Enhanced in vivo antitumor efficacy of dual-functional peptide-modified docetaxel nanoparticles through tumor targeting and Hsp90 inhibition.
Although conventional anticancer drugs exhibit excellent efficacy, serious adverse effects and/or even toxicity have occurred due to their nonselectivity. Moreover, active targeting approaches have not consistently led to successful outcomes. Ligands that simultaneously possess targeting capability and exert a strong influence on intracellular signaling cascades may be expected to improve the therapeutic efficacy of active targeting nanoparticulate carriers. In this study, we screened a targeting peptide, LPLTPLP, which specifically bound to non-small cell lung cancer (NSCLC) specimens in vitro. Surprisingly, this peptide inhibited the expression of Hsp90 and induced apoptosis by preventing autophagy in A549 cells treated with docetaxel. The results suggested that this peptide might be used as a promising dual-functional ligand for cancer treatment. Based on these findings, we designed and developed a novel active targeting delivery system by modifying docetaxel nanoparticles (DNP) with the dual-functional ligand LPLTPLP. We consistently demonstrated that the cellular uptake of nanoparticles (NPs) was significantly enhanced in vitro. Furthermore, the targeting NPs exhibited significantly improved antitumor efficacy and biodistribution compared with nontargeting nanodrug and free docetaxel. These findings demonstrate the feasibility of dual-functional NPs for efficient anticancer therapy.


Facilitators of and barriers to return to work after mild traumatic brain injury: A thematic analysis.
Individuals with mild traumatic brain injury (mTBI) may experience protracted symptoms affecting their ability to work. Several actors may facilitate or act as a barrier to a successful return to work (RTW). This qualitative study used semi-structured in-depth interviews with 22 adults with mTBI at 2-5 years post injury, targeting experiences of how different actors facilitated or acted as a barrier in the RTW process, and encompassed the mTBI trajectory from the acute phase to the post-acute phase. A thematic analysis with a hermeneutical phenomenological approach was used to analyse data. Three main themes emerged. (1) Worker-employer relationship: Workplace accommodations such as decreased working hours, modified working conditions, and support from co-workers were lacking. (2) The role of the general practitioner: The general practitioner was lacking treatment and referral opportunities and failed to provide the patient with relevant and individualized guidance. (3) Municipal case management: Participants perceived being met with distrust by social workers, follow-up assessments were too frequent, unnecessary, and did not target concussion, and rehabilitation was referred too late. Clinical practice guidelines to improve referral to relevant concussion rehabilitation in case of persistent symptoms are needed to inform clinicians, employers, and public institutions to facilitate a successful RTW.


Balancing the gastrointestinal benefits and risks of nonselective NSAIDs.
Nonselective nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most widely used classes of medications to treat pain and inflammation. However, gastrointestinal complications associated with NSAIDs are prevalent, largely due to the frequent use of these agents. Adverse events associated with NSAIDs include minor side effects, such as dyspepsia, as well as serious complications, such as bleeding and perforation. Although the probability that any given individual user of an NSAID will suffer a serious gastrointestinal complication is fairly low, widespread patient exposure can translate into a major national health burden. The increasing use of aspirin in the prevention of cardiovascular events and the availability of select over-the-counter NSAIDs represent additional challenges to clinicians in their efforts to make the most appropriate therapeutic decisions while minimizing the potential gastrointestinal risks associated with the use of these agents. Side effects such as dyspepsia do not provide adequate warning of gastrointestinal complications, because most complications occur without the presence of antecedent symptoms. Therefore, accurate risk assessment and the management of controllable risk factors are crucial to the safe administration of NSAIDs. This review focuses on the gastrointestinal effects of aspirin, acetaminophen, and other nonselective NSAIDs, and discusses those factors that are associated with increased risk for adverse gastrointestinal events in certain individuals.


Dissatisfaction versus over-evaluation in a general population sample of women.
We compared levels of eating disorder psychopathology, general psychological distress and impairment in role functioning among subgroups of women, recruited from a large, general population sample, who reported dissatisfaction with weight or shape but not over-evaluation of weight or shape (n = 482) or over-evaluation but not dissatisfaction (n = 105). Self-report questionnaires that included measures of each outcome, as well as height and weight and socio-demographic information, were completed by all participants. Participants who reported dissatisfaction but not over-evaluation were older, heavier and had higher levels of weight or shape concerns, higher levels of general psychological distress, and poorer physical health than those who reported over-evaluation but not dissatisfaction. However, only differences with respect to weight or shape concerns remained significant after age and BMI were statistically controlled. In multivariable analysis, dissatisfaction with weight or shape made a stronger contribution to variance in all three outcomes-eating disorder psychopathology, general psychological distress and functional impairment-than over-evaluation. Although the findings are consistent with the view that over-evaluation and dissatisfaction are distinct constructs, there was no evidence to support the premise that overevaluation is more "pathological" than dissatisfaction among women in the general population.


High prevalence of occult pulmonary venous hypertension revealed by fluid challenge in pulmonary hypertension.
Determining the cause for pulmonary hypertension is difficult in many patients. Pulmonary arterial hypertension (PAH) is differentiated from pulmonary venous hypertension (PVH) by a wedge pressure (PWP)>15 mm Hg in PVH. Patients undergoing right heart catheterization for evaluation of pulmonary hypertension may be dehydrated and have reduced intravascular volume, potentially leading to a falsely low measurement of PWP and an erroneous diagnosis of PAH. We hypothesized that a fluid challenge during right heart catheterization would identify occult pulmonary venous hypertension (OPVH). We reviewed the results of patients undergoing fluid challenge in our pulmonary hypertension database from 2004 to 2011. Baseline hemodynamics were obtained and repeated after infusion of 0.5 L of normal saline for 5 to 10 minutes. Patients were categorized as OPVH if PWP increased to >15 mm Hg after fluid challenge. Baseline hemodynamics in 207 patients met criteria for PAH. After fluid challenge, 46 patients (22.2%) developed a PWP>15 mm Hg and were reclassified as OPVH. Patients with OPVH had a greater increase in PWP compared with patients with PAH, P<0.001, and their demographics and comorbid illnesses were similar to patients with PVH. There were no adverse events related to fluid challenge. Fluid challenge at the time of right heart catheterization is easily performed, safe, and identifies a large group of patients diagnosed initially with PAH, but for whom OPVH contributes to pulmonary hypertension. These results have implications for therapeutic trials in PAH and support the routine use of fluid challenge during right heart catheterization in patients with risk factors for PVH.


Metabolites of a tobacco-specific nitrosamine, 4-(methylnitrosamino)- 1-(3-pyridyl)-1-butanone (NNK), in the urine of smokeless tobacco users: relationship between urinary biomarkers and oral leukoplakia.
Two metabolites of the carcinogenic tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone were quantified in the urine of smokeless tobacco users. The metabolites are 4-(methylnitrosamino) -1-(3-pyridyl)-1-butanol (NNAL) and [4-(methylnitrosamino)-1-(3-pyridyl) but-1-yl]-beta-O-D-glucosiduronic acid (NNAL-Gluc). The study population consisted of 47 male nonsmokers, of whom 23 were snuff dippers, 13 were tobacco chewers, 3 were users of both products, and 8 were nonusers. The levels of NNAL-Gluc in urine ranged from 0.14-30.3 pmol/mg creatinine with a mean +/- SD of 3.47 +/- 5.86, whereas the levels of NNAL ranged from 0.02-8.73 pmol/mg creatinine with a mean +/- SD of 0.92 +/- 1.59. The mean levels of NNAL-Gluc and NNAL were not significantly different from those measured in a previous study of smokers. The levels of NNAL-Gluc were significantly higher in snuff dippers than in tobacco chewers. The ratio of NNAL-Gluc:NNAL was higher in snuff dippers than in tobacco chewers or smokers. There was no indication of two phenotypes of the NNAL-Gluc:NNAL ratio in smokeless tobacco users, in contrast to previous observations in smokers. Of the 39 smokeless tobacco users in this study, 16 presented with oral leukoplakia. When the total levels of NNAL-Gluc, NNAL, or NNAL-Gluc + NNAL were divided into tertiles, there was a significant association between the presence of leukoplakia and increasing levels of these metabolites; a similar relationship was found between urinary cotinine and leukoplakia. The results of this study demonstrate that there is significant uptake of carcinogenic nitrosamines in smokeless tobacco users, and that such products are not harmless alternatives to cigarettes. Moreover, the urinary biomarkers NNAL-Gluc, NNAL, and cotinine were associated with the presence of leukoplakia, which provides biochemical support for the role of smokeless tobacco products as a cause of oral leukoplakia.


Destruction of ERP responses to deviance in an auditory oddball paradigm in amyloid infusion mice with memory deficits.
The amyloid-β (Aβ) oligomer is considered one of the major pathogens responsible for neuronal and synaptic loss in Alzheimer's disease (AD) brains. Although the neurotoxic mechanisms of Aβ have been widely investigated, experimental evidence for the direct linkage between neural signaling and cognitive impairments in association with peptide oligomers is lacking. Here, we conducted an auditory oddball paradigm utilizing an Aβ-infused Alzheimer's disease mouse model and interpreted the results based on Y-maze behavioral tests. We acutely injected Aβ oligomers into the intracerebroventricular brain region of normal mice to induce Aβ-associated cognitive impairments. During the auditory oddball paradigm, electroencephalograms (EEG) were recorded from frontal and parietal cortex of Aβ-infused and control mice. The event-related potentials (ERPs) elicited by auditory stimuli showed no significant difference in Aβ-infused mice compared to control mice. On the other hand, the differential ERP signature elicited by oddball sound stimuli was destructed in the Aβ-infused mice group. We noticed that ERP traces to standard and deviant tones were not significantly different in the Aβ group, while the control group showed differences in the amplitude of ERP components. In particular, the difference in the first negative component (N1) between standard and deviant tone, which indexes the sensory memory system, was significantly reduced in the parietal cortex of Aβ-infused mice. These findings demonstrate the direct influence of Aβ oligomers on the functional integrity of cortical areas in vivo. Furthermore, the N1 amplitude difference may provide a potential marker of sensory memory deficits in a mouse model of AD and yield additional targets for drug assessment in AD.


