 cycles at 50, 100, 150 and 200 N were calculated and plotted. Fractured samples were analyzed using a stereomicroscope and scanning electron microscope. The crystalline spectra depicted a zirconia system primarily composed of the tetragonal phase. Laboratory aging yielded a 20%- and 37%-increase in the monoclinic content for abutments aged in autoclave and hydrothermal reactor relative to control, respectively. A fully crystalline matrix with a regular grain size was observed in the FEG-SEM for control abutments, with a considerable presence of intergranular defects. While autoclave aging triggered no significant alteration to the microstructure, defect population was reduced after hydrothermal reactor aging. Control abutments presented a significantly higher volume misfit (2.128 ± 0.54 mm3) relative to aged abutments using autoclave (1.244 ± 0.48 mm3) or hydrothermal reactor (1.441 ± 0.41 mm3). The beta (β) values indicated that failures were predominantly controlled by material strength rather than fatigue damage accumulation for all groups, except for straight control abutments. Irrespective of aging, the probability of survival of straight and angled zirconia abutments was up to 95% (95-100%) at 50 and 100 N. A 50N-increase in the load resulted in wider range of survival estimate, with straight autoclave abutments percentage significantly lower probability of survival (77%) than angled hydrothermal reactor abutments (99%). At 200N, angled hydrothermal reactor (97%) or autoclave (82%) aged abutments demonstrated the highest probability of survival, angled control (71%) and straight hydrothermal reactor (69%) abutments intermediate values, and straight autoclave (23%) and control (7%) abutments the lowest estimate. The failure mode predominantly involved abutment and/or abutment screw fracture for both straight and angled abutments. Hydrothermal aging significantly influenced volume misfit, as well as the probability of survival of zirconia abutments at higher loads for both angled and straight abutments.


Dog and human acid beta-D-galactosidases are structurally similar.
The purification of dog liver acid beta-galactosidase is described. The dog enzyme migrated as a single major band on polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate, with a molecular weight of 60000. Antiserum raised against purified human liver acid beta-galactosidase cross-reacted with beta-galactosidase from dog liver, but not with those from cat liver or Escherichia coli. Tryptic peptide maps of the dog and human acid beta-galactosidases indicate that 21 of the 24 peptides observed were homologous; a similar result was obtained after chymotryptic peptide mapping. We conclude that dog and human acid beta-galactosidases are structurally similar, and that canine GM1 gangliosidosis (acid beta-galactosidase deficiency) is an excellent model for the same disease in man.


Antibody-dependent cell-mediated cytotoxicity- and complement-dependent cytotoxicity-independent bactericidal activity of an IgG against Pseudomonas aeruginosa O6ad.
In addition to Ag recognition, some Abs are capable of killing target organisms in the absence of phagocytes and complement. In this study, we report that an anti-Pseudomonas aeruginosa O6ad LPS IgG(1), tobacco-expressed human S20 IgG(1) (te-hS20), as well as its recombinant Fab and single-chain variable fragment (scFv) fragments have cellular- and complement-independent bactericidal activity. te-hS20 and its Fab and scFv significantly reduced viability of P. aeruginosa O6ad in dose- and time-dependent manners in vitro and also showed lower levels of bactericidal activity against P. aeruginosa PAO1, but had no activity against P. aeruginosa O10, Escherichia coli TG1, and Streptococcus agalactiae. The H chain and its Fd fragment both had significant Ag-binding and bactericidal activities against P. aeruginosa O6ad. Bactericidal activity was completely inhibited with specific LPS Ag, suggesting that Ag binding is involved in the bactericidal mechanism. Live/dead cell staining and electron microscopic observations indicate that the bactericidal effect was due to disruption of the cell wall and suggest inhibition of cell division. In addition to te-hS20, the Fab and scFv were also protective in vivo, as leukopenic mice had prolonged and improved survival after administration of these Ab fragments followed by challenge with P. aeruginosa O6ad cells at 80-90% lethal dose, supporting a bactericidal mechanism independent of phagocytes and complement. Understanding of the bactericidal mechanism will allow assessment of the potential for therapeutic application of these Abs.


White matter hyperintensities and their penumbra lie along a continuum of injury in the aging brain.
Aging is accompanied by clinically silent cerebral white matter injury identified through white matter hyperintensities (WMHs) on fluid-attenuated inversion recovery (FLAIR)- and diffusion tensor imaging-based measures of white matter integrity. The temporal course of FLAIR and diffusion tensor imaging changes within WMHs and their less-injured periphery (ie, their penumbra), however, has not been fully studied. We used longitudinal diffusion tensor imaging and FLAIR to explore these changes. One hundred fifteen participants, aged 73.7±6.7 years, received clinical evaluations and MRIs on 2 dates. WMHs and fractional anisotropy (FA) maps were produced from FLAIR and diffusion tensor imaging and coregistered to a standardized space. Each distinct WMH was categorized as growing, stagnant, or noncontiguous incident. The penumbra of each WMH was similarly categorized as corresponding to a stagnant, growing, or noncontiguous incident WMH. Linear mixed-effect models were used to assess whether FA and FLAIR measurements changed between baseline and follow-up and differed between tissue categories. Baseline FA differed significantly by tissue category, with the following ordering of categories from highest to lowest FA: penumbra of noncontiguous incident, then stagnant, then growing WMHs; noncontiguous incident, then stagnant, then growing WMHs. Despite differences in baseline values, all tissue categories experienced declines in FA over time. Only noncontiguous incident WMHs showed significant FLAIR signal increases over time, and FLAIR signal significantly decreased in stagnant WMHs. WMHs and their penumbra vary in severity and together span a continuous spectrum of white matter injury that worsens with time. FLAIR fails to capture this continuous injury process fully but does identify a subclass of lesions that seem to improve over time.


