Comparing in-person, video, and telephonic medical interpretation.
Using trained interpreters to provide medical interpretation services is superior to services provided on an ad hoc basis, but little is known about the effectiveness of providing their services remotely, especially using video. To compare remote medical interpretation services by trained interpreters via telephone and videoconference to those provided in-person. Quasi-randomized control study. Two hundred and forty-one Spanish speaking patient volunteers, twenty-four health providers, and seven interpreters. Patients, providers and interpreters each independently completed scales evaluating the quality of clinical encounters and, optionally, made free text comments. Interviews were conducted with 23 of the providers, the seven interpreters, and a subset of 30 patients. Time data were collected. Encounters with in-person interpretation were rated significantly higher by providers and interpreters, while patients rated all methods the same. There were no significant differences in provider and interpreter ratings of remote methods. Provider and interpreter comments on scales and interview data support the higher in-person ratings, but they also showed a distinct preference for video over the phone. Phone interviews were significantly shorter than in-person. Patients rated interpretation services highly no matter how they were provided but experienced only the method employed at the time of the encounter. Providers and interpreters were exposed to all three methods, were more critical of remote methods, and preferred videoconferencing to the telephone as a remote method. The significantly shorter phone interviews raise questions about the prospects of miscommunication in telephonic interpretation, given the absence of a visual channel, but other factors might have affected time results. Since the patient population studied was Hispanic and predominantly female care must be taken in generalizing these results to other populations.


Challenges in conducting genome-wide association studies in highly admixed multi-ethnic populations: the Generation R Study.
Genome-wide association studies (GWAS) have been successful in identifying loci associated with a wide range of complex human traits and diseases. Up to now, the majority of GWAS have focused on European populations. However, the inclusion of other ethnic groups as well as admixed populations in GWAS studies is rapidly rising following the pressing need to extrapolate findings to non-European populations and to increase statistical power. In this paper, we describe the methodological steps surrounding genetic data generation, quality control, study design and analytical procedures needed to run GWAS in the multiethnic and highly admixed Generation R Study, a large prospective birth cohort in Rotterdam, the Netherlands. Furthermore, we highlight a number of practical considerations and alternatives pertinent to the quality control and analysis of admixed GWAS data.


Recombinant Leishmania mexicana CRK3:CYCA has protein kinase activity in the absence of phosphorylation on the T-loop residue Thr178.
The activity of cyclin-dependent kinases (CDKs), which are key regulators of the eukaryotic cell cycle, is regulated through post-translational mechanisms, including binding of a cyclin and phosphorylation. Previously studies have shown that Leishmania mexicana CRK3 is an essential CDK that is a functional homologue of human CDK1. In this study, recombinant histidine tagged L. mexicana CRK3 and the cyclin CYCA were combined in vitro to produce an active histone H1 kinase that was inhibited by the CDK inhibitors, flavopiridol and indirubin-3'-monoxime. Protein kinase activity was observed in the absence of phosphorylation of the T-loop residue Thr178, but increased 5-fold upon phosphorylation by the CDK activating kinase Civ1 of Saccharomyces cerevisiae. Seven recombinant L. major CRKs (1, 2, 3, 4, 6, 7 and 8) were also expressed and purified, none of which were active as monomers. Moreover, only CRK3 was phosphorylated by Civ1. HA-tagged CYCA expressed in L. major procyclic promastigotes was co-precipitated with CRK3 and exhibited histone H1 kinase activity. These data indicate that in Leishmania CYCA interacts with CRK3 to form an active protein kinase, confirm the conservation of the regulatory mechanisms that control CDK activity in other eukaryotes, but identifies biochemical differences to human CDK1.


Experiences of violence and association with decreased drug abstinence among women in Cape Town, South Africa.
Drug abuse is a contributing factor in women's HIV risk in low-income communities in Cape Town, South Africa. This study assessed whether experiencing violence is associated with reduced drug abstinence among adult women (n = 603) participating in a randomized field trial for an HIV prevention study in Cape Town. In relation to drug abstinence at 12-month follow-up, multivariable regression models were used to assess (1) baseline partner and non-partner victimization, and (2) victimization at 12-month follow-up among participants reporting baseline victimization. Baseline partner (AOR = 0.6; 95 % CI 0.4-0.9) and non-partner victimization (AOR = 0.6; 95 % CI 0.4-0.9) were associated with a reduced likelihood of drug abstinence at follow-up. Among participants who reported victimization at baseline, those no longer reporting victimization at follow-up did not differ significantly in drug abstinence compared with those who reported victimization at follow-up. The study findings highlight the lasting impact of victimization on women's drug use outcomes, persisting regardless of whether violence was no longer reported at follow-up. Overall, the findings support the need for the primary prevention of violence to address the cycle of violence, drug use, and HIV among women in this setting.


Lidocaine increases the anti-inflammatory cytokine IL-10 following mechanical ventilation in healthy mice.
Mechanical ventilation (MV) induces an inflammatory response that may result in (acute) lung injury. Lidocaine, an amide local anesthetic, has anti-inflammatory properties in vitro and in vivo, possibly due to an attenuation of pro-inflammatory cytokines, intracellular adhesion molecule-1 (ICAM-1), and reduction of neutrophils influx. We hypothesized an attenuation of MV-induced inflammatory response with intravenously administered lidocaine. Lidocaine (Lido) (2, 4, and 8 mg/kg/h) was intravenously administered during 4 h of MV with a tidal volume of 8 ml/kg, positive end expiratory pressure 1,5 cmH2O and FiO2 0.4. We used one ventilated control (CON) group receiving vehicle. After MV, mice were euthanized, and lungs and blood were immediately harvested, and cytokine levels and ICAM-1 levels were measured in plasma and lung homogenates. Pulmonary neutrophils influx was determined in LEDER-stained slices of lungs. Anesthetic need was determined by painful hind paw stimulation. Lidocaine-treated animals (Lido 2, 4 and 8 mg/kg/h) showed higher interleukin (IL)-10 plasma levels compared to control animals. Lidocaine treatment with 8 mg/kg/h (Lido 8) resulted in higher IL-10 in lung homogenates. No differences were observed in pro-inflammatory cytokines, ICAM-1, and pulmonary influx between the different ventilated groups. Intravenously administered lidocaine increases levels of plasma IL-10 with infusion from 2, 4, and 8 mg/kg/h and pulmonary levels of IL-10 with 8 mg/kg/h in a murine mechanical ventilation model. Intravenously administered lidocaine appears to reduce anesthetic need in mice.


Cryopreservation of human embryonic stem cells by a programmed freezer with an oscillating magnetic field.
Human embryonic stem cells (hESCs), due to their self-renewal capacity and pluripotency, are an important source of cells for regenerative medicine. The immediate obstacles that need to be addressed are the poor cell survival rate of hESCs and their cell quality after cryopreservation. In this study, we used the Cell Alive System (CAS) which combines a programmed freezer with an oscillating magnetic field to reduce cryo-injury during the freezing process. The hESC clumps suspended in freezing medium were divided into three groups: (i) cells frozen by a conventional freezing container, Mr. Frosty and kept in a -80 °C freezer (MF); (ii) cells frozen to -32 °C by CAS, and then transferred to a -80 °C freezer (CAS); (iii) cells frozen to -32 °C by CAS, and then transferred to a pre-cooled Mr. Frosty and kept in a -80 °C freezer (CAS-MF) for overnight. All cryovials were placed in liquid nitrogen for one week, and hESCs were then thawed and cultured on feeder for 7 days. The results of alkaline phosphatase (AP) staining showed that the attachment efficiency of the cells cryopreserved by CAS and CAS-MF was significantly higher (29.0% and 44.0%) than in the MF method (7.0%). Furthermore, we confirmed the cells cryopreserved using CAS-MF could be subcultured while expressing pluripotent markers, differentiate into three germ layers, and maintain a normal karyotype. These results demonstrate that the use of CAS-MF offers an efficient method of hESC banking.


Irrigated urban vegetable production in Ghana: microbiological contamination in farms and markets and associated consumer risk groups.
Ghana is a typical low-income sub-Saharan African country facing significant sanitation challenges. In Ghana, fresh salads are not part of the normal diet, but have become a common supplement to urban fast food served in streets, canteens and restaurants. In Accra, about 200 000 people consume from such supplements every day. The figure also describes the size of the risk group from contamination, which comprises all income classes including the poor and children. The purpose of this study was to investigate widespread water pollution in urban and peri-urban areas, where 95% of the lettuce consumed in the city is produced. Over 12 months (April 2004-June 2005), lettuce samples from the same production sites in two cities were followed and analyzed along the "farm to fork" pathway for total and faecal coliform (FC) and helminth egg numbers. Questionnaire surveys were conducted among producers, sellers and consumers to quantify lettuce flows to the final risk group. The study identified the farm as the main point of lettuce contamination. Besides the irrigation water, contamination was also attributed to manure application and already contaminated soil. Despite poor sanitary conditions in markets, post-harvest handling and marketing did not further increase the farm-gate contamination levels. To reduce the health risk associated with the consumption of contaminated lettuce; safer farming and irrigation practices are required while the remaining risk could best be addressed where lettuce is prepared for consumption.


Enhancing cardiovascular artificial intelligence (AI) research in the Netherlands: CVON-AI consortium.
Machine learning (ML) allows the exploration and progressive improvement of very complex high-dimensional data patterns that can be utilised to optimise specific classification and prediction tasks, outperforming traditional statistical approaches. An enormous acceleration of ready-to-use tools and artificial intelligence (AI) applications, shaped by the emergence, refinement, and application of powerful ML algorithms in several areas of knowledge, is ongoing. Although such progress has begun to permeate the medical sciences and clinical medicine, implementation in cardiovascular medicine and research is still in its infancy. To lay out the theoretical framework, purpose, and structure of a novel AI consortium. We have established a new Dutch research consortium, the CVON-AI, supported by the Netherlands Heart Foundation, to catalyse and facilitate the development and utilisation of AI solutions for existing and emerging cardiovascular research initiatives and to raise AI awareness in the cardiovascular research community. CVON-AI will connect to previously established CVON consortia and apply a cloud-based AI platform to supplement their planned traditional data-analysis approach. A pilot experiment on the CVON-AI cloud was conducted using cardiac magnetic resonance data. It demonstrated the feasibility of the platform and documented excellent correlation between AI-generated ventricular function estimates as compared to expert manual annotations. The resulting AI solution was then integrated in a web application. CVON-AI is a new consortium meant to facilitate the implementation and raise awareness of AI in cardiovascular research in the Netherlands. CVON-AI will create an accessible cloud-based platform for cardiovascular researchers, demonstrate the clinical applicability of AI, optimise the analytical methodology of other ongoing CVON consortia, and promote AI awareness through education and training.


