Thrombotic thrombocytopenic purpura: a syndrome of intravascular platelet consumption.
In four of five patients with thrombotic thrombocytopenic purpura (TTP) in whom serial tests of hemostatic function were performed, severe thrombocytopenia, normal plasma fibrinogen concentrations and mildly increased concentrations of fibrinogen/fibrin degradation products were observed. Widespread platelet thrombi were found in arterioles and capillaries. Fibrin could be seen around some of the platelet clumps and was the main component in a small number of the thrombi in two patients. The observations show that TTP is a disorder in which intravascular platelet consumption results in disseminated platelet thrombosis. The coagulation system is apparently activated secondarily to platelet aggregation and variable quantities of fibrin are incorporated into the thrombi. Clinical improvement resulted from combined therapy with corticosteroids, heparin and drugs that suppress platelet function.


Expression of the thiazide-sensitive Na-Cl cotransporter by rabbit distal convoluted tubule cells.
A thiazide-sensitive Na-Cl cotransporter contributes importantly to mammalian salt homeostasis by mediating Na-Cl transport along the renal distal tubule. Although it has been accepted that thiazide-sensitive Na-Cl cotransport occurs predominantly along the distal convoluted tubule in rats and mice, sites of expression in the rabbit have been controversial. A commonly accepted model of rabbit distal nephron transport pathways identifies the connecting tubule, not the distal convoluted tubule, as the predominant site of thiazide-sensitive Na-Cl cotransport. The thiazide-sensitive Na-Cl cotransporter has been cloned recently. The present experiments were designed to localize sites of thiazide-sensitive Na-Cl cotransporter mRNA expression along the rabbit distal nephron. Nonradioactive in situ hybridization with a thiazide-sensitive Na-Cl cotransporter probe was combined with immunocytochemistry with an antibody that recognizes distal convoluted tubule cells and with a Na+/Ca2+ exchanger antibody that recognizes only connecting tubule cells. The results indicate that thiazide-sensitive Na-Cl cotransporter mRNA is highly expressed by cells of the distal convoluted tubule and not by connecting tubule cells. Segments that stain with the Na+/Ca2+ exchanger antibody (connecting tubules) do not demonstrate thiazide-sensitive Na-Cl cotransporter mRNA expression. We conclude that the predominant site of thiazide-sensitive Na-Cl cotransporter mRNA expression in rabbit distal nephron is the distal convoluted tubule and that sites of mRNA expression of electroneutral Na and Cl transport are similar in rabbits, rats, and mice.


Precise Dispensing Technology Using Electroformed Tubes for Micro-Volume Blood Diagnosis.
Precise dispensing of droplets is a crucial step for acquiring reliable blood diagnostic results. When the source sample volume is limited, the need for precise dispensing of submicroliter and nanoliter quantities is especially important. In this paper, we developed a positive-displacement high-precision dispensing technique using a nickel electroformed tube with an inner diameter accuracy of [Formula: see text] or less. When dispensing variation of 100 nL was evaluated by using a photometric method, the most stable coefficients of variation (CV) were observed for a tube thickness of [Formula: see text] with hydrophobic treatment, where the average CV value was 1.3%, i.e., 1.3 nL. Furthermore, the glucose concentration of 100- and 200-nL animal-based control serum was colorimetrically measured using enzymatic reactions without drying and mixing reagents. The CV value was approximately 6.36% at 100 nL and 3.23% at 200 nL, suggesting that several biochemical panels can be precisely measured even from less than one drop of blood. This positive-displacement dispenser ensures zero contamination and almost-zero dead volume, thus it would be useful for multi-panel clinical blood testing.


Effects of diets supplemented with branched-chain amino acids on the performance and fatigue mechanisms of rats submitted to prolonged physical exercise.
This study aimed to determine the effects of diets chronically supplemented with branched-chain amino acids (BCAA) on the fatigue mechanisms of trained rats. Thirty-six adult Wistar rats were trained for six weeks. The training protocol consisted of bouts of swimming exercise (one hour a day, five times a week, for six weeks). The animals received a control diet (C) (n = 12), a diet supplemented with 3.57% BCAA (S1) (n = 12), or a diet supplemented with 4.76% BCAA (S2) (n = 12). On the last day of the training protocol, half the animals in each group were sacrificed after one hour of swimming (1H), and the other half after a swimming exhaustion test (EX). Swimming time until exhaustion was increased by 37% in group S1 and reduced by 43% in group S2 compared to group C. Results indicate that the S1 diet had a beneficial effect on performance by sparing glycogen in the soleus muscle (p < 0.05) and by inducing a lower concentration of plasma ammonia, whereas the S2 diet had a negative effect on performance due to hyperammonemia (p < 0.05). The hypothalamic concentration of serotonin was not significantly different between the 1H and EX conditions. In conclusion, chronic BCAA supplementation led to increased performance in rats subjected to a swimming test to exhaustion. However, this is a dose-dependent effect, since chronic ingestion of elevated quantities of BCAA led to a reduction in performance.


Expression profile of FGF receptors in preimplantation ovine embryos and the effect of FGF2 and PD173074.
Fibroblast growth factors (FGFs) and their receptors (FGFRs) are increasingly recognized as important regulators of embryo development in mammals. This study investigated the importance of FGF signaling during in vitro development of ovine embryo. The mRNAs of four FGFR subtypes were detected throughout preimplantation development of in vitro fertilized (IVF) embryos, peaked in abundance at the morula stage, and decreased significantly at the blastocyst stage. To gain insight into the role of these mRNAs in embryo development, IVF embryos were cultured in the presence of FGF2 (100 or 500 ng/ml: beginning from days 1 or 4 to 7) or PD173074 (1 µM: beginning from days 1 to 7) as usual treatments for activation or inhibition of FGFRs, respectively. FGF2-supplementation did not affect the percentage of embryos that developed to the blastocyst, blastocyst cell count and the proportion of cells allocated in inner cell mass (ICM) and trophectoderm (TE) compared to control (p > 0.05). Also, increasing the dosage or duration of FGF2 treatment did not significantly alter blastocyst yield or differential cell count (p > 0.05). PD173074-mediated inhibition of FGFRs did not significantly affect blastocyst yield (p > 0.05). Assessment of expression profiles of lineage-associated markers revealed that FGF2 (500 ng/ml) supplementation: (i) significantly increased expression of putative hypoblast marker (GATA4), (ii) significantly decreased expression of putative epiblast (EPI) marker (NANOG) and (iii) did not change TE markers (CDX2 and IFNT) and pluripotency makers (OCT4, SOX2 and REX1). In summary, FGF2-mediated activation of FGFRs may promote a switch in transcriptional profile of ovine ICM from EPI- to hypoblast-associated gene expression.


Haemodynamics and Flow Modification Stents for Peripheral Arterial Disease: A Review.
Endovascular stents are widely used for the treatment of peripheral arterial disease (PAD). However, the development of in-stent restenosis and downstream PAD progression remain a challenge. Stent revascularisation of PAD causes arterial trauma and introduces abnormal haemodynamics, which initiate complicated biological processes detrimental to the arterial wall. The interaction between stent struts and arterial cells in contact, and the blood flow field created in a stented region, are highly affected by stent design. Spiral flow is known as a normal physiologic characteristic of arterial circulation and is believed to prevent the development of flow disturbances. This secondary flow motion is lost in atheromatous disease and its re-introduction after endovascular treatment of PAD has been suggested as a method to induce stabilised and coherent haemodynamics. Stent designs able to generate spiral flow may support endothelial function and therefore increase patency rates. This review is focused on secondary flow phenomena in arteries and the development of flow modification stent technologies for the treatment of PAD.


Accessing Gold π-Acid Reactivity under Electrochemical Anode Oxidation (EAO) through Oxidation Relay.
The gold π-acid activation under electrochemical condition is achieved for the first time. While EAO allowing easy access to gold(III) intermediates over alternative chemical oxidation under mild conditions, the reported examples so far limited to coupling reactions due to the rapid AuIII reductive elimination. Using aryl hydrazine-HOTf salt as precursors, the π-activation reaction mode was realized through oxidation relay. Both alkene and alkyne di-functionalization were achieved with excellent functional group compatibility and regioselectivity, which extended the versatility and utility of electrochemical gold redox chemistry for future applications to come.


A Review on the Role of the Neuroscience of Flow States in the Modern World.
Flow states have been shown to help people reach peak performance, yet this elusive state is not easily attained. The review describes the current state of literature on flow by addressing the environmental influences as well as the cognitive and neurocognitive elements that underlie the experience. In particular, the research focusses on the transition of cognitive control from an explicit to an implicit process. This is further expanded upon to look at the current, yet related neurocognitive research of high performance associated with the implicit process of automaticity. Finally, the review focusses on transcranial direct current stimulation (tDCS) as a novel method to facilitates an induction of flow states. Implications are aimed at a general technique to improve on skill acquisition and overall performance.


Air traffic controllers' long-term speech-in-noise training effects: A control group study.
Speech perception in noise relies on the capacity of the auditory system to process complex sounds using sensory and cognitive skills. The possibility that these can be trained during adulthood is of special interest in auditory disorders, where speech in noise perception becomes compromised. Air traffic controllers (ATC) are constantly exposed to radio communication, a situation that seems to produce auditory learning. The objective of this study has been to quantify this effect. 19 ATC and 19 normal hearing individuals underwent a speech in noise test with three signal to noise ratios: 5, 0 and -5 dB. Noise and speech were presented through two different loudspeakers in azimuth position. Speech tokes were presented at 65 dB SPL, while white noise files were at 60, 65 and 70 dB respectively. Air traffic controllers outperform the control group in all conditions [P<0.05 in ANOVA and Mann-Whitney U tests]. Group differences were largest in the most difficult condition, SNR=-5 dB. However, no correlation between experience and performance were found for any of the conditions tested. The reason might be that ceiling performance is achieved much faster than the minimum experience time recorded, 5 years, although intrinsic cognitive abilities cannot be disregarded. ATC demonstrated enhanced ability to hear speech in challenging listening environments. This study provides evidence that long-term auditory training is indeed useful in achieving better speech-in-noise understanding even in adverse conditions, although good cognitive qualities are likely to be a basic requirement for this training to be effective. Our results show that ATC outperform the control group in all conditions. Thus, this study provides evidence that long-term auditory training is indeed useful in achieving better speech-in-noise understanding even in adverse conditions.