Differential expression of silent polyketide biosynthesis gene clusters in chemostat cultures of Aspergillus nidulans.
The genome of the fungal model organism Aspergillus nidulans harbors nearly 30 polyketide synthase genes, yet the majority of these genes remain silent in the absence of particular stimuli. In this study, environmental conditions such as low specific microbial growth rate as well as nitrate, orthophosphate and glucose limitations were simulated under a continuous cultivation regime to induce the expression of silent polyketide synthase genes. In addition to offline and online bioprocess parameters, the physiological equilibrium was defined at the transcript level in terms of indicator gene expression. The different cultivation parameters resulted in a differential expression of two polyketide synthase genes coding for the biosynthesis of a variety of phenolic compounds, such as orsellinic acid, lecanoric acid, emodins, chrysophanol, shamixanthone, and sanghaspirodin. Further investigation of the metabolome revealed the formation of a novel prenylated benzophenone derivative designated as pre-shamixanthone. Our data indicate that employing chemostat fermentations in combination with genome mining, transcriptome analysis and metabolic profiling represents a valuable approach for triggering cryptic biosynthetic pathways.


Genetic background influences cataractogenesis, but not lens growth deficiency, in Cx50-knockout mice.
Deletion of connexin (Cx)50 produces microphthalmia with nuclear cataracts. To determine whether these two traits are influenced by genetic background and are dependent on each other, mice carrying the Cx50 deletion in two different strains were generated, and the growth defect and severity of cataracts were analyzed. Cx50-knockout mice were generated in the 129S6 strain, and back-crossed into the C57BL/6J genetic background. To analyze the influence of genetic background on the observed phenotype, postnatal lens growth, lens clarity, lens histology and crystallin solubility were determined and compared between the two strains of Cx50-knockout mice. The growth deficiency persisted, regardless of genetic background, but genetic modifiers that differentially altered the solubility of crystallin proteins influenced the severity of cataracts. Expression levels of Cx46 were similar in all animals, regardless of genetic background, indicating that the differences were not due to a compensatory upregulation of Cx46. Taken together, these data indicate that the two components of the Cx50 phenotype are independent of each other and that cataractogenesis is under the influence of an unidentified genetic modifier.


Adrenergic regulation of blood glucose levels: possible involvement of postsynaptic alpha-2 type adrenergic receptors regulating insulin release.
We have demonstrated that while phenoxybenzamine and prazosin (10(-7)-10(-4) mol/kg i.p.) are ineffective in inhibiting the hyperglycemia induced by alpha adrenergic agonists in fed male mice, phentolamine, yohimbine and dihydroergotamine are effective. These results suggest the involvement of alpha-2 type adrenergic receptors in the regulation of the plasma glucose levels. When attempting to elucidate the mechanism of such differences, we found that the release of immunoreactive insulin induced by epinephrine was markedly stimulated by phentolamine, yohimbine and dihydroergotamine, whereas phenoxybenzamine and prazosin had little effect. The ineffectiveness of these two compounds to inhibit the hyperglycemia induced by alpha adrenergic agonists indicates that the postsynaptic alpha adrenergic receptors which suppress the secretion of immunoreactive insulin from B cells of pancreatic islets are not blocked and that the alpha adrenergic receptors which control immunoreactive insulin secretion differ from the classical postsynaptic alpha adrenergic receptors and more closely resemble presynaptic alpha-2 adrenergic receptors.


APPL proteins link Rab5 to nuclear signal transduction via an endosomal compartment.
Signals generated in response to extracellular stimuli at the plasma membrane are transmitted through cytoplasmic transduction cascades to the nucleus. We report the identification of a pathway directly linking the small GTPase Rab5, a key regulator of endocytosis, to signal transduction and mitogenesis. This pathway operates via APPL1 and APPL2, two Rab5 effectors, which reside on a subpopulation of endosomes. In response to extracellular stimuli such as EGF and oxidative stress, APPL1 translocates from the membranes to the nucleus where it interacts with the nucleosome remodeling and histone deacetylase multiprotein complex NuRD/MeCP1, an established regulator of chromatin structure and gene expression. Both APPL1 and APPL2 are essential for cell proliferation and their function requires Rab5 binding. Our findings identify an endosomal compartment bearing Rab5 and APPL proteins as an intermediate in signaling between the plasma membrane and the nucleus.


[Simultaneous determination of bisphenol A and tetrabromobisphenol A in plastic products using high performance liquid chromatography-mass spectrometry].
A method for the determination of bisphenol A (BPA) and tetrabromobisphenol A (TBBP-A) in plastic products using high performance liquid chromatography-mass spectrometry (HPLC-MS) was developed with the ultrasonic extraction. The conditions of sample pretreatment, chromatography and mass-spectrometry parameters were studied systematically. The research showed that the extraction of the BPA and TBBP-A can reach a good performance with 20 mL dichloromethane at 50 degrees C under ultrasonic extraction for 60 min. The BPA and TBBP-A were separated and detected using HPLC-MS with the mobile phase of methanol and water. The linearity range of the method was 0.1-2.0 mg/L. The detection limits were 0.01 mg/kg for BPA and 0.02 mg/kg for TBBP-A. The recoveries for the spiked samples were 85.4%-97.6%. This method is time-saving, easy-handling, practicable and sensitive, and it can be applied to determine the residues of BPA and TBBP-A in plastic products.


Ultrasound-guided transrectal drainage of deep pelvic abscesses in children: a modified and simplified technique.
Appendicitis is a common cause of acute surgical abdomen in children and often presents after perforation. Intra-abdominal abscesses can be drained percutaneously via transabdominal, transgluteal or, in the case of deep pelvis abscess, via transrectal approach. To describe a modification of previously described techniques for transrectal drainage procedures, which involves the use of a transvaginal probe and a modified enema tip as a guide for the one-step trocar technique. We also aimed to evaluate the safety and effectiveness of this modified technique for drainage of deep pelvic abscesses in children. A retrospective review of medical records was performed to identify all patients who underwent transrectal abscess drainage at our pediatric institution during a 5-year period. Surgical and radiologic procedure notes and imaging studies were evaluated. The data were analyzed to determine technical and clinical success rates, and to evaluate for any procedure-related complication. The study population consisted of 46 patients with a mean age of 10.9 years. Of the 46 children, 20 underwent transrectal abscess drainage solely using transrectal US for guidance; 2 also underwent minimal fluoroscopy at the time of transrectal drainage. Fifteen children required placement of one or more percutaneous transabdominal drains at the same time as transrectal catheter placement, and nine required addition percutaneous drainage catheters placed at another time. All transrectal drainage procedures were technically and clinically successful. There were no procedure-related complications. The described modified technique for US-guided transrectal drainage of deep pelvic abscesses resulting from perforated appendicitis in children is safe, effective and relatively easy to perform, with the added benefit of omitting radiation exposure in children.


Serum levels of advanced glycation end products are associated with in-stent restenosis in diabetic patients.
The formation of advanced glycation end products (AGEs), in various tissues has been known to enhance immunoinflammatory reactions and local oxidant stresses in long standing diabetes. Recently, AGEs have been reported to play a role in neointimal formation in animal models of arterial injury. We attempted to determine whether the serum levels of AGEs are associated with coronary restenosis in diabetic patients. Blood samples were collected from diabetic patients with coronary artery disease undergoing stent implantation and the serum levels of AGEs were analyzed by the fluorescent intensity method. The development of in-stent restenosis (ISR) was evaluated by a 6-month follow-up coronary angiography. A total of 263 target lesions were evaluated, in 203 patients. The ISR rate in the high-AGE (>170 U/ml) group (40.1%) was significantly higher than in the low-AGE group (<or=170 U/ml) (19.6%) (p<0.001). Furthermore, multivariate analysis revealed that a high level of serum AGEs is an independent risk factor for the development of ISR (odds ratio, 2.659; 95% CI, 1.431-4.940; p=0.002). The serum levels of AGEs constitute an excellent predictive factor for ISR, and should be one of the guidelines for medical therapy and interventional strategy to prevent ISR in diabetic patients.


Neuromuscular blockade and inspiratory breath hold during stereotactic body radiation therapy for treatment of heart base tumors in four dogs.
CASE DESCRIPTION 4 dogs were examined because of pleural effusion and ventricular tachycardia, coughing and supraventricular tachycardia, appendicular osteosarcoma, and syncopal episodes. CLINICAL FINDINGS In all 4 dogs, a heart base tumor was identified by means of thoracic CT. TREATMENT AND OUTCOME In all 4 dogs, the heart base tumors were treated by means of stereotactic body radiation therapy. Dogs were anesthetized, and neuromuscular blockade was achieved with atracurium or vecuronium. A circle rebreathing system with 15 m (50 feet) of anesthetic tubing coursing through the vault wall was used to connect the patient to the anesthesia machine, which was located in the control room. After a brief period of hyperventilation, an inspiratory breath was held at 20 cm H2O for the duration of beam delivery. Each beam delivery lasted between 30 and 100 seconds. Immediately following the breath hold, assisted ventilation was resumed. Mean treatment delivery time for each patient was 26 minutes; mean total anesthesia time was 89 minutes. All patients recovered without complications. There was no evidence of hemoglobin desaturation or hypercapnia during the anesthetic procedure. CLINICAL RELEVANCE The technique allowed for control of the respiration cycle from outside the radiation vault and a short overall treatment time. No adverse effects were encountered. This procedure should be considered when delivering radiation to structures within the thoracic cavity.


Can antibiotic use be both just and sustainable... or only more or less so?
Antibiotic resistance threatens the capacity to treat life-threatening infections. If it is accepted that it will be many years (if not decades) until the production of new antibiotics overcomes current concerns with antibiotic resistance then ways to conserve the effectiveness of current antibiotics will have to be found. For many bacterial agents of infection levels of antibiotic resistance are directly dependent on the quantity of antibiotic prescribed. Antibiotics are currently underutilised in many parts of the world. If a just distribution of access to antibiotics requires equal access for individuals with equal need irrespective of wealth then responding to this requirement of justice has the potential to shorten the effective life of currently available antibiotics. Increasing the range and numbers of individuals treated with antibiotics would seem to threaten sustainability and also potentially undermine the access of future generations to cost-effective treatments for bacterial infection. The control of antibiotic resistance requires that the determinants of infectious disease transmission are addressed, such as poor housing, education and nutrition as well as the provision of antibiotics. The apparent tension between intragenerational justice and sustainability diminishes when the account of distributive justice extends beyond access to antibiotics and includes plural entitlements. Controlling antibiotic resistance requires more than the redistribution or reduction (in the overall use) of antibiotics.