Natural SIV hosts: showing AIDS the door.
Many species of African nonhuman primates are naturally infected with simian immunodeficiency viruses (SIVs) in the wild and in captivity. In contrast to HIV-infected humans, these natural SIV hosts typically do not develop AIDS, despite chronic infection with a highly replicating virus. In this Review, we discuss the most recent advances on the mechanisms of protection from disease progression in natural SIV hosts, with emphasis on how they differ from pathogenic HIV/SIV infections of humans and rhesus macaques. These mechanisms include: (i) resolution of immune activation after acute infection, (ii) restricted pattern of target cell infection, and (iii) protection from mother-to-infant transmission. We highlight the areas that should be pursued in future studies, focusing on potential applications for the treatment and prevention of HIV infection.


Noradrenergic inhibition of synaptic transmission between mitral and granule cells in mammalian olfactory bulb cultures.
Noradrenergic modulation of the glutamatergic-GABAergic synapses between mitral/tufted (M/T) and granule cells has been implicated in some forms of olfactory learning (Brennan et al., 1990). Norepinephrine (NE) has been shown to disinhibit mitral cells (Jahr and Nicoll, 1982), but its site of action is not well defined. The effects of NE on synaptic transmission between monosynaptically coupled pairs of mitral and granule cells have been examined using primary culture and whole-cell recording techniques. Intracellular stimulation of M/T cells evoked dual-component EPSPs in granule cells consisting of both NMDA and AMPA (alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic acid) receptor-mediated mechanisms. The EPSPs were reversibly inhibited by approximately 50% during application of 30 microM NE. NE had no effect, however, on the membrane current evoked by exogenous application of glutamate, indicating a presynaptic site of action. The effect of NE on EPSPs was mimicked by the alpha-adrenergic agonist clonidine but not by the beta-adrenergic agonist isoproterenol. NE had no significant effect on either accommodation or macroscopic currents in either M/T or granule cells. NE also inhibited spontaneous GABAergic IPSPs recorded in M/T cells, by a presynaptic alpha-adrenergic-mediated mechanism. These results support previous results suggesting a disinhibitory role for NE in the olfactory bulb. This action, however, is at least in part mediated by a reduction in mitral cell-mediated granule cell excitation.


TGF-β/β2-spectrin/CTCF-regulated tumor suppression in human stem cell disorder Beckwith-Wiedemann syndrome.
Beckwith-Wiedemann syndrome (BWS) is a human stem cell disorder, and individuals with this disease have a substantially increased risk (~800-fold) of developing tumors. Epigenetic silencing of β2-spectrin (β2SP, encoded by SPTBN1), a SMAD adaptor for TGF-β signaling, is causally associated with BWS; however, a role of TGF-β deficiency in BWS-associated neoplastic transformation is unexplored. Here, we have reported that double-heterozygous Sptbn1+/- Smad3+/- mice, which have defective TGF-β signaling, develop multiple tumors that are phenotypically similar to those of BWS patients. Moreover, tumorigenesis-associated genes IGF2 and telomerase reverse transcriptase (TERT) were overexpressed in fibroblasts from BWS patients and TGF-β-defective mice. We further determined that chromatin insulator CCCTC-binding factor (CTCF) is TGF-β inducible and facilitates TGF-β-mediated repression of TERT transcription via interactions with β2SP and SMAD3. This regulation was abrogated in TGF-β-defective mice and BWS, resulting in TERT overexpression. Imprinting of the IGF2/H19 locus and the CDKN1C/KCNQ1 locus on chromosome 11p15.5 is mediated by CTCF, and this regulation is lost in BWS, leading to aberrant overexpression of growth-promoting genes. Therefore, we propose that loss of CTCF-dependent imprinting of tumor-promoting genes, such as IGF2 and TERT, results from a defective TGF-β pathway and is responsible at least in part for BWS-associated tumorigenesis as well as sporadic human cancers that are frequently associated with SPTBN1 and SMAD3 mutations.


Abstinence and relapse rates following a college campus-based quit & win contest.
To conduct and evaluate Quit & Win contests at 2 2-year college and 2 4-year university campuses. During Spring semester, 2006, undergraduates (N = 588) interested in quitting smoking signed up for a Quit & Win 30-day cessation contest for a chance to win a lottery prize. Participants (N = 588) completed a baseline survey, provided a urine sample to verify smoking status before joining the contest, and completed a follow-up survey at contest end to assess abstinence. Participants reporting continuous 30-day abstinence were surveyed again 2 weeks post contest to assess relapse. Participants smoked an average of 9.8 +/- 6.7 cigarettes/day on 26.7 +/- 5.7 days/month. Among participants completing a follow-up survey (74%), 72.1% reported abstinence during the entire contest period (Intent-to-Treat Analysis = 53.2%). 55.3% of those abstinent at the end of contest had resumed smoking 2 weeks post contest. Campus Quit & Win contests appear feasible, acceptable, and effective at facilitating short-term abstinence. Further research is needed to identify strategies to prevent postcontest relapse.


Both genetic and dietary factors underlie individual differences in DNA damage levels and DNA repair capacity.
The interplay between dietary habits and individual genetic make-up is assumed to influence risk of cancer, via modulation of DNA integrity. Our aim was to characterize internal and external factors that underlie inter-individual variability in DNA damage and repair and to identify dietary habits beneficial for maintaining DNA integrity. Habitual diet was estimated in 340 healthy individuals using a food frequency questionnaire and biomarkers of antioxidant status were quantified in fasting blood samples. Markers of DNA integrity were represented by DNA strand breaks, oxidized purines, oxidized pyrimidines and a sum of all three as total DNA damage. DNA repair was characterized by genetic variants and functional activities of base and nucleotide excision repair pathways. Sex, fruit-based food consumption and XPG genotype were factors significantly associated with the level of DNA damage. DNA damage was higher in women (p=0.035). Fruit consumption was negatively associated with the number of all measured DNA lesions, and this effect was mediated mostly by β-cryptoxanthin and β-tocopherol (p<0.05). XPG 1104His homozygotes appeared more vulnerable to DNA damage accumulation (p=0.001). Sex and individual antioxidants were also associated with DNA repair capacity; both the base and nucleotide excision repairs were lower in women and the latter increased with higher plasma levels of ascorbic acid and α-carotene (p<0.05). We have determined genetic and dietary factors that modulate DNA integrity. We propose that the positive health effect of fruit intake is partially mediated via DNA damage suppression and a simultaneous increase in DNA repair capacity.


One protein, multiple pathologies: multifaceted involvement of amyloid β in neurodegenerative disorders of the brain and retina.
Accumulation of amyloid β (Aβ) and its aggregates in the ageing central nervous system is regarded synonymous to Alzheimer's disease (AD) pathology. Despite unquestionable advances in mechanistic and diagnostic aspects of the disease understanding, the primary cause of Aβ accumulation as well as its in vivo roles remains elusive; nonetheless, the majority of the efforts to address pathological mechanisms for therapeutic development are focused towards moderating Aβ accumulation in the brain. More recently, Aβ deposition has been identified in the eye and is linked with distinct age-related diseases including age-related macular degeneration, glaucoma as well as AD. Awareness of the Aβ accumulation in these markedly different degenerative disorders has led to an increasing body of work exploring overlapping mechanisms, a prospective biomarker role for Aβ and the potential to use retina as a model for brain related neurodegenerative disorders. Here, we present an integrated view of current understanding of the retinal Aβ deposition discussing the accumulation mechanisms, anticipated impacts and outlining ameliorative approaches that can be extrapolated to the retina for potential therapeutic benefits. Further longitudinal investigations in humans and animal models will determine retinal Aβ association as a potential pathognomonic, diagnostic or prognostic biomarker.


Coupling of remote alternating-access transport mechanisms for protons and substrates in the multidrug efflux pump AcrB.
Membrane transporters of the RND superfamily confer multidrug resistance to pathogenic bacteria, and are essential for cholesterol metabolism and embryonic development in humans. We use high-resolution X-ray crystallography and computational methods to delineate the mechanism of the homotrimeric RND-type proton/drug antiporter AcrB, the active component of the major efflux system AcrAB-TolC in Escherichia coli, and one most complex and intriguing membrane transporters known to date. Analysis of wildtype AcrB and four functionally-inactive variants reveals an unprecedented mechanism that involves two remote alternating-access conformational cycles within each protomer, namely one for protons in the transmembrane region and another for drugs in the periplasmic domain, 50 Å apart. Each of these cycles entails two distinct types of collective motions of two structural repeats, coupled by flanking α-helices that project from the membrane. Moreover, we rationalize how the cross-talk among protomers across the trimerization interface might lead to a more kinetically efficient efflux system.


United States trends in lumbar fusion surgery for degenerative conditions.
Retrospective cohort study using national sample administrative data. To determine if lumbar fusion rates increased in the 1990s and to compare lumbar fusion rates with those of other major musculoskeletal procedures. Previous studies found that lumbar fusion rates rose more rapidly during the 1980s than did other types of lumbar surgery. We used the Healthcare Cost and Utilization Project Nationwide Inpatient Sample from 1988 through 2001 to examine trends. U.S. Census data were used for calculating age and sex-adjusted population-based rates. We excluded patients with vertebral fractures, cancer, or infection. In 2001, over 122,000 lumbar fusions were performed nationwide for degenerative conditions. This represented a 220% increase from 1990 in fusions per 100,000. The increase accelerated after 1996, when fusion cages were approved. From 1996 to 2001, the number of lumbar fusions increased 113%, compared with 13 to 15% for hip replacement and knee arthroplasty. Rates of lumbar fusion rose most rapidly among patients aged 60 and above. The proportion of lumbar operations involving a fusion increased for all diagnoses. Lumbar fusion rates rose even more rapidly in the 90s than in the 80s. The most rapid increases followed the approval of new surgical implants and were much greater than increases in other major orthopedic procedures. The most rapid increases in fusion rates were among adults aged 60 and above. These increases were not associated with reports of clarified indications or improved efficacy, suggesting a need for better data on the efficacy of various fusion techniques for various indications.