Brief report: Effects of exercise and self-affirmation intervention after traumatic brain injury.
Physical exercise has been proven to be an effective method for improving cognition and mood, but little is known about its benefits among individuals with traumatic brain injury. This pilot study investigated the feasibility of a combined exercise and self-affirmation intervention (IntenSati) for enhancing cognition and mood in individuals with TBI. It was hypothesized that this intervention would improve individuals' cognition and mood following the completion of the program. This intervention was held at an outpatient rehabilitation department in an urban medical center. A wait-list control design was conducted. Twenty-one adult participants-at least 12-months post-TBI-enrolled in the study. Twelve of them completed the study. Assessment was conducted at three time-points throughout the study using neuropsychological and self-report measures to evaluate participants' cognition and mood. Following initial evaluation, participants were assigned into either the immediate intervention group or the waitlist control group. During the intervention, participants attended the program twice a week over the course of 8 weeks. Debriefing was conducted following the completion of the program. Both independent t tests and paired t tests were utilized. Results indicated that the intervention group experienced less depressive symptoms following the completion of the IntenSati program compared to the waitlist control group. Participants also reported having less depressive symptoms, experienced more positive affect, and had a higher quality of life following the completion of the program. Moderate-to-large effect sizes were found on decrease in negative affect. However, results associated with cognitive benefits were mixed. The participants tolerated the program well and reported satisfaction with the program. Results from this study indicated that the IntenSati exercise program is a feasible and beneficial intervention for individuals with TBI as supported by the positive impact on their mood and quality of life.


A comparison of models of the isometric force of locust skeletal muscle in response to pulse train inputs.
Muscle models are an important tool in the development of new rehabilitation and diagnostic techniques. Many models have been proposed in the past, but little work has been done on comparing the performance of models. In this paper, seven models that describe the isometric force response to pulse train inputs are investigated. Five of the models are from the literature while two new models are also presented. Models are compared in terms of their ability to fit to isometric force data, using Akaike's and Bayesian information criteria and by examining the ability of each model to describe the underlying behaviour in response to individual pulses. Experimental data were collected by stimulating the locust extensor tibia muscle and measuring the force generated at the tibia. Parameters in each model were estimated by minimising the error between the modelled and actual force response for a set of training data. A separate set of test data, which included physiological kick-type data, was used to assess the models. It was found that a linear model performed the worst whereas a new model was found to perform the best. The parameter sensitivity of this new model was investigated using a one-at-a-time approach, and it found that the force response is not particularly sensitive to changes in any parameter.


Clinical and laboratory findings in border collies with presumed hereditary juvenile cobalamin deficiency.
Juvenile cobalamin deficiency is a rare disease in border collies and its diagnosis requires a high level of clinical suspicion. The goal of this study was to increase awareness of this disease by describing the clinical and laboratory findings in four young border collies with inherited cobalamin deficiency. The median age of the dogs was 11.5 mo (range, 8-42 mo), and two of the four dogs were full siblings. Clinical signs included intermittent lethargy (n = 4), poor body condition (n = 4), odynophagia (n = 2), glossitis (n = 1), and bradyarrhythmia (n = 1). Pertinent laboratory abnormalities were mild to moderate normocytic nonregenerative anemia (n = 3), increased aspartate aminotransferase (AST) activity (n = 3), and mild proteinuria (n = 3). All of the dogs had serum cobalamin levels below the detection limit of the assay, marked methylmalonic aciduria, and hyperhomocysteinemia. Full clinical recovery was achieved in all dogs with regular parenteral cobalamin supplementation, and laboratory abnormalities resolved, except the proteinuria and elevated AST activity persisted. This case series demonstrates the diverse clinical picture of primary cobalamin deficiency in border collies. Young border collies presenting with ambiguous clinical signs should be screened for cobalamin deficiency.


N-Terminal domain linkage modulates the folding properties of protein S epidermal growth factor-like modules.
Protein S interacts with activated protein C to play a crucial role in blood anticoagulation, and protein S deficiency is associated with increased risk of thrombosis. Despite the large volume of functional data available for this protein, no atomic resolution structure data have yet been reported. This is due at least in part to difficulties encountered when trying to produce fragments dissected from the intact protein; however, a few successful strategies have been described. In this research we have expressed a number of constructs containing protein S epidermal growth factor-like (EGF) domains 1 and 2 in Escherichia coli and Pichia pastoris. None of the proteins produced was stably folded as assayed by solution nuclear magnetic resonance spectroscopy. We therefore constructed a series of non-native protein S EGF concatemers to investigate the role of pairwise domain linkage in domain folding. Our results demonstrate that N-terminal domain linkage can either positively or negatively impact on the refolding of an adjacent domain. Furthermore, analysis of the NMR data for EGF3-4 reveals the expected interdomain NOEs that are characteristic of an extended arrangement of calcium-binding EGF domains and a similar average [(1)H]-(15)N heteronuclear NOE value for each of the two domains. These results provide the first data in support of protein S EGF3-4 adopting the same extended domain orientation as observed for the functionally distinct proteins fibrillin-1 and the low-density lipoprotein receptor. The results also have important implications for future studies, particularly when a dissection approach is used, of tandem EGF domains from protein S and other proteins.


Domestic impact of tuberculosis screening among new immigrants to Ontario, Canada.
All Canadian immigrants undergo screening for tuberculosis (TB) before immigration, and selected immigrants must undergo postimmigration surveillance for the disease. We sought to quantify the domestic health impact of screening for TB in all new immigrants and to identify mechanisms to enhance effectiveness and efficiency of this screening. We linked preimmigration medical examination records from 944,375 immigrants who settled in Ontario between 2002 and 2011 to active TB reporting data in Ontario between 2002 and 2011. Using a retrospective cohort study design, we measured birth country-specific rates of active TB detected through preimmigration screening and postimmigration surveillance. We then quantified the proportion of active TB cases among residents of Ontario born abroad that were detected through postimmigration surveillance. Using Cox regression, we identified independent predictors of active TB postimmigration. Immigrants from 6 countries accounted for 87.3% of active TB cases detected through preimmigration screening, and 10 countries accounted for 80.4% of cases detected through postimmigration surveillance. Immigrants from countries with a TB (all-sites) incidence rate of less than 30 cases per 100 000 persons resulted in pre- and postimmigration detection of 2.4 and 0.9 cases per 100 000 immigrants, respectively. Postimmigration surveillance detected 2.6% of active TB cases in Ontario residents born abroad, and TB was detected a median of 18 days earlier in those undergoing surveillance than in those who were not referred to surveillance or who did not comply. Predictors of active TB postimmigration included radiographic markers of old TB, birth country, immigration category, location of application for residency, immune status and age. Universal screening for TB in new immigrants has a modest impact on the domestic burden of active TB and is highly inefficient. Focusing preimmigration screening in countries with high incidence rates and revising criteria for postimmigration surveillance could increase the effectiveness and efficiency of screening.


Evidence for Functionally Relevant Encounter Complexes in Nitrogenase Catalysis.
Nitrogenase is the only enzyme that can convert atmospheric dinitrogen (N2) into biologically usable ammonia (NH3). To achieve this multielectron redox process, the nitrogenase component proteins, MoFe-protein (MoFeP) and Fe-protein (FeP), repeatedly associate and dissociate in an ATP-dependent manner, where one electron is transferred from FeP to MoFeP per association. Here, we provide experimental evidence that encounter complexes between FeP and MoFeP play a functional role in nitrogenase catalysis. The encounter complexes are stabilized by electrostatic interactions involving a positively charged patch on the β-subunit of MoFeP. Three single mutations (βAsn399Glu, βLys400Glu, and βArg401Glu) in this patch were generated in Azotobacter vinelandii MoFeP. All of the resulting variants displayed decreases in specific catalytic activity, with the βK400E mutation showing the largest effect. As simulated by the Thorneley-Lowe kinetic scheme, this single mutation lowered the rate constant for FeP-MoFeP association 5-fold. We also found that the βK400E mutation did not affect the coupling of ATP hydrolysis with electron transfer (ET) between FeP and MoFeP. These data suggest a mechanism where FeP initially forms encounter complexes on the MoFeP β-subunit surface en route to the ATP-activated, ET-competent complex over the αβ-interface.


Clinical features and outcome of infective endocarditis in yemeni patients treated with empirical antibiotic therapy.
Infective endocarditis is a common disease in Yemen. Although the incidence of rheumatic valvular disease and uncorrected congenital heart disease in adults is high in Yemen, there are few data regarding the pattern, characteristic features and outcome of infective endocarditis in Yemen. The aim was to study the characteristic, clinical features and diagnostic criteria of infective endocarditis in Yemeni patients and the outcome in patients treated with medical therapy. Seventy-two consecutive patients admitted to Kuwait teaching hospital in Sana'a with suspected infective endocarditis between June 1, 2005 and June 1, 2007 were included in this study. A questionnaire including history, clinical findings, and result of requested investigations, treatment, complications and outcome was used. The diagnosis was based on Dukes criteria, which proposed two major or five minor criteria. In our study, we included raised erythrocyte sedimentation rate (ESR) as a minor criteria. The patients were classified as definite, possible and rejected cases. All patients received empirical antibiotic therapy. The mean age was 28.56 ± 14.5 years. Men were 30 (42.2%) while women were 42 (57.7%). 59 (81.9%) of the patients had been admitted due to fever. Past history of rheumatic heart disease was positive in 38 (53.3%) of the patients. Mitral regurgitation was the commonest form of valvular affection 54 (82.1%) of patients followed by aortic regurgitation in 45 (63%). Anemia was present in 53 (74%) while raised erythrocyte sedimentation rate was high in all patients. Vegetations were detected by transthorasic echocardiography in 51 (70.83%) of the patients while blood culture was positive in only 7 (9.6%). After adding high ESR to the criteria, definite infective endocarditis were found in 34 (47.2%); possible in 38 (52.7%) and no rejected patients. Sixty-two (87.5%) of the patients improved and were discharged in good general condition. Nine patients died, an In hospital mortality of 12.5 %. patients with IE in Yemen were younger than those patients in western countries, RHD is the commonest predisposing factor. Duke's criteria in addition to high ESR as minor criteria improved the diagnostic possibilities and using empirical antibiotic therapy improves the outcome.


