Criterion Validity of Catapult ClearSky T6 Local Positioning System for Measuring Inter-Unit Distance.
The validity of a local positioning system (LPS) to measure inter-unit distance was investigated during a team sport movement circuit. Eight recreationally active, female indoor team-sport players completed a circuit, comprising seven types of movements (walk, jog, jump, sprint, 45° change of direction and shuffle), on an indoor court. Participants wore a receiver tag (ClearSky T6, Catapult Sports) and seven reflective markers, to allow for a comparison with the reference system (©Vicon Motion Systems, Oxford Metrics, UK). Inter-unit distance was collected for each combination of participants. Validity was assessed via root mean square error, mean bias and percentage of variance accounted for, both as an overall dataset and split into distance bands. The results presented a mean root mean square error of 0.20 ± 0.05 m, and mean bias detected an overestimation for all distance bands. The LPS shows acceptable accuracy for measuring inter-unit distance, opening up opportunities to utilise player tracking for tactical variables indoors.


Specific receptors for phorbol diesters on freshly isolated human myeloid and lymphoid leukemia cells: comparable binding characteristics despite different cellular responses.
Freshly isolated human leukemia cells have been shown in the past to display varying in vitro responses to phorbol diesters, depending on their cell type. Specific receptors for the phorbol diesters have been demonstrated on numerous different cells. This study was designed to characterize the receptors for phorbol diesters on leukemia cells freshly isolated from patients with different kinds of leukemia and to determine if differences in binding characteristics for tritium-labeled phorbol 12,13-dibutyrate (3H-PDBu) accounted for the different cellular responses elicited in vitro by phorbol diesters. Cells from 26 patients with different kinds of leukemia were studied. PDBu or phorbol 12-myristate 13-acetate (PMA) caused cells from patients with acute myeloblastic leukemia (AML), acute promyelocytic (APML), acute myelomonocytic (AMML), acute monocytic (AMoL), acute erythroleukemia (AEL), chronic myelocytic leukemia (CML) in blast crisis (myeloid), acute undifferentiated leukemia (AUL), and hairy cell leukemia (HCL) (n = 15) to adhere to plastic and spread. However, they caused no adherence or spreading and only slight aggregation of cells from patients with acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), or CML-blast crisis (lymphoid) (n = 11). All leukemia cells studied, irrespective of cellular type, displayed specific receptors for 3H-PDBu. The time courses for binding by all leukemia types were similar, with peak binding at 5-10 min at 37 degrees C and 120 min at 4 degrees C. The binding affinities were similar for patients with ALL (96 +/- 32 nM, n = 4), CLL (126 +/- 32 nM, n = 6), and acute nonlymphoid leukemia (73 +/- 14 nM, n = 11). Likewise, the numbers of specific binding sites/cell were comparable for the patients with ALL (6.2 +/- 1.3 X 10(5) sites/cell, n = 4), CLL (5.0 +/- 2.0 X 10(5) sites/cell, n = 6), and acute nonlymphoid leukemia (4.4 +/- 1.9 X 10(5) sites/cell, n = 11). Thus, the differing responses to phorbol diesters of various types of freshly isolated leukemia cells appear to be due to differences other than initial ligand-receptor binding.


Development and Implementation: B'More Healthy Communities for Kid's Store and Wholesaler Intervention.
Higher rates of obesity and obesity-related chronic disease are prevalent in communities where there is limited access to affordable, healthy food. The B'More Healthy Communities for Kids (BHCK) trial worked at multiple levels of the food environment including food wholesalers and corner stores to improve the surrounding community's access to healthy food. The objective of this article is to describe the development and implementation of BHCK's corner store and wholesaler interventions through formal process evaluation. Researchers evaluated each level of the intervention to assess reach, dose delivered, and fidelity. Corner store and wholesaler reach, dose delivered, and fidelity were measured by number of interactions, promotional materials distributed, and maintenance of study materials, respectively. Overall, the corner store implementation showed moderate reach, dose delivered, and high fidelity. The wholesaler intervention was implemented with high reach, dose, and fidelity. The program held 355 corner store interactive sessions and had 9,347 community member interactions, 21% of which were with children between the ages of 10 and 14 years. There was a 15% increase in corner store promoted food stocking during Wave 1 and a 17% increase during Wave 2. These findings demonstrate a successfully implemented food retailer intervention in a low-income urban setting.


Design and application of bispecific splice-switching oligonucleotides.
Targeting of pre-mRNA by short splice-switching oligonucleotides (SSOs) is increasingly being used as a therapeutic modality, one rationale being to disrupt splicing so as to remove exons containing premature termination codons, or to restore the translation reading frame around out-of-frame deletion mutations. The aim of this study was to investigate the effect of chemically linking individual SSOs so as to ascertain equimolar cellular uptake that would provide for more defined drug formulations. In contrast to conventional bispecific SSOs generated by conjugation in solution, here we describe a protocol for synthesis of bispecific SSOs on solid phase. These SSOs comprised of either a non-cleavable hydrocarbon linker or disulfide-based cleavable linkers. To assess the efficacy of these SSOs we have utilized splice switching to bypass a disease-causing mutation in the DMD gene concurrent with disruption of the reading frame of the myostatin gene (Mstn). The premise of this approach is that disruption of myostatin expression is known to induce muscle hypertrophy and so for Duchenne muscular dystrophy (DMD) could be expected to have a better outcome than dystrophin restoration alone. All tested SSOs mediated simultaneous robust exon removal from mature Dmd and Mstn transcripts in myotubes. Our results also demonstrate that using cleavable SSOs is preferred over the non-cleavable counterparts and that these are equally efficient at inducing exon skipping as cocktails of monospecific versions. In conclusion, we have developed a protocol for solid-phase synthesis of single molecule cleavable bispecific SSOs that can be efficiently exploited for targeting of multiple RNA transcripts.


Misexpression of the opaque-phase-specific gene PEP1 (SAP1) in the white phase of Candida albicans confers increased virulence in a mouse model of cutaneous infection.
Candida albicans WO-1 switches reversibly and at high frequency between a white and an opaque colony-forming phenotype that includes dramatic changes in cell morphology and physiology. A misexpression strategy has been used to investigate the role of the opaque-phase-specific gene PEP1 (SAP1), which encodes a secreted aspartyl proteinase, in the expression of the unique opaque-phase phenotype and phase-specific virulence in two animal models. The PEP1 (SAP1) open reading frame was inserted downstream of the promoter of the white-phase-specific gene WH11 in the transforming vector pCPW7, and the resulting transformants were demonstrated to misexpress PEP1 (SAP1) in the white phase. Misexpression did not confer any of the unique morphological characteristics of the opaque phase to cells in the white phase and had no effect on the switching process. However, misexpression conferred upon white-phase cells the increased capacity of opaque-phase cells to grow in medium in which protein was the sole nitrogen source. Misexpression of PEP1 (SAP1) had no effect on the virulence of white-phase cells in a systemic mouse model, in which white-phase cells were already more virulent than opaque-phase cells. Misexpression did, however, confer upon white-phase cells the dramatic increase in colonization of skin in a cutaneous mouse model that was exhibited by opaque-phase cells. Misexpression of PEP1 (SAP1) conferred upon white-phase cells two dissociable opaque-phase characteristics: increased adhesion and the capacity to cavitate skin. The addition of pepstatin A to the cutaneous model inhibited the latter, but not the former, suggesting that the latter is effected by released enzyme, while the former is effected by cell-associated enzyme.


Applying the model of diffusion of innovations to understand facilitators for the implementation of maternal and neonatal health programmes in rural Uganda.
In Uganda, more than 336 out of every 100,000 women die annually during childbirth. Pregnant women, particularly in rural areas, often lack the financial resources and means to access health facilities in a timely manner for quality antenatal, delivery, and post-natal services. For nearly the past decade, the Makerere University School of Public Health researchers, through various projects, have been spearheading innovative interventions, embedded in implementation research, to reduce barriers to access to care. In this paper, we describe two of projects that were initially conceived to tackle the financial barriers to access to care - through a voucher program in the community - on the demand side - and a series of health systems strengthening activities at the district and facility level - on the supply side. Over time, the projects diverged in the content of the intervention and the modality in which they were implemented, providing an opportunity for reflection on innovation and scaling up. In this short report, we used an adaptation of Greenhalgh's Model of Diffusion to reflect on these projects' approaches to implementing innovative interventions, with the ultimate goal of reducing maternal and neonatal mortality in rural Uganda. We found that the adapted model of diffusion of innovations facilitated the emergence of insights on barriers and facilitators to the implementation of health systems interventions. Health systems research projects would benefit from analyses beyond the implementation period, in order to better understand how adoption and diffusion happen, or not, over time, after the external catalyst departs.