Prospective assessment of health-related quality of life in pediatric patients with beta-thalassemia following hematopoietic stem cell transplantation.
Although hematopoietic stem cell transplantation (HSCT) has been widely used to treat pediatric patients with beta-thalassemia major, evidence showing whether this treatment improves health-related quality of life (HRQoL) is lacking. We used child-self and parent-proxy reports to prospectively evaluate HRQoL in 28 children with beta-thalassemia from Middle Eastern countries who underwent allogeneic HSCT in Italy. The PedsQL 4.0 Generic Core Scales were administered to patients and their parents 1 month before and 3, 6, and 18 months after transplantation. Two-year overall survival, thalassemia-free survival, mortality, and rejection were 89.3%, 78.6%, 10.9% and 14.3%, respectively. The cumulative incidence of acute and chronic graft-versus-host disease (GVHD) was 36% and 18%, respectively. Physical functioning declined significantly from baseline to 3 months after HSCT (median PedsQL score, 81.3 vs 62.5; P = .02), but then increased significantly up to 18 months after HSCT (median score, 93.7; P = .04). Agreement between child-self and parent-proxy ratings was high. Chronic GVHD was the most significant factor associated with lower HRQoL scores over time (P = .02). The child-self and parent-proxy reports showed improved HRQoL in the children with beta-thalassemia after HSCT. Overall, our study provides preliminary evidence-based data to further support clinical decision making in this area.


ETV1-Positive Cells Give Rise to BRAFV600E -Mutant Gastrointestinal Stromal Tumors.
Gastrointestinal stromal tumor (GIST) is the most common subtype of sarcoma. Despite clinical advances in the treatment of KIT/PDGFRA-mutant GIST, similar progress against KIT/PDGFRA wild-type GIST, including mutant BRAF-driven tumors, has been limited by a lack of model systems. ETV1 is a master regulator in the intestinal cells of Cajal (ICC), thought to be the cells of origin of GIST. Here, we present a model in which the ETV1 promoter is used to specifically and inducibly drive Cre recombinase in ICC as a strategy to study GIST pathogenesis. Using a conditional allele for BrafV600E , a mutation observed in clinical cases of GIST, we observed that BrafV600E activation was sufficient to drive ICC hyperplasia but not GIST tumorigenesis. In contrast, combining BrafV600E activation with Trp53 loss was sufficient to drive both ICC hyperplasia and formation of multifocal GIST-like tumors in the mouse gastrointestinal tract with 100% penetrance. This mouse model of sporadic GIST model was amenable to therapeutic intervention, and it recapitulated clinical responses to RAF inhibition seen in human GIST. Our work offers a useful in vivo model of human sporadic forms of BRAF-mutant GIST to help unravel its pathogenesis and therapeutic response to novel experimental agents. Cancer Res; 77(14); 3758-65. ©2017 AACR.


On the automated definition of mobile target volumes from 4D-CT images for stereotactic body radiotherapy.
Stereotactic body radiotherapy (SBRT) can be used to treat small lesions in the chest. A vacuum-based immobilization system is used in our clinic for SBRT, and a motion envelope is used in treatment planning. The purpose of this study is to automatically derive motion envelopes using deformable image registration of 4D-CT images, and to assess the effect of abdominal pressure on the motion envelopes. 4D-CT scans at ten phases were acquired prior to treatment for both free and restricted breathing using a vacuum-based immobilization system that includes an abdominal pressure pillow. To study the stability of the motion envelope over the course of treatment, a mid-treatment 4D-CT scan was obtained after delivery of the third fraction for two patients. The planning target volume excluding breathing motion (PTV(ex)) was defined on the image set at full exhalation phase and transformed into all other phases using displacement maps from deformable image registration. The motion envelope was obtained as the union of PTV(ex) masks of all phases. The ratios of the motion envelope to PTV(ex) volume ranged from 1.3 to 2.5. When pressure was applied, the ratios were reduced by as much as 29% compared to free breathing for some patients, but increased by up to 9% for others. The abdominal pressure pillow has more motion restriction effects on the anterior/inferior region of the lung. For one of the two patients for whom the 4D-CT scan was repeated at mid-treatment, the motion envelope was reproducible. However, for the other patient the tumor location and lung motion pattern significantly changed due to changes in the anatomy surrounding the tumor during the course of treatment, indicating that an image-guided approach to SBRT may increase the efficacy of this treatment.


Computational modeling of the EGFR network elucidates control mechanisms regulating signal dynamics.
The epidermal growth factor receptor (EGFR) signaling pathway plays a key role in regulation of cellular growth and development. While highly studied, it is still not fully understood how the signal is orchestrated. One of the reasons for the complexity of this pathway is the extensive network of inter-connected components involved in the signaling. In the aim of identifying critical mechanisms controlling signal transduction we have performed extensive analysis of an executable model of the EGFR pathway using the stochastic pi-calculus as a modeling language. Our analysis, done through simulation of various perturbations, suggests that the EGFR pathway contains regions of functional redundancy in the upstream parts; in the event of low EGF stimulus or partial system failure, this redundancy helps to maintain functional robustness. Downstream parts, like the parts controlling Ras and ERK, have fewer redundancies, and more than 50% inhibition of specific reactions in those parts greatly attenuates signal response. In addition, we suggest an abstract model that captures the main control mechanisms in the pathway. Simulation of this abstract model suggests that without redundancies in the upstream modules, signal transduction through the entire pathway could be attenuated. In terms of specific control mechanisms, we have identified positive feedback loops whose role is to prolong the active state of key components (e.g., MEK-PP, Ras-GTP), and negative feedback loops that help promote signal adaptation and stabilization. The insights gained from simulating this executable model facilitate the formulation of specific hypotheses regarding the control mechanisms of the EGFR signaling, and further substantiate the benefit to construct abstract executable models of large complex biological networks.


Tunable phase-only optical filters with a uniaxial crystal.
A novel method of fabricating phase-only optical filters that is based on the properties of a uniaxial crystal is proposed. With these optical filters, the phase differences are tunable among the different filter zones. Many focal patterns can be obtained if these optical filters are placed in front of a lens; furthermore, these optical filters can also be used to make up for the distortions in fabrications in which they were used only as untunable optical filters.


Psychological stress and short-term hospitalisations or death in patients with heart failure.
Standard predictors do not fully explain variations in the frequency and timing of heart failure (HF) adverse events (AEs). Psychological stress can trigger acute cardiovascular (CV) events, but it is not known whether stress can precipitate AEs in patients with HF. We investigated prospective associations of psychological stress with AEs in patients with HF. 144 patients with HF (77% male; 57.5±11.5, range 23-87 years, left ventricular ejection fraction ≤40%) were longitudinally evaluated for psychological stress (Perceived Stress Scale) and AEs (CV hospitalisations/death) at 2-week intervals for 3 months and at 9-month follow-up. 42 patients (29.2%) had at least one CV hospitalisation and nine (6.3%) died. Patients reporting high average perceived stress across study measurements had a higher likelihood of AEs during the study period compared with those with lower stress (odds ratio=1.10, 95% confidence interval=1.04 to 1.17). In contrast to average levels, increases in stress did not predict AEs (p=0.96). Perceived stress was elevated after a CV hospitalisation (B=2.70, standard error (SE)=0.93, p=0.004) suggesting that CV hospitalisations increase stress. Subsequent analysis indicated that 24 of 38 (63%) patients showed a stress increase following hospitalisation. However, a prospective association between stress and AEs was present when accounting for prior hospitalisations (B=2.43, SE=1.23, p=0.05). Sustained levels of perceived stress are associated with increased risk of AEs, and increased distress following hospitalisation occurs in many, but not all, patients with HF. Patients with chronically high stress may be an important target group for HF interventions aimed at reducing hospitalisations.


Phylogenetic position and in situ identification of ectosymbiotic spirochetes on protists in the termite gut.
Phylogenetic relationships, diversity, and in situ identification of spirochetes in the gut of the termite Neotermes koshunensis were examined without cultivation, with an emphasis on ectosymbionts attached to flagellated protists. Spirochetes in the gut microbial community investigated so far are related to the genus Treponema and divided into two phylogenetic clusters. In situ hybridizations with a 16S rRNA-targeting consensus oligonucleotide probe for one cluster (known as termite Treponema cluster I) detected both the ectosymbiotic spirochetes on gut protists and the free-swimming spirochetes in the gut fluid of N. koshunensis. The probe for the other cluster (cluster II), which has been identified as ectosymbionts on gut protists of two other termite species, Reticulitermes speratus and Hodotermopsis sjoestedti, failed to detect any spirochete population. The absence of cluster II spirochetes in N. koshunensis was confirmed by intensive 16S ribosomal DNA (rDNA) clone analysis, in which remarkably diverse spirochetes of 45 phylotypes were identified, almost all belonging to cluster I. Ectosymbiotic spirochetes of the three gut protist species Devescovina sp., Stephanonympha sp., and Oxymonas sp. in N. koshunensis were identified by their 16S rDNA and by in situ hybridizations using specific probes. The probes specific for these ectosymbionts did not receive a signal from the free-swimming spirochetes. The ectosymbionts were dispersed in cluster I of the phylogeny, and they formed distinct phylogenetic lineages, suggesting multiple origins of the spirochete attachment. Each single protist cell harbored multiple spirochete species, and some of the spirochetes were common among protist species. The results indicate complex relationships of the ectosymbiotic spirochetes with the gut protists.


Radial keratotomy in India untoward consequences and complications.
R.K. is a surgery of tomorrow. It is still in the stage of evolution. It should be under-taken only for precise indications. Case evaluation and honest advice to the patient is important. The patient should be explained that the worst can happen. The surgery should be limited to moderate degrees of myopia, not less than -3.0 D sphere and not more than 8 to 10 D. It has no place for myopia with degenerative retinal changes. Surgery should be interrupted at the first sign of a significant operative complication. It will be unfortunate if a healthy eye is lost due to complications of R.K. done with indiscriminate approach and ignorance of possible post-operative consequences.