Coordinated concentration changes of transcripts and metabolites in Saccharomyces cerevisiae.
Metabolite concentrations can regulate gene expression, which can in turn regulate metabolic activity. The extent to which functionally related transcripts and metabolites show similar patterns of concentration changes, however, remains unestablished. We measure and analyze the metabolomic and transcriptional responses of Saccharomyces cerevisiae to carbon and nitrogen starvation. Our analysis demonstrates that transcripts and metabolites show coordinated response dynamics. Furthermore, metabolites and gene products whose concentration profiles are alike tend to participate in related biological processes. To identify specific, functionally related genes and metabolites, we develop an approach based on Bayesian integration of the joint metabolomic and transcriptomic data. This algorithm finds interactions by evaluating transcript-metabolite correlations in light of the experimental context in which they occur and the class of metabolite involved. It effectively predicts known enzymatic and regulatory relationships, including a gene-metabolite interaction central to the glycolytic-gluconeogenetic switch. This work provides quantitative evidence that functionally related metabolites and transcripts show coherent patterns of behavior on the genome scale and lays the groundwork for building gene-metabolite interaction networks directly from systems-level data.


Focal adhesion kinase is critical for entry of Kaposi's sarcoma-associated herpesvirus into target cells.
Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 (KSHV/HHV-8) interacts with cell surface alpha3beta1 integrin early during in vitro infection of human endothelial cells and fibroblasts and activates the focal adhesion kinase (FAK) that is immediately downstream in the outside-in signaling pathway by integrins, leading to the activation of several downstream signaling molecules. In this study, using real-time DNA and reverse transcription-PCR assays to measure total internalized viral DNA, viral DNA associated with infected nuclei, and viral gene expression, we examined the stage of infection at which FAK plays the most significant role. Early during KSHV infection, FAK was phosphorylated in FAK-positive Du17 mouse embryonic fibroblasts. The absence of FAK in Du3 (FAK(-/-)) cells resulted in about 70% reduction in the internalization of viral DNA, suggesting that FAK plays a role in KSHV entry. Expression of FAK in Du3 (FAK(-/-)) cells via an adenovirus vector augmented the internalization of viral DNA. Expression of the FAK dominant-negative mutant FAK-related nonkinase (FRNK) in Du17 cells significantly reduced the entry of virus. Virus entry in Du3 cells, albeit in reduced quantity, delivery of viral DNA to the infected cell nuclei, and expression of KSHV genes suggested that in the absence of FAK, another molecule(s) may be partially compensating for FAK function. Infection of Du3 cells induced the phosphorylation of the FAK-related proline-rich tyrosine kinase (Pyk2) molecule, which has been shown to complement some of the functions of FAK. Expression of an autophosphorylation site mutant of Pyk2 in which Y402 is mutated to F (F402 Pyk2) reduced viral entry in Du3 cells, suggesting that Pyk2 facilitates viral entry moderately in the absence of FAK. These results suggest a critical role for KSHV infection-induced FAK in the internalization of viral DNA into target cells.


California pediatricians' knowledge of and response to recommendations for universal infant hepatitis B immunization.
To assess California pediatricians' level of agreement with and implementation of universal infant hepatitis B immunization. Mailed questionnaire in the summer and fall of 1993 to a random sample of 1030 California pediatricians to determine whether they universally immunize infants with hepatitis B vaccine. The response rate was 71%. More than 81% said they universally immunize infants against hepatitis B infection; however, 18.3% stated that they do not. Pediatricians were more likely to immunize infants universally against hepatitis B if they (1) agreed with the recommendations (90% vs 54%, P < .01), (2) practiced in a health maintenance organization setting vs private group practice (92% vs 79%, P < .05), or (3) practiced in settings with predominantly low-income patients (89% vs 78%, P = .01). Pediatricians who universally immunize infants against hepatitis B virus were in practice an average of 1.7 years longer than those who do not (13.3 years vs 11.6 years, P = .01). In multivariate analyses, the most powerful predictor of universal immunization was agreement with the recommendations. Among those who disagreed with the recommendations, the most common reasons for disagreement included the following: (1) the long-term efficacy of hepatitis B vaccine is not proved--54%; (2) an additional immunization would make the visits too costly--53%; and (3) three shots would be too much at one visit--53%. A majority (81.6%) of our sample of California pediatricians universally immunize infants against hepatitis B; however, a significant minority (18.4%) do not agree with the recommendations and do not plan to implement them. Many of their objections are being addressed through combination vaccine development and ongoing research; however, alternative policies and additional research should also be considered.


microRNA involvement in human cancer.
When, ∼20 years ago, investigators first determined that components of the genome considered nonfunctional had, in fact, gene regulatory capacity, they probably had no idea of their potential in controlling cell fate and were forced to revise and somehow reorganize their view of the molecular biology. Indeed, it is currently well documented how a class of small non-coding RNAs, microRNAs, are conserved among the species, expressed in different tissues and cell types and involved in almost every biological process, including cell cycle, growth, apoptosis, differentiation and stress response, exerting a finely tuned regulation of gene expression by targeting multiple molecules. As a consequence of the widespread range of processes they are able to influence, it is not surprising that miRNA deregulation is a hallmark of several pathological conditions, including cancer. Indeed, the aberrant expression of these tiny molecules in human tumors is not just a casual association, but they can exert a causal role, as oncogenes or tumor suppressors, in different steps of the tumorigenic process, from initiation and development to progression toward the acquisition of a metastatic phenotype. An increasing body of evidence has indeed proved the importance of miRNAs in cancer, suggesting their possible use as diagnostic, prognostic and predictive biomarkers and leading to exploit miRNA-based anticancer therapies, either alone or in combination with current targeted therapies, with the goal to improve disease response and increase cure rates. Here, we review our current knowledge about miRNA involvement in cancer.


Transformation-optical design of adaptive beam bends and beam expanders.
We describe the design of adaptive beam bends and beam splitters with arbitrary bend and split angles by use of finite embedded coordinate transformations. The devices do not exhibit reflection at the entrance or exit surfaces. It is shown that moderate and practically achievable values of the relative permittivity and permeability can be obtained for beam bends and splitters with both small and large bend radius. The devices are also discussed in the context of reconfigurable metamaterials, in which the bend and split angles can be dynamically tuned. The performance of adaptive beam bends and splitters is demonstrated in full wave simulations based on a finite-element method. Furthermore, the design of an adaptively adjustable transformation-optical beam expander/compressor is presented. It is observed that a pure transformation-optical design cannot result in a reflectionless beam expander/compressor.


"Every Newborn-INDEPTH" (EN-INDEPTH) study protocol for a randomised comparison of household survey modules for measuring stillbirths and neonatal deaths in five Health and Demographic Surveillance sites.
Under-five and maternal mortality were halved in the Millennium Development Goals (MDG) era, with slower reductions for 2.6 million neonatal deaths and 2.6 million stillbirths. The Every Newborn Action Plan aims to accelerate progress towards national targets, and includes an ambitious Measurement Improvement Roadmap. Population-based household surveys, notably Demographic and Health Surveys (DHS) and Multiple Indicator Cluster Surveys, are major sources of population-level data on child mortality in countries with weaker civil registration and vital statistics systems, where over two-thirds of global child deaths occur. To estimate neonatal/child mortality and pregnancy outcomes (stillbirths, miscarriages, birthweight, gestational age) the most common direct methods are: (1) the standard DHS-7 with Full Birth History with additional questions on pregnancy losses in the past 5 years (FBH+) or (2) a Full Pregnancy History (FPH). No direct comparison of these two methods has been undertaken, although descriptive analyses suggest that the FBH+ may underestimate mortality rates particularly for stillbirths. This is the protocol paper for the Every Newborn-INDEPTH study (INDEPTH Network, International Network for the Demographic Evaluation of Populations and their Health Every Newborn, Every Newborn Action Plan), aiming to undertake a randomised comparison of FBH+ and FPH to measure pregnancy outcomes in a household survey in five selected INDEPTH Network sites in Africa and South Asia (Bandim in urban and rural Guinea-Bissau; Dabat in Ethiopia; IgangaMayuge in Uganda; Kintampo in Ghana; Matlab in Bangladesh). The survey will reach >68 000 pregnancies to assess if there is ≥15% difference in stillbirth rates. Additional questions will capture birthweight, gestational age, birth/death certification, termination of pregnancy and fertility intentions. The World Bank's Survey Solutions platform will be tailored for data collection, including recording paradata to evaluate timing. A mixed methods assessment of barriers and enablers to reporting of pregnancy and adverse pregnancy outcomes will be undertaken. This large-scale study is the first randomised comparison of these two methods to capture pregnancy outcomes. Results are expected to inform the evidence base for survey methodology, especially in DHS, regarding capture of stillbirths and other outcomes, notably neonatal deaths, abortions (spontaneous and induced), birthweight and gestational age. In addition, this study will inform strategies to improve health and demographic surveillance capture of neonatal/child mortality and pregnancy outcomes.


Ocular surface cytokine profile in chronic Stevens-Johnson syndrome and its response to mucous membrane grafting for lid margin keratinisation.
To study the tear cytokine and the conjunctival and oral mucosal marker profile in chronic ocular Stevens-Johnson syndrome (SJS) and their alteration following mucous membrane grafting (MMG) for lid margin keratinisation (LMK). In a 1-year prospective study, SJS cases (n=25) and age-matched/sex-matched healthy controls (n=25) were recruited. Tear specimen (Schirmer's strip), conjunctival and oral mucosal imprints were collected from controls and SJS cases pre-MMG and post-MMG (at first follow-up, n=17). Tear cytokines were profiled using 27-bioplex array. Transforming growth factor-beta (TGF-β)-mediated extracellular matrix changes in conjunctival and oral mucosal cells were analysed by gene expression studies. 30 RESULTS: Tear cytokine profiling of chronic SJS cases at pre-MMG stage revealed significant upregulation of cytokines granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-8, IL-1β, monocyte chemoattractant protein-1, IL-15, IL-2, IL-17A and basic fibroblast growth factor (bFGF) with downregulation of IP-10 (interferon gamma-induced protein 10), tumour necrosis factor-α, interferon-γ, IL-10, vascular endothelial growth factor, regulated upon activation normal T-cell expressed and secreted (RANTES), IL-7, IL-12p70 and IL-13, with maximal increase in GM-CSF and maximal downregulation of IP-10, respectively. Of these, IL-2, IL-15, bFGF and IL-17A showed significant correlation with disease severity, pre-MMG. Conjunctival cells pre-MMG showed increase in TGF-β1, TGF-βRII, connective tissue growth factor and collagen-III gene expression by 10, 67, 173 and 184 folds, respectively, which dropped to 1.3, 11, 13.5 and 19 folds correspondingly, post-MMG. However, their expressions in oral mucosa were negligible. A proinflammatory, profibrotic, antiapoptotic ocular surface milieu characterises chronic ocular SJS. IP-10, an antifibrotic cytokine was noted to be maximally downregulated, unlike in other forms of chronic dry eye disease. The alterations in the ocular surface are seen to reverse largely with MMG for LMK.