Integrating phenotypic data for depression.
The golden era of molecular genetic research brings about an explosion of phenotypic, genotypic and sequencing data. Building on the common aims to exploit understanding of human diseases, it also opens up an opportunity for scientific communities to share and combine research data. Genome-wide association studies (GWAS) have been widely used to locate genetic variants, which are susceptible for common diseases. In the field of medical genetics, many international collaborative consortiums have been established to conduct meta-analyses of GWAS results and to combine large genotypic data sets to perform mega genetic analyses. Having an integrated phenotype database is significant for exploiting the full potential of extensive genotypic data. In this paper, we aim to share our experience gained from integrating four heterogeneous sets of major depression phenotypic data onto the MySQL platform. These data sets constitute clinical data which had been gathered for various genetic studies for the past decade. We also highlight in this report some generic data handling techniques, the costs and benefits regarding the use of integrated phenotype database within our own institution and under the consortium framework.


Management of gallstones in diabetic patients.
The management of gallstones in diabetic patients has traditionally been considered problematic. Autopsy findings and uncontrolled studies have documented a higher prevalence of cholelithiasis in diabetics, and early reports showed dramatically increased perioperative morbidity and mortality for treatment of diabetics with acute cholecystitis. As a result, some authorities have recommended prophylactic cholecystectomy for diabetic patients with asymptomatic gallstones, which is in contrast to recommendations for nondiabetics. More recent investigators have shown comparable rates of operative morbidity and mortality for biliary surgery in diabetics when compared with the general population. Recent studies have questioned whether diabetes is an independent risk factor for gallstone formation. Decision analyses using these new data have shown that prophylactic cholecystectomy is not of clear benefit and should not be routinely recommended for diabetics with asymptomatic gallstones. We believe that available data, although limited, indicate that asymptomatic patients with diabetes do not benefit from screening for gallstones and that cholecystectomy should only be performed in cases of symptomatic cholelithiasis, as is the case in the general population.


Gut Microbial Divergence between Two Populations of the Hadal Amphipod Hirondellea gigas.
Hadal environments sustain diverse microorganisms. A few studies have investigated hadal microbial communities consisting of free-living or particle-associated bacteria and archaea. However, animal-associated microbial communities in hadal environments remain largely unexplored, and comparative analyses of animal gut microbiota between two isolated hadal environments have never been done so far. In the present study, 228 Gb of gut metagenomes of the giant amphipod Hirondellea gigas from two hadal trenches, the Mariana Trench and Japan Trench, were sequenced and analyzed. Taxonomic analysis identified 49 microbial genera commonly shared by the gut microbiota of the two H. gigas populations. However, the results of statistical analysis, in congruency with the alpha and beta diversity analyses, revealed significant differences in gut microbial composition across the two trenches. Abundance variation of Psychromonas, Propionibacterium, and Pseudoalteromonas species was observed. Microbial cooccurrence was demonstrated for microbes that were overrepresented in the Mariana trench. Comparison of functional potential showed that the percentage of carbohydrate metabolic genes among the total microbial genes was significantly higher in the guts of H. gigas specimens from the Mariana Trench. Integrating carbon input information and geological characters of the two hadal trenches, we propose that the differences in the community structure might be due to several selective factors, such as environmental variations and microbial interactions.IMPORTANCE The taxonomic composition and functional potential of animal gut microbiota in deep-sea environments remain largely unknown. Here, by performing comparative metagenomics, we suggest that the gut microbial compositions of two Hirondellea gigas populations from the Mariana Trench and the Japan Trench have undergone significant divergence. Through analyses of functional potentials and microbe-microbe correlations, our findings shed light on the contributions of animal gut microbiota to host adaptation to hadal environments.


Mean-field beyond mean-field: the single particle view for moderately to strongly coupled charged fluids.
In a counter-ion only charged fluid, Coulomb coupling is quantified by a single dimensionless parameter. Yet, the theoretical treatment of moderately to strongly coupled charged fluids is a difficult task, central to the understanding of a wealth of soft matter problems, including biological systems. We show that the corresponding coupling regime can be remarkably well described by a single particle treatment, which, at variance with previous works, takes due account of inter-ionic interactions. To this end, the prototypical problem of a planar charged dielectric interface is worked out. Testing our predictions against Monte Carlo simulation data reveals an excellent agreement.


Troponin C in different insect muscle types: identification of two isoforms in Lethocerus, Drosophila and Anopheles that are specific to asynchronous flight muscle in the adult insect.
The indirect flight muscles (IFMs) of Lethocerus (giant water bug) and Drosophila (fruitfly) are asynchronous: oscillatory contractions are produced by periodic stretches in the presence of a Ca(2+) concentration that does not fully activate the muscle. The troponin complex on thin filaments regulates contraction in striated muscle. The complex in IFM has subunits that are specific to this muscle type, and stretch activation may act through troponin. Lethocerus and Drosophila have an unusual isoform of the Ca(2+)-binding subunit of troponin, troponin C (TnC), with a single Ca(2+)-binding site near the C-terminus (domain IV); this isoform is only in IFMs, together with a minor isoform with an additional Ca(2+)-binding site in the N-terminal region (domain II). Lethocerus has another TnC isoform in leg muscle which also has two Ca(2+)-binding sites. Ca(2+) binds more strongly to domain IV than to domain II in two-site isoforms. There are four isoforms in Drosophila and Anopheles (malarial mosquito), three of which are also in adult Lethocerus. A larval isoform has not been identified in Lethocerus. Different TnC isoforms are expressed in the embryonic, larval, pupal and adult stages of Drosophila; the expression of the two IFM isoforms is increased in the pupal stage. Immunoelectron microscopy shows the distribution of the major IFM isoform with one Ca(2+)-binding site is uniform along Lethocerus thin filaments. We suggest that initial activation of IFM is by Ca(2+) binding to troponin with the two-site TnC, and full activation is through the action of stretch on the complex with the one-site isoform.


The applications of closure phase to astronomical imaging.
Closure phase is a number measured by triplets of Michelson interferometers that is completely independent of certain types of otherwise severe instrumental errors. In the 30 years since closure phase was invented, it has been applied to a diverse number of different problems in astronomical imaging. Methods based on the closure phase now allow imaging of complex objects in the presence of severe aberrations and are vital to the success of modern, high-resolution astronomical imaging both at radio and at optical wavelengths. Over the past 10 to 15 years, the concept of closure phase has been extended and generalized. One of the most important advances has been the development of automatic or self-calibration techniques. This article reviews closure phase methods and some of the many spin-offs and related ideas.


Using in silico perturbational approach to identify critical areas in schizophrenia.
Schizophrenia is a debilitating neuropsychiatric disorder whose underlying correlates remain unclear despite decades of neuroimaging investigation. One contentious topic concerns the role of global signal (GS) fluctuations and how they affect more focal functional changes. Moreover, it has been difficult to pinpoint causal mechanisms of circuit disruption. Here, we analyzed resting-state fMRI data from 47 schizophrenia patients and 118 age-matched healthy controls and used dynamical analyses to investigate how global fluctuations and other functional metastable states are affected by this disorder. We found that brain dynamics in the schizophrenia group were characterized by an increased probability of globally coherent states and reduced recurrence of a substate dominated by coupled activity in the default mode and limbic networks. We then used the in silico perturbation of a whole-brain model to identify critical areas involved in the disease. Perturbing a set of temporo-parietal sensory and associative areas in a model of the healthy brain reproduced global pathological dynamics. Healthy brain dynamics were instead restored by perturbing a set of medial fronto-temporal and cingulate regions in the model of pathology. These results highlight the relevance of GS alterations in schizophrenia and identify a set of vulnerable areas involved in determining a shift in brain state.


Tomographic 3D reconstruction of quick-frozen, Ca2+-activated contracting insect flight muscle.
Motor actions of myosin were directly visualized by electron tomography of insect flight muscle quick-frozen during contraction. In 3D images, active cross-bridges are usually single myosin heads, bound preferentially to actin target zones sited midway between troponins. Active attached bridges (approximately 30% of all heads) depart markedly in axial and azimuthal angles from Rayment's rigor acto-S1 model, one-third requiring motor domain (MD) tilting on actin, and two-thirds keeping rigor contact with actin while the light chain domain (LCD) tilts axially from approximately 105 degrees to approximately 70 degrees. The results suggest the MD tilts and slews on actin from weak to strong binding, followed by swinging of the LCD through an approximately 35 degrees axial angle, giving an approximately 13 nm interaction distance and an approximately 4-6 nm working stroke.


Quantitative assessment of paravalvular leakage after transcatheter aortic valve replacement using a patient-specific pulsatile flow model.
Quantitative assessment of post-transcatheter aortic valve replacement (TAVR) aortic regurgitation (AR) remains challenging. We developed patient-specific anatomical models with pulsatile flow circuit and investigated factors associated with AR after TAVR. Based on pre-procedural computed tomography (CT) data of the six patients who underwent transfemoral TAVR using a 23-mm SAPIEN XT, anatomically and mechanically equivalent aortic valve models were developed. Forward flow and heart rate of each patient in two days after TAVR were duplicated under mean aortic pressure of 80mmHg. Paravalvular leakage (PVL) volume in basal and additional conditions was measured for each model using an electromagnetic flow sensor. Incompletely apposed tract between the transcatheter and aortic valves was examined using a micro-CT. PVL volume in each patient-specific model was consistent with each patient's PVL grade, and was affected by hemodynamic conditions. PVL and total regurgitation volume increased with the mean aortic pressure, whereas closing volume did not change. In contrast, closing volume increased proportionately with heart rate, but PVL did not change. The minimal cross-sectional gap had a positive correlation with the PVL volumes (r=0.89, P=0.02). The gap areas typically occurred in the vicinity of the bulky calcified nodules under the native commissure. PVL volume, which could be affected by hemodynamic conditions, was significantly associated with the minimal cross-sectional gap area between the aortic annulus and the stent frame. These data may improve our understanding of the mechanism of the occurrence of post-TAVR PVL.