SU-E-T-483: Delay Treatment or Switch to Different Machines? Dosimetric Effect and Tumor Control Probability.
If the Linac is unavailable during the IMRT treatment schedule, the patient can be switched to a different Linac or prostpond treatment until the origonal Linac is available. The resulting dosimetric difference is estimated and the compromise in the TCP is estimated for both scenarios. This work investigates the feasibility and rationale of switching patients between different accelerators for IMRT in contrast to prostponing the treatment. We performed Monte Carlo simulations of photon beams from different Linac models and vendors. Prostate and head and neck treatment plans for Siemens Primus, Primart, Artiste and Varian-21Ex/IX accelerators are studied in this work. Dose distributions for given plans are recalculated using different beam data with the same nominal energy from different Linacs. We have compared DVHs, the maximum, the minimum and the mean dose to the target and critical structures due to switching accelerators. In the process of switching a treatment plan to a different accelerator, there are issues, such as optimum penumbra compensation, dose distribution at the boundary of target and critical structures and multileaf collimator (MLC) leaf width effects, needed to be considered and verified with measurements. In making the final decision whether to switch machines, the TCP based on a linear-quadratic model with time factor is considered. Two DVHs of two plans from Varian and Siemens models are delivered on different machines. Slight dose coverage differences have been observed. TCP estimation with both delayed and without delayed treatments is calculated. Undesired drop of TCP is observed with treatment gap. Based on the analyses done in this work, it is therapeutically more beneficial to switch a patient to a different machine than to postpone a treatment until the original machine is available, especially for fast growing tumors such as head and neck cancers.


A single-institution evaluation of factors important in fallopian tube carcinoma recurrence and survival.
The aim of this study was to identify prognostic factors and markers that influence clinical outcomes in patients with primary fallopian tube carcinoma at a single tertiary health care center. These prognostic factors may be of clinical importance and can subsequently be included in future clinical trials. A retrospective review of our Tumor Registry and Gynecologic Oncology database was conducted to include any patients with a diagnosis of fallopian tube carcinoma between the years 1994 and 2005. We identified clinicopathological data to evaluate factors important in recurrence, disease-specific and overall survival. Kaplan-Meier curves were generated, and log-rank tests were used to evaluate survival differences. Thirty-six patients had a diagnosis with primary fallopian tube carcinoma at a median age of 69 years. Patients most frequently presented with abdominal pain (19%) and a palpable mass (14%). The most common histological subtype was papillary serous adenocarcinoma in 56% of cases. Stage III disease (39%) and poorly differentiated tumors (81%) were most common. The median follow-up was 39.6 months. The 5-year cancer-specific survival was 42%, and the overall survival rate was 34%. Factors important in disease-free survival were International Federation of Gynecology and Obstetrics stage, tumor laterality, and serum CA-125, whereas International Federation of Gynecology and Obstetrics stage, serum CA-125, and residual disease were prognostic factors for overall survival. The most common locations of recurrence were pelvis and abdomen (63%) as opposed to distant sites. Factors associated with recurrence were stage, tumor laterality, and serum CA-125. Fallopian tube malignancies are rare. We have identified factors associated with recurrence, disease specific survival, and overall survival that could be further examined and included in larger clinical trials involving this uncommon malignancy.


Sustained Secretion of the Antimicrobial Peptide S100A7 Is Dependent on the Downregulation of Caspase-8.
Antimicrobial peptides (AMPs) are the body's natural innate immune defense against a spectrum of pathogens and can also modulate cell proliferation, chemotaxis, angiogenesis, wound healing, and immune cell activity. Harnessing these diverse functions for prophylactic use is contingent upon understanding the regulatory mechanisms governing their unconventional secretion from cells. Analysis of the secretion of S100A7 (Psoriasin), an abundant AMP stored in differentiated keratinocytes of the skin, has revealed an unexpected biphasic secretory response to bacterial exposure. The core components regulating S100A7 secretion are NFκB/p38MAPK, caspase-1, and interleukin (IL)-1α. The initial activation of this core machinery is mediated by Toll-like receptor signaling, whereas the chronic response is mediated by Caspase-8 downregulation. Interestingly, there is a concomitant downregulation of Caspase-8 in inflammatory skin diseases wherein S100A7 is constitutively released. These results highlight the potential of targeting these components to control the release of AMPs from the skin in both homeostatic and disease conditions.


A Herpesviral induction of RAE-1 NKG2D ligand expression occurs through release of HDAC mediated repression.
Natural Killer (NK) cells are essential for control of viral infection and cancer. NK cells express NKG2D, an activating receptor that directly recognizes NKG2D ligands. These are expressed at low level on healthy cells, but are induced by stresses like infection and transformation. The physiological events that drive NKG2D ligand expression during infection are still poorly understood. We observed that the mouse cytomegalovirus encoded protein m18 is necessary and sufficient to drive expression of the RAE-1 family of NKG2D ligands. We demonstrate that RAE-1 is transcriptionally repressed by histone deacetylase inhibitor 3 (HDAC3) in healthy cells, and m18 relieves this repression by directly interacting with Casein Kinase II and preventing it from activating HDAC3. Accordingly, we found that HDAC inhibiting proteins from human herpesviruses induce human NKG2D ligand ULBP-1. Thus our findings indicate that virally mediated HDAC inhibition can act as a signal for the host to activate NK-cell recognition.


Laser monitoring of chest wall displacement.
We developed a prototype laser monitor, consisting of a single laser sensor, to observe chest wall displacement during respiration. With this monitor, respiratory waveforms are expressed as an anterioposterior motion of the chest wall. The purpose of this study was to examine the characteristics and performance of this prototype. Performance was assessed: 1) under static conditions; 2) using a lung model ventilated in both conventional and high frequency oscillation (HFOV) modes; and 3) during spontaneous breathing in normal adults. In vitro, the monitor performed well both under static conditions and during mechanical ventilation. Reliable "respiratory" wave forms, with no frequency-dependent change in the relationship between displacement and volume, were produced during both conventional ventilation and HFOV at 15 Hz. In vivo, abdominal displacement, measured in the midline, was linearly correlated with the tidal volume signal integrated from flow. The waveforms produced by the monitor were adequate for monitoring respiration and for calculating respiratory timing variables. While a single laser sensor is unlikely to be sufficient for monitoring respiration in spontaneously breathing subjects, the performance of the prototype monitor was sufficiently impressive to encourage further development and further study of this type of truly noninvasive respiratory monitor.


Deinococcus radiodurans: what belongs to the survival kit?
Deinococcus radiodurans, one of the most radioresistant organisms known to date, is able to repair efficiently hundreds of DNA double- and single-strand breaks as well as other types of DNA damages promoted by ionizing or ultraviolet radiation. We review recent discoveries concerning several aspects of radioresistance and survival under high genotoxic stress. We discuss different hypotheses and possibilities that have been suggested to contribute to radioresistance and propose that D. radiodurans combines a variety of physiological tools that are tightly coordinated. A complex network of regulatory proteins may be discovered in the near future that might allow further understanding of radioresistance.


Communicating Climate Change: Lessons Learned from a Researcher-Museum Collaboration.
The need for science education and outreach is great. However, despite the ever-growing body of available scientific information, facts are often misrepresented to or misunderstood by the general public. This can result in uninformed decisions that negatively impact society at both individual and community levels. One solution to this problem is to make scientific information more available to the public through outreach programs. Most outreach programs, however, focus on health initiatives, STEM programs, or young audiences exclusively. This article describes a collaboration between the Research and Learning Center at the Fort Worth Museum of Science and History and an interdisciplinary team of researchers from the Dallas-Fort Worth (DFW) metroplex area. The collaboration was a pilot effort of a science communication fellowship and was designed to train researchers to effectively convey current science information to the public with a focus on lifelong learning. We focus on the broader idea of a university-museum collaboration that bridges the science communication gap as we outline the process of forming this collaboration, lessons we learned from the process, and directions that can support future collaborations.


Sign language: its history and contribution to the understanding of the biological nature of language.
The development of conceptualization of a biological basis of language during the 20th century has come about, in part, through the appreciation of the central nervous system's ability to utilize varied sensory inputs, and particularly vision, to develop language. Sign language has been a part of the linguistic experience from prehistory to the present day. Data suggest that human language may have originated as a visual language and became primarily auditory with the later development of our voice/speech tract. Sign language may be categorized into two types. The first is used by individuals who have auditory/oral language and the signs are used for special situations, such as communication in a monastery in which there is a vow of silence. The second is used by those who do not have access to auditory/oral language, namely the deaf. The history of the two forms of sign language and the development of the concept of the biological basis of language are reviewed from the fourth century BC to the present day. Sign languages of the deaf have been recognized since at least the fourth century BC. The codification of a monastic sign language occurred in the seventh to eighth centuries AD. Probable synergy between the two forms of sign language occurred in the 16th century. Among other developments, the Abbey de L'Epée introduced, in the 18th century, an oral syntax, French, into a sign language based upon indigenous signs of the deaf and newly created signs. During the 19th century, the concept of a "critical" period for the acquisition of language developed; this was an important stimulus for the exploration of the biological basis of language. The introduction of techniques, e.g. evoked potentials and functional MRI, during the 20th century allowed study of the brain functions associated with language.