Integration of clinical decision support with on-line encounter documentation for well child care at the point of care.
Electronic medical record systems and clinical practice guideline (CPG) support applications are emerging in the clinical environment to document and support care. Applications which integrate online documentation with CPG are often complex systems bound to a proprietary infrastructure and as such, can be difficult to adapt to changing care guidelines. This paper describes integration of point-of-care clinical documentation to an Internet-based CPG system that was easily modified, utilized available software resources, and separated patient information from CPG. The system combined a text-based encounter documentation tool, Inbox, with a web-based CPG system, SIEGFRIED (System for Interactive Electronic Guidelines with Feedback and Resources for Instructional and Educational Development), which interactively presented care guidelines to providers. Age-specific well child care documentation templates were developed using Inbox for point-of-care documentation. SIEGFRIED contained the knowledge base of child safety education guidelines and executed independent of the program presenting the guidelines. The CPG were accessed from within the documentation template via an Internet hyperlink. Patient chart evaluation indicated that 77% of safety topics were reviewed and 32% of the charts contained documentation indicating all the safety topics were reviewed. Last, routine use of the Inbox-SIEGFRIED system was not realized due to the clinical time constraints and workload of the medical providers, and lack of data entry experience. A user survey indicated time cost (network access and software execution) were negative aspects of the system. However, the system function was highly regarded and the Internet-based patient education materials were described as useful and accurate. In summary, the system was functional, met original development goals, and provided valuable patient education materials; however, routine system use was prevented by time requirements. We recommend further development be oriented towards integrating the identified beneficial components of the system into clinician workflow.


Children under 10 years of age were more affected by the 2018/19 influenza A(H1N1)pdm09 epidemic in Canada: possible cohort effect following the 2009 influenza pandemic.
IntroductionFindings from the community-based Canadian Sentinel Practitioner Surveillance Network (SPSN) suggest children were more affected by the 2018/19 influenza A(H1N1)pdm09 epidemic.AimTo compare the age distribution of A(H1N1)pdm09 cases in 2018/19 to prior seasonal influenza epidemics in Canada.MethodsThe age distribution of unvaccinated influenza A(H1N1)pdm09 cases and test-negative controls were compared across A(H1N1)pdm09-dominant epidemics in 2018/19, 2015/16 and 2013/14 and with the general population of SPSN provinces. Similar comparisons were undertaken for influenza A(H3N2)-dominant epidemics.ResultsIn 2018/19, more influenza A(H1N1)pdm09 cases were under 10 years old than controls (29% vs 16%; p < 0.001). In particular, children aged 5-9 years comprised 14% of cases, greater than their contribution to controls (4%) or the general population (5%) and at least twice their contribution in 2015/16 (7%; p < 0.001) or 2013/14 (5%; p < 0.001). Conversely, children aged 10-19 years (11% of the population) were under-represented among A(H1N1)pdm09 cases versus controls in 2018/19 (7% vs 12%; p < 0.001), 2015/16 (7% vs 13%; p < 0.001) and 2013/14 (9% vs 12%; p = 0.12).ConclusionChildren under 10 years old contributed more to outpatient A(H1N1)pdm09 medical visits in 2018/19 than prior seasonal epidemics in Canada. In 2018/19, all children under 10 years old were born after the 2009 A(H1N1)pdm09 pandemic and therefore lacked pandemic-induced immunity. In addition, more than half those born after 2009 now attend school (i.e. 5-9-year-olds), a socio-behavioural context that may enhance transmission and did not apply during prior A(H1N1)pdm09 epidemics.


Role of the rfe gene in the biosynthesis of the Escherichia coli O7-specific lipopolysaccharide and other O-specific polysaccharides containing N-acetylglucosamine.
We report that rfe mutants of wild-type strains of Escherichia coli O7, O18, O75, and O111 did not express O-specific polysaccharide unless the rfe mutation was complemented by a cloned rfe gene supplied in a plasmid. The O polysaccharides in these strains are known to have N-acetylglucosamine (GlcNAc) in their O repeats. In addition, in vitro transferase assays with bacterial membranes from either the O7 wild-type strain or its isogenic rfe mutant showed that GlcNAc is the first carbohydrate added onto the lipid acceptor in the assembly of the O7 repeat and that this function is inhibited by tunicamycin. Our results indicate that the rfe gene product is a general requirement for the synthesis of O polysaccharides containing GlcNAc.


Chronic Dialysis Patients Are Depleted of Creatine: Review and Rationale for Intradialytic Creatine Supplementation.
There is great need for the identification of new, potentially modifiable risk factors for the poor health-related quality of life (HRQoL) and of the excess risk of mortality in dialysis-dependent chronic kidney disease patients. Creatine is an essential contributor to cellular energy homeostasis, yet, on a daily basis, 1.6-1.7% of the total creatine pool is non-enzymatically degraded to creatinine and subsequently lost via urinary excretion, thereby necessitating a continuous supply of new creatine in order to remain in steady-state. Because of an insufficient ability to synthesize creatine, unopposed losses to the dialysis fluid, and insufficient intake due to dietary recommendations that are increasingly steered towards more plant-based diets, hemodialysis patients are prone to creatine deficiency, and may benefit from creatine supplementation. To avoid problems with compliance and fluid balance, and, furthermore, to prevent intradialytic losses of creatine to the dialysate, we aim to investigate the potential of intradialytic creatine supplementation in improving outcomes. Given the known physiological effects of creatine, intradialytic creatine supplementation may help to maintain creatine homeostasis among dialysis-dependent chronic kidney disease patients, and consequently improve muscle status, nutritional status, neurocognitive status, HRQoL. Additionally, we describe the rationale and design for a block-randomized, double-blind, placebo-controlled pilot study. The aim of the pilot study is to explore the creatine uptake in the circulation and tissues following different creatine supplementation dosages.


Mobilisation of data to stakeholder communities. Bridging the research-practice gap using a commercial shellfish species model.
Knowledge mobilisation is required to "bridge the gap" between research, policy and practice. This activity is dependent on the amount, richness and quality of the data published. To understand the impact of a changing climate on commercial species, stakeholder communities require better knowledge of their past and current situations. The common cockle (Cerastoderma edule) is an excellent model species for this type of analysis, as it is well-studied due to its cultural, commercial and ecological significance in west Europe. Recently, C. edule harvests have decreased, coinciding with frequent mass mortalities, due to factors such as a changing climate and diseases. In this study, macro and micro level marine historical ecology techniques were used to create datasets on topics including: cockle abundance, spawning duration and harvest levels, as well as the ecological factors impacting those cockle populations. These data were correlated with changing climate and the Atlantic Multidecadal Oscillation (AMO) index to assess if they are drivers of cockle abundance and harvesting. The analyses identified the key stakeholder communities involved in cockle research and data acquisition. It highlighted that data collection was sporadic and lacking in cross-national/stakeholder community coordination. A major finding was that local variability in cockle populations is influenced by biotic (parasites) and abiotic (temperature, legislation and harvesting) factors, and at a global scale by climate (AMO Index). This comprehensive study provided an insight into the European cockle fishery but also highlights the need to identify the type of data required, the importance of standardised monitoring, and dissemination efforts, taking into account the knowledge, source, and audience. These factors are key elements that will be highly beneficial not only to the cockle stakeholder communities but to other commercial species.


Metal-amplified Density Assays, (MADAs), including a Density-Linked Immunosorbent Assay (DeLISA).
This paper reports the development of Metal-amplified Density Assays, or MADAs - a method of conducting quantitative or multiplexed assays, including immunoassays, by using Magnetic Levitation (MagLev) to measure metal-amplified changes in the density of beads labeled with biomolecules. The binding of target analytes (i.e. proteins, antibodies, antigens) to complementary ligands immobilized on the surface of the beads, followed by a chemical amplification of the binding in a form that results in a change in the density of the beads (achieved by using gold nanoparticle-labeled biomolecules, and electroless deposition of gold or silver), translates analyte binding events into changes in density measureable using MagLev. A minimal model based on diffusion-limited growth of hemispherical nuclei on a surface reproduces the dynamics of the assay. A MADA - when performed with antigens and antibodies - is called a Density-Linked Immunosorbent Assay, or DeLISA. Two immunoassays provided a proof of principle: a competitive quantification of the concentration of neomycin in whole milk, and a multiplexed detection of antibodies against Hepatitis C virus NS3 protein and syphilis T. pallidum p47 protein in serum. MADAs, including DeLISAs, require, besides the requisite biomolecules and amplification reagents, minimal specialized equipment (two permanent magnets, a ruler or a capillary with calibrated length markings) and no electrical power to obtain a quantitative readout of analyte concentration. With further development, the method may be useful in resource-limited or point-of-care settings.


Identification of blood protein biomarkers that aid in the clinical assessment of patients with malignant glioma.
Analyzing molecular biomarkers using blood is an important approach for clinical assessment of malignant glioma. We investigated a molecular proteomic biomarker-based approach for glioblastoma using patients' blood samples. The expression levels of a list of candidate proteins were quantified in plasma and serum samples from two different cohorts of patients with malignant glioma and normal controls. The biological function was studied for one of the identified markers. Additionally, the prognostic significance of protein marker expression was measured by survival analysis. As a result, protein biomarkers associated with malignant glioma were identified from the blood specimens and five of the protein biomarkers were common to both cohorts. Immunohistochemical analysis demonstrated that many of the protein biomarkers identified in peripheral blood specimens were expressed in malignant gliomas. Staining levels for one of the biomarkers, MIP-1α, was found to correlate with WHO grade among invasive gliomas, and we demonstrate that MIP-1α promotes human glioblastoma cell proliferation and migration. Additionally, four prognostic protein biomarkers were identified. In conclusion, we demonstrate that both peripheral blood plasma and serum specimens are highly valuable and complementary to each other in the quest for protein biomarkers of malignant glioma. Sets of novel protein biomarkers were identified that may aid in the diagnosis and prognosis of patients with malignant glioma.