Temporal prediction changes motor-related EEG phase synchronization and network centrality in alpha and beta band.
Much neurophysiological evidence revealed motor system is involved in temporal prediction. However, It remains unknown how temporal prediction influences motor-related neural representations. Thus, more neural evidence is needed to understand better how temporal prediction influences the motor. This study designed a rhythmic finger-tap task and formed three temporal prediction conditions, i.e., 1000ms temporal prediction, 1500ms temporal prediction, and no temporal prediction. Behavioral and EEG data from 24 healthy subjects were recorded. The weighted phase lag index was calculated to measure the degree of phase synchronization. Eigenvector centrality and betweenness centrality were used to measure brain connectivity. Behavioral results showed that tap-visual asynchronies were decreased when temporal prediction existed. Phase synchronization results showed, compared to no temporal prediction, the alpha-band phase synchronization between the frontal and central area was reduced in 1000ms temporal prediction, and the beta-band phase synchronization between the frontal and parietal area was decreased in 1500ms temporal prediction. As to the brain connectivity, compared to no temporal prediction condition, the eigenvector centrality of the left frontal in 1500ms temporal prediction was decreased in the alpha band, and the betweenness centrality of the right temporal in 1000ms temporal prediction was reduced in the alpha-band. These results can provide new neural evidence for a better understanding of temporal prediction and motor interactions.


Deficient pulmonary IFN-β expression in COPD patients.
COPD patients are prone to acute infectious exacerbations that impair their quality of life and hamper prognosis. The purpose of the present study was to investigate the in situ IFN-β response in the lungs of stable COPD and non-COPD patients. Lung samples from 70 subjects (9 control never smokers, 19 control smokers without COPD, 21 patients with moderate COPD and 21 patients with very severe COPD) were studied for the expression of IFN-β, its main transcription factor, IRF-7, and two products of its autocrine function, namely RIG-I and MDA-5, by immunohistochemical techniques and quantitative real-time PCR. IFN-β, IRF-7, RIG-I and MDA-5 were widely detected in most lung cell types. In epithelial tissues and alveolar macrophages, IFN-β and IRF-7 labeling scores were decreased up to 65% and 74%, respectively, for COPD patients, paralleling an analogous reduction (43% and 65%, respectively) in the amount of their lung mRNA. Moreover, this decreased production of IFN-β in COPD patients correlated with a similar decrease in the expression of RIG-I and MDA-5, two essential members of the innate immune system. Our study indicates that most lung cells from stable COPD patients show a constitutive decreased expression of IFN-β, IRF-7, RIG-I and MDA-5, suggesting that this deficiency is the main cause of their acute viral exacerbations.


Three-dimensional telegrapher's equation and its fractional generalization.
We derive the three-dimensional telegrapher's equation out of a random walk model. The model is a three-dimensional version of the multistate random walk where the number of different states form a continuum representing the spatial directions that the walker can take. We set the general equations and solve them for isotropic and uniform walks which finally allows us to obtain the telegrapher's equation in three dimensions. We generalize the isotropic model and the telegrapher's equation to include fractional anomalous transport in three dimensions.


High-grade dysplasia in genital warts from two patients infected with the human immunodeficiency virus.
Cancer-associated human papillomavirus (HPV) types are detected in genital warts removed from immunosuppressed individuals more commonly than from those occurring in otherwise healthy individuals. The prognosis of genital warts containing cancer-associated HPV types is not known. Because it is assumed that genital warts are benign lesions, they are usually treated by destructive therapies without prior knowledge of histopathology. The aim of the present study was to determine whether genital warts from individuals with or without human immunodeficiency virus (HIV) contain high-risk HPV types or areas of dysplasia. The study design was a nonrandomized analysis of genital warts removed by excision biopsy from 15 HIV-infected patients and 15 HIV-negative patients. The tissue was analyzed for HPV DNA by hybrid capture, and microscopic sections of each biopsy were examined for areas of dysplasia. Genital warts from HIV-infected patients contained cancer-associated ("high risk") HPV types in 9 of 15 cases, including 1 that contained only a high-risk type. High-grade dysplastic abnormalities were present in 2 of the 15 lesions from this group, both of which contained high-risk HPV types. Four genital warts removed from HIV-negative patients contained high-risk HPV types, but none contained dysplastic abnormalities. It is concluded that genital warts from HIV-infected patients often contain high-risk HPV types. Such lesions may exhibit dysplastic changes. The frequency of dysplastic changes in genital warts from HIV-infected patients is not known. Biopsy of genital warts may be indicated prior to additional therapy in HIV-infected patients, and surgical removal should be considered as a preferred treatment option in these patients.


Impact of Bacteriophage-Supplemented Drinking Water on the E. coli Population in the Chicken Gut.
Among intestinal coliform microbes in the broiler gut, there are potentially pathogenic Escherichia (E.) coli that can cause avian colibacillosis. The treatment with antibiotics favors the selection of multidrug-resistant bacteria and an alternative to this treatment is urgently required. A chicken model of intestinal colonization with an apathogenic model strain of E. coli was used to test if oral phage application can prevent or reduce the gut colonization of extraintestinal pathogenic E. coli variants in two individual experiments. The E. coli strain E28 was used as a model strain, which could be differentiated from other E. coli strains colonizing the broiler gut, and was susceptible to all cocktail phages applied. In the first trial, a mixture of six phages was continuously applied via drinking water. No reduction of the model E. coli strain E28 occurred, but phage replication could be demonstrated. In the second trial, the applied mixture was limited to the four phages, which showed highest efficacy in vitro. E. coli colonization was reduced in this trial, but again, no reduction of the E. coli strain E28 was observed. The results of the trials presented here can improve the understanding of the effect of phages on single strains in the multi-strain microbiota of the chicken gut.