Semiempirical self-consistent polarization description of bulk water, the liquid-vapor interface, and cubic ice.
We have applied an efficient electronic structure approach, the semiempirical self-consistent polarization neglect of diatomic differential overlap (SCP-NDDO) method, previously parametrized to reproduce properties of water clusters by Chang, Schenter, and Garrett [ J. Chem. Phys. 2008 , 128 , 164111 ] and now implemented in the CP2K package, to model ambient liquid water at 300 K (both the bulk and the liquid-vapor interface) and cubic ice at 15 and 250 K. The SCP-NDDO potential retains its transferability and good performance across the full range of conditions encountered in the clusters and the bulk phases of water. In particular, we obtain good results for the density, radial distribution functions, enthalpy of vaporization, self-diffusion coefficient, molecular dipole moment distribution, and hydrogen bond populations, in comparison to experimental measurements.


Evaluation of an Artificial Neural Network Retention Index Model for Chemical Structure Identification in Nontargeted Metabolomics.
Liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) is a major analytical technique used for nontargeted identification of metabolites in biological fluids. Typically, in LC-ESI-MS/MS based database assisted structure elucidation pipelines, the exact mass of an unknown compound is used to mine a chemical structure database to acquire an initial set of possible candidates. Subsequent matching of the collision induced dissociation (CID) spectrum of the unknown to the CID spectra of candidate structures facilitates identification. However, this approach often fails because of the large numbers of potential candidates (i.e., false positives) for which CID spectra are not available. To overcome this problem, CID fragmentation predication programs have been developed, but these also have limited success if large numbers of isomers with similar CID spectra are present in the candidate set. In this study, we investigated the use of a retention index (RI) predictive model as an orthogonal method to help improve identification rates. The model was used to eliminate candidate structures whose predicted RI values differed significantly from the experimentally determined RI value of the unknown compound. We tested this approach using a set of ninety-one endogenous metabolites and four in silico CID fragmentation algorithms: CFM-ID, CSI:FingerID, Mass Frontier, and MetFrag. Candidate sets obtained from PubChem and the Human Metabolite Database (HMDB) were ranked with and without RI filtering followed by in silico spectral matching. Upon RI filtering, 12 of the ninety-one metabolites were eliminated from their respective candidate sets, i.e., were scored incorrectly as negatives. For the remaining seventy-nine compounds, we show that RI filtering eliminated an average of 58% from PubChem candidate sets. This resulted in an approximately 2-fold improvement in average rankings when using CFM-ID, Mass Frontier, and MetFrag. In addition, RI filtering slightly increased the occurrence of number one rankings for all 4 fragmentation algorithms. However, RI filtering did not significantly improve average rankings when HMDB was used as the candidate database, nor did it significantly improve average rankings when using CSI:FingerID. Overall, we show that the current RI model incorrectly eliminated more true positives (12) than were expected (4-5) on the basis of the filtering method. However, it slightly improved the number of correct first place rankings and improved overall average rankings when using CFM-ID, Mass Frontier, and MetFrag.


PTI-125 Reduces Biomarkers of Alzheimer's Disease in Patients.
The most common dementia worldwide, Alzheimer's disease is often diagnosed via biomarkers in cerebrospinal fluid, including reduced levels of Aβ1-42, and increases in total tau and phosphorylated tau-181. Here we describe results of a Phase 2a study of a promising new drug candidate that significantly reversed all measured biomarkers of Alzheimer's disease, neurodegeneration and neuroinflammation. PTI-125 is an oral small molecule drug candidate that binds and reverses an altered conformation of the scaffolding protein filamin A found in Alzheimer's disease brain. Altered filamin A links to the α7-nicotinic acetylcholine receptor to allow Aβ42's toxic signaling through this receptor to hyperphosphorylate tau. Altered filamin A also links to toll-like receptor 4 to enable Aβ-induced persistent activation of this receptor and inflammatory cytokine release. Restoring the native shape of filamin A prevents or reverses filamin A's linkages to the α7-nicotinic acetylcholine receptor and toll-like receptor 4, thereby blocking Aβ42's activation of these receptors. The result is reduced tau hyperphosphorylation and neuroinflammation, with multiple functional improvements demonstrated in transgenic mice and postmortem Alzheimer's disease brain. Safety, pharmacokinetics, and cerebrospinal fluid and plasma biomarkers were assessed following treatment with PTI-125 for 28 days. Target engagement and mechanism of action were assessed in patient lymphocytes by measuring 1) the reversal of filamin A's altered conformation, 2) linkages of filamin A with α7-nicotinic acetylcholine receptor or toll-like receptor 4, and 3) levels of Aβ42 bound to α7-nicotinic acetylcholine receptor or CD14, the co-receptor for toll-like receptor 4. This was a first-in-patient, open-label Phase 2a safety, pharmacokinetics and biomarker study. Five clinical trial sites in the U.S. under an Investigational New Drug application. This study included 13 mild-to-moderate Alzheimer's disease patients, age 50-85, Mini Mental State Exam ≥16 and ≤24 with a cerebrospinal fluid total tau/Aβ42 ratio ≥0.30. PTI-125 oral tablets (100 mg) were administered twice daily for 28 consecutive days. Safety was assessed by electrocardiograms, clinical laboratory analyses and adverse event monitoring. Plasma levels of PTI-125 were measured in blood samples taken over 12 h after the first and last doses; cerebrospinal fluid levels were measured after the last dose. Commercial enzyme linked immunosorbent assays assessed levels of biomarkers of Alzheimer's disease in cerebrospinal fluid and plasma before and after treatment with PTI-125. The study measured biomarkers of pathology (pT181 tau, total tau and Aβ42), neurodegeneration (neurofilament light chain and neurogranin) and neuroinflammation (YKL-40, interleukin-6, interleukin-1β and tumor necrosis factor α). Plasma levels of phosphorylated and nitrated tau were assessed by immunoprecipitation of tau followed by immunoblotting of three different phospho-epitopes elevated in AD (pT181-tau, pS202-tau and pT231-tau) and nY29-tau. Changes in conformation of filamin A in lymphocytes were measured by isoelectric focusing point. Filamin A linkages to α7-nicotinic acetylcholine receptor and toll-like receptor 4 were assessed by immunoblot detection of α7-nicotinic acetylcholine receptor and toll-like receptor 4 in anti-filamin A immunoprecipitates from lymphocytes. Aβ42 complexed with α7-nicotinic acetylcholine receptor or CD14 in lymphocytes was also measured by co-immunoprecipitation. The trial did not measure cognition. Consistent with the drug's mechanism of action and preclinical data, PTI-125 reduced cerebrospinal fluid biomarkers of Alzheimer's disease pathology, neurodegeneration and neuroinflammation from baseline to Day 28. All patients showed a biomarker response to PTI-125. Total tau, neurogranin, and neurofilament light chain decreased by 20%, 32% and 22%, respectively. Phospho-tau (pT181) decreased 34%, evidence that PTI-125 suppresses tau hyperphosphorylation induced by Aβ42's signaling through α7-nicotinic acetylcholine receptor. Cerebrospinal fluid biomarkers of neuroinflammation (YKL-40 and inflammatory cytokines) decreased by 5-14%. Biomarker effects were similar in plasma. Aβ42 increased slightly - a desirable result because low Aβ42 indicates Alzheimer's disease. This increase is consistent with PTI-125's 1,000-fold reduction of Aβ42's femtomolar binding affinity to α7-nicotinic acetylcholine receptor. Biomarker reductions were at least p ≤ 0.001 by paired t test. Target engagement was shown in lymphocytes by a shift in filamin A's conformation from aberrant to native: 93% was aberrant on Day 1 vs. 40% on Day 28. As a result, filamin A linkages with α7-nicotinic acetylcholine receptor and toll-like receptor 4, and Aβ42 complexes with α7-nicotinic acetylcholine receptor and CD14, were all significantly reduced by PTI-125. PTI-125 was safe and well-tolerated in all patients. Plasma half-life was 4.5 h and approximately 30% drug accumulation was observed on Day 28 vs. Day 1. PTI-125 significantly reduced biomarkers of Alzheimer's disease pathology, neurodegeneration, and neuroinflammation in both cerebrospinal fluid and plasma. All patients responded to treatment. The magnitude and consistency of reductions in established, objective biomarkers imply that PTI-125 treatment counteracted disease processes and reduced the rate of neurodegeneration. Based on encouraging biomarker data and safety profile, approximately 60 patients with mild-to-moderate AD are currently being enrolled in a Phase 2b randomized, placebo-controlled confirmatory study to assess the safety, tolerability and efficacy of PTI-125.