A preliminary validation of the Swedish version of the Critical-Care Pain Observation Tool in adults.
Assessing pain in critically ill patients can be complicated, especially for those unable to communicate. A recently developed pain assessment tool, the Critical-Care Pain Observation Tool (CPOT), has been shown to be a reliable tool for pain assessment in the Intensive Care Unit (ICU). The aim of the study was to validate the Swedish version of the CPOT. Conscious and unconscious adults were observed during two procedures: one non-nociceptive procedure (NNP) (arm- and face wash) and one nociceptive procedure (NP) (turning). In total, there were 240 patient assessments pre-, per- and post-procedure performed by two independent staff members at rest, during and 15 min after the different procedures. Measures of interrater reliability, internal consistency and discriminant validity of the CPOT were obtained to examine the properties of the Swedish version of CPOT. The results provide indications of good agreement between the independent raters (ICC=0.84). There was an adequate discriminant validity of the Swedish version of CPOT established by a significant peak for CPOT scores during the NP (per-procedure). There was also a consistent pattern of significant correlations between CPOT and the mean artery pressure (ρ=0.32-0.45). The Swedish version of the CPOT is a suitable instrument for assessing pain in critically ill adults. The overall reliability and validity measures converge with findings from previous studies of the CPOT, but in order to achieve enhanced generalizability of the CPOT, we encourage further evaluation of CPOT in broader groups of critically ill patients.


Successive Emergence of Ceftazidime-Avibactam Resistance through Distinct Genomic Adaptations in blaKPC-2-Harboring Klebsiella pneumoniae Sequence Type 307 Isolates.
Ceftazidime-avibactam (CAZ-AVI) is a promising novel treatment for infections caused by carbapenem-resistant Enterobacteriaceae (CRE). Despite improved treatment outcomes compared to those achieved with aminoglycoside- and colistin-based regimens, the rapid evolution of CAZ-AVI resistance during treatment has previously been reported in Klebsiella pneumoniae sequence type 258 (ST258) blaKPC-3-harboring isolates. Here, we report the stepwise evolution and isolation of two phenotypically distinct CAZ-AVI-resistant Klebsiella pneumoniae isolates from a patient with pancreatitis. All susceptible (n = 3) and resistant (n = 5) isolates were of the ST307 clonal background, a rapidly emerging clone. Taking advantage of short-read Illumina and long-read Oxford Nanopore sequencing and full-length assembly of the core chromosome and plasmids, we demonstrate that CAZ-AVI resistance first occurred through a 532G → T blaKPC-2 point mutation in blaKPC-2 (D179Y protein substitution) following only 12 days of CAZ-AVI exposure. While subsequent isolates exhibited substantially decreased meropenem (MEM) MICs (≤2 μg/ml), later cultures demonstrated a second CAZ-AVI resistance phenotype with a lower CAZ-AVI MIC (12 μg/ml) but also MEM resistance (MIC > 128 μg/ml). These CAZ-AVI- and MEM-resistant isolates showed evidence of multiple genomic adaptations, mainly through insertions and deletions. This included amplification and transposition of wild-type blaKPC-2 into a novel plasmid, an IS1 insertion upstream of ompK36, and disruption of the rfb gene locus in these isolates. Our findings illustrate the potential of CAZ-AVI resistance to emerge in non-K. pneumoniae ST258 clonal backgrounds and alternative blaKPC variants. These results raise concerns about the strong selective pressures incurred by novel carbapenemase inhibitors, such as avibactam, on isolates previously considered invulnerable to CAZ-AVI resistance. There is an urgent need to further characterize non-KPC-mediated modes of carbapenem resistance and the intrinsic bacterial factors that facilitate the rapid emergence of resistance during treatment.


Unconventional therapies for cancer: 1. Essiac. The Task Force on Alternative Therapies of the Canadian Breast Cancer Research Initiative.
Physicians and patients have been frustrated by the lack of reliable information on unconventional therapies. To help fill this gap in the area of breast cancer therapy, the Canadian Breast Cancer Research Initiative formed a task force to advise it on how best to promote research into unconventional therapies. As part of the work of the task force, a review of the available literature was carried out for each of the following products: Essiac, green tea, Iscador, hydrazine sulfate, vitamins A, C and E, and 714-X. The first article in this series on unconventional therapies for cancer describes the methodology used to obtain and evaluate the information and provides a summary of the findings on Essiac. Subsequent articles will cover the other products. For most of the products reviewed, there has been some indication of possible benefit but no definitive evidence. Innovative and collaborative research needed to meet the information needs of growing numbers of patients and their physicians is now being sponsored by the Canadian Breast Cancer Research Initiative. Open communication between patients and physicians is also necessary for the maintenance of an appropriate therapeutic partnership and for the identification and control of side effects. The Ontario Division of the Canadian Cancer Society, a partner in the Canadian Breast Cancer Research Initiative, supported the preparation of a patient-information piece on unconventional therapies to accompany the series. This item will assist patients who are considering such therapies and will promote open communication between patients and their physicians.


The influence of surgical excision margins on keloid prognosis.
Keloid disease is known to have variable clinical behavior in response to therapy and there is no clinicopathologic classification that predicts such varied behavior. The aim of this study was to study the effect of excision margins and other histopathologic characteristics on keloid prognosis.Seventy-five multiethnic patients presenting with keloid scars at a department of plastic and reconstructive surgery during an 11-year period were included in this study. Clinical data was collected and detailed histologic analyses using light microscopy were carried out on archived patient specimens.A detailed histopathologic examination of all tissue samples identified keloid border or margin characteristics which were classified into "circumscribed" (borders clearly-demarcated) and "infiltrative" (borders not clearly-demarcated and not easily-definable). The specific histologic findings were correlated with keloid recurrence which revealed that incomplete peripheral (P < 0.001) and deep excision margins (P < 0.001), as well as infiltrative borders (P < 0.05) were associated with higher 1-year reported recurrence rates.This study has given evidence that incomplete surgical excision are associated with higher recurrence and this may justify the practice of routine histopathologic reporting of keloid excision margins.


Effect of substrate features and mutagenesis of active site tyrosine residues on the reaction course catalysed by Trypanosoma brucei sterol C-24-methyltransferase.
TbSMT [Trypanosoma brucei 24-SMT (sterol C-24-methyltransferase)] synthesizes an unconventional 24-alkyl sterol product set consisting of Δ24(25)-, Δ24(28)- and Δ25(27)-olefins. The C-methylation reaction requires Si(β)-face C-24-methyl addition coupled to reversible migration of positive charge from C-24 to C-25. The hydride shifts responsible for charge migration in formation of multiple ergostane olefin isomers catalysed by TbSMT were examined by incubation of a series of sterol acceptors paired with AdoMet (S-adenosyl-L-methionine). Results obtained with zymosterol compared with the corresponding 24-2H and 27-13C derivatives revealed isotopic-sensitive branching in the hydride transfer reaction on the path to form a 24-methyl-Δ24(25)-olefin product (kinetic isotope effect, kH/kD=1.20), and stereospecific CH3→CH2 elimination at the C28 branch and C27 cis-terminal methyl to form Δ24(28) and Δ25(27) products respectively. Cholesta-5,7,22,24-tetraenol converted into ergosta-5,7,22,24(28)-tetraenol and 24β-hydroxy ergosta-5,7,23-trienol (new compound), whereas ergosta-5,24-dienol converted into 24-dimethyl ergosta-5,25(27)-dienol and cholesta-5,7,24-trienol converted into ergosta-5,7,25(27)trienol, ergosta-5,7,24(28)-trienol, ergosta-5,7,24-trienol and 24 dimethyl ergosta-5,7,25(27)-trienol. We made use of our prior research and molecular modelling of 24-SMT to identify contact amino acids that might affect catalysis. Conserved tyrosine residues at positions 66, 177 and 208 in TbSMT were replaced with phenylalanine residues. The substitutions generated variable loss of activity during the course of the first C-1-transfer reaction, which differs from the corresponding Erg6p mutants that afforded a gain in C-2-transfer activity. The results show that differences exist among 24-SMTs in control of C-1- and C-2-transfer activities by interactions of intermediate and aromatic residues in the activated complex and provide an opportunity for rational drug design of a parasite enzyme not synthesized by the human host.


Different HLA haplotypes in Mexican Americans with IDDM.
The study of HLA histocompatibility antigens and insulin-dependent diabetes mellitus (IDDM) in non-White populations may provide a unique opportunity to more accurately define the diabetes susceptibility gene(s) located within the HLA region. To determine whether HLA haplotypes differ between ethnic groups, we compared 105 HLA haplotypes from 55 Mexican-American IDDM patients with 272 haplotypes from 136 IDDM patients of non-Hispanic White descent. The accurate determination of genotypes and haplotypes requires the study of family units. Therefore, all diabetic patients in this study were from studies of families having one or more siblings with IDDM. In the Mexican-American group, HLA-DR3 and -DR4 were the most common HLA-DR alleles and were present in comparable frequencies in the non-Hispanic White group (HLA-DR3, 27% of Mexican-American and 29% of non-Hispanic White haplotypes; DR4, 46% of Mexican-American and 43% of non-Hispanic White haplotypes). However, the HLA-B/DR-containing haplotypes and haplotype frequencies differed between the two groups. Several common haplotypes (B8/DR3, B15/DR4) in the non-Hispanic White group occurred less frequently in the Mexican-American group. In contrast, uncommon haplotypes in the non-Hispanic White group comprised nearly 50% of the DR4-containing haplotypes (B35/DR4, B40/DR4, B44/DR4) in the Mexican-American group. Although both DR3- and DR4-haplotype frequencies differed significantly between the two groups, the relative frequency of DR3- but not DR4-containing haplotypes was similar in both ethnic groups. This adds to the evidence suggesting that different susceptibilities are provided by the haplotypes carrying the DR3 and DR4 alleles.