Antiviral RNA interference responses induced by Semliki Forest virus infection of mosquito cells: characterization, origin, and frequency-dependent functions of virus-derived small interfering RNAs.
RNA interference (RNAi) is an important mosquito defense mechanism against arbovirus infection. In this paper we study the processes underlying antiviral RNAi in Aedes albopictus-derived U4.4 mosquito cells infected with Semliki Forest virus (SFV) (Togaviridae; Alphavirus). The production of virus-derived small interfering RNAs (viRNAs) from viral double-stranded RNA (dsRNA) is a key event in this host response. dsRNA could be formed by RNA replication intermediates, by secondary structures in RNA genomes or antigenomes, or by both. Which of these dsRNAs is the substrate for the generation of viRNAs is a fundamental question. Here we used deep sequencing of viRNAs and bioinformatic analysis of RNA secondary structures to gain insights into the characteristics and origins of viRNAs. An asymmetric distribution of SFV-derived viRNAs with notable areas of high-level viRNA production (hot spots) and no or a low frequency of viRNA production (cold spots) along the length of the viral genome with a slight bias toward the production of genome-derived viRNAs over antigenome-derived viRNAs was observed. Bioinformatic analysis suggests that hot spots of viRNA production are rarely but not generally associated with putative secondary structures in the SFV genome, suggesting that most viRNAs are derived from replicative dsRNA. A pattern of viRNAs almost identical to those of A. albopictus cells was observed for Aedes aegypti-derived Aag2 cells, suggesting common mechanisms that lead to viRNA production. Hot-spot viRNAs were found to be significantly less efficient at mediating antiviral RNAi than cold-spot viRNAs, pointing toward a nucleic acid-based viral decoy mechanism to evade the RNAi response.


A shift from oral to blood pH is a stimulus for adaptive gene expression of Streptococcus gordonii CH1 and induces protection against oxidative stress and enhanced bacterial growth by expression of msrA.
Viridans group streptococci (VS) from the oral cavity entering the bloodstream may initiate infective endocarditis (IE). We aimed to identify genes expressed in response to a pH increase from slightly acidic (pH 6.2) to neutral (pH 7.3) as encountered by VS entering the bloodstream from the oral cavity. Using a recently developed promoter-screening vector, we isolated five promoter fragments from the genomic DNA of Streptococcus gordonii CH1 responding to this stimulus. No common regulatory sequences were identified in these promoter fragments that could account for the coordinate expression of the corresponding genes. One of the isolated fragments contained the promoter region and 5' end of a gene highly homologous to the methionine sulfoxide reductase gene (msrA) of various bacterial and eukaryotic species. This gene has been found to be activated in S. gordonii strain V288 in a rabbit model of IE (A. O. Kiliç, M. C. Herzberg, M. W. Meyer, X. Zhao, and L. Tao, Plasmid 42:67-72, 1999). We isolated and characterized the msrA gene of S. gordonii CH1 and constructed a chromosomal insertion mutant. This mutant was more sensitive to hydrogen peroxide, suggesting a role for the streptococcal MsrA in protecting against oxidative stress. Moreover, MsrA appeared to be important for the growth of S. gordonii CH1 under aerobic and anaerobic conditions. Both these properties of MsrA may contribute to the ability of S. gordonii to cause IE.


Modulation of whey protein-kappa carrageenan hydrogel properties via enzymatic protein modification.
Treatment of whey protein isolate (WPI; 1 to 25% w/w) in heated κ-carrageenan (KC; 2% w/w) slurries with protease and/or transglutaminase modulated the properties of the hydrogels formed after cooling. Observation of peak compression stress and strain at gel rupture showed WPI incorporation at 1, 5 and 10% (w/w) significantly reduced the strength and deformability of 2% (w/w) KC gels. Treatment of WPI solutions in KC slurries with Alcalase 2.4L was shown by both SDS-Page and size exclusion HPLC to reduce protein/peptide molecular weight distributions below 10 kDa, with large portions below 1 kDa. This peptide size reduction within the KC matrix produced more translucent gels with a more organized wall and cell structure as observed by SEM, which resulted in gels with observed rupture stress/strain levels similar to 2% KC alone. Transglutaminase treatment of WPI-KC slurries showed the reverse behavior, reducing gel translucency, strength and deformability. At these loadings, WPI-KC gel strength/deformability appears to relate decreasing peptide size to gel behavior trending towards KC-only gels; suggesting peptide size modulation in protein-carbohydrate complexes will allow significant tailoring of texture for the delivery of protein/peptide rich gelled products.


Global validation of the WSES Sepsis Severity Score for patients with complicated intra-abdominal infections: a prospective multicentre study (WISS Study).
To validate a new practical Sepsis Severity Score for patients with complicated intra-abdominal infections (cIAIs) including the clinical conditions at the admission (severe sepsis/septic shock), the origin of the cIAIs, the delay in source control, the setting of acquisition and any risk factors such as age and immunosuppression. The WISS study (WSES cIAIs Score Study) is a multicenter observational study underwent in 132 medical institutions worldwide during a four-month study period (October 2014-February 2015). Four thousand five hundred thirty-three patients with a mean age of 51.2 years (range 18-99) were enrolled in the WISS study. Univariate analysis has shown that all factors that were previously included in the WSES Sepsis Severity Score were highly statistically significant between those who died and those who survived (p < 0.0001). The multivariate logistic regression model was highly significant (p < 0.0001, R2 = 0.54) and showed that all these factors were independent in predicting mortality of sepsis. Receiver Operator Curve has shown that the WSES Severity Sepsis Score had an excellent prediction for mortality. A score above 5.5 was the best predictor of mortality having a sensitivity of 89.2 %, a specificity of 83.5 % and a positive likelihood ratio of 5.4. WSES Sepsis Severity Score for patients with complicated Intra-abdominal infections can be used on global level. It has shown high sensitivity, specificity, and likelihood ratio that may help us in making clinical decisions.


Intestinal stromal cells in mucosal immunity and homeostasis.
A growing body of evidence suggests that non-hematopoietic stromal cells of the intestine have multiple roles in immune responses and inflammation at this mucosal site. Despite this, many still consider gut stromal cells as passive structural entities, with past research focused heavily on their roles in fibrosis, tumor progression, and wound healing, rather than their contributions to immune function. In this review, we discuss our current knowledge of stromal cells in intestinal immunity, highlighting the many immunological axes in which stromal cells have a functional role. We also consider emerging data that broaden the potential scope of their contribution to immunity in the gut and argue that these so-called "non-immune" cells are reclassified in light of their diverse contributions to intestinal innate immunity and the maintenance of mucosal homeostasis.


Underrepresentation and Symbolic Annihilation of Socially Disenfranchised Groups ("Out Groups") in Animated Cartoons.
For many years, the mass media have been accused of providing negative and potentially-damaging messages to viewers. Some have complained that the media are replete with too much violence while others have lamented on media stereotyping of various groups. In this article, the authors examine the issues of underrepresentation and symbolic annihilation as they apply to one particular medium-namely, animated cartoons-to which people are exposed early in life, typically on a regular basis for many years. Our principal research questions are (a) To what extent do cartoons underrepresent and/or symbolically annihilate social groups that are not considered desirable in society-at-large? (b) Has underrepresentation and/or symbolic annihilation changed over time? and (c) When social "out groups" are shown, how are they depicted vis-a-vis "in groups"? To examine these questions, the authors examine portrayals based on gender, age, race, and sexual orientation. The data revealed that animated cartoons have a long history of underrepresenting and symbolically annihilating socially devalued "out groups" and that little has changed over the course of the past 65+ years. When "out group" members are included in cartoons, however, their portrayals tend not to be dramatically different-not better and not much worse-than those typical of their "in group" counterparts.


Genotype-phenotype correlations in recessive RYR1-related myopathies.
RYR1 mutations are typically associated with core myopathies and are the most common overall cause of congenital myopathy. Dominant mutations are most often associated with central core disease and malignant hyperthermia, and genotype-phenotype patterns have emerged from the study of these mutations that have contributed to the understanding of disease pathogenesis. The recent availability of genetic testing for the entire RYR1 coding sequence has led to a dramatic expansion in the identification of recessive mutations in core myopathies and other congenital myopathies. To date, no clear patterns have been identified in these recessive mutations, though no systematic examination has yet been performed. In this study, we investigated genotype-phenotype correlations in a large combined cohort of unpublished (n = 14) and previously reported (n = 92) recessive RYR1 cases. Overall examination of this cohort revealed nearly 50% of cases to be non-core myopathy related. Our most significant finding was that hypomorphic mutations (mutations expected to diminish RyR1 expression) were enriched in patients with severe clinical phenotypes. We also determined that hypomorphic mutations were more likely to be encountered in non-central core myopathies. With analysis of the location of non-hypomorphic mutations, we found that missense mutations were generally enriched in the MH/CCD hotspots and specifically enriched in the selectivity filter of the channel pore. These results support a hypothesis that loss of protein function is a key predictive disease parameter. In addition, they suggest that decreased RyR1 expression may dictate non-core related pathology though, data on protein expression was limited and should be confirmed in a larger cohort. Lastly, the results implicate abnormal ion conductance through the channel pore in the pathogenesis in recessive core myopathies. Overall, our findings represent a comprehensive analysis of genotype-phenotype associations in recessive RYR1-myopathies.


Variation in positron emission tomography use after colon cancer resection.
Colon cancer surveillance guidelines do not routinely include positron emission tomography (PET) imaging; however, its use after surgical resection has been increasing. We evaluated the secular patterns of PET use after surgical resection of colon cancer among elderly patients and identified factors associated with its increasing use. We used the SEER-linked Medicare database (July 2001 through December 2009) to establish a retrospective cohort of patients age ≥ 66 years who had undergone surgical resection for colon cancer. Postoperative PET use was assessed with the test for trends. Patient, tumor, and treatment characteristics were analyzed using univariable and multivariable logistic regression analyses. Of the 39,221 patients with colon cancer, 6,326 (16.1%) had undergone a PET scan within 2 years after surgery. The use rate steadily increased over time. The majority of PET scans had been performed within 2 months after surgery. Among patients who had undergone a PET scan, 3,644 (57.6%) had also undergone preoperative imaging, and 1,977 (54.3%) of these patients had undergone reimaging with PET within 2 months after surgery. Marriage, year of diagnosis, tumor stage, preoperative imaging, postoperative visit to a medical oncologist, and adjuvant chemotherapy were significantly associated with increased PET use. PET use after colon cancer resection is steadily increasing, and further study is needed to understand the clinical value and effectiveness of PET scans and the reasons for this departure from guideline-concordant care.