Neurofibromatosis-Noonan syndrome or LEOPARD Syndrome? A clinical dilemma.
Neurofibromatosis (NF), Noonan syndrome (NS), and LEOPARD syndrome are all autosomal dominant conditions, each being a distinct clinical entity by itself. Rarely, one encounters cases with features of NF and NS and is termed as the 'Neurofibromatosis-Noonan syndrome' (NF-NS). The authors report a clinical dilemma with major clinical features of the NF-NS syndrome and LEOPARD syndrome co-existing in the same patient. Also, features of Noonan syndrome and LEOPARD syndrome are compared with the case reported.


Low cardiac output syndrome requiring extracorporeal membrane oxygenation following pericardiocentesis in an adolescent with Hodgkin Lymphoma: a case report.
We present a case of a 16-year-old male with large pericardial effusion due to Hodgkin Lymphoma (HL). Shortly after drainage of pericardial effusion he developed a low cardiac output syndrome which had to be treated with extracorporeal membrane oxygenation (ECMO). This 9-day ECMO support helped the patient to recover his cardiac function, and thereafter a remission of his primary disease was successfully achieved with chemotherapy. It is a matter of discussion whether a large pericardial effusion with moderate symptoms in patients with HL should be evacuated or just observed since the effusion should ameliorate with chemotherapy. But based upon our experience in this case of hemodynamic instability due to a large effusion requiring evacuation, we propose that pericardiocentesis procedure should be performed with caution at a slow drainage rate of 0.5-1 ml/kg/hour with a maximum rate of 50 ml/hour, to help avoid the low cardiac output syndrome in patients with similar disease conditions.


Significantly mutated genes and regulatory pathways in SCLC-a meta-analysis.
Small cell lung cancer (SCLC) accounts for approximately 15% of all lung cancers and demands effective targeted therapeutic strategies. In this meta-analysis study, we aim to identify significantly mutated genes and regulatory pathways to help us better understand the progression of SCLC and to identify potential biomarkers. Besides ranking genes based on their mutation frequencies, we sought to identify statistically significant mutations in SCLC with the MutSigCV software. Our analysis identified several genes with relatively low mutation frequency, including PTEN, as highly significant (p < 0.001), suggesting these genes may play an important role in the progression of SCLC. Our results also indicated mutations in genes involved in the axon guidance pathways likely play an important role in SCLC progression. In addition, we observed that the mutation rate was significantly higher in samples with RB1 gene mutated when compared to samples with wild type RB1, suggesting that RB1 status has significant impact on the mutation profile and disease progression in SCLC.


Using community volunteers to promote exclusive breastfeeding in Sokoto State, Nigeria.
Exclusive Breastfeeding (EBF) refers to the practice of feeding breast milk only, (including expressed breast milk) to infants; and excluding water, other liquids, breast milk substitutes, and solid foods. Inadequately breastfed infants are likely to be undernourished and have childhood infections. EBF knowledge and infant feeding practices have not been studied sufficiently in Sokoto State, Nigeria. We describe the results of a randomized community trial to promote Exclusive Breastfeeding (EBF) in two local government areas Kware and Bodinga selected as intervention and control groups respectively. During advocacy meetings with community leaders, a Committee was formed. Members of the Committee were consulted for informed consent and selection of ten female volunteers who would educate mothers about breastfeeding during home visits. Participants comprised mothers of infants who were breastfeeding at the time of the study. A total of 179 mothers were recruited through systematic random sampling from each community. Volunteers conducted in-person interviews using a structured questionnaire and counseled mothers in the intervention group only. At baseline, intervention and control groups differed significantly regarding maternal occupation (P=0.07), and age of the index child (P=0.07). 42% of infants in the intervention group were up to 6 months old and about 30% of them were exclusively breastfed. Intention to EBF was significantly associated with maternal age (P=0.01), education (P=0.00) and women who were exclusively breastfeeding (P=0.00). After counseling, all infants up to 6 months of age were exclusively breastfed. The proportion of mothers with intention to EBF increased significantly with maternal age (P=0.00), occupation (P=0.00) and women who were exclusively breastfeeding (P=0.01). Post-intervention surveys showed that source of information and late initiation of breastfeeding was not significantly associated with intention to EBF. Mothers who reported practicing EBF for 6 months, were older (P=0.00) multi-parous (P=0.05) and more educated (P=0.00) compared to those who did not practice EBF. Among them, significantly increased proportion of women agreed that EBF should be continued during the night (P=0.03), infant should be fed on demand (P=0.05), sick child could be given medication (P=0.02), EBF offered protection against childhood diarrhea (P=0.01), and helped mothers with birth spacing (P=0.00). This study shows that there is a need for reaching women with reliable information about infant nutrition in Sokoto State. The results show decreased EBF practice among working mothers, young women, mothers with poor education and fewer than five children. Counseling is a useful strategy for promoting the duration of EBF for six months and for developing support systems for nursing mothers. Working mothers may need additional resources in this setting to enable them to practice EBF.


Contrasting effects of defaunation on aboveground carbon storage across the global tropics.
Defaunation is causing declines of large-seeded animal-dispersed trees in tropical forests worldwide, but whether and how these declines will affect carbon storage across this biome is unclear. Here we show, using a pan-tropical data set, that simulated declines of large-seeded animal-dispersed trees have contrasting effects on aboveground carbon stocks across Earth's tropical forests. In our simulations, African, American and South Asian forests, which have high proportions of animal-dispersed species, consistently show carbon losses (2-12%), but Southeast Asian and Australian forests, where there are more abiotically dispersed species, show little to no carbon losses or marginal gains (±1%). These patterns result primarily from changes in wood volume, and are underlain by consistent relationships in our empirical data (∼2,100 species), wherein, large-seeded animal-dispersed species are larger as adults than small-seeded animal-dispersed species, but are smaller than abiotically dispersed species. Thus, floristic differences and distinct dispersal mode-seed size-adult size combinations can drive contrasting regional responses to defaunation.