Quality of care in a chain of walk-in centers.
Several aspects of quality of care at one chain of freestanding ambulatory health care walk-in centers were evaluated. Those areas for which data were available--medical record review, physician credentials, and patient satisfaction--suggest that for primary acute episodic care, HSMMI walk-in centers provided care comparable to that which would have been received in traditional health care settings. In fact, HSMMI offices were found to be similar in organizational structure and appeared very much like private physicians' offices in management, staffing, patient flow, and physician performance. The corporation's QA program evolved from a series of informal managers' meetings (in 1986), to a detailed and structured program involving a CEO and regional medical directors by the end of 1990. The exact way in which HSMMI's new emphasis on formal quality-of-care assessment and assurance will affect patient care will be seen as the program fully matures. The issue of the proper relationship between money and medicine remains a problem. As health services organizations are increasingly influenced by market forces and consumers have ever-higher expectations, patients and physicians alike want to be sure that high-quality health care remains the first priority. Although this is an issue for all providers, it is much more visible in proprietary offices, making physicians as well as the public uneasy. For this reason, HSMMI's burden of proof regarding quality may be higher--particularly within the medical community--than the burden on traditional private practice physicians.(ABSTRACT TRUNCATED AT 250 WORDS)


Implementing a third-year emergency medicine medical student curriculum.
Emergency medicine (EM) is commonly introduced in the fourth year of medical school because of a perceived need to have more experienced students in the complex and dynamic environment of the emergency department. However, there is no evidence supporting the optimal time or duration for an EM rotation, and a number of institutions offer third-year rotations. A recently published syllabus provides areas of knowledge, skills, and attitudes that third-year EM rotation directors can use to develop curricula. This article expands on that syllabus by providing a comprehensive curricular guide for the third-year medical student rotation with a focus on implementation. Included are consensus-derived learning objectives, discussion of educational methods, considerations for implementation, and information on feedback and evaluation as proposed by the Clerkship Directors in Emergency Medicine Third-Year Curriculum Work Group. External validation results, derived from a survey of third-year rotation directors, are provided in the form of a content validity index for each content area. This consensus-derived curricular guide can be used by faculty who are developing or revising a third-year EM medical student rotation and provide guidance for implementing this curriculum at their institution.


Predictors of Outcomes of COVID-19 in Patients With Chronic Liver Disease: US Multi-center Study.
Chronic liver disease (CLD) represents a major global health burden. We undertook this study to identify the factors associated with adverse outcomes in patients with CLD who acquire the novel coronavirus-2019 (COVID-19). We conducted a multi-center, observational cohort study across 21 institutions in the United States (US) of adult patients with CLD and laboratory-confirmed diagnosis of COVID-19 between March 1, 2020 and May 30, 2020. We performed survival analysis to identify independent predictors of all-cause mortality and COVID-19 related mortality, and multivariate logistic regression to determine the risk of severe COVID-19 in patients with CLD. Of the 978 patients in our cohort, 867 patients (mean age 56.9 ± 14.5 years, 55% male) met inclusion criteria. The overall all-cause mortality was 14.0% (n = 121), and 61.7% (n = 535) had severe COVID-19. Patients presenting with diarrhea or nausea/vomiting were more likely to have severe COVID-19. The liver-specific factors associated with independent risk of higher overall mortality were alcohol-related liver disease (ALD) (hazard ratio [HR] 2.42, 95% confidence interval [CI] 1.29-4.55), decompensated cirrhosis (HR 2.91 [1.70-5.00]) and hepatocellular carcinoma (HCC) (HR 3.31 [1.53-7.16]). Other factors were increasing age, diabetes, hypertension, chronic obstructive pulmonary disease and current smoker. Hispanic ethnicity (odds ratio [OR] 2.33 [1.47-3.70]) and decompensated cirrhosis (OR 2.50 [1.20-5.21]) were independently associated with risk for severe COVID-19. The risk factors which predict higher overall mortality among patients with CLD and COVID-19 are ALD, decompensated cirrhosis and HCC. Hispanic ethnicity and decompensated cirrhosis are associated with severe COVID-19. Our results will enable risk stratification and personalization of the management of patients with CLD and COVID-19. Clinicaltrials.gov number NCT04439084.


Baseline arterial pressure affects sympathoexcitatory responses to ventricular premature beats.
The seconds to minutes before sudden cardiac death are characterized by fluctuations of arterial pressure, cardiac rhythm, and probably sympathetic nerve activity. We explored the interrelations among these factors in seven patients undergoing clinical electrophysiological testing. We measured muscle sympathetic nerve activity (SNA) and arterial pressure responses to ventricular premature beats induced throughout the cardiac cycle under three conditions: 1) lowered arterial pressure and elevated SNA produced by intravenous nitroprusside, 2) baseline arterial pressure and SNA during saline infusion, and 3) elevated arterial pressure and decreased SNA activity produced by intravenous phenylephrine. Sympathetic responses to premature beats were inversely related to diastolic pressure. The magnitude of the sympathetic response was directly related to the prevailing arterial pressure and inversely related to baseline SNA. These data demonstrate that sympathoexcitation evoked by ventricular dysrhythmias is determined importantly by the prevailing arterial pressure and possibly by the background R-R interval and level of sympathetic activity. This effect may influence hemodynamic and electrophysiological stability during dysrhythmias.


Egg consumption and risk of incident type 2 diabetes: a dose-response meta-analysis of prospective cohort studies.
Experimental data suggest that egg intake could have a beneficial impact on several risk factors for type 2 diabetes. In contrast, some recent epidemiological studies have concluded that egg consumption may increase diabetes risk. We performed a dose-response meta-analysis of prospective cohorts on the relation of egg consumption with incident type 2 diabetes. We searched for cohort studies that assessed egg consumption and diabetes risk up to June 2015. We identified 416 articles and extracted data independently and in duplicate from ten eligible studies. We used random-effects generalised least squares models for pooled dose-response estimation based on thirteen estimates. Our study included 251 213 individuals and 12 156 incident type 2 diabetes cases. Egg intake was associated with incident type 2 diabetes (risk ratio (RR)/egg per d 1·13; 95 % CI 1·04, 1·22). We identified study location as a major source of heterogeneity. For studies conducted in the USA, we observed a stronger association (RR 1·47; 95 % CI 1·32, 1·64), whereas results were null for studies conducted elsewhere. Studies considered to be of high quality yielded null findings (RR 0·94; 95 % CI 0·74, 1·19). The association of egg intake with increased risk of incident type 2 diabetes may be restricted to US cohort studies. There are limited data to support a biological mechanism that could underlie this association; thus, the possibility that these results may be due to residual confounding by dietary behaviours restricted to certain populations cannot be excluded.


Multiple regions of G alpha 16 contribute to the specificity of activation by the C5a receptor.
The C5a chemoattractant factor receptor, when expressed in COS-7 cells, can stimulate phosphoinositide-specific phospholipase C activity through the activation of the G16 isoform of the heterotrimeric G protein, but not through the G11 isoform. To identify the regions of the G alpha 16 subunit protein that are responsible for its activation by the C5a receptor, a series of chimeras between G alpha 16 and G alpha 11 were constructed and tested for their ability to be activated by the C5a receptor. Co-transfection experiments with chimeras in which the carboxyl-terminal regions of G alpha 11 were replaced with the corresponding regions of G alpha 16 demonstrated that changes in the carboxyl terminus, e.g., replacement of 134 amino acids, were not sufficient to confer receptor specificity. An additional segment encompassing residues 220-240 of G alpha 16 was required to confer C5a-induced activation. Testing of a reciprocal series of chimeras composed of G alpha 16 sequences at the amino terminus and G alpha 11 sequences at the carboxyl terminus revealed that certain sequences extending from the amino terminus to amino acid 209 of G alpha 16 were sufficient to endow the chimera with much of the specificity for C5a-induced activation. These results suggest that receptor specificity may involve specific conformations of the G protein stabilized by concerted interactions of multiple amino acid sequences distributed throughout the G alpha protein.


Alanine aminotransferase-based algorithms of liver stiffness measurement by transient elastography (Fibroscan) for liver fibrosis in chronic hepatitis B.
The aim of this study is to know the liver stiffness measurement (LSM) cutoffs for different stages of liver fibrosis in chronic hepatitis B (CHB) and to investigate the effect of alanine aminotransferase (ALT) on LSM. We prospectively studied consecutive CHB patients undergoing liver biopsy and transient elastography examinations. Diagnostic performance of LSM for different degrees of liver fibrosis was evaluated. One hundred and sixty-one CHB patients with adequate liver biopsy sample size were studied. Area under receiver operating characteristics curves of LSM for no fibrosis (F0 vs F1-4), bridging fibrosis (F0-2 vs F3-4) and liver cirrhosis (F0-3 vs F4) was 0.80 (95% CI: 0.68-0.92), 0.87 (95% CI: 0.82-0.93) and 0.93 (95% CI: 0.89-0.97) respectively. For liver cirrhosis, these optimal cutoff values were 8.4 kPa (98% sensitivity), 9.0 kPa (maximum sum of sensitivity and specificity), 13.4 kPa (94% specificity) and 13.4 kPa (maximum diagnostic accuracy, 85%) respectively. Patients with the same fibrosis staging but higher ALT levels tend to have higher LSM, and the diagnostic performance for low stage fibrosis was most seriously affected when ALT was elevated. Different LSM cutoff values and algorithms were derived for normal and elevated ALT levels. Based on these algorithms, liver biopsy can be avoided in 62% and 58% of patients with normal and elevated ALT respectively. In conclusion, transient elastography is a reasonable noninvasive tool to substitute liver biopsy among the lowest and highest risk patients for the assessment of liver fibrosis.