Crystal structures, binding interactions, and ADME evaluation of brain penetrant N-substituted indazole-5-carboxamides as subnanomolar, selective monoamine oxidase B and dual MAO-A/B inhibitors.
The pharmacological and physicochemical analysis of structurally optimized N-alkyl-substituted indazole-5-carboxamides, developed as potential drug and radioligand candidates for the treatment and diagnosis of Parkinson's disease (PD) and other neurological disorders, is reported. Recent efforts have been focused on the development of subnanomolar potent, selective MAO-B (N1-alkyl-substituted compounds 12a-14a and 15) and dual active MAO-A/B (N2-methylated compounds 12b-14b) inhibitors with nanomolar potency towards MAO-B and moderately active against MAO-A enzyme, respectively. The most promising drug-like derivatives in both series were N-(3-chloro-4-fluorophenyl)-1-methyl-1H-indazole-5-carboxamide (13a, NTZ-1441, IC50 hMAO-B 0.662 nM, >15000-fold selective versus MAO-A) and N-(3-chloro-4-fluorophenyl)-2-methyl-2H-indazole-5-carboxamide (13b, NTZ-1442, IC50 hMAO-B 8.08 nM, IC50 hMAO-A 0.56 μM, SI = 70). Moreover, compounds 13a and 13b were predicted to cross both the gastrointestinal tract (at pH 2.0, 5.5, and 7,4) and the blood-brain barrier (BBB) in vitro with appropriate drug-like properties required for CNS active drugs. Combined single X-ray/molecular modeling studies provided insights into the enzyme-inhibitor interactions within both MAO isoforms and the rationale for their inhibitory activity with controlled MAO-A/B selectivity - despite their small structural differences. The binding modes of 12a,b and 13a,b confirmed that the major interactions with hMAO-B were established via the flexible carbonyl group of the carboxamide linkage and the electron-donating nitrogens N1 or N2 of the indazole moiety, allowing further exploration of the alkyl side chain for next step lead optimization efforts.


Greater magnocellular saccadic suppression in high versus low autistic tendency suggests a causal path to local perceptual style.
Saccadic suppression-the reduction of visual sensitivity during rapid eye movements-has previously been proposed to reflect a specific suppression of the magnocellular visual system, with the initial neural site of that suppression at or prior to afferent visual information reaching striate cortex. Dysfunction in the magnocellular visual pathway has also been associated with perceptual and physiological anomalies in individuals with autism spectrum disorder or high autistic tendency, leading us to question whether saccadic suppression is altered in the broader autism phenotype. Here we show that individuals with high autistic tendency show greater saccadic suppression of low versus high spatial frequency gratings while those with low autistic tendency do not. In addition, those with high but not low autism spectrum quotient (AQ) demonstrated pre-cortical (35-45 ms) evoked potential differences (saccade versus fixation) to a large, low contrast, pseudo-randomly flashing bar. Both AQ groups showed similar differential visual evoked potential effects in later epochs (80-160 ms) at high contrast. Thus, the magnocellular theory of saccadic suppression appears untenable as a general description for the typically developing population. Our results also suggest that the bias towards local perceptual style reported in autism may be due to selective suppression of low spatial frequency information accompanying every saccadic eye movement.


Teaching resources. Cell signaling systems: a course for graduate students.
This course uses the primary literature to develop a systems-level understanding of the information flow through the various cell signaling pathways and networks. Current areas of research activities in the experimental and theoretical understanding of cell signaling research are highlighted. This Teaching Resource describes a sample syllabus and organization for this course. Each lecture is 2 hours. After each set of lectures, there is a 2-hour discussion period during which students present an in-depth analysis of a primary literature article, selected in consultation with the lecturer.


[Research reveals a market for a veterinary behaviour clinic].
An enquiry into the requirement of a university veterinary behaviour clinic in The Netherlands revealed that there is a clear call for such a service. The specific demands and wishes of first line practicing veterinarians and companion animal owners were investigated. The research revealed that veterinarians are regular confronted with behaviour problems in companion animals and that they are willing to refer these cases to the University. They also expressed their need for access to continuing professional development opportunities in the field of veterinary behavioural medicine (which is something that most veterinary behaviour clinics associated with veterinary faculties provide). The demand from companion animal owners was also examined. It can be concluded that a large number of them had animals with behaviour problems and that they were willing to seek veterinary advice on these matters. In response to the above mentioned demands the University of Utrecht will open a veterinary behaviour clinic, providing high quality service for animals, their owners and the referring veterinarians. This service will be based on sound scientific practice and delivered by both veterinarians specialised in this field and recognised animal behaviour therapists.


Molecular Profiling in Metastatic Colorectal Cancer.
Colorectal cancer (CRC) is a commonly diagnosed malignancy. Although chemotherapy remains the backbone of treatment, the landscape of treating metastatic CRC (mCRC) is changing with the understanding of its heterogeneity and molecular blueprint. Colon cancer sidedness has proven to hold prognostic implications, with right-sided tumors having higher incidence of BRAF and KRAS mutations and being microsatellite instability-high (MSI-H); overall, they have a worse prognosis compared with left sided-tumors. Results of molecular research have demonstrated the need to profile each mCRC patient for RAS and BRAF mutations, MSI-H status, HER2 amplifications, and NTRK fusions. Ongoing clinical trials using targeted agents aim to further improve survival outcomes. We emphasize the epidemiology, knowledge of primary tumor location, and mutational landscape of mCRC, as well as novel treatment options for patients harboring unique subtypes of these characteristics.


Air pollution and mortality: results from a study of Santiago, Chile.
In 1986, the U.S. EPA issued an air quality standard for particulate matter that included only particulates below 10 microns in diameter (PM10). Unfortunately, epidemiological research investigating the health effects associated with PM10 has been limited by the lack of available daily data from outdoor monitoring stations. Evidence of high concentrations of PM10 in Eastern Europe and in metropolitan areas such as Mexico City and Santiago, Chile underscores the need to evaluate the association between air pollution and mortality. Over the last few years, daily measures of ambient PM10 have been collected in Santiago. Our analysis examines the relationship between PM10 and daily mortality between 1989 and 1991. In addition to total daily mortality, the data were compiled to record total mortality for all males, all females, and those over 65, and mortality from either respiratory disease or cardiovascular disease. Multiple regression analysis was used to explain mortality, with particular attention to controlling for the influence of season and temperature. The results suggest a strong association between PM10 and all of the alternative measures of mortality. The association persists after controlling for daily minimum temperature and binary variables indicating temperature extremes, the day of the week, the month, and the year. Additional sensitivity analyses suggest a fairly robust relationship. In general, a 10 micrograms/m3 change in daily PM10 was associated with a 1% increase in mortality. This relative risk is consistent with the results of recent studies undertaken in the United States.


Introduction: revisiting the "negrito" hypothesis: a transdisciplinary approach to human prehistory in southeast Asia.
The "negrito" hypothesis predicts that a shared phenotype among various contemporary groups of hunter-gatherers in Southeast Asia--dark skin, short stature, tight curly hair--is due to common descent from a region-wide, pre-Neolithic substrate of humanity. The alternative is that their distinctive phenotype results from convergent evolution. The core issues of the negrito hypothesis are today more relevant than ever to studies of human evolution, including the out-of-Africa migration, admixture with Denisovans, and the effects of environment and ecology on life-history traits. Understanding the current distribution of the negrito phenotype dictates a wide-ranging remit for study, including the articulation of the relationship between foragers and farmers in the present, the development of settled agriculture in the mid-Holocene, and terminal Pleistocene population expansions. The consensus reached by the contributors to this special double issue of Human Biology is that there is not yet conclusive evidence either for or against the negrito hypothesis. Nevertheless, the process of revisiting the problem will benefit the knowledge of the human prehistory of Southeast Asia. Whether the term negrito accurately reflects the all-encompassing nature of the resulting inquiry is in itself questionable, but the publication of this double issue is testament to the enduring ability of this hypothesis to unite disparate academic disciplines in a common purpose.


Surgery versus nonsurgical treatment of lumbar spinal stenosis: a randomized trial.
Primary care management decisions for patients with symptomatic lumbar spinal stenosis (LSS) are challenging, and nonsurgical guidance is limited by lack of evidence. To compare surgical decompression with physical therapy (PT) for LSS and evaluate sex differences. Multisite randomized, controlled trial. (ClinicalTrials.gov: NCT00022776). Neurologic and orthopedic surgery departments and PT clinics. Surgical candidates with LSS aged 50 years or older who consented to surgery. Surgical decompression or PT. Primary outcome was physical function score on the Short Form-36 Health Survey at 2 years assessed by masked testers. The study took place from November 2000 to September 2007. A total of 169 participants were randomly assigned and stratified by surgeon and sex (87 to surgery and 82 to PT), with 24-month follow-up completed by 74 and 73 participants in the surgery and PT groups, respectively. Mean improvement in physical function for the surgery and PT groups was 22.4 (95% CI, 16.9 to 27.9) and 19.2 (CI, 13.6 to 24.8), respectively. Intention-to-treat analyses revealed no difference between groups (24-month difference, 0.9 [CI, -7.9 to 9.6]). Sensitivity analyses using causal-effects methods to account for the high proportion of crossovers from PT to surgery (57%) showed no significant differences in physical function between groups. Without a control group, it is not possible to judge success attributable to either intervention. Surgical decompression yielded similar effects to a PT regimen among patients with LSS who were surgical candidates. Patients and health care providers should engage in shared decision-making conversations that include full disclosure of evidence involving surgical and nonsurgical treatments for LSS. National Institutes of Health and National Institute of Arthritis and Musculoskeletal and Skin Diseases.


Prospective controlled study to evaluate laboratory and clinical outcomes of oocyte vitrification obtained in in vitro fertilization patients aged 30 to 39 years.
To determine whether the process of oocyte vitrification affects oocyte viability in in vitro fertilization (IVF) patients between 30 and 39 years of age. Prospective controlled study. Private IVF practice. A total of 30 women assigned and 22 qualified. Denudation of oocytes, cryopreservation of oocytes using vitrification method in a medium with 15% ethylene glycol (EG), 15% dimethylsulfoxide (DMSO), and 0.5 M sucrose. Oocyte survival, fertilization, day-3 embryo quality, blastocyst formation, clinical pregnancy, implantation, and live-birth rates. After denudation of oocytes, mature sibling oocytes were randomly allocated to the fresh and vitrified groups. The survival rate was 79.6% after vitrification/warming. Overall, no statistically significant differences were found in fertilization, day-3 embryo quality, or blastocyst formation rates between the fresh and vitrified groups. The positive β-human chorionic gonadotropin, clinical pregnancy rate, and implantation rate were 13 (59.0%) of 22, 10 (45.4%) of 22, and 16 (30.1%) of 53 for the vitrified group. The overall efficiency in achieving a live birth was 11 (5.9%) of 186 per vitrified oocyte. The impact of vitrification can be reduced to a minimal level, making it possible to achieve high pregnancy and implantation rates in this age group of IVF patients.