Mimicking red blood cell lipid membrane to enhance the hemocompatibility of large-pore mesoporous silica.
Mesoporous silica nanoparticles (MSNs) have been repeatedly demonstrated as potential drug-delivery devices. The study of biocompatibility and interaction of these materials with the various cell types is of great interest with regard to the development of viable pharmaceutical products. By mimicking the cholesterol, phosphatidylcholine, and phosphatidylethanolamine composition of the outer leaflet of a human red blood cell (RBC), lipid-bilayer-coated mesoporous silica particles show considerably improved hemocompatibility over phosphatidylcholine-coated and uncoated large-pore MSN (l-MSN). These inorganic/organic composite nanomaterials are shown to be capable of interfacing with RBCs without damaging the cells even at relatively high concentrations, as observed through electron microscopy, UV-vis spectroscopy, and flow cytometry analyses. Interestingly, the absence of cholesterol in the outer bilayer composition is shown to produce toxic effects without resulting in hemolysis. By maintaining the ζ potential of lipid-bilayer-functionalized MSNs similar to that of the hemolytic l-MSNs, we demonstrate that the bilayer composition, and not the surface charge, plays a significant role in determining the hemocompatibility of MSN-based materials.


Inteleukin-23 promotes interferon-α responsiveness in hepatitis C virus/HIV-coinfected patients.
Patients coinfected with HIV and hepatitis C virus (HCV) have poor to modest rates of response with interferon-based therapies, which remain a backbone of the treatment in HIV/HCV-coinfected patients. The mechanisms responsible for poor responsiveness to interferon are not well described. In this study a targeted proteomic analysis of plasma from 42 patients infected with both HIV and HCV and undergoing therapy for HCV with peginterferon and ribavirin was performed. Higher baseline plasma levels of interleukin (IL)-23 were associated with sustained virologic response. Further investigation of how IL-23 facilitates interferon (IFN) responsiveness, as evidenced by a >2-fold increase in most interferon-stimulated genes (ISGs), revealed that IL-23 indirectly enhances IFN signaling in peripheral blood mononuclear cells and HCV continuous culture system by preventing the down-regulation of the IFNAR2 receptor after exposure to IFN-α. These findings suggest a unique role of the IL-23 pathway in enhancing host response to type I interferons, thereby facilitating eradication of HCV. Low levels of IL-23 present in plasma of nonresponders may reflect an impaired immune state that in the case of HIV/HCV-coinfected subjects could potentially lead to disruption of TH17 CD4(+) T cells. This study suggests a major role for HIV-associated immune dysregulation present in HIV-infected subjects that subsequently determines the overall responsiveness to exogenous interferon-α-based HCV therapy.


An unresponsive progressive pustular and crusting dermatitis with acantholysis in nine cats.
Between 2000 and 2012, nine cats were examined with a visually distinctive, progressive crusting dermatitis that was poorly responsive to all attempted therapies. Documentation of clinical and histopathological findings of this disease. Nine privately owned cats. Retrospective study. Eight neutered males and one (presumably spayed) female ranging in age from two to eight years, presented for a progressive, well-demarcated, crusting dermatitis with variable pruritus of 1.5 months to five years duration. All cats lived in northern California, USA; seven lived within a 30 mile radius. Two males were littermates. Histopathological investigation showed both parakeratotic and orthokeratotic crusts, intraepidermal pustules and superficial folliculitis with rare to frequent acantholytic cells. Bacterial and fungal cultures were performed in six cats: meticillin-susceptible Staphylococcus pseudintermedius was isolated in three cats, two colonies of Trichophyton terrestre and three of Malassezia pachydermatis were isolated from one cat each. Treatment with various antibiotics, antifungal and a variety of immunosuppressive medications did not alter the progressive nature of the skin disease. The described disease shares some clinical and histopathological features with pemphigus foliaceus, but the lack of response to treatment, its progressive nature and the possible relatedness of some of the cats set it apart. The aetiology of this acantholytic dermatitis remains unknown.


Effect of temperature on the assembly of tight junctions and on the mobility of lipids in membranes of HT29 cells.
In the human colon adenocarcinoma cell line HT29, tight junctions can be induced by treatment with appropriate proteases or salt solutions. The temperature dependence of induced tight junction formation is characterized by a marked sigmoidal behavior. The different methods of induction used in this study were characterized by threshold temperatures ranging from 15 to 32 degrees C. Fluorescence photobleaching recovery measurements of the lateral diffusion of a fluorescent phospholipid probe in the cellular plasma membrane gave no evidence for a phase transition or for alteration in the organization of membrane lipids in lateral domains in the temperature range between 0 and 37 degrees C. Moreover, dynamic parameters of the probe in the plasma membrane did not change substantially on mild treatment with trypsin. Thus, the temperature dependence of tight junction formation is not dictated by the bulk properties of the cytoplasmic membrane lipids. The observed temperature dependence suggests that the assembly of tight junctions is a cooperative process, which may involve conformational rearrangement in a protein precursor subsequent to its proteolytic activation.