In vivo quantification of glucose uptake and conversion to glycogen in individual muscles of the rat following exercise.
Glycogen depletion is thought to be a potent stimulus for the substantially increased glucose fluxes observed in skeletal muscle following exercise. The aim of this study was to establish the relationships between the glycogen mass and the rates of glucose uptake (Rg') and glucose incorporation into glycogen (Rgly) in individual muscles of conscious adult Wistar rats following moderate nonexhausting treadmill exercise (15 m/min at a 10 degree slope for 45 minutes, approximately 65% VO2max). Muscle glycogen content was determined at 0, 20, 45, 90, or 135 minutes following exercise and compared with Rg' and Rgly measurements at matched times. Muscle types varied in the rate of glycogen resynthesis. Glycogen depots of glycolytic muscle (white gastrocnemius) were still significantly (P < .01) lower than preexercise levels after 135 minutes; red oxidative muscles (soleus and red gastrocnemius) were essentially repleted by 90 minutes. Immediately following exercise, Rg' and Rgly in red gastrocnemius and soleus were 42 +/- 4 and 42 +/- 5 and 36 +/- 2 and 33 +/- 7 micromol/(min . 100 g), greater than the rates induced by maximal insulin stimulation in previous studies. In red muscles, there was a strong inverse relationship between Rgly and tissue glycogen content, consistent with a dominant role for the glycogen mass in the regulation of glycogen resynthesis.


Duffy negative antigen is no longer a barrier to Plasmodium vivax--molecular evidences from the African West Coast (Angola and Equatorial Guinea).
Plasmodium vivax shows a small prevalence in West and Central Africa due to the high prevalence of Duffy negative people. However, Duffy negative individuals infected with P. vivax have been reported in areas of high prevalence of Duffy positive people who may serve as supply of P. vivax strains able to invade Duffy negative erythrocytes. We investigated the presence of P. vivax in two West African countries, using blood samples and mosquitoes collected during two on-going studies. Blood samples from a total of 995 individuals were collected in seven villages in Angola and Equatorial Guinea, and 820 Anopheles mosquitoes were collected in Equatorial Guinea. Identification of the Plasmodium species was achieved by nested PCR amplification of the small-subunit rRNA genes; P. vivax was further characterized by csp gene analysis. Positive P. vivax-human isolates were genotyped for the Duffy blood group through the analysis of the DARC gene. Fifteen Duffy-negative individuals, 8 from Equatorial Guinea (out of 97) and 7 from Angola (out of 898), were infected with two different strains of P. vivax (VK210 and VK247). In this study we demonstrated that P. vivax infections were found both in humans and mosquitoes, which means that active transmission is occurring. Given the high prevalence of infection in mosquitoes, we may speculate that this hypnozoite-forming species at liver may not be detected by the peripheral blood samples analysis. Also, this is the first report of Duffy negative individuals infected with two different strains of P. vivax (VK247 and classic strains) in Angola and Equatorial Guinea. This finding reinforces the idea that this parasite is able to use receptors other than Duffy to invade erythrocytes, which may have an enormous impact in P. vivax current distribution.


Establishing a regional, multisite database for quality improvement and service planning in community-based palliative care and hospice.
Outpatient palliative care, an evolving delivery model, seeks to improve continuity of care across settings and to increase access to services in hospice and palliative medicine (HPM). It can provide a critical bridge between inpatient palliative care and hospice, filling the gap in community-based supportive care for patients with advanced life-limiting illness. Low capacities for data collection and quantitative research in HPM have impeded assessment of the impact of outpatient palliative care. In North Carolina, a regional database for community-based palliative care has been created through a unique partnership between a HPM organization and academic medical center. This database flexibly uses information technology to collect patient data, entered at the point of care (e.g., home, inpatient hospice, assisted living facility, nursing home). HPM physicians and nurse practitioners collect data; data are transferred to an academic site that assists with analyses and data management. Reports to community-based sites, based on data they provide, create a better understanding of local care quality. The data system was developed and implemented over a 2-year period, starting with one community-based HPM site and expanding to four. Data collection methods were collaboratively created and refined. The database continues to grow. Analyses presented herein examine data from one site and encompass 2572 visits from 970 new patients, characterizing the population, symptom profiles, and change in symptoms after intervention. A collaborative regional approach to HPM data can support evaluation and improvement of palliative care quality at the local, aggregated, and statewide levels.


Developmental pattern formation: insights from physics and biology.
The spatial organization of cell fates during development involves the interpretation of morphogen gradients by cellular signaling cascades and transcriptional networks. Recent studies use biophysical models, genetics, and quantitative imaging to unravel how tissue-level morphogen behavior arises from subcellular events. Moreover, data from several systems show that morphogen gradients, downstream signaling, and the activity of cell-intrinsic transcriptional networks change dynamically during pattern formation. Studies from Drosophila and now also vertebrates suggest that transcriptional network dynamics are central to the generation of gene expression patterns. Together, this leads to the view that pattern formation is an emergent behavior that results from the coordination of events occurring across molecular, cellular, and tissue scales. The development of novel approaches to study this complex process remains a challenge.


The Prevalence of Tree Nut Allergy: A Systematic Review.
Tree nuts are one of the most common foods causing acute allergic reactions and nearly all tree nuts have been associated with fatal allergic reactions. Despite their clinical importance, tree nut allergy epidemiology remains understudied and the prevalence of tree nut allergy in different regions of the world has not yet been well characterised. We aimed to systematically review the population prevalence of tree nut allergy in children and adults. We searched three electronic databases (OVID MEDLINE, EMBASE and PubMed) from January 1996 to December 2014. Eligible studies were categorised by age, region and method of assessment of tree nut allergy. Of the 36 studies identified most were in children (n = 24) and from Europe (n = 18), UK (n = 8) or USA (n = 5). Challenge-confirmed IgE-mediated tree nut allergy prevalence was less than 2 % (although only seven studies used this gold standard) while probable tree nut allergy prevalence ranged from 0.05 to 4.9 %. Prevalence estimates that included oral allergy syndrome (OAS) reactions to tree nut were significantly higher (8-11.4 %) and were predominantly from Europe. Prevalence of individual tree nut allergies varied significantly by region with hazelnut the most common tree nut allergy in Europe, walnut and cashew in the USA and Brazil nut, almond and walnut most commonly reported in the UK. Monitoring time trends of tree nut allergy prevalence (both overall and by individual nuts) as well as the prevalence of OAS should be considered given the context of the overall recent rise in IgE-mediated food allergy prevalence in the developed world.


Gaining insight into the role of FoxO1 in the progression of disuse-induced skeletal muscle atrophy.
Expression of FoxO transcription factors increases during certain forms of atrophy. In a dephosphorylated state, FoxOs participate in ubiquitin-mediated proteasomal degradation through the transcriptional activation of E3-ubiquitin ligases such as MAFbx/atrogin-1 and MuRF1. There is exhaustive research demonstrating that FoxO3a is sufficient to induce MAFbx/atrogin-1 and MuRF-1 expressions. In contrast, the data are conflicting on the requirement of FoxO1 signaling in the activation of the E3-ubiquitin ligases. Moreover, no reports currently exist on the particular role of FoxO1 in the molecular mechanisms involved in the progression of physiological muscle wasting. Here, we have applied the most extensively used rodent model of microgravity/functional unloading to stimulate disuse-induced skeletal muscle atrophy such as rat hindlimb suspension (HS). We showed that inhibition of FoxO1 activity by a selective inhibitor AS1842856 completely reversed an increase in expression of MuRF-1, but not MAFbx/atrogin-1, observed upon HS. Furthermore, we demonstrated that FoxO1 induced upregulation of another E3-ubiquitin-ligase of a MuRF protein family MuRF-2 in skeletal muscle subjected to disuse. Prevention of the MuRF increase upon HS impeded upregulation of transcript expression of a negative regulator of NFATc1 pathway calsarcin-2, which was associated with a partial reversion of MyHC-IId/x and MyHC-IIb mRNA expressions. Importantly, FoxO1 inhibition induced a marked increase in p70S6k phosphorylation, an important stage in the initiation of protein translation, concomitant with the restoration of global protein synthesis in the skeletal muscle of the HS rats. Examination of eIF3f expression and the eEF2k/eEF2 pathway, other factors controlling translation initiation and elongation respectively, did not reveal any impact of FoxO1 on their activity. Lastly, we observed a decrease in transcript levels of Sesn3, but not Sesn1 and Sesn2, upon disuse, which was completely reversed by FoxO1 inhibition. These data demonstrate that FoxO1 signaling contributes to the development of disuse-induced skeletal muscle atrophy, including slow to fast MyHC isoform shift, mostly through upregulation of MuRF-1 and MuRF-2 expression. Furthermore, FoxO1 inhibition is required to recover Sesn3 mRNA expression in atrophic conditions, which likely contributes to the enhanced p70S6k activity and restoration of the protein synthesis rate.


Exploration of pathomechanisms triggered by a single-nucleotide polymorphism in titin's I-band: the cardiomyopathy-linked mutation T2580I.
Missense single-nucleotide polymorphisms (mSNPs) in titin are emerging as a main causative factor of heart failure. However, distinguishing between benign and disease-causing mSNPs is a substantial challenge. Here, we research the question of whether a single mSNP in a generic domain of titin can affect heart function as a whole and, if so, how. For this, we studied the mSNP T2850I, seemingly linked to arrhythmogenic right ventricular cardiomyopathy (ARVC). We used structural biology, computational simulations and transgenic muscle in vivo methods to track the effect of the mutation from the molecular to the organismal level. The data show that the T2850I exchange is compatible with the domain three-dimensional fold, but that it strongly destabilizes it. Further, it induces a change in the conformational dynamics of the titin chain that alters its reactivity, causing the formation of aberrant interactions in the sarcomere. Echocardiography of knock-in mice indicated a mild diastolic dysfunction arising from increased myocardial stiffness. In conclusion, our data provide evidence that single mSNPs in titin's I-band can alter overall muscle behaviour. Our suggested mechanisms of disease are the development of non-native sarcomeric interactions and titin instability leading to a reduced I-band compliance. However, understanding the T2850I-induced ARVC pathology mechanistically remains a complex problem and will require a deeper understanding of the sarcomeric context of the titin region affected.