A self-report home environment screening tool identified older women at risk of falls.
To evaluate a self-report version of the Home Falls and Accidents Screening Tool (HOME FAST-SR). The HOME FAST-SR was designed using expert review, pretesting and piloting. Agreement of self-reported home hazard ratings and ratings by health professionals was evaluated using the kappa statistic. The HOME FAST-SR was validated in a cross-sectional survey of 568 older community-living Australian women using endorsement rates for HOME FAST-SR items and the association of scores with self-reported falls. The 87-item HOME FAST-SR was constructed, and a scoring system was devised to calculate equivalent scores for the health professional version of the HOME FAST (HOME FAST-HP). Endorsement rates demonstrated that no items needed to be removed. Agreement between self-reported and professional ratings was moderate with therapists under-reporting hazards compared with older people. The mean HOME FAST score for the 568 cross-sectional participants was 9.39 (95% confidence interval: 9.1, 9.7) with a range from 2 to 23 out of a possible 25. Fallers had a significantly higher HOME FAST score (P=0.02). The HOME FAST-SR is a viable alternative to the HOME FAST-HP, and scores were associated with falls experienced by older women in a cross-sectional study.


Recombinant human C1 inhibitor for the prophylaxis of hereditary angioedema attacks: a pilot study.
Hereditary angioedema (HAE) is a disease characterized by recurrent tissue swelling affecting various body locations. Recent literature shows that patients with frequent attacks may benefit from long-term prophylaxis. This study evaluated the safety and prophylactic effect of weekly administrations of recombinant C1INH (rhC1INH). Patients with a history of HAE attacks occurring ≥every 2 weeks received a once weekly administration of 50 U/kg rhC1INH. Hereditary angioedema attack history was collected at screening. Breakthrough attacks during the study were recorded at each visit. Following a 2-week run-in period, HAE patients received 8 weekly rhC1INH administrations and were followed-up for an additional 6 weeks. Efficacy was evaluated by comparing the HAE attack incidence during the treatment period to the historical attacks over the previous 2 years. Safety evaluation was based on clinical laboratory and adverse events (AEs) reports. The 25 participants reported a mean of 0.9 attacks/week over the past 2 years. The mean breakthrough attack rate during the treatment period was 0.4 attacks/week (95% CI 0.28-0.56). A total of 30 treatment-emergent-AEs were reported in 13 patients, all mild to moderate. One patient died from a laryngeal attack 25 days after last study drug administration. The only possible drug related AEs reported were dry mouth, dizziness and anxiety in one patient and hypotension in another. There were no allergic AEs and no neutralizing antibodies observed. Weekly administrations of 50 U/kg rhC1INH appeared to reduce the frequency of HAE attacks and were generally safe and well tolerated.


Insurance adjudication favoring prophylactic surgery in hereditary breast-ovarian cancer syndrome.
Insurance payment to cover costs of prophylactic surgery is occasionally denied to patients who may be at inordinately high risk for hereditary cancer. We describe a 43-year-old woman from a hereditary breast-ovarian cancer (HBOC) family which showed linkage to BRCA1 and who was at inordinately high risk for cancer but who was denied insurance coverage for prophylactic oophorectomy, despite strong recommendations by her gynecologist and a cancer geneticist-medical oncologist. The insurance company claimed that the surgery was not medically necessary because the woman's condition (hereditary cancer predisposition) was not an illness. A summary judgment issued by the Douglas County (Nebraska) District Court ruled in favor of the denial decision of Blue Cross/Blue Shield. But on appeal, the Nebraska Supreme Court offered the conclusion that the patient did in fact require this prophylactic surgery and overruled the District Court's decision for denial. The litigation provides the basis for this case report and may provide a precedent for insurance coverage for patients at high risk for hereditary forms of cancer.


Cholestane-3β, 5α, 6β-triol suppresses proliferation, migration, and invasion of human prostate cancer cells.
Oxysterols are oxidation products of cholesterol. Cholestane-3β, 5α, 6β-triol (abbreviated as triol) is one of the most abundant and active oxysterols. Here, we report that triol exhibits anti-cancer activity against human prostate cancer cells. Treatment of cells with triol dose-dependently suppressed proliferation of LNCaP CDXR-3, DU-145, and PC-3 human prostate cancer cells and reduced colony formation in soft agar. Oral administration of triol at 20 mg/kg daily for three weeks significantly retarded the growth of PC-3 xenografts in nude mice. Flow cytometric analysis revealed that triol treatment at 10-40 µM caused G1 cell cycle arrest while the TUNEL assay indicated that triol treatment at 20-40 µM induced apoptosis in all three cell lines. Micro-Western Arrays and traditional Western blotting methods indicated that triol treatment resulted in reduced expression of Akt1, phospho-Akt Ser473, phospho-Akt Thr308, PDK1, c-Myc, and Skp2 protein levels as well as accumulation of the cell cycle inhibitor p27(Kip). Triol treatment also resulted in reduced Akt1 protein expression in PC-3 xenografts. Overexpression of Skp2 in PC-3 cells partially rescued the growth inhibition caused by triol. Triol treatment suppressed migration and invasion of DU-145, PC-3, and CDXR-3 cells. The expression levels of proteins associated with epithelial-mesenchymal transition as well as focal adhesion kinase were affected by triol treatment in these cells. Triol treatment caused increased expression of E-cadherin protein levels but decreased expression of N-cadherin, vimentin, Slug, FAK, phospho-FAK Ser722, and phospho-FAK Tyr861 protein levels. Confocal laser microscopy revealed redistribution of β-actin and α-tubulin at the periphery of the CDXR-3 and DU-145 cells. Our observations suggest that triol may represent a promising therapeutic agent for advanced metastatic prostate cancer.


Mapping patterns of long-term settlement in Northern Mesopotamia at a large scale.
The landscapes of the Near East show both the first settlements and the longest trajectories of settlement systems. Mounding is a characteristic property of these settlement sites, resulting from millennia of continuing settlement activity at distinguished places. So far, however, this defining feature of ancient settlements has not received much attention, or even been the subject of systematic evaluation. We propose a remote sensing approach for comprehensively mapping the pattern of human settlement at large scale and establish the largest archaeological record for a landscape in Mesopotamia, mapping about 14,000 settlement sites--spanning eight millennia--at 15-m resolution in a 23,000-km(2) area in northeastern Syria. To map both low- and high-mounded places--the latter of which are often referred to as "tells"--we develop a strategy for detecting anthrosols in time series of multispectral satellite images and measure the volume of settlement sites in a digital elevation model. Using this volume as a proxy to continued occupation, we find a dependency of the long-term attractiveness of a site on local water availability, but also a strong relation to the relevance within a basin-wide exchange network that we can infer from our record and third millennium B.C. intersite routes visible on the ground until recent times. We believe it is possible to establish a nearly comprehensive map of human settlements in the fluvial plains of northern Mesopotamia and beyond, and site volume may be a key quantity to uncover long-term trends in human settlement activity from such a record.


Wavelength-tunable bidirectional passively Q-switched Er-doped fiber laser incorporating a single-walled carbon nanotube and tunable bandpass filter.
We demonstrated a continuously wavelength-tunable bidirectional passively $ Q $Q-switched fiber laser based on a single-walled carbon nanotube saturable absorber and a bandpass filter (BPF). By tuning the commercial BPF, the $ Q $Q-switched pulse can be obtained in the same cavity at the same pump power in clockwise (CW) and counter-clockwise (CCW) directions. The central wavelength can be continuously tuned from 1520.88 to 1568.56 nm in the CW direction, and from 1520.96 to 1568.64 nm in the CCW direction. A wide tuning range of 48 nm is achieved for two directions in the passively $ Q $Q-switched fiber laser. During the tuning process of the fiber laser, the output pulses with the same central lasing wavelength can be obtained in both oscillation directions owing to the BPF. By increasing the pump power from 130 to 350 mW, the cavity delivers a $ Q $Q-switched pulse with the central wavelength of 1560 nm whose repetition rate changes from 9.64 to 59.18 kHz for the CW direction. In the CCW direction, the repetition rate of the $ Q $Q-switched pulse changes from 10.26 to 61.03 kHz. To the best of our knowledge, it is the first time that continuous wavelength-tunable passively $ Q $Q-switched pulses have been achieved in a bidirectional erbium-doped fiber laser.


How people achieve their amazing temporal precision in interception.
People can hit rapidly moving balls with amazing precision. To determine how they manage to do so, we explored how various factors that we could manipulate influenced people's precision when intercepting virtual targets. We found that temporal precision was highest for fast targets that subjects were free to intercept wherever they wished. Temporal precision was much poorer when the point of interception was specified in advance. Examining responses to abrupt perturbations of the target's motion revealed that people adjusted where rather than when they would hit the target if given the choice. A model that combines judging how long it will take to reach the target's path with estimating the target's position at that time from its visually perceived position and velocity could account for the observed precision with reasonable values for all the parameters. The model considers all relevant sources of errors, together with the delays with which the various aspects can be adjusted. Our analysis provides a biologically plausible explanation for how light falling on the eye can guide the hand to intercept a moving ball with such high precision.


Testing Dietary Hypotheses of East African Hominines Using Buccal Dental Microwear Data.
There is much debate on the dietary adaptations of the robust hominin lineages during the Pliocene-Pleistocene transition. It has been argued that the shift from C3 to C4 ecosystems in Africa was the main factor responsible for the robust dental and facial anatomical adaptations of Paranthropus taxa, which might be indicative of the consumption of fibrous, abrasive plant foods in open environments. However, occlusal dental microwear data fail to provide evidence of such dietary adaptations and are not consistent with isotopic evidence that supports greater C4 food intake for the robust clades than for the gracile australopithecines. We provide evidence from buccal dental microwear data that supports softer dietary habits than expected for P. aethiopicus and P. boisei based both on masticatory apomorphies and isotopic analyses. On one hand, striation densities on the buccal enamel surfaces of paranthropines teeth are low, resembling those of H. habilis and clearly differing from those observed on H. ergaster, which display higher scratch densities indicative of the consumption of a wide assortment of highly abrasive foodstuffs. Buccal dental microwear patterns are consistent with those previously described for occlusal enamel surfaces, suggesting that Paranthropus consumed much softer diets than previously presumed and thus calling into question a strict interpretation of isotopic evidence. On the other hand, the significantly high buccal scratch densities observed in the H. ergaster specimens are not consistent with a highly specialized, mostly carnivorous diet; instead, they support the consumption of a wide range of highly abrasive food items.