Lack of efficacy of topotecan in the treatment of metastatic or recurrent squamous carcinoma of the head and neck: an Eastern Cooperative Oncology Group Trial (E3393).
Recurrent or metastatic squamous carcinoma of the head and neck (RMSCHN) is a modestly chemoresponsive tumor; however, currently available agents have failed to improve survival. New active agents are needed for the treatment of this disease. Topotecan is a topoisomerase inhibitor that demonstrated initial promising activity in squamous carcinoma of the head and neck. The Eastern Cooperative Oncology Group conducted a phase II trial of topotecan to determine the efficacy and toxicity of a weekly treatment schedule in patients with RMSCHN. Patients with metastatic or locally recurrent squamous carcinoma of the head and neck were treated with topotecan 1.5 mg/m2 x 24 hours by continuous infusion on days 1, 8, 15, and 22 of each 35-day cycle. Patients were stratified in two cohorts: chemonaive and previously treated. Sixteen chemonaive and 16 previously treated patients were registered on study. Grade III/IV neutropenia and anemia occurred in 16% and 18% of patients, respectively. No responses were observed in either cohort. Median survival for previously untreated patients was 4.6 months and 3.2 months for previously treated patients. Topotecan failed to demonstrate efficacy in patients with RMSCHN. Further evaluation of this agent is not planned.


Asymmetry and variation in the normal hill of vision.
This analysis describes the symmetry and variation in the shape and height of the "hill of vision" obtained from automated static visual field testing of 146 normal eyes of 81 people aged 20 to 78 years. Threshold values dropped from the center to the 30 degrees periphery, with the greatest decrease occurring in the superior quadrant. Variability also increased with distance from the center of fixation, with the greatest variability occurring in the superior quadrant. Threshold values were lower throughout the field in individuals over age 60 years compared with those who were younger, with the greatest decrease in thresholds occurring in the superior quadrant. This age effect was not explained by differences in pupil diameter or central acuity. Variability was higher in the older group, but only in the periphery (greater than 24 degrees from fixation).


A novel microscopic assay for oral malodor-related microorganisms.
Previous research has shown that the production of volatile sulfide compounds (VSC) by oral bacteria is associated with oral malodor. In the present study, we report a novel technique (microscopic sulfide assay (MSA)) for the quantification of VSC-producing oral microorganisms. The MSA was performed by overnight incubation of saliva samples in the presence of ferrous sulfate and sodium thiosulfate, followed by digital analysis of cells stained black due to cell-associated precipitation of ferric sulfide. This method was found to correlate significantly with oral malodor parameters, including mean odor judge scores (two judges, r = 0.48 and p = 0.001) and Halimeter® readings (r = 0.53 and p < 0.001), in a group of 42 subjects. As compared with odor judge scores as the gold standard, the new MSA technique yielded a diagnostic accuracy of 0.7 (ROC, p = 0.023). Results indicate that the MSA may serve as a diagnostic technique for assessing oral malodor levels and aid in identifying the particular bacteria involved in this condition.


Allosteric inhibition of topoisomerase I by pinostrobin: Molecular docking, spectroscopic and topoisomerase I activity studies.
Cancer, the second major cause of mortality trailing the cardiovascular diseases, is a multifactorial heterogeneous disease and growing public health problem worldwide. Owing to severe adverse effects of currently available therapies, there is a growing interest in natural compounds present in our daily diet. Among various natural products, flavonoids-polyphenolic compounds have attracted much attention and have been well-documented for their biological activities. Some flavonoids may inhibit cancer cell proliferation by modulating the action of different enzymes and signal transduction pathways. In the present study, we have evaluated the effect of pinostrobin, a natural flavonoid, on the catalytic activity of topoisomerase I, an essential enzyme for normal DNA replication. Catalytic inhibition of topoisomerase I activity would impair DNA replication of rapidly dividing cancer cells and hence inhibits tumor progression. Pinostrobin interaction with the topoisomerase I and DNA assessed in silico indicated it to form a ternary complex with both. In silico data also suggested pinostrobin to be an effective allosteric inhibitor for topoisomerase I. Further, in vitro investigations such as ethidium bromide displacement assay and spectroscopic studies supported in silico results on the binding of pinostrobin at the interface of topoisomerase I and DNA. Pinostrobin effectively inhibited topoisomerase I activity in vitro further confirming our in silico and in vitro findings. Since topoisomerase I is essential for DNA replication, inhibition of its activity by pinostrobin highlights the therapeutic potential of pinostrobin as an anti-proliferative agent.


Antibiotic prophylaxis for orthopedic prostheses and GI procedures: report of a survey.
To determine the practice recommendations of Program Directors of infectious disease training programs with regard to infection prophylaxis for patients with prosthetic orthopedic devices who undergo gastrointestinal procedures. We surveyed Program Directors of infectious disease training programs to determine what they recommend when asked about antibiotic prophylaxis for patients with orthopedic prostheses who undergo gastrointestinal procedures. More than 50% of the respondents agreed that prophylaxis is not indicated at any time for these procedures, although there was an almost even split when confronted with colonoscopy and polypectomy within 6 months of prosthesis insertion. Most Program Directors agree with the recommendations of the American Society for Gastrointestinal Endoscopy and do not recommend prophylactic antibiotics for these patients. If antibiotics are chosen, they should be the same ones that are recommended for infectious endocarditis by The American Heart Association.


[Proximal humerus fracture - what to do?].
Fracture of the proximal humerus, representing 5 % of all extremity fractures, is a common fracture in everyday clinical life. Apart from the distal fracture of the radius and fractures adjacent to the hip joint, the proximal humerus fracture is the most common fracture in elderly people. From the point of view of evidence-based medicine it is still not possible to define the "gold standard" for stabilisation of fractures of the humeral head. Operative stabilisation of fractures of the humeral head is still a surgical challenge and remains the subject of many clinical and experimental investigations. New findings about the biomechanics and pathophysiology of bones and soft tissue have, in recent years, influenced and pointed the way in the design and function of new forms of osteosynthesis. In particular, the development of locking implants has strongly influenced modern surgical techniques. The aim of this manuscript was to describe the special features of the proximal humerus fracture and its treatment.