The State of the World's Beaches.
Coastal zones constitute one of the most heavily populated and developed land zones in the world. Despite the utility and economic benefits that coasts provide, there is no reliable global-scale assessment of historical shoreline change trends. Here, via the use of freely available optical satellite images captured since 1984, in conjunction with sophisticated image interrogation and analysis methods, we present a global-scale assessment of the occurrence of sandy beaches and rates of shoreline change therein. Applying pixel-based supervised classification, we found that 31% of the world's ice-free shoreline are sandy. The application of an automated shoreline detection method to the sandy shorelines thus identified resulted in a global dataset of shoreline change rates for the 33 year period 1984-2016. Analysis of the satellite derived shoreline data indicates that 24% of the world's sandy beaches are eroding at rates exceeding 0.5 m/yr, while 28% are accreting and 48% are stable. The majority of the sandy shorelines in marine protected areas are eroding, raising cause for serious concern.


Perceptions of quality of life among Ugandan patients living with HIV: a qualitative study.
Ugandans have endured the HIV epidemic for three decades. Now, with the availability of antiretroviral therapy (ART) and early diagnosis, those living with HIV can live longer and can enjoy the same life expectancy as the rest of the Ugandan population. This emerging trend necessitates the assessment of quality of life, alongside other patient outcomes, of those undergoing therapy, alongside other patient outcomes. While major strides have been made in developing measures of quality of life in the developed world, there remains a paucity of evidence from resource-limited settings. This challenge is further complicated by the contentious definition of quality of life, which is highly subjective and varies between individuals. In this paper, we aim to identify the determinants of quality of life for people living with HIV in a Ugandan context to contribute to the chronic care model for persons living with HIV/AIDS. Twenty HIV-positive participants took part in in-depth interviews at an urban clinic, with follow-ups at three and six months. Ten patients were on ART and ten not on ART. All interviews were transcribed and translated for analysis. Data were analysed manually using the framework approach to content analysis. Individuals reported on four aspects of quality of life: liveability of the environment, utility of life, life ability of a person and appreciation of life. Respondents described multiple expectations and expressed hope for their future. However, many still suffered from stigma, fears of disclosure and poverty, which negatively affected their quality of life. Individuals living with HIV receiving treatment or in care experienced an improved quality of life in this setting, although the situation for many remains precarious.


New hosts for the mite Ornithonyssus bursa in Argentina.
The mite Ornithonyssus bursa (Berlese) (Mesostigmata: Macronyssidae) is considered a poultry pest causing important infestations in chickens and it is considered a potential vector of arbovirus. Despite being considered a common parasite in wild birds, there is scarce published information about its potential hosts and effects on them. Here we present new bird hosts for O. bursa, assess the presence of Alphavirus, Flavivirus and Bunyavirus in mites from three host species, and discuss its potential impact on wild bird populations. We found O. bursa infecting five raptor and six passerine wild bird species. For nine of these species, this is the first record of infection by O. bursa. Although all analysed mites were negative for the examined arboviruses, the small sample size of mites does not allow further conclusions at the present moment. Because of the general nature of this ectoparasite, its presence in migratory long dispersal and endangered bird species, and the seropositivity for arboviruses in some of the species studied here, we consider it critical to assess the role of O. bursa and other ectoparasites as vectors and reservoirs of pathogens and as potential deleterious agents in wild bird populations.


The NAADP receptor: commentary on Billington et al.
NAADP is a recently described calcium-mobilizing messenger. First discovered as a potent calcium-releasing molecule in sea urchin eggs, its actions have now been reported in several mammalian cell types. In the sea urchin egg, NAADP-sensitive calcium release channels appear distinct from inositol trisphosphate or ryanodine receptors, and are mainly localized to acidic compartments. In this study, Billington et al. extend the pharmacology of the putative NAADP receptor utilizing molecules unrelated to NAADP itself. This work may provide an important step in developing selective NAADP receptor modulators that will help define the role of NAADP in cell signalling.


Disorders of Consciousness and Disability Law.
In 2018, the American Academy of Neurology, the American Congress of Rehabilitation Medicine, and the National Institute on Disability, Independent Living, and Rehabilitation Research published a systematic evidence-based review and an associated practice guideline for improved assessment, treatment, and rehabilitation of patients with disorders of consciousness. Patients with disorders of consciousness include individuals in the vegetative and minimally conscious states, as well as others with covert consciousness and cognitive motor dissociation. These landmark publications (concurrently published in Neurology and Archives of Physical Medicine and Rehabilitation) supplant the 1994 New England Journal of Medicine Multi-Society Task Force report on the vegetative state and the 2002 criteria establishing minimally conscious states. The guideline re-designates the permanent vegetative state as chronic. In our article, we consider the legal and ethical implications of the practice guideline for clinical practice and explain the vulnerability of these patients who suffer from high rates of misdiagnosis, inadequate medical surveillance, undertreatment of pain, inadequate rehabilitation, and segregation in chronic care. We argue that these deficiencies in medical care are inconsistent with our growing appreciation of the dynamic nature of these brain states and an emerging standard of care as articulated by the national guideline. These deficiencies also violate domestic and international disability law. To substantiate this latter claim, we apply disability law to this population, focusing on key Americans with Disabilities Act mandates, the relevance of the 1999 Supreme Court, Olmstead v. L.C., and the utility of Olmstead enforcement actions to integrate the care of these individuals into the medical mainstream.


Hierarchical cobalt-nitride and -oxide co-doped porous carbon nanostructures for highly efficient and durable bifunctional oxygen reaction electrocatalysts.
Here we report the preparation of hollow microspheres with a thin shell composed of mixed cobalt nitride (Co-N) and cobalt oxide (Co-O) nanofragments encapsulated in thin layers of nitrogen-doped carbon (N-C) nanostructure (Co-N/Co-O@N-C) arrays with enhanced bifunctional oxygen electrochemical performance. The hybrid structures are synthesized via heat treatment of N-doped hollow carbon microspheres with cobalt nitrate, and both the specific ratio of these precursors and the selected annealing temperature are found to be the key factors for the formation of the unique hybrid structure. The as-obtained product (Co-N/Co-O@N-C) presents a large specific surface area (493 m2 g-1), high-level heteroatom doping (Co-N, Co-O, and N-C), and hierarchical porous nanoarchitecture containing macroporous frameworks and mesoporous walls. Electronic interaction between the thin N-C layers and the encapsulated Co-N and Co-O nanofragments efficiently optimizes oxygen adsorption properties on the Co-N/Co-O@N-C and thereby triggers bifunctional oxygen electrochemical activity at the surface. The Co-N/Co-O@N-C nanohybrid exhibited a high onset potential of 0.93 V, and a limiting current density of 5.6 mA cm-2 indicating 4-electron oxygen reduction reaction (ORR), afforded high catalytic activity for the oxygen evolution reaction (OER) and even exceeded the catalytic stability of the commercial precious electrocatalysts; furthermore, when integrated into the oxygen electrode of a regenerative fuel cell device, it exhibited high-performance oxygen electrodes for both the ORR and the OER.


Excessive Hypoxia-Inducible Factor-1α Expression Induces Cardiac Rupture via p53-Dependent Apoptosis After Myocardial Infarction.
Background Apoptosis plays a pivotal role in cardiac rupture after myocardial infarction (MI), and p53 is a key molecule in apoptosis during cardiac rupture. Hif-1α (hypoxia-inducible factor-1α), upregulated under hypoxia, is a known p53 inducer. However, the role of Hif-1α in the regulatory mechanisms underlying p53 upregulation, apoptosis, and cardiac rupture after MI is unclear. Methods and Results We induced MI in mice by ligating the left anterior descending artery. Hif-1α and p53 expressions were upregulated in the border zone at day 5 after MI, accompanied by apoptosis. In rat neonatal cardiomyocytes, treatment with cobalt chloride (500 μmol/L), which mimics severe hypoxia by inhibiting PHD (prolyl hydroxylase domain-containing protein), increased Hif-1α and p53, accompanied by myocyte death with caspase-3 cleavage. Silencing Hif-1α or p53 inhibited caspase-3 cleavage, and completely prevented myocyte death under PHD inhibition. In cardiac-specific Hif-1α hetero-knockout mice, expression of p53 and cleavage of caspase-3 and poly (ADP-ribose) polymerase were reduced, and apoptosis was suppressed on day 5. Furthermore, the cleavage of caspase-8 and IL-1β (interleukin-1β) was also suppressed in hetero knockout mice, accompanied by reduced macrophage infiltration and matrix metalloproteinase/tissue inhibitor of metalloproteinase activation. Although there was no intergroup difference in infarct size, the cardiac rupture and survival rates were significantly improved in the hetero knockout mice until day 10 after MI. Conclusions Hif-1α plays a pivotal role in apoptosis, inflammation, and cardiac rupture after MI, in which p53 is a critical mediator, and may be a prospective therapeutic target for preventing cardiac rupture.


A role for CSLD3 during cell-wall synthesis in apical plasma membranes of tip-growing root-hair cells.
In plants, cell shape is defined by the cell wall, and changes in cell shape and size are dictated by modification of existing cell walls and deposition of newly synthesized cell-wall material. In root hairs, expansion occurs by a process called tip growth, which is shared by root hairs, pollen tubes and fungal hyphae. We show that cellulose-like polysaccharides are present in root-hair tips, and de novo synthesis of these polysaccharides is required for tip growth. We also find that eYFP-CSLD3 proteins, but not CESA cellulose synthases, localize to a polarized plasma-membrane domain in root hairs. Using biochemical methods and genetic complementation of a csld3 mutant with a chimaeric CSLD3 protein containing a CESA6 catalytic domain, we provide evidence that CSLD3 represents a distinct (1→4)-β-glucan synthase activity in apical plasma membranes during tip growth in root-hair cells.


Light microscopic changes in Plasmodium falciparum from Gabonese children treated with artemether.
The changes in Plasmodium falciparum in four Gabonese children suffering from severe malaria and treated with pure artemether were observed in thin blood smears fixed and stained with Giemsa and examined by light microscopy. Peripheral blood samples were taken every 8 hr up to 72 hr from three children and every 3 hr up to 9 hr from the other child. The morphologic changes involved all development stages (trophozoites, schizonts, and gametocytes); they were first seen 3 hr after the start of treatment and all parasites were abnormal after 24 hr. After two days of treatment, all infected erythrocytes disappeared except for a few with necrotic trophozoites. The morphologic changes were similar to the ultrastructural changes previously described in vivo and in vitro in experimental models. They confirm the rapid effect of artemisinin derivatives on parasite clearance and clinical recovery, particularly in cases of cerebral malaria.