The effectiveness of traditional methods and altered auditory feedback in improving speech rate and intelligibility in speakers with Parkinson's disease.
Communication problems are a frequent symptom for people with Parkinson's disease (PD) which can have a significant impact on their quality-of-life. Deciding on the right management approach can be problematic though, as, with the exception of LSVT, very few studies have been published demonstrating the effectiveness of treatment techniques. The aim of this study was to compare traditional rate reduction methods with altered auditory feedback (AAF) with respect to their effectiveness to reduce speech rate and improve intelligibility in speakers with PD. Ten participants underwent both types of treatments in once weekly sessions for 6 weeks. Outcomes measures were speech rate for passage reading as well as intelligibility on both a passage reading and a monologue task. The results showed that, as a group, there was no significant change in either speech rate or intelligibility resulting from either treatment type. However, individual speakers showed improvements in speech performance as a result of each therapy technique. In most cases, these benefits persisted for at least 6 months post-treatment. Possible reasons for the variable response to treatment, as well as issues to consider when planning to use AAF devices in treatment are discussed.


Stimulation of the subthalamic nucleus improves velocity of ballistic movements in Parkinson's disease.
High-frequency stimulation of the subthalamic nucleus can markedly improve motor function in patients with Parkinson's disease. However, the underlying mechanisms mediating these improvements are not well understood. In particular, whether motor function is differentially improved in distal or proximal movements is not fully determined. Also, whether reaction time is improved along with other motor parameters is still a matter of debate. Here, we test patients OFF and ON subthalamic nucleus stimulation by capturing simple ballistic movements across four joints using kinematic motion analysis. We show that velocity, but not reaction time, is significantly improved with stimulation. There was no strong differential effect between joints. These results add evidence that deep brain stimulation of the subthalamic nucleus can enhance performance of ballistic movements in Parkinson's disease, and demonstrate that the subthalamic nucleus may be important in driving parameters of motor control after the response has been initiated.


3-[211At]astato-4-fluorobenzylguanidine: a potential therapeutic agent with prolonged retention by neuroblastoma cells.
An analogue of meta-iodobenzylguanidine (MIBG) in which an aromatic hydrogen was replaced with fluorine has been found to possess many properties similar to those of the parent compound. Moreover, 4-fluoro-3-iodobenzylguanidine (FIBG) was retained in vitro by human neuroblastoma cells to a much greater extent than MIBG itself. Since alpha-emitters such as 211At could be valuable for the treatment of micrometastatic disease, an FIBG analogue in which the iodine atom is replaced by 211At would be of interest. In this study, we have evaluated the in vitro and in vivo properties of 3-[211At]astato-4-fluorobenzylguanidine ([211At]AFBG). The specific binding of [211At]AFBG to SK-N-SH human neuroblastoma cells remained fairly constant over 2- to 3-log activity range and was similar to that of [131I]MIBG. The uptake of [211At]AFBG by this cell line was reduced by desipramine, ouabain, 4 degrees C incubation, noradrenaline, unlabelled MIBG and FIBG, suggesting that its uptake is specifically mediated through an active uptake-1 mechanism. Over the 16 h period studied, the amount of [211At]AFBG retained was similar to that of [131I]FIBG, whereas the per cent of retained meta-[211At]astatobenzylguanidine ([211At]MABG) was considerably less than that of [131I]FIBG (53% vs 75%; P < 0.05). The IC50 values for the inhibition of uptake of [131I]MIBG, [211At]MABG, [125I]FIBG and [211At]AFBG by unlabelled MIBG were 209, 300, 407 and 661 nM respectively, suggesting that the affinities of these tracers for the noradrenaline transporter in SK-N-SH cells increase in that order. Compared with [211At]MABG, higher uptake of [211At]AFBG was seen in vivo in normal mouse target tissues such as heart and, to a certain extent, in adrenals. That the uptake of [211At]AFBG in these tissues was related to the uptake-1 mechanism was demonstrated by its reduction when mice were pretreated with desipramine. However, the stability of [211At]AFBG towards in vivo dehalogenation was less than that of [211At]MABG, as evidenced by the higher uptake of 211At in thyroid, spleen, lungs and stomach.


Recent updates on the molecular network of elastic fiber formation.
Elastic fibers confer elasticity and recoiling to tissues and organs and play an essential role in induction of biochemical responses in a cell against mechanical forces derived from the microenvironment. The core component of elastic fibers is elastin (ELN), which is secreted as the monomer tropoelastin from elastogenic cells, and undergoes self-aggregation, cross-linking and deposition on to microfibrils, and assemble into insoluble ELN polymers. For elastic fibers to form, a microfibril scaffold (primarily formed by fibrillin-1 (FBN1)) is required. Numerous elastic fiber-associated proteins are involved in each step of elastogenesis and they instruct and/or facilitate the elastogenesis processes. In this review, we designated five proteins as key molecules in elastic fiber formation, including ELN, FBN1, fibulin-4 (FBLN4), fibulin-5 (FBLN5), and latent TGFβ-binding protein-4 (LTBP4). ELN and FBN1 serve as building blocks for elastic fibers. FBLN5, FBLN4 and LTBP4 have been demonstrated to play crucial roles in elastogenesis through knockout studies in mice. Using these molecules as a platform and expanding the elastic fiber network through the generation of an interactome map, we provide a concise review of elastogenesis with a recent update as well as discuss various biological functions of elastic fiber-associated proteins beyond elastogenesis in vivo.


Estrogens, progestogens and thrombosis.
Hundreds of millions of women worldwide use either oral contraceptives or postmenopausal hormone replacement. The use of oral contraceptives leads to an increased risk of venous thrombosis, of myocardial infarction, of stroke and of peripheral artery disease, the risks of which are highest during the first year of use. Women with coagulation abnormalities have a higher risk of venous thrombosis when they use oral contraceptives (or postmenopausal hormones) than women without these abnormalities. The risk of venous thrombosis is also higher for preparations containing desogestrel or gestodene (third-generation progestogens) than for those containing levonorgestrel (second-generation progestogens). A previous thrombosis as well as obesity also increase the risk of oral contraceptive-related thrombosis. Hormone replacement therapy increases the risk of venous thrombosis, and has no beneficial, and possibly even a detrimental, effect on the risk of arterial disease. The risk of arterial disease in oral contraceptive users and users of hormone replacement therapy is at most weakly affected by the presence of prothrombotic abnormalities.


Nanoscale Amperometry Reveals that Only a Fraction of Vesicular Serotonin Content is Released During Exocytosis from Beta Cells.
Recent work has shown that chemical release during the fundamental cellular process of exocytosis in model cell lines is not all-or-none. We tested this theory for vesicular release from single pancreatic beta cells. The vesicles in these cells release insulin, but also serotonin, which is detectible with amperometric methods. Traditionally, it is assumed that exocytosis in beta cells is all-or-none. Here, we use a multidisciplinary approach involving nanoscale amperometric chemical methods to explore the chemical nature of insulin exocytosis. We amperometrically quantified the number of serotonin molecules stored inside of individual nanoscale vesicles (39 317±1611) in the cell cytoplasm before exocytosis and the number of serotonin molecules released from single cells (13 310±1127) for each stimulated exocytosis event. Thus, beta cells release only one-third of their granule content, clearly supporting partial release in this system. We discuss these observations in the context of type-2 diabetes.


Organization of ribosomal genes in Paramecium tetraurelia.
The macronuclear ribosomal DNA (rDNA) of the ciliated protozoan Paramecium tetraurelia (stock 51) was analyzed by digestion with restriction endonucleases. The fragments which contained ribosomal RNA (rRNA) coding sequences and spacer sequences were identified. The spacer sequences exhibited some heterogeneity in size. The genes coding for 5.8S RNA, but not for 5S RNA, are linked to the 17S and 25S rRNA genes. Complementary RNA, synthesized from rDNA of stock 51, was hybridized with restriction digests of whole cell DNA from six other allopatric stocks of this species. The restriction patterns of the rDNA from these seven stocks were, in general, very similar, and the sizes of the coding sequences were identical in all seven stocks. Only the restriction pattern of rDNA from stock 127 differed significantly from that of stock 51. The rDNA from stock 127 was isolated and characterized, and with the exception of the restriction pattern of its spacer, it resembled the rDNA from stock 51. It is concluded that the rDNA repeat in Paramecium, including the spacer, has, in general, been conserved during the course of evolution. It is suggested that in some species, even in the absence of genetic exchange among geographically separated populations, selection pressure may act to conserve spacers of tandemly repeated rDNA. The conservation may be related to the number of rDNA copies in the germinal nucleus.