Hypoxanthine levels in human urine serve as a screening indicator for the plasma total cholesterol and low-density lipoprotein modulation activities of fermented red pepper paste.
Fermented red pepper paste (FRPP) is one of the most well-known traditional foods in Korea. The effects of FRPP in experimental animals and adipocytes have been previously reported. However, the biochemical effects have not yet been validated in humans with various genetic backgrounds and environmental factors. In this study, 28 female volunteers (body mass index, more than 23 kg/m(2)) aged 19 to 60 years were treated with either FRPP or a placebo for 12 weeks. Marked cholesterol modulation was observed in the FRPP-treated group compared with the placebo group. Although the baseline (pretreatment) total cholesterol and low-density lipoprotein levels and body mass index of the volunteers did not differ significantly between the placebo- and FRPP-treated groups, FRPP caused a modulation of cholesterol levels not seen in the placebo group, causing either no variation or a decrease in low-density lipoprotein and total cholesterol levels. Thus, urinary metabolomic profiling of pretreatment samples was carried out in these 2 FRPP-treated groups using (1)H-nuclear magnetic resonance-based metabolomic techniques. These 2 groups, with their opposing cholesterol-modulation tendencies, could be clearly differentiated by orthogonal projections to latent structures-discriminant analysis-derived score plots. In addition, their levels of hypoxanthine differed markedly. We propose that urinary hypoxanthine levels can be used as a screening biomarker to predict the efficacy of the cholesterol-modulating activity of FRPP in human subjects.


Diastolic wall strain: a simple marker of abnormal cardiac mechanics.
Diastolic wall strain (DWS), defined using posterior wall thickness (PWT) measurements from standard echocardiographic images (DWS = [PWT(systole)-PWT(diastole)]/PWT(systole)), has been proposed as a marker of left ventricular (LV) diastolic stiffness. However, the equation for DWS is closely related to systolic radial strain, and whether DWS is associated with abnormal cardiac mechanics (reduced systolic strains and diastolic tissue velocities) is unknown. We sought to determine the relationship between DWS and systolic and diastolic cardiac mechanics. We calculated DWS and performed speckle-tracking analysis in a large population- and family-based study (Hypertension Genetic Epidemiology Network [HyperGEN]; N=1907 after excluding patients with ejection fraction [EF] <50% or posterior wall motion abnormalities). We measured global longitudinal, circumferential, and radial strain (GLS, GCS, and GRS, respectively) and early diastolic (e') tissue velocities, and we determined the independent association of DWS with cardiac mechanics using linear mixed effects models to account for relatedness among study participants. We also prospectively performed receiver-operating characteristic (ROC) analysis of DWS for the detection of abnormal cardiac mechanics in a separate, prospective validation study (N=35). In HyperGEN (age 51 ± 14 years, 59% female, 45% African-American, 57% hypertensive), mean DWS was 0.38 ± 0.05. DWS decreased with increasing comorbidity burden (β-coefficient -0.013 [95% CI -0.015, -0.011]; P<0.0001). DWS was independently associated with GLS, GCS, GRS, and e' velocity (adjusted P<0.05) but not LV chamber compliance (EDV20, P=0.97). On prospective speckle-tracking analysis, DWS correlated well with GLS, GCS, and GRS (R=0.61, 0.57, and 0.73, respectively; P<0.001 for all comparisons). C-statistics for DWS as a diagnostic test for abnormal GLS, GCS, and GRS were: 0.78, 0.79, and 0.84, respectively. DWS, a simple parameter than can be calculated from routine 2D echocardiography, is closely associated with systolic strain parameters and early diastolic (e') tissue velocities but not LV chamber compliance.


Simultaneous repetitive transcranial magnetic stimulation does not speed fine movement in PD.
To reevaluate the effect of subthreshold repetitive transcranial magnetic stimulation (rTMS) on concurrent fine movement in PD. A previous study showed a beneficial effect of rTMS on the performance of six patients with PD on the Grooved Pegboard test. The authors repeated this experiment in 11 patients with idiopathic PD who performed the test while the stimulating coil discharged continuously at 5 Hz, either over the contralateral motor cortex at just below the threshold for movement, or in the air near the head as a control. Patients were tested twice under both conditions. Although some patients performed faster with rTMS, others showed the opposite effect. There was no significant effect of rTMS in the group, nor did baseline performance or the order of conditions interact with the effect of rTMS. Based on this larger and more comprehensive study, simultaneous, 5-Hz subthreshold rTMS over the motor cortex does not have consistent or potentially therapeutic effects on movement in PD.


Long-Term In Vitro Expansion of Salivary Gland Stem Cells Driven by Wnt Signals.
Adult stem cells are the ultimate source for replenishment of salivary gland (SG) tissue. Self-renewal ability of stem cells is dependent on extrinsic niche signals that have not been unraveled for the SG. The ductal compartment in SG has been identified as the location harboring stem cells. Here, we report that rare SG ductal EpCAM(+) cells express nuclear β-catenin, indicating active Wnt signaling. In cell culture experiments, EpCAM(high) cells respond potently to Wnt signals stimulating self-renewal and long-term expansion of SG organoids, containing all differentiated SG cell types. Conversely, Wnt inhibition ablated long-term organoid cultures. Finally, transplantation of cells pre-treated with Wnt agonists into submandibular glands of irradiated mice successfully and robustly restored saliva secretion and increased the number of functional acini in vivo. Collectively, these results identify Wnt signaling as a key driver of adult SG stem cells, allowing extensive in vitro expansion and enabling restoration of SG function upon transplantation.