Harmonin-b, an actin-binding scaffold protein, is involved in the adaptation of mechanoelectrical transduction by sensory hair cells.
We assessed the involvement of harmonin-b, a submembranous protein containing PDZ domains, in the mechanoelectrical transduction machinery of inner ear hair cells. Harmonin-b is located in the region of the upper insertion point of the tip link that joins adjacent stereocilia from different rows and that is believed to gate transducer channel(s) located in the region of the tip link's lower insertion point. In Ush1c (dfcr-2J/dfcr-2J) mutant mice defective for harmonin-b, step deflections of the hair bundle evoked transduction currents with altered speed and extent of adaptation. In utricular hair cells, hair bundle morphology and maximal transduction currents were similar to those observed in wild-type mice, but adaptation was faster and more complete. Cochlear outer hair cells displayed reduced maximal transduction currents, which may be the consequence of moderate structural anomalies of their hair bundles. Their adaptation was slower and displayed a variable extent. The latter was positively correlated with the magnitude of the maximal transduction current, but the cells that showed the largest currents could be either hyperadaptive or hypoadaptive. To interpret our observations, we used a theoretical description of mechanoelectrical transduction based on the gating spring theory and a motor model of adaptation. Simulations could account for the characteristics of transduction currents in wild-type and mutant hair cells, both vestibular and cochlear. They led us to conclude that harmonin-b operates as an intracellular link that limits adaptation and engages adaptation motors, a dual role consistent with the scaffolding property of the protein and its binding to both actin filaments and the tip link component cadherin-23.


Unbiased pro-thrombotic features at diagnosis in 977 thrombocythemic patients with Philadelphia-negative chronic myeloproliferative neoplasms.
In patients with Philadelphia-negative chronic myeloproliferative neoplasms (MPNs), the anti-thrombotic and/or cytoreductive treatment in the follow-up may affect the evaluation of the pro-thrombotic weight of the clinical and biological characteristics at diagnosis. In order to avoid this potential confounding effect, we investigated the relationship between prior thrombosis (PrTh: thrombosis occurred before diagnosis and before treatment) and the characteristics at diagnosis in 977 thrombocythemic patients with MPN, reclassified according to the WHO 2008 criteria. PrTh occurred in 194 (19.9%) patients, with similar rates in the different MPNs. In multivariate analysis, PrTh rate was significantly related to minor thrombocytosis (platelets ≤700×10(9)/L), leukocytosis (leukocytes >10×10(9)/L), higher hematocrit (HCT >45%), JAK2 V617F mutation, older age, and cardiovascular risk factors (CVRFs). The highest PrTh rate (33.9%) was associated with the coexistence of minor thrombocytosis and leukocytosis. Of note, the inverse relationship between PrTh rate and platelet count is consistent with the hemostatic paradox of thrombocytosis. In conclusion, this analysis in MPN patients disclosed the unbiased characteristics at diagnosis with a pro-thrombotic effect. Moreover, it suggests that the optimal control of blood cells counts, and CVRFs might be of utmost importance in the prevention of thrombosis during the follow-up.


Bacterial phylogenetic reconstruction from whole genomes is robust to recombination but demographic inference is not.
Phylogenetic inference in bacterial genomics is fundamental to understanding problems such as population history, antimicrobial resistance, and transmission dynamics. The field has been plagued by an apparent state of contradiction since the distorting effects of recombination on phylogeny were discovered more than a decade ago. Researchers persist with detailed phylogenetic analyses while simultaneously acknowledging that recombination seriously misleads inference of population dynamics and selection. Here we resolve this paradox by showing that phylogenetic tree topologies based on whole genomes robustly reconstruct the clonal frame topology but that branch lengths are badly skewed. Surprisingly, removing recombining sites can exacerbate branch length distortion caused by recombination. Phylogenetic tree reconstruction is a popular approach for understanding the relatedness of bacteria in a population from differences in their genome sequences. However, bacteria frequently exchange regions of their genomes by a process called homologous recombination, which violates a fundamental assumption of phylogenetic methods. Since many researchers continue to use phylogenetics for recombining bacteria, it is important to understand how recombination affects the conclusions drawn from these analyses. We find that whole-genome sequences afford great accuracy in reconstructing evolutionary relationships despite concerns surrounding the presence of recombination, but the branch lengths of the phylogenetic tree are indeed badly distorted. Surprisingly, methods to reduce the impact of recombination on branch lengths can exacerbate the problem.


Normative data for the Rey-Osterrieth and the Taylor complex figure tests in Quebec-French people.
The Rey-Osterrieth (ROCF) and Taylor (TCF) complex figure tests are widely used to assess visuospatial and constructional abilities as well as visual/non-verbal memory. Normative data adjusted to the cultural and linguistic reality of older Quebec-French individuals is still nonexistent for these tests. In this article, we report the results of two studies that aimed to establish normative data for Quebec-French people (aged at least 50 years) for the copy, immediate recall, and delayed recall trials of the ROCF (Study 1) and the TCF (Study 2). For both studies, the impact of age, education, and sex on test performance was examined. Moreover, the impact of copy time on test performance, the impact of copy score on immediate and delayed recall score, and the impact of immediate recall score on delayed recall performance were examined. Based on regression models, equations to calculate Z scores for copy and recall scores are provided for both tests.


Prevalence of Serum IgG Antibodies to Cystic Echinococcus Antigen among Patients in an Uzbekistan Emergency Hospital.
Cystic echinococcosis (CE) is one of the most widespread zoonotic helminthiases, which can last an asymptomatic infection for several years. The purpose of this study was to demonstrate serum antibody prevalence of CE among asymptomatic people in Uzbekistan using ELISA. A total of 2,547 serum samples were collected, 66 from confirmed CE patients and 2,481 of patients with other diseases than CE at a hospital in Tashkent, Uzbekistan. The serum samples were screened for CE specific IgG antibodies by ELISA using cystic fluid antigen obtained from sheep. The serum antibody positive rate was 89.4% (59/66) in CE and 3.6% (89/2,481) in other disease patients. The present ELISA recognized 89.4% sensitivity and 96.4% specificity. The ELISA absorbance of positive samples was distributed 0.271-0.971 for CE and 0.273-0.887 for other disease patients. The other disease patients with high absorbance over 0.3 were 50 (2.0%) who were presumed to be active CE patients. The patients in their 40s showed the highest positive rate of 5.2% (P=0.181), and women were 4.4% while men were 3.1% positive (P=0.136). The data confirmed that there are many asymptomatic patients of CE in Tashkent. It is indicated that CE is an endemic disease of public health importance in Uzbekistan.


A molecular view of DNA-conjugated nanoparticle association energies.
Nanoparticles functionalized with short sequences of DNA represent a promising platform for customizable self assembly. Though much recent research has focused on the phase behavior and assembly of these structures, little has been done to precisely characterize the pairwise interaction between particles. Here we present a detailed calculation of the association between DNA-nanoparticle conjugates using 3SPN.2, a coarse-grained model of DNA that accounts for molecular structure and base-pairing. We compare our results to those obtained experimentally using μm sized particles and analyze the free energy surfaces that characterize interparticle hybridization. Next, we study the importance of three-body effects and their impact on particle association and melting. Lastly, we explore the observation by Park et al. [Nature, 451, 553 (2008)] that DNA-nanoparticle crystallization can be inhibited by the deletion of a single nucleotide. Using our model, we suggest that the role of this nucleotide is to disrupt frustration.


Risk factors for the development of invasive cancer in unresected ductal carcinoma in situ.
The natural history of ductal carcinoma in situ (DCIS) remains uncertain. The risk factors for the development of invasive cancer in unresected DCIS are unclear. Women diagnosed with DCIS on needle biopsy after 1997 who did not undergo surgical resection for ≥1 year after diagnosis were identified by breast centres and the cancer registry and outcomes were reviewed. Eighty-nine women with DCIS diagnosed 1998-2010 were identified. The median age at diagnosis was 75 (range 44-94) years with median follow-up (diagnosis to death, invasive disease or last review) of 59 (12-180) months. Twenty-nine women (33%) developed invasive breast cancer after a median interval of 45 (12-144) months. 14/29 (48%) with high grade, 10/31 (32%) with intermediate grade and 3/17 (18%) with low grade DCIS developed invasive cancer after median intervals of 38, 60 and 51 months. The cumulative incidence of invasion was significantly higher in high grade DCIS than other grades (p = .0016, log-rank test). Invasion was more frequent in lesions with calcification as the predominant feature (23/50 v. 5/25; p = .042) and in younger women (p = .0002). Endocrine therapy was associated with a lower rate of invasive breast cancer (p = .048). High cytonuclear grade, mammographic microcalcification, young age and lack of endocrine therapy were risk factors for DCIS progression to invasive cancer. Surgical excision of high grade DCIS remains the treatment of choice. Given the uncertain long-term natural history of non-high grade DCIS, the option of active surveillance of women with this condition should be offered within a clinical trial.


Enhancing Nab-Paclitaxel Delivery Using Microbubble-Assisted Ultrasound in a Pancreatic Cancer Model.
A combination of microbubbles (MBs) and ultrasound (US) is an emerging method for noninvasive and targeted enhancement of anti-cancer drug uptake. This method showed an increase local drug extravasation in tumor tissue while reducing the systemic adverse effects in various tumor models. The present study aims to evaluate the effectiveness of this approach for Nab-paclitaxel delivery in a pancreatic tumor model. US and MBs of different types in combination with Nab-paclitaxel showed a loss in cell viability of pancreatic cancer cells in comparison with Nab-paclitaxel treatment alone in in vitro scenario. The in vivo data revealed that US and MBs in combination with Nab-paclitaxel induced a significant decrease in the tumor volume in a subcutaneous pancreatic adenocarcinoma mouse model in comparison to tumors treated with Nab-paclitaxel alone. The postmortem anatomopathological analyses of tumor tissues partially confirmed these results. In conclusion, this study demonstrates that MB-assisted US is a relevant technology to increase the therapeutic effectiveness of Nab-paclitaxel in a pancreatic cancer model.