Khat Use, PTSD and Psychotic Symptoms among Somali Refugees in Nairobi - A Pilot Study.
In East-African and Arab countries, khat leaves are traditionally chewed in social settings. They contain the amphetamine-like alkaloid cathinone. Especially among Somali refugees, khat use has been associated with psychiatric symptoms. We assessed khat-use patterns and psychiatric symptoms among male Somali refugees living in a disadvantaged urban settlement area in Kenya, a large group that has not yet received scientific attention. We wanted to explore consume patterns and study the associations between khat use, traumatic experiences, and psychotic symptoms. Using privileged access sampling, we recruited 33 healthy male khat chewers and 15 comparable non-chewers. Based on extensive preparatory work, we assessed khat use, khat dependence according to DSM-IV, traumatic experiences, posttraumatic stress disorder (PTSD), and psychotic symptoms using standardized diagnostic instruments that had been adapted to the Somali language and culture. Hazardous use patterns like chewing for more than 24 h without interruption were frequently reported. All khat users fulfilled the DSM-IV-criteria for dependence and 85% reported functional khat use, i.e., that khat helps them to forget painful experiences. We found that the studied group was heavily burdened by traumatic events and posttraumatic symptoms. Khat users had experienced more traumatic events and had more often PTSD than non-users. Most khat users experience khat-related psychotic symptoms and in a quarter of them we found true psychotic symptoms. In contrast, among control group members no psychotic symptoms could be detected. We found first evidence for the existence and high prevalence of severely hazardous use patterns, comorbid psychiatric symptoms, and khat use as a self-medication of trauma-consequences among male Somali refugees in urban Kenyan refugee settlements. There is a high burden by psychopathology and adequate community-based interventions urgently need to be developed.


Update on the natural history of cavernous malformations and factors predicting aggressive clinical presentation.
Cavernous malformations (CMs) are angiographically occult, low-pressure neurovascular lesions with distinct imaging and clinical characteristics. They present with seizure, neurological compromise due to lesion hemorrhage or expansion, or as incidental findings on neuroimaging studies. Treatment options include conservative therapy, medical management of seizures, surgical intervention for lesion resection, and in select cases stereotactic radiosurgery. Optimal management requires a thorough understanding of the natural history of CMs including consideration of issues such as mode of presentation, lesion location, and genetics that may impact the associated neurological risk. Over the past 2 decades, multiple studies have been published, shedding valuable light on the clinical characteristics and natural history of these malformations. The purpose of this review is to provide the reader with a concise consolidation of this published material such that they may better understand the risks associated with CMs and their implications on patient treatment.


DNA methylation in an engineered heart tissue model of cardiac hypertrophy: common signatures and effects of DNA methylation inhibitors.
DNA methylation affects transcriptional regulation and constitutes a drug target in cancer biology. In cardiac hypertrophy, DNA methylation may control the fetal gene program. We therefore investigated DNA methylation signatures and their dynamics in an in vitro model of cardiac hypertrophy based on engineered heart tissue (EHT). We exposed EHTs from neonatal rat cardiomyocytes to a 12-fold increased afterload (AE) or to phenylephrine (PE 20 µM) and compared DNA methylation signatures to control EHT by pull-down assay and DNA methylation microarray. A 7-day intervention sufficed to induce contractile dysfunction and significantly decrease promoter methylation of hypertrophy-associated upregulated genes such as Nppa (encoding ANP) and Acta1 (α-skeletal actin) in both intervention groups. To evaluate whether pathological consequences of AE are affected by inhibiting de novo DNA methylation we applied AE in the absence and presence of DNA methyltransferase (DNMT) inhibitors: 5-aza-2'-deoxycytidine (aza, 100 µM, nucleosidic inhibitor), RG108 (60 µM, non-nucleosidic) or methylene disalicylic acid (MDSA, 25 µM, non-nucleosidic). Aza had no effect on EHT function, but RG108 and MDSA partially prevented the detrimental consequences of AE on force, contraction and relaxation velocity. RG108 reduced AE-induced Atp2a2 (SERCA2a) promoter methylation. The results provide evidence for dynamic DNA methylation in cardiac hypertrophy and warrant further investigation of the potential of DNA methylation in the treatment of cardiac hypertrophy.


Calcium oxalate crystals in the thyroid. Their identification, prevalence, origin, and possible significance.
Calcium oxalate crystals are not encountered in normal animal tissues, except for the human thyroid, where they were found in 79 of 100 routine consecutive autopsies. They appear during childhood, and numbers of crystals increase with age. In diffuse hyperplasia, prevalence was higher, but crystals were fewer than expected. In adenomas and carcinomas, crystals were decreased except for three cases with a striking focal increase. None was found in 22 adult primate thyroids. After Clorox digestion of human thyroids, calcium oxalate dihydrate was identified by x-ray diffraction and infrared spectroscopy. Origin, tissue and species localization are discussed in relation to ascorbate metabolism, thyroperoxidase, and calcitonin. Possible metabolic roles are suggested. Calcium oxalate crystals injected in animals and humans initiate a foreign body reaction with giant cells. In Hashimoto's thyroiditis, crystals disappear but occasionally remain with giant cell reaction. In subacute thyroiditis, granulomas are related more to colloid than to crystals.


Cognitive and mood improvements of caffeine in habitual consumers and habitual non-consumers of caffeine.
The cognitive and mood effects of caffeine are well documented. However, the majority of studies in this area involve caffeine-deprived, habitual caffeine users. It is therefore unclear whether any beneficial findings are due to the positive effects of caffeine or to the alleviation of caffeine withdrawal. The present placebo-controlled, double-blind, balanced crossover study investigated the acute cognitive and mood effects of caffeine in habitual users and habitual non-users of caffeine. Following overnight caffeine withdrawal, 24 habitual caffeine consumers (mean=217 mg/day) and 24 habitual non-consumers (20 mg/day) received a 150 ml drink containing either 75 or 150 mg of caffeine or a matching placebo, at intervals of > or =48 h. Cognitive and mood assessments were undertaken at baseline and 30 min post-drink. These included the Cognitive Drug Research computerised test battery, two serial subtraction tasks, a sentence verification task and subjective visual analogue mood scales. There were no baseline differences between the groups' mood or performance. Following caffeine, there were significant improvements in simple reaction time, digit vigilance reaction time, numeric working memory reaction time and sentence verification accuracy, irrespective of group. Self-rated mental fatigue was reduced and ratings of alertness were significantly improved by caffeine independent of group. There were also group effects for rapid visual information processing false alarms and spatial memory accuracy with habitual consumers outperforming non-consumers. There was a single significant interaction of group and treatment effects on jittery ratings. Separate analyses of each groups' responses to caffeine revealed overlapping but differential responses to caffeine. Caffeine tended to benefit consumers' mood more while improving performance more in the non-consumers. These results do not support a withdrawal alleviation model. Differences in the patterns of responses to caffeine by habitual consumers and habitual non-consumers may go some way to explaining why some individuals become caffeine consumers.


Microbial community structure and biodegradation activity of particle-associated bacteria in a coal tar contaminated creek.
The Chattanooga Creek Superfund site (Chattanooga, TN) is one of the most polluted waterways in the southeastern U.S. with high polycyclic aromatic hydrocarbon (PAH) concentrations in the sediments. PAHs associate with suspended solids in the water column, and may be redeposited onto the floodplain. These suspended particles represent an interesting but understudied environment for PAH-degrading microbial communities. This study tested the hypotheses that particle-associated bacterial (PAB) communities have genotypic potential (PAH-dioxygenase genes) and activity (naphthalene and pyrene mineralization), and can contribute to natural attenuation of PAHs in Chattanooga Creek. Upstream of the Superfund site, mineralization ranged from 0.2 to 2.0% of added 14C-naphthalene and 0 to 0.1% 14C-pyrene (after 40 h), with first order biodegradation rate constants (k1) ranging from 1.09 to 9.18 x 10(-5) h(-1) and 0 to 1.13 x 10(-6) h(-1), respectively. Mineralization was significantly greater in PAB communities within the contaminated zone, with 11.8 to 31.2% 14C-naphthalene (k1 5.34 to 14.2 x 10(-4) h(-1)) and 1.3 to 6.6% 14C-pyrene mineralized (k1 2.89 to 15.0 x 10(-5) h(-1)). Abundances of nagAc (naphthalene dioxygenase) and nidA (pyrene dioxygenase) genes indicated that PAB communities harbored populations with genetic potential for both low- and high-molecularweight PAH degradation, and quantification of Mycobacterium 16S rDNA genes indicated that PAH-degrading mycobacteria are also prevalent in this environment. Phylogenetic comparisons (T-RFLPs) between PAB and sediments indicated these microbial communities were taxonomically distinct, but shared some functional similarities, namely PAH catabolic genotypes, mineralization capabilities, and community structuring along a contamination gradient