Sugar-containing beverage intake at the age of 1 year and cardiometabolic health at the age of 6 years: the Generation R Study.
Consumption of sugar-containing beverages (SCBs) in adults has been associated with an increased risk of metabolic syndrome. Although the effect of SCB on body weight in children is well established, little is known about the cardiometabolic effects in young children. We studied the associations of SCB intake at the age of 1 year with cardiometabolic health at age 6 years. This study was performed among 2,045 Dutch children from a population based prospective birth cohort. SCB intake was assessed with a semi-quantitative food frequency questionnaire at the age of 13 months and sex-specific tertiles were created. Children visited the research center at the age of 6 years. We created a continuous cardiometabolic risk factor score including: body fat percentage, blood pressure, insulin, HDL-cholesterol and triglycerides. Age-and sex-specific standard deviation (SD) scores were created for all outcomes. Multivariable linear regression was performed with adjustment for socio-demographic and lifestyle variables of mother and child. In the total population, we observed an association between higher SCB intake at 13 months of age and a higher cardiometabolic risk factor score at the age of 6 years (0.13SD (95 % CI 0.01; 0.25), highest vs. lowest tertile) After stratification by sex, we found that boys in the highest tertile of SCB intake had a higher cardiometabolic risk factor score (0.18 SD (95 % CI 0.01; 0.34)), as compared to boys in the lowest tertile of SCB intake. There was no significant association in girls. We did not find associations of SCB intake with the individual cardiometabolic risk factors in the total population, or in the stratified analyses. Higher SCB intake at 1 year of age was associated with a higher cardiometabolic risk factor score at age 6 years in boys, but not in girls. Further research on sex-specific effects of SCBs is needed.


Prognostic and therapeutic implications of mTORC1 and Rictor expression in human breast cancer.
The mammalian target of rapamycin (mTOR) plays a key role in the regulation of cellular metabolism, growth and proliferation. It forms two multi-protein complexes known as complex 1 (mTORC1) and 2 (mTORC2). Raptor and Rictor are the core proteins for mTORC1 and mTORC2, respectively. This study examines the relationship between mTORC1, Rictor and Raptor mRNA expression and human breast cancer. Furthermore, the correlation between mTORC1 and hTERT was investigated. Breast cancer tissues (n=150) and normal tissues (n=31) were analysed using reverse transcription and quantitative PCR. Transcript levels were correlated with clinicopathological data. Higher mTOR expression was noted in breast cancer tissue (P=0.0018), higher grade tumours (grade 2 vs. 3, P=0.047), in ductal tumours (P=0.0014), and was associated with worse overall survival (P=0.01). Rictor expression was significantly higher in background breast tissues compared with tumours and was inversely related to the Nottingham Prognostic Index (NPI1 vs. 2, P=0.03) and tumour grade (grade 1 vs. 3, P=0.01) and was associated with better overall (P=0.037) and disease-free survival (P=0.048). The mRNA expression of Raptor was higher in tumours compared with normal tissues. Furthermore, the expression of Raptor was associated with a higher tumour grade (grade 1 vs. 3, P=0.027). A highly significant positive correlation between mTOR and hTERT (P<0.00001) was observed. These observations are consistent with the role of mTORC1 in the anti-apoptosis pathway and suggest that selective inhibitors of mTORC1 may be more efficacious in human breast cancer. Our findings support the hypothesis that mTORC1 is an important upregulator of telomerase in breast cancer.


Brown adipose TRX2 deficiency activates mtDNA-NLRP3 to impair thermogenesis and protect against diet-induced insulin resistance.
Brown adipose tissue (BAT), a crucial heat-generating organ, regulates whole-body energy metabolism by mediating thermogenesis. BAT inflammation is implicated in the pathogenesis of mitochondrial dysfunction and impaired thermogenesis. However, the link between BAT inflammation and systematic metabolism remains unclear. Herein, we use mice with BAT deficiency of thioredoxin-2 (TRX2), a protein that scavenges mitochondrial reactive oxygen species (ROS), to evaluate the impact of BAT inflammation on metabolism and thermogenesis and its underlying mechanism. Our results show that BAT-specific TRX2 ablation improves systematic metabolic performance via enhancing lipid uptake, which protects mice from diet-induced obesity, hypertriglyceridemia, and insulin resistance. TRX2 deficiency impairs adaptive thermogenesis by suppressing fatty acid oxidation. Mechanistically, loss of TRX2 induces excessive mitochondrial ROS, mitochondrial integrity disruption, and cytosolic release of mitochondrial DNA, which in turn activate aberrant innate immune responses in BAT, including the cGAS/STING and the NLRP3 inflammasome pathways. We identify NLRP3 as a key converging point, as its inhibition reverses both the thermogenesis defect and the metabolic benefits seen under nutrient overload in BAT-specific Trx2-deficient mice. In conclusion, we identify TRX2 as a critical hub integrating oxidative stress, inflammation, and lipid metabolism in BAT, uncovering an adaptive mechanism underlying the link between BAT inflammation and systematic metabolism.


Physical activity, sedentary behavior time and lipid levels in the Observation of Cardiovascular Risk Factors in Luxembourg study.
Recently attention has been drawn to the health impacts of time spent engaging in sedentary behaviors. While many studies have investigated general physical activity (PA) in relation to blood lipid levels, the current study aimed to examine the intensity of activity, including sedentary behavior time, and time spent engaging in moderate and intense PA, with concentrations of HDL and LDL-cholesterol, total cholesterol, and triglycerides. Participants comprised 1331 individuals, aged 18 to 70 years, from the Observation of Cardiovascular Risk Factors in Luxembourg (ORISCAV-LUX) study, who underwent objective cardiovascular health assessments and completed the International Physical Activity Questionnaire (IPAQ). Time spent engaging in sedentary behaviors (screen time on a workday and a day off, and total sitting time on a work day), and moderate and intense PA, were related to levels of HDL and LDL-cholesterol, total cholesterol, and triglycerides. Analyses were conducted in the whole sample, and then with stratification according to BMI (normal weight versus overweight/obese). Both lower screen time during days off and higher intense PA time were significantly associated with higher HDL-cholesterol after full adjustment for socio-demographic factors, dietary factors and smoking (both p < 0.05). In normal weight individuals, consistent positive relations between triglycerides, LDL, and total cholesterol with all sedentary behavior time variables were observed (all p < 0.05; adjusted for age, education, gender). There were no statistically significant associations between any intensity level of PA or sedentary behavior time variable and lipid levels in those overweight or obese. Spending less time in sedentary behaviors, and engaging in medium levels of intense physical activity may be associated with a more favorable blood lipid profile, particularly with regard to levels of HDL and triglycerides.


[Current microbiological aspects of community respiratory infection beyond COVID-19].
From a microbiological point of view, both empirical and targeted antimicrobial treatment in respiratory infection is based on the sensitivity profile of isolated microorganisms and the possible resistance mechanisms that they may present. The latter may vary in different geographic areas according to prescription profiles and vaccination programs. Beta-lactam antibiotics, fluoroquinolones, and macrolides are the most commonly used antimicrobials during the exacerbations of chronic obstructive pulmonary disease and community-acquired pneumonia. In their prescription, different aspects such as intrinsic activity, bactericidal effect or their ability to prevent the development of resistance must be taken into account. The latter is related to the PK/PD parameters, the mutant prevention concentration and the so-called selection window. More recently, the potential ecological impact has grown in importance, not only on the intestinal microbiota, but also on the respiratory one. Maintaining the state of eubiosis requires the use of antimicrobials with a low profile of action on anaerobic bacteria. With their use, the resilience of the bacterial populations belonging to the microbiota, the state of resistance of colonization and the collateral damage related to the emergence of resistance to the antimicrobials in pathogens causing the infections and in the bacterial populations integrating the microbiota.


The associations of earlier trauma exposures and history of mental disorders with PTSD after subsequent traumas.
Although earlier trauma exposure is known to predict posttraumatic stress disorder (PTSD) after subsequent traumas, it is unclear whether this association is limited to cases where the earlier trauma led to PTSD. Resolution of this uncertainty has important implications for research on pretrauma vulnerability to PTSD. We examined this issue in the World Health Organization (WHO) World Mental Health (WMH) Surveys with 34 676 respondents who reported lifetime trauma exposure. One lifetime trauma was selected randomly for each respondent. DSM-IV (Diagnostic and Statistical Manual of Mental Disorders, 4th Edition) PTSD due to that trauma was assessed. We reported in a previous paper that four earlier traumas involving interpersonal violence significantly predicted PTSD after subsequent random traumas (odds ratio (OR)=1.3-2.5). We also assessed 14 lifetime DSM-IV mood, anxiety, disruptive behavior and substance disorders before random traumas. We show in the current report that only prior anxiety disorders significantly predicted PTSD in a multivariate model (OR=1.5-4.3) and that these disorders interacted significantly with three of the earlier traumas (witnessing atrocities, physical violence victimization and rape). History of witnessing atrocities significantly predicted PTSD after subsequent random traumas only among respondents with prior PTSD (OR=5.6). Histories of physical violence victimization (OR=1.5) and rape after age 17 years (OR=17.6) significantly predicted only among respondents with no history of prior anxiety disorders. Although only preliminary due to reliance on retrospective reports, these results suggest that history of anxiety disorders and history of a limited number of earlier traumas might usefully be targeted in future prospective studies as distinct foci of research on individual differences in vulnerability to PTSD after subsequent traumas.


An update to the HIV-TRePS system: the development of new computational models that do not require a genotype to predict HIV treatment outcomes.
The optimal individualized selection of antiretroviral drugs in resource-limited settings is challenging because of the limited availability of drugs and genotyping. Here we describe the development of the latest computational models to predict the response to combination antiretroviral therapy without a genotype, for potential use in such settings. Random forest models were trained to predict the probability of a virological response to therapy (<50 copies HIV RNA/mL) following virological failure using the following data from 22,567 treatment-change episodes including 1090 from southern Africa: baseline viral load and CD4 cell count, treatment history, drugs in the new regimen, time to follow-up and follow-up viral load. The models were assessed during cross-validation and with an independent global test set of 1000 cases including 100 from southern Africa. The models' accuracy [area under the receiver-operating characteristic curve (AUC)] was evaluated and compared with genotyping using rules-based interpretation systems for those cases with genotypes available. The models achieved AUCs of 0.79-0.84 (mean 0.82) during cross-validation, 0.80 with the global test set and 0.78 with the southern African subset. The AUCs were significantly lower (0.56-0.57) for genotyping. The models predicted virological response to HIV therapy without a genotype as accurately as previous models that included a genotype. They were accurate for cases from southern Africa and significantly more accurate than genotyping. These models will be accessible via the online treatment support tool HIV-TRePS and have the potential to help optimize antiretroviral therapy in resource-limited settings where genotyping is not generally available.