Staphylococcus aureus Shifts toward Commensalism in Response to Corynebacterium Species.
Staphylococcus aureus-human interactions result in a continuum of outcomes from commensalism to pathogenesis. S. aureus is a clinically important pathogen that asymptomatically colonizes ~25% of humans as a member of the nostril and skin microbiota, where it resides with other bacteria including commensal Corynebacterium species. Commensal Corynebacterium spp. are also positively correlated with S. aureus in chronic polymicrobial diabetic foot infections, distinct from acute monomicrobial S. aureus infections. Recent work by our lab and others indicates that microbe-microbe interactions between S. aureus and human skin/nasal commensals, including Corynebacterium species, affect S. aureus behavior and fitness. Thus, we hypothesized that S. aureus interactions with Corynebacterium spp. diminish S. aureus virulence. We tested this by assaying for changes in S. aureus gene expression during in vitro mono- versus coculture with Corynebacterium striatum, a common skin and nasal commensal. We observed a broad shift in S. aureus gene transcription during in vitro growth with C. striatum, including increased transcription of genes known to exhibit increased expression during human nasal colonization and decreased transcription of virulence genes. S. aureus uses several regulatory pathways to transition between commensal and pathogenic states. One of these, the quorum signal accessory gene regulator (agr) system, was strongly inhibited in response to Corynebacterium spp. Phenotypically, S. aureus exposed to C. striatum exhibited increased adhesion to epithelial cells, reflecting a commensal state, and decreased hemolysin activity, reflecting an attenuation of virulence. Consistent with this, S. aureus displayed diminished fitness in experimental in vivo coinfection with C. striatum when compared to monoinfection. These data support a model in which S. aureus shifts from virulence toward a commensal state when exposed to commensal Corynebacterium species.


Structural dynamics of the box C/D RNA kink-turn and its complex with proteins: the role of the A-minor 0 interaction, long-residency water bridges, and structural ion-binding sites revealed by molecular simulations.
Kink-turns (K-turns) are recurrent elbow-like RNA motifs that participate in protein-assisted RNA folding and contribute to RNA dynamics. We carried out a set of molecular dynamics (MD) simulations using parm99 and parmbsc0 force fields to investigate structural dynamics of the box C/D RNA and its complexes with two proteins: native archaeal L7ae protein and human 15.5 kDa protein, originally bound to very similar structure of U4 snRNA. The box C/D RNA forms K-turn with A-minor 0 tertiary interaction between its canonical (C) and noncanonical (NC) stems. The local K-turn architecture is thus different from the previously studied ribosomal K-turns 38 and 42 having A-minor I interaction. The simulations reveal visible structural dynamics of this tertiary interaction involving altogether six substates which substantially contribute to the elbow-like flexibility of the K-turn. The interaction can even temporarily shift to the A-minor I type pattern; however, this is associated with distortion of the G/A base pair in the NC-stem of the K-turn. The simulations show reduction of the K-turn flexibility upon protein binding. The protein interacts with the apex of the K-turn and with the NC-stem. The protein-RNA interface includes long-residency hydration sites. We have also found long-residency hydration sites and major ion-binding sites associated with the K-turn itself. The overall topology of the K-turn remains stable in all simulations. However, in simulations of free K-turn, we observed instability of the key C16(O2')-A7(N1) H-bond, which is a signature interaction of K-turns and which was visibly more stable in simulations of K-turns possessing A-minor I interaction. It may reflect either some imbalance of the force field or it may be a correct indication of early stages of unfolding since this K-turn requires protein binding for its stabilization. Interestingly, the 16(O2')-7(N1) H- bond is usually not fully lost since it is replaced by a water bridge with a tightly bound water, which is adenine-specific similarly as the original interaction. The 16(O2')-7(N1) H-bond is stabilized by protein binding. The stabilizing effect is more visible with the human 15.5 kDa protein, which is attributed to valine to arginine substitution in the binding site. The behavior of the A-minor interaction is force-field-dependent because the parmbsc0 force field attenuates the A-minor fluctuations compared to parm99 simulations. Behavior of other regions of the box C/D RNA is not sensitive to the force field choice. Simulation with net-neutralizing Na(+) and 0.2 M excess salt conditions appear in all aspects equivalent. The simulations show loss of a hairpin tetraloop, which is not part of the K-turn. This was attributed to force field limitations.


[Heart valve disease evaluated with MRI].
Patients with heart valve disease are evaluated with non-invasive imaging techniques notably echocardiography. Cardiovascular MRI (CMR) is increasingly applied, not only in case of a poor echocardiographic window, but also as a consequence of CMR's possibilities of precise determination of regurgitation volumes in insufficiency and precise determination of the impact of valve disease on corres-ponding heart chambers including chamber volumes, myocardial masse and fibrosis. In this paper, the emerging role of CMR in evaluation of patients with heart valve disease is reviewed.


Crystal structures and properties of de novo circularly permuted 1,3-1,4-beta-glucanases.
The 1,3-1,4-beta-glucanases from Bacillus macerans and Bacillus licheniformis, as well as related hybrid enzymes, are stable proteins comprised of one compact jellyroll domain. Their structures are studied in an effort to reveal the degree of redundancy to which the three-dimensional structure of protein domains is encoded by the amino acid sequence. For the hybrid 1,3-1,4-beta-glucanase H(A16-M), it could be shown recently that a circular permutation of the sequence giving rise to the variant cpA16M-59 is compatible with wildtype-like enzymatic activity and tertiary structure (Hahn et al., Proc. Natl. Acad. Sci. USA 91:10417-10421, 1994). Since the circular permutation yielding cpA16M-59 mimicks that found in the homologous enzyme from Fibrobacter succinogenes, the question arose whether de novo circular permutations, not guided by molecular evolution of the 1,3-1,4-beta-glucanases, could also produce proteins with native-like fold. The circularly permuted variants cpA16M-84, cpA16M-127, and cpA16M-154 were generated by PCR mutagenesis of the gene encoding H(A16-M), synthesized in Escherichia coli and shown to be active in beta-glucan hydrolysis. CpA16M-84 and cpA16M-127 were crystallized in space groups P2(1) and P1, respectively, and their crystal structures were determined at 1.80 and 2.07 A resolution. In both proteins the main parts of the beta-sheet structure remain unaffected by the circular permutation as is evident from a root-mean-square deviation of main chain atoms from the reference structure within the experimental error. The only major structural perturbation occurs near the novel chain termini in a surface loop of cpA16M-84, which becomes destabilized and rearranged. The results of this study are interpreted to show that: (1) several circular permutations in the compact jellyroll domain of the 1,3-1,4-beta-glucanases are tolerated without radical change of enzymatic activity or tertiary structure, (2) the three-dimensional structures of simple domains are encoded by the amino acid sequence with sufficient redundancy to tolerate a change in the sequential order of secondary structure elements along the sequence, and (3) the native N-terminal region is not needed to guide the folding polypeptide chain toward its native conformation.