Pacific lamprey (Entosphenus tridentatus) ammocoetes exposed to contaminated Portland Harbor sediments: Method development and effects on survival, growth, and behavior.
Many anthropogenic disturbances have contributed to the decline of Pacific lampreys (Entosphenus tridentatus), but potential negative effects of contaminants on lampreys are unclear. Lamprey ammocoetes are the only detritivorous fish in the lower Willamette River, Oregon, USA, and have been observed in Portland Harbor sediments. Their long benthic larval stage places them at risk from the effects of contaminated sediment. The authors developed experimental methods to assess the effects of contaminated sediment on the growth and behavior of field-collected ammocoetes reared in a laboratory. Specifically, they developed methods to assess individual growth and burrowing behavior. Burrowing performance demonstrated high variability among contaminated sediments; however, ammocoetes presented with noncontaminated reference sediment initiated burrowing more rapidly and completed it faster. Ammocoete reemergence from contaminated sediments suggests avoidance of some chemical compounds. The authors conducted long-term exposure experiments on individually held ammocoetes using sediment collected from their native Siletz River, which included the following: contaminated sediments collected from 9 sites within Portland Harbor, 2 uncontaminated reference sediments collected upstream, 1 uncontaminated sediment with characteristics similar to Portland Harbor sediments, and clean sand. They determined that a 24-h depuration period was sufficient to evaluate weight changes and observed no mortality or growth effects in fish exposed to any of the contaminated sediments. However, the effect on burrowing behavior appeared to be a sensitive endpoint, with potentially significant implications for predator avoidance. Environ Toxicol Chem 2016;35:2092-2102. © 2016 SETAC.


Color management of porcelain veneers: influence of dentin and resin cement colors.
Porcelain veneers have become an interesting treatment option to correct the shape and color of anterior teeth. Because of their limited thickness and high translucency, achieving a good color match is influenced by several variables. The aim of this work was to investigate the influence of natural dentin and resin cement colors on final color match of porcelain veneers. A preselected shade tab (A1) was chosen as the target color for a maxillary central incisor, and its color parameters (L*a*b*) were measured using a digital spectrophotometer (SpectroShade, MHT). Nine natural dentin colors (Natural Die Material, Ivoclar Vivadent) representing a wide range of tooth colors were used to prepare resin replicas of the maxillary central incisor with a standard preparation for porcelain veneers. The prepared porcelain veneers (IPS Empress Esthetic, A1, 0.6 mm thick, Ivoclar Vivadent) were cemented on the resin dies (nine groups of natural dentin colors) using seven shades of resin cement (Variolink Veneers, Ivoclar Vivadent). The L*a*b* values of the cemented veneers were measured, and DE values were calculated against the preselected target color (A1). DE greater than 3.3 was considered as a significant color mismatch detectable by the human eye. The seven shades of resin cement had no significant influence on the final color of the veneers, as the measured DE values were almost identical for every test group. On the other hand, the color of natural dentin was a significant factor that influenced final color match. None of the 63 tested combinations (nine natural dentin colors and seven resin cement colors) produced an acceptable color match. Thin porcelain veneers cannot mask underlying tooth color even when different shades of resin cement are used. Incorporation of opaque porcelain (high chroma) may improve final color match.


Review of pain content in three-year preregistration pediatric nursing courses in the United Kingdom.
The results of several studies suggest that gaps remain in nurses' knowledge about pain in children, and particularly regarding pain assessment, analgesic drugs, and nondrug methods, suggesting a need to explore the pain content of preregistration nursing curricula. Over the past decade, the way nurse education is delivered has changed with the advent of the internet and the use of virtual learning environments. This study, therefore, explored the depth and breadth of pain content in 3-year preregistration pediatric nursing courses across the U.K. as well as ascertaining how this content is delivered. The leads for pediatric nursing at all 56 U.K. higher education institutions running the 3-year preregistration pediatric nursing course were e-mailed a questionnaire. Participants were asked to indicate whether topics are covered in the program, and if so how it was delivered. Results suggest that preregistration pediatric nursing courses in the U.K. do not always equip students to manage pain effectively in clinical practice. There appears to be limited content on pain in nursing curricula, with several institutions not covering key topics. Most teaching is delivered face to face with limited workbooks and virtual learning activities. Further research is needed in this area to ascertain the impact of this on the quality of care provided.


Varying uses of the ABCD2 scoring system in primary and secondary care: a qualitative study.
To explore the usage of the ABCD2 risk stratification score by general practitioners (GPs) and hospital staff during the referral of patients with suspected transient ischaemic attack (TIA) or minor stroke. Qualitative study using semistructured interviews. Nine general practices and two hospital sites in England (Birmingham and Cambridge). Nine GPs and nine hospital staff (two consultants, four nurses, two ultrasonographers and one administrator). In both sites, clinicians used a referral proforma based around the ABCD2 scoring system for a range of purposes including self-education, to assist emphasising urgency to the patient, as a referral pathway facilitator and as a diagnostic tool. Negative views of its role included potential medicolegal threats, that it was a barrier to appropriate care, and led to misdiagnoses. Despite having differing uses by different clinicians, the ABCD2 proforma was the central means of interprofessional communication in TIA referrals across both sites. Understanding how prediction rules are used in practice is key to determining their impact on processes of care and clinical outcomes. In practice, GPs and their colleagues use the ABCD2 score in subtly different ways and it functions as a 'boundary object' by both accommodating these multiple purposes, yet still successfully aiding communication between them.


Sustainability and cost of a community-based strategy against Aedes aegypti in northern and central Vietnam.
We previously reported a new community-based mosquito control that resulted in the elimination of Aedes aegypti in 40 of 46 communes in northern and central Vietnam. During 2007 and 2008, we revisited Nam Dinh and Khanh Hoa provinces in northern and central Vietnam, respectively, to evaluate whether or not these programs were still being maintained 7 years and 4.5 years after formal project activities had ceased, respectively. Using a previously published sustainability framework, we compared 13 criteria from Tho Nghiep commune in Nam Dinh where the local community had adopted our community-based project model using Mesocyclops from 2001. These data were compared against a formal project commune, Xuan Phong, where our successful intervention activities had ceased in 2000 and four communes operating under the National Dengue Control Program with data available. In Khanh Hoa province, we compared 2008 data at Ninh Xuan commune with data at project completion in 2003 and benchmarked these, where possible, against an untreated control commune, Ninh Binh, where few control activities had been undertaken. The three communes where the above community-based strategy had been adopted were rated as well-sustained with annual recurrent total costs (direct and indirect) of $0.28-0.89 international dollars per person.


Cartilage oligomeric matrix protein forms protein complexes with synovial lubricin via non-covalent and covalent interactions.
Understanding the cartilage surface structure, lost in arthritic disease, is essential for developing strategies to effectively restore it. Given that adherence of the lubricating protein, lubricin, to the cartilage surface is critical for boundary lubrication, an interaction with cartilage oligomeric matrix protein (COMP) was investigated. COMP, an abundant cartilage protein, is known to be important for matrix formation. Synovial fluid (SF) from arthritic patients was used to detect possible COMP-lubricin complexes by immunological methods. Recombinant (RC) COMP and lubricin fragments were expressed to characterize this bonding and mass spectrometry employed to specifically identify the cysteines involved in inter-protein disulfide bonds. COMP-lubricin complexes were identified in the SF of arthritic patients by Western blot, co-immunoprecipitation and sandwich ELISA. RC fragment solid-phase binding assays showed that the C-terminal (amino acids (AA) 518-757) of COMP bound non-covalently to the N-terminal of lubricin (AA 105-202). Mass spectrometry determined that although cysteines throughout COMP were involved in binding with lubricin, the cysteines in lubricin were primarily focused to an N-terminal region (AA 64-86). The close proximity of the non-covalent and disulfide binding domains on lubricin suggest a two-step mechanism to strongly bind lubricin to COMP. These data demonstrate that lubricin forms a complex network with COMP involving both non-covalent and covalent bonds. This complex between lubricin and the cartilage protein COMP can be identified in the SF of patients with arthritis conditions including osteoarthritis (OA) and rheumatoid arthritis (RA).


Use of Active-Play Video Games to Enhance Aerobic Fitness in Schizophrenia: Feasibility, Safety, and Adherence.
Active-play video games have been used to enhance aerobic fitness in various clinical populations, but their use among individuals with schizophrenia has been limited. Feasibility, acceptability, safety, and adherence data were obtained for use of aerobic exercise (AE) equipment by 16 individuals with schizophrenia during a 12-week AE program consisting of three one-hour exercise sessions per week. Equipment included exercise video games for Xbox 360 with Kinect motion sensing devices and traditional exercise equipment. Most participants (81%) completed the training, attending an average of 79% of sessions. The proportion of time spent playing Xbox (39%) exceeded time spent on any other type of equipment. When using Xbox, participants played 2.24±1.59 games per session and reported high acceptability and enjoyment ratings, with no adverse events. Measures of feasibility, acceptability, adherence, and safety support the integration of active-play video games into AE training for people with schizophrenia.