Insights from Bcl-2 and Myc: malignancy involves abrogation of apoptosis as well as sustained proliferation.
The chromosome translocations typifying Burkitt's lymphoma and follicular lymphoma deregulate very different oncogenes, myc and bcl-2. Transgenic mouse models have illuminated how each contributes to lymphomagenesis. Constitutive myc expression provokes sustained cell proliferation and retards differentiation. However, the resulting expansion in cell number is self-limiting, because the cells remain dependent on cytokines and undergo apoptosis when these become limiting. In contrast, bcl-2 is the prototype of a new class of oncogene that enhances cell survival but does not promote proliferation. Coexpression of these genes leads to the rapid transformation of lymphocytes, probably because each can counter an antioncogenic aspect of the other. Several close homologues of Bcl-2 also enhance cell survival and are thus potential oncogenes; each is essential for maintenance of particular major organs. More distant Bcl-2 relatives instead promote apoptosis and can be regarded as tumor suppressors. For many but not all apoptic signals, the balance between these competing activities determines cell survival. Learning how to adjust the apoptotic threshold in cancer cells should promote development of more effective therapeutic strategies.


Lenvatinib for the treatment of unresectable hepatocellular carcinoma: evidence to date.
During the last 10 years, the multikinase inhibitor sorafenib has emerged as the only systemic treatment for unresectable hepatocellular carcinoma (HCC). More recently, data from the Phase III REFLECT trial showed that another multikinase inhibitor, namely, lenvatinib, was non-inferior to sorafenib in terms of overall survival (OS). In contrast, with respect to OS, previous randomized Phase III trials have been negative, and several agents tested have failed to prove non-inferiority (or superiority) when compared with sorafenib in a first-line setting. Furthermore, the REFLECT trial demonstrated that lenvatinib, in comparison with sorafenib, significantly increased progression-free survival, time to progression, and objective response rate. Overall, the incidence of grade ≥3 treatment-emergent adverse events (TEAEs) was similar in the two treatment arms of the trial, with a higher incidence of serious TEAEs in the lenvatinib arm. Encouraging efficacy signals had already been reported for immune checkpoint inhibitors in HCC, and different synergisms have been postulated in the frame of interplay between vascular endothelial growth factor receptor-2 inhibitors and immunotherapy. Given these premises, future approaches are being developed in Phase I trials testing lenvatinib in combination with pembrolizumab or nivolumab. As the treatment landscape of HCC is expanding with novel agents being approved for patients who are intolerant or are progressing on prior sorafenib, we will discuss current challenges pertaining to the optimal sequencing of active agents in first- and second-line setting.


Socio-economic and age variations in response to supermarket-led checkout food policies: a repeated measures analysis.
Dietary inequalities between population groups are common with older and more affluent individuals tending to have healthier diets. Differential responses to health interventions may exacerbate inequalities. Changing what foods are displayed at supermarket checkouts is one intervention that has the potential to change diets. The aim of this study was to assess whether differences in purchases of common checkout foods from supermarkets with different checkout food policies varied according to age group and social grade. We analysed annual household purchase data for 2013-17 from nine leading UK supermarkets, split according to age of the main household shopper and household social grade. Checkout food policies were categorised as clear and consistent, vague or inconsistent, and none. Policies were heterogeneous but all included removal of confectionery and/or chocolate from checkouts. Mixed effects linear regression models were used to assess differences in purchases of common checkout foods (sugary confectionery, chocolate and potato crisps) by checkout food policy and whether these varied by age group or occupational social grade. Relative to supermarkets with no checkout food policy, 14% (95% CI: 4-22%) fewer purchases of common checkout foods per household per percentage market share were made in supermarkets with a clear and consistent policy. Adjusted mean numbers of purchases were higher in older age groups than the youngest, but there were no differences between the highest and other social grades. There were significant interactions between checkout food policy and both age group and social grade. In supermarkets with clear and consistent policies, 23% (6-36%), 20% (2-34%), and 23% (7-37%) fewer purchases were made in age groups 45-54, 55-64 and 65+ years respectively, compared to all groups combined. In supermarkets with clear and consistent policies, there were 21% (4-35%), 26% (9-39%) and 21% (3-35%) fewer purchases made by households in the highest two and lowest social grades respectively, compared to all groups combined. Households with older main shoppers and those in the most and least affluent social grades may be most responsive to supermarket checkout food policies. As older and more affluent groups tend to have healthier diets overall, it is unlikely that supermarket checkout food policies contribute to greater dietary equity.


Postnatal development of organic cation transport in the rat liver.
Previous studies have demonstrated that the small permanently charged organic cation 1-methyl-4-phenylpyridinium (MPP+) is avidly taken up by rat hepatocytes. The aim of this study was to characterise the postnatal development of hepatic uptake of organic cations, using the model compound MPP+. Accumulation of [3H]MPP+ by liver slices obtained from rats ranging from 1 day to 7 weeks was measured, and the effect of a series of compounds on [3H]MPP+ uptake was examined. The accumulation of [3H]MPP+ by liver slices was similar in adult (87.5 +/- 19.9 pmol g-1; n = 7) and neonatal rats (110.6 +/- 11.5 pmol g-1; n = 15). Verapamil, quinidine (100 microM) and progesterone (200 microM) produced very marked reductions on [3H]MPP+ uptake at all ages, and the inhibitory effect of verapamil and quinidine was maximum in livers from 1-day-old rats. Bilirubin (200 microM) significantly reduced [3H]MPP+ uptake by liver slices from 1 day, 1 week and 7-week-old rats. However, [3H]MPP+ accumulation was reduced by cimetidine, vinblastine and daunomycin (100 microM) in 1-day-old rats, but the effect of these drugs disappeared as the animals age increased. These findings demonstrate that hepatic organic cation uptake capacity is remarkably high shortly after birth and suggest that at least two distinct uptake mechanisms are involved in this process. These uptake systems are the type I hepatic transporter of organic cations, active from birth to adulthood, and P-glycoprotein, active only in very young rats.


Nitric oxide selectively suppresses IH currents mediated by HCN1-containing channels.
Hyperpolarization-activated non-specific cation-permeable channels (HCN) mediate I(H) currents, which are modulated by cGMP and cAMP and by nitric oxide (NO) signalling. Channel properties depend upon subunit composition (HCN1-4 and accessory subunits) as demonstrated in expression systems, but physiological relevance requires investigation in native neurons with intact intracellular signalling. Here we use the superior olivary complex (SOC), which exhibits a distinctive pattern of HCN1 and HCN2 expression, to investigate NO modulation of the respective I(H) currents, and compare properties in wild-type and HCN1 knockout mice. The medial nucleus of the trapezoid body (MNTB) expresses HCN2 subunits exclusively, and sends inhibitory projections to the medial and lateral superior olives (MSO, LSO) and the superior paraolivary nucleus (SPN). In contrast to the MNTB, these target nuclei possess an I(H) with fast kinetics, and they express HCN1 subunits. NO is generated in the SOC following synaptic activity and here we show that NO selectively suppresses HCN1, while enhancing IH mediated by HCN2 subunits. NO hyperpolarizes the half-activation of HCN1-mediated currents and slows the kinetics of native IH currents in the MSO, LSO and SPN. This modulation was independent of cGMP and absent in transgenic mice lacking HCN1. Independently, NO signalling depolarizes the half-activation of HCN2-mediated I(H) currents in a cGMP-dependent manner. Thus, NO selectively suppresses fast HCN1-mediated I(H) and facilitates a slow HCN2-mediated I(H) , so generating a spectrum of modulation, dependent on the local expression of HCN1 and/or HCN2.


Self-monitoring in hypertension: a web-based survey of primary care physicians.
Although self-monitoring of blood pressure is common among people with hypertension, little is known about how general practitioners (GPs) use such readings. This survey aimed to ascertain current views and practice on self-monitoring of UK primary care physicians. An internet-based survey of UK GPs was undertaken using a provider of internet services to UK doctors. The hyperlink to the survey was opened by 928 doctors, and 625 (67%) GPs completed the questionnaire. Of them, 557 (90%) reported having patients who self-monitor, 191 (34%) had a monitor that they lend to patients, 171 (31%) provided training in self-monitoring for their patients and 52 (9%) offered training to other GPs. Three hundred and sixty-seven GPs (66%) recommended at least two readings per day, and 416 (75%) recommended at least 4 days of monitoring at a time. One hundred and eighty (32%) adjusted self-monitored readings to take account of lower pressures in out-of-office settings, and 10/5 mm Hg was the most common adjustment factor used. Self-monitoring of blood pressure was widespread among the patients of responding GPs. Although the majority used appropriate schedules of measurement, some GPs suggested much more frequent home measurements than usual. Further, interpretation of home blood pressure was suboptimal, with only a minority recognising that values for diagnosis and on-treatment target are lower than those for clinic measurement. Subsequent national guidance may improve this situation but will require adequate implementation.


Genome sequence diversity and clues to the evolution of variola (smallpox) virus.
Comparative genomics of 45 epidemiologically varied variola virus isolates from the past 30 years of the smallpox era indicate low sequence diversity, suggesting that there is probably little difference in the isolates' functional gene content. Phylogenetic clustering inferred three clades coincident with their geographical origin and case-fatality rate; the latter implicated putative proteins that mediate viral virulence differences. Analysis of the viral linear DNA genome suggests that its evolution involved direct descent and DNA end-region recombination events. Knowing the sequences will help understand the viral proteome and improve diagnostic test precision, therapeutics, and systems for their assessment.


What rheumatologists in the United States think of complementary and alternative medicine: results of a national survey.
We aimed to describe prevailing attitudes and practices of rheumatologists in the United States toward complementary and alternative medicine (CAM) treatments. We wanted to determine whether rheumatologists' perceptions of the efficacy of CAM therapies and their willingness to recommend them relate to their demographic characteristics, geographic location, or clinical practices. A National Institutes of Health-sponsored cross-sectional survey of internists and rheumatologists was conducted regarding CAM for treatment of chronic back pain or joint pain. In this study we analyzed responses only from rheumatologists. Response items included participant characteristics and experience with 6 common CAM categories, as defined by the National Institutes of Health. Descriptive statistics were used to describe attitudes to CAM overall and to each CAM category. Composite responses were devised for respondents designating 4 or more of the 6 CAM therapies as "very" or "moderately" beneficial or "very likely" or "somewhat likely" to recommend. Of 600 rheumatologists who were sent the questionnaire, 345 responded (58%); 80 (23%) were women. Body work had the highest perceived benefit, with 70% of respondents indicating benefit. Acupuncture was perceived as beneficial by 54%. Most were willing to recommend most forms of CAM. Women had significantly higher composite benefit and recommend responses than men. Rheumatologists not born in North America were more likely to perceive benefit of select CAM therapies. In this national survey of rheumatologists practicing in the United States, we found widespread favorable opinion toward many, but not all, types of CAM. Further research is required to determine to what extent CAM can or should be integrated into the practice of rheumatology in the United States.