Medicaid funding for abortion: providers' experiences with cases involving rape, incest and life endangerment.
The Hyde Amendment bans federal Medicaid funding for abortion in the United States except if a pregnancy resulted from rape or incest or endangers the life of the woman. Some evidence suggests that providers do not always receive Medicaid reimbursement for abortions that should qualify for funding. From October 2007 to February 2008, semistructured in-depth interviews about experiences with Medicaid reimbursement for qualifying abortions were conducted with 25 respondents representing abortion providers in six states. A thematic analysis approach was used to explore respondents' knowledge of and experiences seeking Medicaid reimbursement for qualifying abortions, as well as individual, clinical and structural influences on reimbursement. The numbers of qualifying cases that were and were not reimbursed were assessed. More than half of Medicaid-eligible cases reported by respondents in the past year were not reimbursed. Respondents reported that filing for reimbursement takes excessive staff time and is hampered by bureaucratic claims procedures and ill-informed Medicaid staff, and that reimbursements are small. Many had stopped seeking Medicaid reimbursement and relied on nonprofit abortion funds to cover procedure costs. Respondents reporting receiving reimbursement said that streamlined forms, a statewide education intervention and a legal intervention to ensure that Medicaid reimbursed claims facilitated the process. The policy governing federal funding of abortion is inconsistently implemented. Eliminating administrative burdens, educating providers about women's rights to obtain Medicaid reimbursement for abortion in certain circumstances and holding Medicaid accountable for reimbursing qualifying cases are among the steps that may facilitate Medicaid reimbursement for qualifying abortions.


Analysis of fluorescently labeled glycosphingolipid-derived oligosaccharides following ceramide glycanase digestion and anthranilic acid labeling.
Interest in cellular glycosphingolipid (GSL) function has necessitated the development of a rapid and sensitive method to both analyze and characterize the full complement of structures present in various cells and tissues. An optimized method to characterize oligosaccharides released from glycosphingolipids following ceramide glycanase digestion has been developed. The procedure uses the fluorescent compound anthranilic acid (2-aminobenzoic acid; 2-AA) to label oligosaccharides prior to analysis using normal-phase high-performance liquid chromatography. The labeling procedure is rapid, selective, and easy to perform and is based on the published method of Anumula and Dhume [Glycobiology 8 (1998) 685], originally used to analyze N-linked oligosaccharides. It is less time consuming than a previously published 2-aminobenzamide labeling method [Anal. Biochem. 298 (2001) 207] for analyzing GSL-derived oligosaccharides, as the fluorescent labeling is performed on the enzyme reaction mixture. The purification of 2-AA-labeled products has been improved to ensure recovery of oligosaccharides containing one to four monosaccharide units, which was not previously possible using the Anumula and Dhume post-derivatization purification procedure. This new approach may also be used to analyze both N- and O-linked oligosaccharides.


Trajectories of physical dating violence from middle to high school: association with relationship quality and acceptability of aggression.
Although research on dating violence is growing, little is known about the distinct developmental trajectories of dating violence during adolescence. The current study identifies trajectories of physical dating violence victimization and perpetration that boys and girls follow from sixth to twelfth grade, examines the overlap of these trajectories, and characterizes them by perceptions of a caring dating relationship and acceptability of dating aggression. The sample consisted of randomly selected sixth graders from nine schools in Northeast Georgia (n = 588; 52 % boys; 49 % White, 36 % African American, 12 % Latino) who completed yearly surveys from Grades 6-12. We used latent class mixture modeling to identify the trajectories and generalized estimating equations models to examine the acceptability of dating aggression by dating violence trajectories. Participants followed two trajectories of dating violence victimization (boys: low and high; girls: low and increasing) and two of perpetration (boys and girls: low and increasing). When examining the joint trajectories of victimization and perpetration, a similar proportion of boys (62 %) and girls (65 %) were in the low victimization and low perpetration group and reported the lowest acceptance of dating aggression. The same proportion of boys and girls (27 %) were in the high/increasing victimization and perpetration group, and reported the highest acceptance of dating aggression. However, acceptance of dating aggression decreased from Grade 6-12 for all groups, even for those whose trajectory of dating violence increased. Victimization and perpetration were associated with reporting a less caring dating relationship. Results highlight the importance of focusing prevention efforts early for adolescents who follow this increasing probability of physical dating violence.


The factor structure of the 12-item General Health Questionnaire in a literate Kenyan population.
The study evaluated the factorial structure of the 12-item General Health Questionnaire (GHQ-12) in a population of Kenyan adults and adolescents. A sample of 1216 people aged 12-60 years completed the English version of the GHQ-12. To evaluate the factor structure of the GHQ-12, a confirmatory analysis using MPLUS was carried out. A unidimensional model of GHQ-12 as originally conceptualized did not provide a good fit for the data (Comparative Fit Index = 0.843, Tuckler Lewis Index = 0.841, Root Mean Square of Error Approximation = 0.078, Standardized Root Mean Square Residual = 0.063). Unidimensional models, which partial out the effects of negative wording showed an adequate fit to the data. Additionally, multidimensional models showed an excellent fit to the data. Based on the principles of parsimony, pattern of item loading and the correlation between identified factors in the multidimensional models, it was concluded that the GHQ-12 is a unidimensional measure whose structure is significantly influenced by wording effects. The GHQ-12, when applied to a literate population in Africa, showed similar factor structure as in other regions of the world; hence, it can be recommended for use as a screening tool for general symptoms of psychological distress.


Residue Trp-48 of Tva is critical for viral entry but not for high-affinity binding to the SU glycoprotein of subgroup A avian leukosis and sarcoma viruses.
Previously, mutant Tva receptors were classified as either partially or completely defective in mediating subgroup A avian leukosis and sarcoma virus (ALSV-A) entry (C. Bélanger, K. Zingler, and J. A. T. Young, J. Virol. 69:1019-1024, 1995; K. Zingler, C. Bélanger, R. Peters, D. Agard, and J. A. T. Young, J. Virol. 69:4261-4266, 1995). To specifically test the abilities of these mutant Tva proteins to bind ALSV-A surface (SU) protein, binding studies were performed with a subgroup A SU-immunoadhesin. This fusion protein is composed of the subgroup A Schmidt-Ruppin SU protein fused in frame to a rabbit immunoglobulin constant region. This reagent was conjugated to fluorescein isothiocyanate and used for flow cytometric analysis with transfected human 293 cells expressing different forms of Tva. The SU-immunoadhesin bound the wild-type Tva protein with a KD of approximately 1.5 nM. Amino acid substitutions that reduced viral entry at Asp-46 and at Cys-35 and Cys-50, which are predicted to form an intrachain disulfide bond in Tva, drastically reduced the binding affinity for the SU-immunoadhesin. Thus, the effects on viral entry of some mutations could be explained solely by changes in the binding affinity for ALSV-A SU. However, this was not true for other mutations tested, especially those with amino acid substitutions that replaced Trp-48. Compared with the wild-type receptor, these latter mutations led to approximately 43- to 200-fold reductions in viral infectivity but only to approximately 2.5- to 3.4-fold reductions in the binding affinity for the SU-immunoadhesin. These results support a role for Trp-48 of Tva in mediating steps of viral entry subsequent to binding ALSV-A SU.


Materials for solar fuels and chemicals.
The conversion of sunlight into fuels and chemicals is an attractive prospect for the storage of renewable energy, and photoelectrocatalytic technologies represent a pathway by which solar fuels might be realized. However, there are numerous scientific challenges in developing these technologies. These include finding suitable materials for the absorption of incident photons, developing more efficient catalysts for both water splitting and the production of fuels, and understanding how interfaces between catalysts, photoabsorbers and electrolytes can be designed to minimize losses and resist degradation. In this Review, we highlight recent milestones in these areas and some key scientific challenges remaining between the current state of the art and a technology that can effectively convert sunlight into fuels and chemicals.


De novo membranoproliferative glomerulonephritis in hepatitis C virus-infected renal allograft recipients.
Hepatitis C virus (HCV) is the leading cause of non-A, non-B hepatitis among renal allograft recipients. We sought to identify and describe a proteinuric renal disease occurring in our HCV-infected renal transplant patients. Patients with proteinuria exceeding 1 g/day were identified from a cohort of 98 HCV-infected kidney recipients. Qualitative and quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and restriction fragment-length polymorphism of the amplified RT-PCR product was performed to detect circulating HCV RNA, viral titer, and strain type, respectively. An immune complex nephritis (ICN) of the membranoproliferative pattern (MPGN) was found on five of eight biopsies. Two patients infected with the Hutch strain-type developed nephrotic-range proteinuria within three months posttransplant while the remaining three MPGN patients had been transplanted greater than 5 years prior to the onset of proteinuria. Testing for rheumatoid factors, cryoglobulins, hypocomplementemia, and circulating immune complexes failed to show a consistent pattern. Sucrose density gradient (SDG) equilibrium centrifugation was used to determine the buoyant-density of HCV virions from control (HCV-infected nonproteinuric recipients; n = 5) and nephrotic patients (n = 5). Whereas HCV virions from the control patients had a low buoyant density on sucrose gradients, a substantial percentage of the circulating HCV RNA from the MPGN patients was present in the high-density fractions in association with IgM and IgG. Treatment of the pooled high-density layers with NP40 followed by recentrifugation resulted in a shift of the HCV RNA to the medium-density layers. In conclusion, MPGN developed in five HCV-infected kidney recipients despite pharmacologic immunosuppression. Both the physicochemical properties of the HCV virions on SDG and their association with IgG and IgM in the high-density layers provide indirect evidence for the presence of circulating complexes of anti-HCV antibody and HCV antigen(s).


Deriving appropriate boundary conditions, and accelerating position-jump simulations, of diffusion using non-local jumping.
In this paper we explore lattice-based position-jump models of diffusion, and the implications of introducing non-local jumping; particles can jump to a range of nearby boxes rather than only to their nearest neighbours. We begin by deriving conditions for equivalence with traditional local jumping models in the continuum limit. We then generalize a previously postulated implementation of the Robin boundary condition for a non-local process of arbitrary maximum jump length, and present a novel implementation of flux boundary conditions, again generalized for a non-local process of arbitrary maximum jump length. In both these cases we validate our results using stochastic simulation. We then proceed to consider two variations on the basic diffusion model: a hybrid local/non-local scheme suitable for models involving sharp concentration gradients, and the implementation of biased jumping. In all cases we show that non-local jumping can deliver substantial time savings for stochastic simulations.