[The anodically moving thermolabile antigen (ATA) of gram-negative bacteria: its molecular heterogeneity and its chemical composition].
The anodically-moving thermolabile antigen (ATA) of gram-negative bacteria represents a glycolipoprote in. In aqueous solution ATA exhibits heterogeneity of the molecular size, the range varying between 2.10(4) and 8.10(5) Daltons. In the molecular range between 5.10(4) and 8.10(5) all the molecular states give precipitation lines with the corresponding antisera. It could be shown that all the molecular states of ATA are formed by different aggregation of the same subunit with a molecular weight of approximatively 22000.


Disturbed Laminar Blood Flow Vastly Augments Lipoprotein Retention in the Artery Wall: A Key Mechanism Distinguishing Susceptible From Resistant Sites.
Atherosclerosis develops initially at branch points and in areas of high vessel curvature. Moreover, experiments in hypercholesterolemic mice have shown that the introduction of disturbed flow in straight, atherosclerosis-resistant arterial segments turns them highly atherosclerosis susceptible. Several biomechanical mechanisms have been proposed, but none has been demonstrated. In the present study, we examined whether a causal link exists between disturbed laminar flow and the ability of the arterial wall to retain lipoproteins. Lipoprotein retention was detected at natural predilection sites of the murine thoracic aorta 18 hours after infusion of fluorescently labeled low-density lipoprotein. To test for causality between blood flow and the ability of these areas to retain lipoproteins, we manipulated blood flow in the straight segment of the common carotid artery using a constrictive collar. Disturbed laminar flow did not affect low-density lipoprotein influx, but increased the ability of the artery wall to bind low-density lipoprotein. Concordantly, disturbed laminar flow led to differential expression of genes associated with phenotypic modulation of vascular smooth muscle cells, increased expression of proteoglycan core proteins associated with lipoprotein retention, and of enzymes responsible for chondroitin sulfate glycosaminoglycan synthesis and sulfation. Blood flow regulates genes associated with vascular smooth muscle cell phenotypic modulation, as well as the expression and post-translational modification of lipoprotein-binding proteoglycan core proteins, and the introduction of disturbed laminar flow vastly augments the ability of a previously resistant, straight arterial segment to retain lipoproteins.


Effect of increasing blood flow on distribution of pulmonary transit times in man.
The architecture of the vascular network is a major determinant of the distribution of transit times in the organ. Changes in pulmonary blood volumes, pulmonary transit times and the heterogeneity (coefficient of variation) of pulmonary transit times in man were measured using the first-pass radiotracer technique both at rest and during muscular exercise. Dichotomously branching and dispersive flow models of the lung vasculature were applied. The results showed that total pulmonary blood volume increased by 29%, total transit time decreased by 55% and the heterogeneity of pulmonary transit times decreased by 31% when blood flow increased threefold. All changes obeyed power law functions. The changes were greatest when blood flow increased from baseline to intermediate levels. Further increases in total blood flow were associated with decreases in transit times but not with more homogeneous distribution of transit times. The simulated results suggest that the transit time through the arterial vessels and the large veins have a negligible heterogeneity. Most of the variability of transit times through the lungs is due to the heterogeneity of the transit times through the capillaries and venules.


The DGAT1 K232A polymorphism and feeding modify milk fat triacylglycerol composition.
In the present study, we aimed to investigate the changes in triacylglycerol (TAG) composition as affected by alterations in the cows' diet due to seasonal variations and genetic factors. For this study, 50 milk fat samples in winter and 50 in summer were used from 25 cows with the DGAT1 KK genotype and 25 cows with the DGAT1 AA genotype. The samples were analyzed for milk fat content (%), fat composition, and TAG composition. We found that the content of TAG species CN54 was higher and that of CN34 and CN36 lower in summer than in winter. This seasonal variation in TAG profile was related to seasonal changes in the fatty acids C14:0, C16:0, C18:0, C18:1 cis-9, total unsaturated fatty acids, and total long-chain fatty acids, most likely resulting from dietary differences between seasons. Furthermore, we quantified the effect of DGAT1 K232A polymorphism on TAG profile and detected a significant effect on TAG species CN36, with higher values for the DGAT1 KK genotype. When adjusting for differences in fat content, we found no significant effects of the DGAT1 K232A polymorphism on TAG profile. We detected a significant interaction between DGAT1 K232A polymorphism and season for TAG species CN42 and CN52; in summer, the KK genotype was associated with higher levels for CN42 than the AA genotype, whereas in winter, the difference between the genotypes was small. For CN52, in summer the AA genotype was associated with higher levels than the KK genotype. In winter, the difference between the genotypes was also small. We show that, regardless of preference for DGAT1 genotype (AA or KK) and depending on the availability of FA according to season, UFA (C18:1 cis-9), short-chain FA (C6:0 and C10:0), and medium-chain FA might be esterified on the glycerol backbone of the TAG, keeping the structure characteristics of each TAG species. To our knowledge, this is the first report on the interaction effect of DGAT1 K232A polymorphism and season on the TAG composition in milk fat.