Effect of Zinc on Spermatogenesis and Sperm Chromatin Condensation in Bleomycin, Etoposide, Cisplatin Treated Rats.
The incidence rate of testicular cancer among young males is high. Co-administration of bleomycin, etoposide and cisplatin (BEP) has increased survival rate of patients with testicular cancer. Although BEP is one of the most effective treatment for testicular cancer, but it severely affects the reproductive system that ultimately leads to infertility. In addition to its antioxidant activity, zinc has an important role in progression of spermiogenesis. This study aimed to evaluate the effect of zinc on sperm parameters, chromatin condensation and testicular structure after BEP treatment. In this experimental study, 40 male rats were divided into 4 groups (control, BEP, BEP+ zinc and zinc) and examined for 2 spermatogenesis periods (i.e. 18 weeks). The rats in BEP and BEP+ zinc group were treated with BEP at appropriate doses (0.75, 7.5, and 1.5 mg/kg) for three cycles of three weeks. Zinc at a dose of 10 mg/kg/day was administered to BEP+ zinc and zinc groups. After 18 weeks, we assessed sperm parameters, and excessive histone in sperm chromatin using aniline blue staining, as well as testicular structure and germ line cells using periodic acid-Schiff staining. After BEP treatment, significant decreases were observed in normal sperm morphology, motility, and concentration, as well as alterations in rat sperm chromatin condensation and testicular tissue (P<0.001). Furthermore, after zinc consumption for 9 weeks, we observed significant improvements of sperm parameters and chromatin condensation as well as a significant retrieval of spermatogonia, leydig cells and tubular architecture (P<0.05). Zinc administration after chemotherapy with BEP in testicular cancer might be potentially useful in declining the off target consequence associated with oxidative stress.


UV wavelengths experienced during development affect larval newt visual sensitivity and predation efficiency.
We experimentally investigated the influence of developmental plasticity of ultraviolet (UV) visual sensitivity on predation efficiency of the larval smooth newt, Lissotriton vulgaris. We quantified expression of SWS1 opsin gene (UV-sensitive protein of photoreceptor cells) in the retinas of individuals who had developed in the presence (UV+) or absence (UV-) of UV light (developmental treatments), and tested their predation efficiency under UV+ and UV- light (testing treatments). We found that both SWS1 opsin expression and predation efficiency were significantly reduced in the UV- developmental group. Larvae in the UV- testing environment displayed consistently lower predation efficiency regardless of their developmental treatment. These results prove for the first time, we believe, functional UV vision and developmental plasticity of UV sensitivity in an amphibian at the larval stage. They also demonstrate that UV wavelengths enhance predation efficiency and suggest that the magnitude of the behavioural response depends on retinal properties induced by the developmental lighting environment.


Characterization of the microtubule binding domain of microtubule actin crosslinking factor (MACF): identification of a novel group of microtubule associated proteins.
MACF (microtubule actin cross-linking factor) is a large, 608-kDa protein that can associate with both actin microfilaments and microtubules (MTs). Structurally, MACF can be divided into 3 domains: an N-terminal domain that contains both a calponin type actin-binding domain and a plakin domain; a rod domain that is composed of 23 dystrophin-like spectrin repeats; and a C-terminal domain that includes two EF-hand calcium-binding motifs, as well as a region that is homologous to two related proteins, GAR22 and Gas2. We have previously demonstrated that the C-terminal domain of MACF binds to MTs, although no homology was observed between this domain and other known microtubule-binding proteins. In this report, we describe the characterization of this microtubule-binding domain of MACF by transient transfection studies and in vitro binding assays. We found that the C-terminus of MACF contains at least two microtubule-binding regions, a GAR domain and a domain containing glycine-serine-arginine (GSR) repeats. In transfected cells, the GAR domain bound to and partially stabilized MTs to depolymerization by nocodazole. The GSR-containing domain caused MTs to form bundles that are still sensitive to nocodazole-induced depolymerization. When present together, these two domains acted in concert to bundle MTs and render them stable to nocodazole treatment. Recently, a study has shown that the N-terminal half of the plakin domain (called the M1 domain) of MACF also binds MTs. We therefore examined the microtubule binding ability of the M1 domain in the context of the entire plakin domain with and without the remaining N-terminal regions of two different MACF isoforms. Interestingly, in the presence of the surrounding sequences, the M1 domain did not bind MTs. In addition to MACF, cDNA sequences encoding the GAR and GSR-containing domains are also found in the partial human EST clone KIAA0728, which has high sequence homology to the 3' end of the MACF cDNA; hence, we refer to it as MACF2. The C-terminal domain of mouse MACF2 was cloned and characterized. The microtubule-binding properties of MACF2 C-terminal domain are similar to that of MACF. The GAR domain was originally found in Gas 2 protein and here we show that it can associate with MTs in transfected cells. Plectin and desmoplakin have GSR-containing domains at their C-termini and we further demonstrate that the GSR-containing domain of plectin, but not desmoplakin, can bind to MTs in vivo.


Pancreas transplantation in crossmatch-positive recipients.
Prolonged cold preservation time can unfavorably affect outcome in pancreas transplantation. To reduce this ischemic time, cadaver pancreas grafts, in selected cases, are sometimes transplanted before crossmatch results are known. We report our experience with pancreas transplants in recipients with either current or historically positive T- or B-cell crossmatches. Crossmatch-positive pancreas transplants were identified using a computerized database. T-cell crossmatches were performed using an antihuman-globulin-augmented complement-dependent cytotoxicity (CDC) test; B-cell crossmatches were performed using an extended incubation CDC test. All patients received anti-T-cell induction therapy and either cyclosporine (1987-1993) or tacrolimus-based (1994-2001) immunosuppression. More recent recipients (2000-2001) also received intravenous gamma globulin and postoperative plasmapheresis. Between October 1, 1987 and March 31, 2001, of a total of 1076 pancreas transplants performed, 59 (5.48%) were crossmatch-positive. Of these, 8 had a current T-cell-positive crossmatch and 15 had a current B-cell-positive crossmatch. One recipient was both current B- and T-positive, and the rest were past B- and/or T-cell positive. One-year pancreas graft survival for current T- and B-cell crossmatch-positive transplants was 63% and 67%, respectively. T- or B-cell crossmatch-negative transplants had a 1-yr survival of 70%. In the T-cell crossmatch-positive group, four grafts are still functioning (follow-up range, 2-12 yr), one patient died with a functioning graft at 4 months, and four grafts failed (one each from pancreatitis, infection, primary nonfunction, and vascular thrombosis). No grafts were lost to rejection. In the B-cell crossmatch-positive group, six grafts are still functioning (follow-up range, 2-11 yr) and nine have failed (four from chronic rejection, three from vascular thromboses, and two from pancreatitis). Crossmatch-positive cases were significantly more likely to be retransplants (70.8%) than crossmatch-negative cases (14.8%, p < 0.0001). In a multivariate analysis, crossmatch positivity did not affect pancreas graft outcome, whereas retransplants had a significant impact on outcome (relative risk 1.84, p < 0.0001). (: i) Pancreas transplants performed in the setting of a positive current crossmatch may have long-term function. (ii) With current immunosuppressive protocols, graft loss from hyperacute and acute rejection may be prevented in current crossmatch-positive pancreas transplants. Chronic rejection was only seen in B-cell crossmatch-positive cases. (iii) High rates of technical graft loss in crossmatch-positive cases may reflect a high frequency of retransplants in this group.


Third-order dispersion in femtosecond fiber lasers.
We experimentally investigate the effects of third-order dispersion (TOD) on ultrafast sources, using dispersion-compensated femtosecond fiber lasers. By in situ measurement of the intracavity dispersion, we identify spectral resonances that are due to coupling of the cavity pulse energy to dispersive wave components at the phase velocity and group-velocity-matched wavelengths. We obtain pulse widths as short as 91 fs by partially compensating the TOD. We relate the observed effects to consequences for soliton transmission and operation of other solitary-pulse lasers.


Quantitative assessment of dictionary-based protein named entity tagging.
Natural language processing (NLP) approaches have been explored to manage and mine information recorded in biological literature. A critical step for biological literature mining is biological named entity tagging (BNET) that identifies names mentioned in text and normalizes them with entries in biological databases. The aim of this study was to provide quantitative assessment of the complexity of BNET on protein entities through BioThesaurus, a thesaurus of gene/protein names for UniProt knowledgebase (UniProtKB) entries that was acquired using online resources. We evaluated the complexity through several perspectives: ambiguity (i.e., the number of genes/proteins represented by one name), synonymy (i.e., the number of names associated with the same gene/protein), and coverage (i.e., the percentage of gene/protein names in text included in the thesaurus). We also normalized names in BioThesaurus and measures were obtained twice, once before normalization and once after. The current version of BioThesaurus has over 2.6 million names or 2.1 million normalized names covering more than 1.8 million UniProtKB entries. The average synonymy is 3.53 (2.86 after normalization), ambiguity is 2.31 before normalization and 2.32 after, while the coverage is 94.0% based on the BioCreAtive data set comprising MEDLINE abstracts containing genes/proteins. The study indicated that names for genes/proteins are highly ambiguous and there are usually multiple names for the same gene or protein. It also demonstrated that most gene/protein names appearing in text can be found in BioThesaurus.


Metformin as an anti-cancer agent: actions and mechanisms targeting cancer stem cells.
Metformin, a first line medication for type II diabetes, initially entered the spotlight as a promising anti-cancer agent due to epidemiologic reports that found reduced cancer risk and improved clinical outcomes in diabetic patients taking metformin. To uncover the anti-cancer mechanisms of metformin, preclinical studies determined that metformin impairs cellular metabolism and suppresses oncogenic signaling pathways, including receptor tyrosine kinase, PI3K/Akt, and mTOR pathways. Recently, the anti-cancer potential of metformin has gained increasing interest due to its inhibitory effects on cancer stem cells (CSCs), which are associated with tumor metastasis, drug resistance, and relapse. Studies using various cancer models, including breast, pancreatic, prostate, and colon, have demonstrated the potency of metformin in attenuating CSCs through the targeting of specific pathways involved in cell differentiation, renewal, metastasis, and metabolism. In this review, we provide a comprehensive overview of the anti-cancer actions and mechanisms of metformin, including the regulation of CSCs and related pathways. We also discuss the potential anti-cancer applications of metformin as mono- or combination therapies.