Sensitization of TRAIL-induced cell death by 20(S)-ginsenoside Rg3 via CHOP-mediated DR5 upregulation in human hepatocellular carcinoma cells.
The TRAIL pathway is a potential therapeutic target for anticancer drugs due to selective cytotoxicity in cancer cells. Despite considerable promise, TRAIL or TRAIL receptor agonists have been used thus far with limited success in multiple clinical trials, in part due to acquired TRAIL resistance during chemotherapeutic treatment. Hepatocellular carcinoma (HCC) is a common solid tumor and the third leading cause of cancer-related death worldwide. Classical chemotherapy is not effective for HCC treatment and targeted therapy is limited to sorafenib. Isolated from Panax ginseng CA Meyer, 20(S)-ginsenoside Rg3 is a steroidal saponin with high pharmacologic activity that has been shown to sensitize cells to some chemotherapeutic agents. We investigated the sensitizing effect of Rg3 on TRAIL-induced cell death in HCC cells. We show Rg3 is capable of promoting TRAIL-induced apoptosis in a number of HCC cell lines, including HepG2, SK-Hep1, Huh-7, and Hep3B, but not in normal HL-7702 hepatocytes, indicating that Rg3 sensitization to TRAIL may be specific to cancer cells. Mechanistically, we found that Rg3 upregulates DR5 expression at the transcriptional level. DR5 upregulation in this case is mediated by C/EBP homology protein (CHOP), an important endoplasmic reticulum stress responsive protein. Furthermore, Rg3 is well tolerated and enhances the therapeutic efficacy of TRAIL in mouse xenograft models, suggesting that chemosensitization also occurs in vivo. Taken together, our study identifies Rg3 as a novel anticancer therapeutic agent and supports the further development of Rg3 as a chemosensitizer in combined therapy with TRAIL.


An overview of zirconia dental implants: basic properties and clinical application of three cases.
Due to the possible aesthetic problems of titanium implants, the developments in ceramic implant materials are increasing. Natural tooth colored ceramic implants may be an alternative to overcome aesthetic problems. The purpose of this article is to give information about the basic properties of dental zirconia implants and present 3 cases treated with two-piece zirconia implants. Two-piece zirconia dental implants, 4.0 mm diameter and 11.5 mm in length, were inserted into maxillary incisor region. They were left for 6 months to osseointegrate. Panoramic and periapical radiographs were obtained and examined for bone-implant osseointegration. During the follow-up period the patients were satisfied with their prosthesis and no complication was observed.


Derivation of optical constants of metals from thin-film measurements at oblique incidence.
The reflectance R and transmittance T of thin absorbing films deposited on a transparent substrate are calculated for normal and oblique incidence, s and p polarization, and different film thicknesses. The results are presented as contours of constant R and T on the ñ plane, where ñ is the complex refractive index. The conditions for sensitive dependence of measured quantities on ñ are examined. A computer-based method of finding ñ from chosen combinations of measured R or T values is described. Oblique incidence measurements on thin films can give accurate results in some regions where other methods may be less sensitive. Accurate film-thickness values can be obtained from the optical measurements.


Ocelots on Barro Colorado Island are infected with feline immunodeficiency virus but not other common feline and canine viruses.
Transmission of pathogens from domestic animals to wildlife populations (spill-over) has precipitated local wildlife extinctions in multiple geographic locations. Identifying such events before they cause population declines requires differentiating spillover from endemic disease, a challenge complicated by a lack of baseline data from wildlife populations that are isolated from domestic animals. We tested sera collected from 12 ocelots (Leopardus pardalis) native to Barro Colorado Island, Panama, which is free of domestic animals, for antibodies to feline herpes virus, feline calicivirus, feline corona virus, feline panleukopenia virus, canine distemper virus, and feline immunodeficiency virus (FIV), typically a species-specific infection. Samples also were tested for feline leukemia virus antigens. Positive tests results were only observed for FIV; 50% of the ocelots were positive. We hypothesize that isolation of this population has prevented introduction of pathogens typically attributed to contact with domestic animals. The high density of ocelots on Barro Colorado Island may contribute to a high prevalence of FIV infection, as would be expected with increased contact rates among conspecifics in a geographically restricted population.


Ancient DNA reveals lack of continuity between neolithic hunter-gatherers and contemporary Scandinavians.
The driving force behind the transition from a foraging to a farming lifestyle in prehistoric Europe (Neolithization) has been debated for more than a century [1-3]. Of particular interest is whether population replacement or cultural exchange was responsible [3-5]. Scandinavia holds a unique place in this debate, for it maintained one of the last major hunter-gatherer complexes in Neolithic Europe, the Pitted Ware culture [6]. Intriguingly, these late hunter-gatherers existed in parallel to early farmers for more than a millennium before they vanished some 4,000 years ago [7, 8]. The prolonged coexistence of the two cultures in Scandinavia has been cited as an argument against population replacement between the Mesolithic and the present [7, 8]. Through analysis of DNA extracted from ancient Scandinavian human remains, we show that people of the Pitted Ware culture were not the direct ancestors of modern Scandinavians (including the Saami people of northern Scandinavia) but are more closely related to contemporary populations of the eastern Baltic region. Our findings support hypotheses arising from archaeological analyses that propose a Neolithic or post-Neolithic population replacement in Scandinavia [7]. Furthermore, our data are consistent with the view that the eastern Baltic represents a genetic refugia for some of the European hunter-gatherer populations.


Validation and evaluation of the Mandarin version of the oral health literacy adult questionnaire in Taiwan.
This study aimed to translate the Oral Health Literacy Adult Questionnaire into a Mandarin version (MOHL-AQ) and to examine its psychometric properties. A methodological research using psychometric testing and evaluation of a translated instrument. A convenience sample of 402 participants from northern Taiwan were recruited for the validation of the MOHL-AQ. Internal consistency reliability, split-half reliability, inter-rater reliability, face validity, content validity, and construct validity were evaluated. The value of internal consistency and split-half reliability of the MOHL-AQ were 0.77 and 0.78, respectively. Content validity reported a high content validity index (CVI = 95%). Exploratory factor analysis (EFA) and parallel analysis (PA) were used to determine a unidimensional model and confirmatory factor analysis (CFA) was employed to confirm the model. The indices of good fit model were achieved at GFI = 0.93, AGFI = 0.92, RMSEA = 0.04, CFI = 0.90, PGFI = 0.73, χ2 /df = 1.86 (p < .001). Most of the item-total correlations indicated adequate and acceptable convergent validity (r > .30). MOHL-AQ demonstrates adequate psychometric properties for measuring the oral health literacy in Mandarin-speaking population. Public health nurses can use MOHL-AQ to assess oral health literacy in the community settings and further screen potential population with inadequate oral health literacy.


Treatment of Alzheimer's disease with a cholinesterase inhibitor combined with antioxidants.
A formula (formula F) was prepared to counteract oxidative stress (OS) in the brain. The formula contained the most common antioxidants and was intended to: (a) protect proteins, lipids, DNA and proteoglycans from oxidation (carnosine, coenzyme Q(10), vitamin E, vitamin C, beta-carotene, selenium, L-cysteine and ginkgo biloba); (b) reduce homocysteine (HCy) blood levels (vitamins B(6), B(9) and B(12)), and (c) sustain the pentose phosphate cycle in circulating cells (vitamins B(1), B(2) and B(3)). Formula F contained low doses of each antioxidant component and was administered in a two-phase ampoule. A cohort of 52 patients (21 males and 31 females) affected with moderate probable AD (according to NINCDS-ARDA and NINCS-AIREN criteria) already being treated with donepezil (5 mg/day for at least two months) was randomly divided into two groups, and followed for 6 months. A double-blind design was used in which 26 cases were treated once a day with formula F plus donepezil, and the other 26 with placebo plus donepezil. The level of OS was measured on the basis of a d-ROMs test (which measures plasma hydroperoxides), plasma HCy and glutathione, and percentage of sickle erythrocytes. The two patient groups were comparable for all variables (age, sex, concomitant diseases, ApoE epsilon4, MMSE II score, OS, antioxidant reserve and sickle erythrocytes). Forty-eight subjects completed the trial. Significant decreases in OS and HCy were only observed when there was an increase in glutathione (in erythrocytes) and a decrease in sickle erythrocytes in patients treated with formula F. The MMSE II score remained almost the same in the group treated with donepezil and placebo, whereas some significant improvements were found in the group treated with donepezil plus formula F.


Efficacy and Safety of Tirzepatide in Adults With Type 2 Diabetes: A Perspective for Primary Care Providers.
This article reviews the efficacy and safety data of tirzepatide, a once-weekly, novel glucose-dependent insulinotropic polypeptide and glucagon-like peptide 1 (GLP-1) receptor agonist approved in the United States, the European Union, and other regions for the treatment of type 2 diabetes. All doses of tirzepatide demonstrated superiority in reducing A1C and body weight from baseline versus placebo or active comparators. The safety profile of tirzepatide was consistent with that of the GLP-1 receptor agonist class, with mild to moderate and transient gastrointestinal side effects being the most common adverse events. With clinically and statistically significant reductions in A1C and body weight without increased risk of hypoglycemia in various populations, tirzepatide has demonstrated potential as a first-in-class treatment option for many people with type 2 diabetes.


Regulation of PTEN (phosphatase and tensin homolog deleted on chromosome 10) expression by estradiol and progesterone in human endometrium.
PTEN (phosphatase and tensin homolog deleted on chromosome 10) is a tumor suppressor gene, mutated frequently in a variety of human tumors. PTEN regulates cell growth, apoptosis, and proliferation. Phosphorylation in PTEN tail causes its inactivation and decreases its degradation. There is little known about the regulation of PTEN by ovarian steroids. We hypothesized that PTEN expression in human endometrium is variable throughout the menstrual cycle and early pregnancy, and that ovarian steroids regulate PTEN expression because PTEN is critical in many steroid-sensitive tissues such as endometrium, prostate, and breast. In the present study, we have observed a direct regulation of PTEN by ovarian steroids. Estradiol increased PTEN phosphorylation at 5-15 min. After 24-h treatment, progesterone induced a significant increase in PTEN protein levels, assessed by Western blot. Furthermore, we evaluated for the first time a comparison between menstrual cycle and early pregnancy, immunohistochemically. Endometrial PTEN expression revealed temporal and spatial changes throughout the menstrual cycle and during early pregnancy. We conclude that estradiol may down-regulate PTEN activity by increasing its phosphorylation, but progesterone is likely to regulate the PTEN pool by decreasing its phosphorylation and increasing its protein level. Presented data, therefore, suggest that ovarian steroids regulate the endometrial PTEN pool. We propose that PTEN might be one of the signaling proteins that estrogen and progesterone are acting to affect endometrial cell proliferation and/or apoptosis.