Performance, Fermentation Characteristics and Composition of the Microbiome in the Digest of Piglets Kept on a Feed With Humic Acid-Rich Peat.
The transition from breast milk to solid feed is a dramatic change in the nutrition of piglets, frequently necessitating antibiotic treatment. In efforts to reduce the use of antibiotics, dietetic concepts based on natural feed additives are becoming more and more important. In the present study, experiments were carried out with 15 rearing piglets (days 28-56) divided into three groups that were offered different diets (Ctr [0% peat]; H1.5 [1.5% peat]; and H3.0 [3.0% peat] based on a commercial weaner recipe; all ~178 g CP, 13.7 MJ ME, 13.3 g Lys, as-fed). The contents of cecal and colon digesta were removed at necropsy. The gas formation (4 h) in colon digesta was measured using in vitro batch fermenters. For microbiome studies, 16S rRNA amplification was performed within the hypervariable region V 4 and sequenced with Illumina MiSeq platform. DNA read mapping and statistical analysis were performed using QIIME (version 1.8.0), MicrobiomeAnalyst, RStudio, and SAS Enterprise Guide. The mean body weight of the animals at the end of the trial did not show statistical differences (in kg: Ctr: 26.1 ± 4.85, H1.5: 28.5 ± 3.41, H3.0: 26.2 ± 4.92). The daily weight gains were high for this age (in g/day; Ctr: 607 ± 157; H1.5: 692 ± 101; H3.0: 615 ± 113) and the feed to gain ratio low (in kg/kg; Ctr: 1.538; H1.5: 1.462; H3.0: 1.462). Concentrations of short-chain fatty acids in the cecal content were significantly lower when peat was used (mmol/kg wet weight; Ctr: 173 ± 30.0; H1.5:134 ± 15.0; H3.0:133 ± 17.3). Numerical differences were found in the gas formation (in mL gas per 10 mL batch in 4 h; Ctr: 7.9 ± 2.2; H1.5: 7.4 ± 2.4; H3.0: 6.6 ± 1.1). The microbiome analyses in the cecal content showed significantly higher values for alpha diversity Chao 1 index for samples from the control group. Significant differences were found for bacterial relative abundance for Tenericutes at phylum level and Mollicutes at class level (p < 0.05) in cecal microbiota. Therefore, there was initial evidence that peat influences intestinal microflora causing a shift in the overall concentration of fermentation products in both, the cecal and the colon content.


Regulation of macrophage alpha 2-macroglobulin receptor/low density lipoprotein receptor-related protein by lipopolysaccharide and interferon-gamma.
alpha 2-Macroglobulin receptor/low density lipoprotein receptor-related protein (alpha 2M-R/LRP) is a broad specificity receptor that may function in lipoprotein metabolism, proteinase regulation, and growth factor regulation. In this study, we demonstrated that alpha 2M-R/LRP expression in macrophages can be markedly decreased by LPS and by IFN-gamma. Regulation of alpha 2M-R/LRP in RAW 264.7 cells was demonstrated at the mRNA, antigen, and receptor-function levels. In receptor-function studies, the decrease in alpha 2M-R/LRP expression was detected as a 90% decrease in the Bmax or maximum receptor binding capacity for activated alpha 2M after treatment with LPS or IFN-gamma. Western blot analysis of whole cell lysates demonstrated significant loss of alpha 2M-R/LRP heavy-chain. Northern blot analysis of poly(A)+ RNA revealed a marked decrease in alpha 2M-R/LRP mRNA after treatment with LPS (79% decrease) or IFN-gamma (70% decrease). Other cytokines, including tumor necrosis factor-alpha, transforming growth factor-beta-1, and interleukin-6 did not regulate alpha 2M-R/LRP. The ability of LPS and IFN-gamma to regulate alpha 2M-R/LRP was confirmed in experiments with primary cultures of murine bone marrow macrophages. These studies demonstrate that macrophage alpha 2M-R/LRP is subject to significant downregulation by physiologically significant cytokines and signaling macromolecules.


Cardiovascular consequences of life-long exposure to dietary isoflavones in the rat.
Dietary soy intake in man is proposed to provide cardiovascular protection, but it is not established whether this property is attributable to the soy protein per se or to associated dietary isoflavones. This investigation aimed to establish whether the dietary isoflavones in soy protein affect cardiovascular function. Ten days prior to mating, male and female Wistar rats were habituated to either a soy based isoflavone rich diet (plasma concentration 1.87 micromol l(-1) isoflavones) or the same diet after isoflavone elution (plasma isoflavone not detectable). Offspring were weaned onto and maintained on the same diet as their dam and sire for 6 months. Blood pressure, and constrictor and dilator responses in the aorta and mesenteric resistance arteries were assessed at 3 and 6 months of age. There was no effect of isoflavone removal from the diet on blood pressure, heart rate, aortic function or mesenteric artery contractile function, at either 3 or 6 months of age. Resistance mesenteric arteries from 6-month-old female rats fed the isoflavone rich diet demonstrated a modest increase in arterial distensibility compared with those fed the depleted diet, and mesenteric arteries from male and female rats fed the isoflavone rich diet showed increased sensitivity to acetylcholine. In summary, the isoflavone content of soy protein has no influence on blood pressure in healthy rats fed a diet based on soy protein, but influences small artery function.


Cornea Thermography: Optimal Evaluation of the Outcome and the Resulting Reproducibility.
The aim of the study was to establish a standardized quantitative evaluation of corneal temperature (CT) that includes anchoring reference points in the topography and minimization of artifacts. We further investigated the distribution and the short- and long-term reproducibility of the CT values, as well as the influence of the core temperatures. The CT values in both eyes of 40 healthy subjects were measured through thermography. These examinations took place over the course of four visits within 2 consecutive weeks. At each visit, the CTs were measured twice in both eyes with intervals of 15 minutes between measurements. CT values were not significantly different between the right and left eyes and their distribution was nearly normal. The CTs increased slightly when measured twice over the 15-minute intervals (short-term reproducibility) but remained stable over a period of 2 weeks (long-term reproducibility). In addition, the CT values depended on the core temperatures. Ocular surface thermography is a fast and noninvasive examination. The methods of optimized and standardized evaluation of the CT values facilitate comparisons and follow-ups. Thermography can be used clinically and scientifically only if both the measurement and its evaluation are efficient and standardized and if the outcomes are highly reproducible.


Random walk properties from lattice bond enumeration: Steady-state diffusion on two- and three-dimensional lattices with traps.
We have applied the lattice bond enumeration method to the calculation of the steady-state diffusion in a lattice with fixed traps. We show that, to first order in density of traps, our random walk calculations for the effective diffusion constant in lattices with periodically arrayed traps are in exact agreement with calculations carried out previously for randomly arrayed traps embedded in a three-dimensional continuum medium (fluid). Our lattice random walk results are independent of dimension for d > 1, and we conjecture that this is also true for the continuum diffusion model.


Serum proteomic analysis identifies interleukin 16 as a biomarker for clinical response during early treatment of rheumatoid arthritis.
To conduct a comprehensive quantitative proteomics analysis of novel serum protein biomarkers based on synovitis status associated with matrix metalloproteinase-3 (MMP-3) and to determine the clinical significance of these biomarkers in rheumatoid arthritis (RA). Patients with untreated RA (n=28), primary Sjogren's syndrome (pSS) (n=30), and healthy controls (HCs) (n=30) were enrolled for the screening assay. A total of 1128 serum proteins were analyzed using the SOMAscan™ assay. Serum levels of MMP-3 and interleukin (IL)-16 were measured using a latex turbidimetric immunoassay and ELISA at baseline and 12weeks after treatment with methotrexate (MTX) for MTX-naïve RA patients (n=28) or with the biologics tocilizumab (TCZ) (n=7), abatacept (ABT) (n=11) or infliximab (n=22) for MTX-inadequate response (IR) RA patients. Correlation analysis was conducted using Spearman's rank correlation method. Proteomics showed that serum IL-16 levels were most positively correlated with those of MMP-3 (ρ=0.51, p<0.01) and were significantly increased in patients with untreated active RA compared to HCs (p<0.01) or those with pSS (p<0.01). IL-16 levels decreased following treatment in both the MTX-naïve and MTX-IR groups. Regarding clinical response, fluctuations in IL-16 levels were positively associated with changes in clinical indicators, particularly the Clinical Disease Activity Index (ρ=0.89, p<0.01) in the TCZ and ABT-treated group. However, no similar correlation was noted in MMP-3 and acute phase reactants in any groups. IL-16 was a more effective clinical parameter than MMP-3, C-reactive protein, or erythrocyte sedimentation rate in both MTX-naive and MTX-IR RA patients. IL-16 might be a useful biomarker for evaluating clinical response in RA patients.


Exploratory longitudinal cohort study of associations of fatigue after stroke.
The pathogenesis of poststroke fatigue is unclear. In this prospective study, we explored whether reduced physical activity might contribute to poststroke fatigue or be a consequence of it. Patients with a recent acute stroke were assessed at 1, 6, and 12 months with, Fatigue Assessment Scale (FAS), a fatigue case definition, Hospital Anxiety and Depression Score, sleepiness, quality of life, and accelerometry (ActivPAL). Bivariate analyses determined associations between fatigue and step count at each time point. Multiple linear regression tested whether 1-month step count independently predicted 6- and 12-month FAS. A total of 136 participants (mean age, 72 years; 64% men) attended ≥1 assessment. ActivPAL data were available for 84 (64%), 69 (66%), and 58 (64%) participants at 1, 6, and 12 months, respectively. At 6 and 12 months, a positive fatigue case definition was associated with lower daily step counts (P=0.014 and 0.013, respectively). At 1, 6, and 12 months, higher FAS (more fatigue) was associated with lower step count (P<0.001, 0.01, and 0.007), higher depression (P<0.001), anxiety scores (P<0.001) and sleepiness (P<0.001), and poorer quality of life (P<0.001). Lower daily step count (P<0.002 and 0.006) and greater anxiety (P<0.001 for both) at 1 month independently predicted higher FAS at 6 and 12 months. Lower step counts at 1 month independently predicted greater FAS for ≤12 months. Physical activity might be a therapeutic target for poststroke fatigue.