Complications and Route of Delivery in a Large Cohort Study of HIV-1-Infected Women-IMPAACT P1025.
To investigate complications of cesarean section in a cohort of HIV-infected pregnant women. IMPAACT P1025 is a prospective cohort study of HIV-1-infected women and infants, enrolled 2002-2013, at clinical sites in the United States and Puerto Rico. Demographic, medical, and obstetric data were collected and analyzed including cesarean indications. The delivery route was categorized as elective cesarean (ECS) (before labor and <5 minutes before membrane rupture), nonelective cesarean (NECS) (all other cesareans) or vaginal delivery. Logistic regression models evaluated associations between delivery route and maternal intrapartum/postpartum morbidities. Composite morbidity of vaginal delivery was compared with ECS and NECS. This study included 2297 women. Of note, 99% used antiretroviral medication and 89% were on a combination antiretroviral therapy regimen; 84% had a HIV-1 viral load ≤400 copies per milliliter before delivery; 46% (1055) delivered vaginally, 35% (798) by ECS, and 19% (444) by NECS. Although interruption of HIV-1 infection was the second most frequent indication for cesarean after repeat cesarean, it decreased as an indication over time. There were no delivery-related maternal mortalities. Overall, 19% of women had ≥1 complication(s)-primarily wound complications (14%) or other infections (11%). Vaginal delivery had the lowest complication rate (13%), followed by ECS (23%), and highest NECS (28%) with an overall P < 0.001. HIV-1 mother-to-child transmission rates were low and did not differ by delivery mode group. HIV interruption as cesarean indicator declined during the study. Morbidity was more common in HIV-infected women delivering by NECS than ECS and lowest with vaginal delivery. Prenatal and Postnatal Studies of Interventions for Prevention of Mother-To-Child Transmission https://clinicaltrials.gov/ct2/show/NCT00028145?term=impaact+1025&rank=2 NCT00028145.


Effect of bio-optical parameter variability on the remote estimation of chlorophyll-a concentration in turbid productive waters: experimental results.
The analytical development and underlying hypothesis of a three-band algorithm for estimating chlorophyll-a concentration ([Chla]) in turbid productive waters are presented. The sensitivity of the algorithm to the spectral location of the bands used is analyzed. A large set of experimental observations ([Chla] varied between 4 and 217 mg m(-3) and turbidity between 2 and 78 nephelometric turbidity units) was used to calibrate and validate the algorithm. It was found that the variability of the chlorophyll-a fluorescence quantum yield and of the chlorophyll-a specific absorption coefficient can reduce considerably the accuracy of remote predictions of [Chla]. Instead of parameterizing these interferences, their effects were minimized by tuning the spectral regions used in the algorithm. This allowed us to predict [Chla] with a relative root-mean-square error of less than 30%.


Modal analysis of linear Talbot-cavity semiconductor lasers.
A modal analysis at threshold is presented for linear Talbot-cavity semiconductor lasers with a finite number of array elements. The analysis self-consistently calculates the array supermode profiles and the loss each incurs owing to imperfect Talbot imaging, including edge diffraction losses. It is found that the decrease in mode discrimination between supermodes incurred by fabricating larger arrays can be offset by reducing the near-field fill factor.


Impact of tissue inhomogeneity on dose distribution in the canine carpal and tarsal regions for cobalt and 6 MV photons.
We quantified the effect of tissue inhomogeneity on dose distribution in a canine distal extremity resulting from treatment with cobalt photons and photons from a 6MV accelerator. Monitor units for a typical distal extremity treatment were calculated by two methods, using equally weighted, parallel-opposed fields. The first method was a computed tomography (CT)-based, computerized treatment plan, calculated without inhomogeneity correction. The second method was a manual point dose calculation to the isocenter. A computerized planning system was then used to assess the dose distribution achieved by these two methods when tissue inhomogeneity was taken into account. For cobalt photons, the median percentage of the planning target volume (PTV) that received < 95% of the prescribed dose was 4.5% for the CT-based treatment plan, and 26.2% for the manually calculated plan. For 6 MV photons, the median percentage of the PTV that received < 95% of the prescribed dose was < 1% for both planning methods. The PTV dose achieved without using inhomogeneity correction for cobalt photons results in potentially significant under dosing of portions of the PTV.


Heligmosomoides polygyrus elicits a dominant nonprotective antibody response directed against restricted glycan and peptide epitopes.
Heligmosomoides polygyrus is a widely used gastrointestinal helminth model of long-term chronic infection in mice, which has not been well-characterized at the antigenic level. We now identify the major targets of the murine primary Ab response as a subset of the secreted products in H. polygyrus excretory-secretory (HES) Ag. An immunodominant epitope is an O-linked glycan (named glycan A) carried on three highly expressed HES glycoproteins (venom allergen Ancylostoma-secreted protein-like [VAL]-1, -2, and -5), which stimulates only IgM Abs, is exposed on the adult worm surface, and is poorly represented in somatic parasite extracts. A second carbohydrate epitope (glycan B), present on both a non-protein high molecular mass component and a 65-kDa molecule, is widely distributed in adult somatic tissues. Whereas the high molecular mass component and 65-kDa molecules bear phosphorylcholine, the glycan B epitope itself is not phosphorylcholine. Class-switched IgG1 Abs are found to glycan B, but the dominant primary IgG1 response is to the polypeptides of VAL proteins, including also VAL-3 and VAL-4. Secondary Ab responses include the same specificities while also recognizing VAL-7. Although vaccination with HES conferred complete protection against challenge H. polygyrus infection, mAbs raised against each of the glycan epitopes and against VAL-1, VAL-2, and VAL-4 proteins were unable to do so, even though these specificities (with the exception of VAL-2) are also secreted by tissue-phase L4 larvae. The primary immune response in susceptible mice is, therefore, dominated by nonprotective Abs against a small subset of antigenic epitopes, raising the possibility that these act as decoy specificities that generate ineffective humoral immunity.


Achilles tendon allograft reconstruction of the fibular collateral ligament and posterolateral corner.
The purpose of this study was to investigate the functional and clinical outcomes of a consecutive series of patients who underwent fibular collateral ligament (FCL) and posterolateral corner (PLC) reconstruction by use of a single Achilles tendon allograft construct. Patients were identified through prospective sports medicine databases at 2 academic institutions. Only patients who had undergone FCL and PLC reconstruction (as opposed to repair) were included. All patients followed a standard postoperative rehabilitation protocol. Various patterns of combined ligament injuries were included and divided into 2 groups (2 ligament v multiligament). Functional and clinical outcomes were assessed by clinical examination, Lysholm scores, and International Knee Documentation Committee subjective scores. Statistical analysis was performed by use of Wilcoxon paired rank-sum tests and multivariate regression. We identified 16 knees in 16 patients, with a minimum of 2 years' follow-up. There were 13 men and 3 women. The mean age was 30 years (range, 19 to 61 years). The mean length of clinical follow-up was 30 months (range, 24 to 75 months). The mean International Knee Documentation Committee subjective scores were 80 points and 80 points (P = .79) in the 2-ligament and multiligament groups, respectively, and the mean Lysholm scores were 90 points and 89 points (P = .96), respectively. Age (P = .41), gender (P = .84), and interval between injury and surgery (P = .72) did not affect the clinical and functional outcomes between the 2 groups. Arthrofibrosis requiring manipulation developed in 1 patient. Residual varus laxity (1+) was noted in 4 patients, none of whom displayed functional instability. No patient has required revision reconstruction to date. We describe a novel technique that takes into account the main static PLC stabilizers (FCL, popliteofibular ligament, and posterolateral capsule) that has not been previously reported. Our series showed no significant differences in clinical and functional outcomes between 2-ligament and multiligament PLC-based reconstructions. However, given the heterogeneity and small sample size of our study group, it is difficult to draw qualitative conclusions. Level IV, therapeutic case series.


Downregulating aspen xylan biosynthetic GT43 genes in developing wood stimulates growth via reprograming of the transcriptome.
Xylan is one of the main compounds determining wood properties in hardwood species. The xylan backbone is thought to be synthesized by a synthase complex comprising two members of the GT43 family. We downregulated all GT43 genes in hybrid aspen (Populus tremula × tremuloides) to understand their involvement in xylan biosynthesis. All three clades of the GT43 family were targeted for downregulation using RNA interference individually or in different combinations, either constitutively or specifically in developing wood. Simultaneous downregulation in developing wood of the B (IRX9) and C (IRX14) clades resulted in reduced xylan Xyl content relative to reducing end sequence, supporting their role in xylan backbone biosynthesis. This was accompanied by a higher lignocellulose saccharification efficiency. Unexpectedly, GT43 suppression in developing wood led to an overall growth stimulation, xylem cell wall thinning and a shift in cellulose orientation. Transcriptome profiling of these transgenic lines indicated that cell cycling was stimulated and secondary wall biosynthesis was repressed. We suggest that the reduced xylan elongation is sensed by the cell wall integrity surveying mechanism in developing wood. Our results show that wood-specific suppression of xylan-biosynthetic GT43 genes activates signaling responses, leading to increased growth and improved lignocellulose saccharification.


Effect of Pilocarpine on impaired salivary secretion in patients with Sjögren's syndrome.
The aim of this study was to analyze resting whole saliva (RWS) and stimulated whole saliva (SWS) flow rates before and after administration of Pilocarpine in secondary Sjögren's Syndrome patients. Fifty-one patients (49 women, 2 men, mean age 61 years, range 38-85), all with a resting saliva < or =0.1 ml/min, participated. Volumes of RWS and SWS collected over periods of 15 and 5 min, respectively, using standardized protocols were measured and the same procedure was repeated after oral administration of Pilocarpine (0.7 mg per 10 kg body weight). The sample was then divided into two groups, according to those in whom Pilocarpine stimulation had caused RWS flow to reach >0.1 ml/min (responders) and those who remained at values < or =0.1 ml/min (non-responders). All participants completed a questionnaire related to general and oral health status, as well as their subjective intraoral complaints before and after administration of Pilocarpine. Thirteen patients (25%) were classified as non-responders and the remaining 38 (75%) as responders. No statistically significant differences between the non-responders and responders were detected as regards general health parameters or intake of medicines with anticholinergic affect. As regards intraoral subjective complaints, no difference between the groups was found before Pilocarpine administration. After administration of Pilocarpine, complaints were significantly fewer among the responders (p<0.01). Both groups exhibited a significant decrease of intraoral symptoms after administration of Pilocarpine (responders P<0.001 and non-responders P<0.05) compared to baseline. For the whole group, more severe intraoral complaints were significantly associated with a lower SWS (P<0.05), but not a RWS, rate at baseline. It is concluded that a subgroup of Sjögren patients do not respond to Pilocarpine stimulation. The clinical implications of this finding need further investigation.