Management of ankle sprains.
Without adequate care, acute ankle trauma can result in chronic joint instability. Use of a standardized protocol enhances the management of ankle sprains. In patients with grades I or II sprains, emphasis should be placed on accurate diagnosis, early use of RICE (rest, ice, compression and elevation), maintenance of range of motion and use of an ankle support. Sprains with complete ligament [corrected] tears (grade III) may require surgical intervention. Although early motion and mobility are recommended, ligamentous strength does not return until months after an ankle sprain.


Odorant-dependent generation of nitric oxide in Mammalian olfactory sensory neurons.
The gaseous signalling molecule nitric oxide (NO) is involved in various physiological processes including regulation of blood pressure, immunocytotoxicity and neurotransmission. In the mammalian olfactory bulb (OB), NO plays a role in the formation of olfactory memory evoked by pheromones as well as conventional odorants. While NO generated by the neuronal isoform of NO synthase (nNOS) regulates neurogenesis in the olfactory epithelium, NO has not been implicated in olfactory signal transduction. We now show the expression and function of the endothelial isoform of NO synthase (eNOS) in mature olfactory sensory neurons (OSNs) of adult mice. Using NO-sensitive micro electrodes, we show that stimulation liberates NO from isolated wild-type OSNs, but not from OSNs of eNOS deficient mice. Integrated electrophysiological recordings (electro-olfactograms or EOGs) from the olfactory epithelium of these mice show that NO plays a significant role in modulating adaptation. Evidence for the presence of eNOS in mature mammalian OSNs and its involvement in odorant adaptation implicates NO as an important new element involved in olfactory signal transduction. As a diffusible messenger, NO could also have additional functions related to cross adaptation, regeneration, and maintenance of MOE homeostasis.


Bacterial c-di-GMP is an immunostimulatory molecule.
Cyclic diguanylate (c-di-GMP) is a bacterial intracellular signaling molecule. We have shown that treatment with exogenous c-di-GMP inhibits Staphylococcus aureus infection in a mouse model. We now report that c-di-GMP is an immodulator and immunostimulatory molecule. Intramammary treatment of mice with c-di-GMP 12 and 6 h before S. aureus challenge gave a protective effect and a 10,000-fold reduction in CFUs in tissues (p < 0.001). Intramuscular vaccination of mice with c-di-GMP coinjected with S. aureus clumping factor A (ClfA) Ag produced serum with significantly higher anti-ClfA IgG Ab titers (p < 0.001) compared with ClfA alone. Intraperitoneal injection of mice with c-di-GMP activated monocyte and granulocyte recruitment. Human immature dendritic cells (DCs) cultured in the presence of c-di-GMP showed increased expression of costimulatory molecules CD80/CD86 and maturation marker CD83, increased MHC class II and cytokines and chemokines such as IL-12, IFN-gamma, IL-8, MCP-1, IFN-gamma-inducible protein 10, and RANTES, and altered expression of chemokine receptors including CCR1, CCR7, and CXCR4. c-di-GMP-matured DCs demonstrated enhanced T cell stimulatory activity. c-di-GMP activated p38 MAPK in human DCs and ERK phosphorylation in human macrophages. c-di-GMP is stable in human serum. We propose that cyclic dinucleotides like c-di-GMP can be used clinically in humans and animals as an immunomodulator, immune enhancer, immunotherapeutic, immunoprophylactic, or vaccine adjuvant.


Demonstration of the use of windows of operation to visualize the effects of fouling on the performance of a chromatographic step.
The high resolution afforded by packed bed chromatography makes it an indispensable operation in the downstream processing of therapeutic molecules. Packed bed performance is however inherently susceptible to changes in feed stream characteristics and fouling processes. The impact of fouling is seldom considered during the early stages of bioprocess development which is concerned with the selection of purification conditions. Instead these are performed with rigorously clarified feeds. Under such conditions, chromatography is effectively treated as an isolated step, independent from its preceding unit operations. In this study, we demonstrate how windows of operation could be used to visualize the impact of changes in the preceding clarification step on the fouling response of a subsequent cation exchange capture step. Laboratory columns (2,5 and 12 cm height) were subjected to varying fouling challenges of Escherichia coli lysate containing different amounts of solids carried over from the previous step. Changes in trans-column pressure drop and breakthrough of the target protein (Fab') were monitored. The limits of operability of the resin were determined with respect to the process material's properties. This information was used to extract the parameters for the adsorption kinetics used in the general rate (GR) model to create windows of operation for manufacturing scale operation.


Evidence for Ca(2+)-regulated ATP release in gastrointestinal stromal tumors.
Gastrointestinal stromal tumors (GISTs) are thought to originate from the electrically active pacemaker cells of the gastrointestinal tract. Despite the presence of synaptic-like vesicles and proteins involved in cell secretion it remains unclear whether GIST cells possess regulated release mechanisms. The GIST tumor cell line GIST882 was used as a model cell system, and stimulus-release coupling was investigated by confocal microscopy of cytoplasmic free Ca(2+) concentration ([Ca(2+)]i), flow cytometry, and luminometric measurements of extracellular ATP. We demonstrate that GIST cells have an intact intracellular Ca(2+)-signaling pathway that regulates ATP release. Cell viability and cell membrane integrity was preserved, excluding ATP leakage due to cell death and suggesting active ATP release. The stimulus-secretion signal transduction is at least partly dependent on Ca(2+) influx since exclusion of extracellular Ca(2+) diminishes the ATP release. We conclude that measurements of ATP release in GISTs may be a useful tool for dissecting the signal transduction pathway, mapping exocytotic components, and possibly for the development and evaluation of drugs. Additionally, release of ATP from GISTs may have importance for tumor tissue homeostasis and immune surveillance escape.


Hyperextension injury to the PIP joint or to the MP joint of the thumb--a clinical study.
We present a prospective study of the diagnosis and clinical course of 60 patients with 57 pure hyperextension injuries to the proximal interphalangeal (PIP) joint of the long fingers (fingers 2-5) and seven injuries to the metacarpophalangeal (MP) joint of the thumb. Thirty four of the injuries (57%) were related to ball sports, and the ulnar fingers of the non-dominant hand were usually affected. There were 24 avulsion fractures at the site of the insertion of the volar plate on to the middle phalanx. Twelve (20%) initially presented with hyperextension instability, and this was usually associated with an avulsion fracture. Thirty four of the patients (57%) had symptoms for less than one month, while 10 (17%) complained of symptoms six months after the injury. Severe complications such as daily pain and stiffness were encountered in three cases. The triad sign (pain on extreme flexion and extension) was of no use as a diagnostic or prognostic factor, nor did the radiographic stress-view help to identify acute instability of the joint.


Language outcomes among ELBW infants in early childhood.
Limited data are available evaluating language outcomes of preterm infants in early childhood. Furthermore, the relationship between language outcomes, medical morbidities and developmental trajectory in early infancy is unclear. The goal of this study was to evaluate language outcomes among extremely low birth weight (ELBW) infants at 30months adjusted age (AA). The Bayley Scales of Infant Development II and the Peabody Picture Vocabulary Test or Expressive One Word Picture Vocabulary Test/Receptive One Word Picture Vocabulary Test were administered at 30months AA to a prospective cohort of ELBW infants who participated in the NICHD Neonatal Network Glutamine Trial and Neurodevelopmental Follow-Up Study. A standardized history and physical examination and query regarding feeding behaviors were performed at 18months AA and 30months AA. Of the 467 infants evaluated, 55% had receptive language delay at 30months with 23% having severe delays. Fewer (26%) had expressive language delays, with 16% of those being severe delays. Non-English speaking infants had poorer performance on all language measures compared to English-speaking infants. Forty-seven percent of the cohort required assistance with feeds at 18months. These children were more likely to have language delay at the 30month assessment compared to infants who could feed themselves. ELBW infants are at risk of language delay in early childhood. Additional research is needed to further explore the relationship between early predictors of language delay and the use of monolingual language assessments in non-English speaking patients with a history of prematurity.


Early telomerase inactivation accelerates aging independently of telomere length.
Telomerase is required for long-term telomere maintenance and protection. Using single budding yeast mother cell analyses we found that, even early after telomerase inactivation (ETI), yeast mother cells show transient DNA damage response (DDR) episodes, stochastically altered cell-cycle dynamics, and accelerated mother cell aging. The acceleration of ETI mother cell aging was not explained by increased reactive oxygen species (ROS), Sir protein perturbation, or deprotected telomeres. ETI phenotypes occurred well before the population senescence caused late after telomerase inactivation (LTI). They were morphologically distinct from LTI senescence, were genetically uncoupled from telomere length, and were rescued by elevating dNTP pools. Our combined genetic and single-cell analyses show that, well before critical telomere shortening, telomerase is continuously required to respond to transient DNA replication stress in mother cells and that a lack of telomerase accelerates otherwise normal aging.


Comprehensive analysis of imaging quality degradation of an airborne optical system for aerodynamic flow field around the optical window.
We investigated the influences exerted by the nonuniform aerodynamic flow field surrounding the optical window on the imaging quality degradation of an airborne optical system. The density distribution of flow fields around three typical optical windows, including a spherical window, an ellipsoidal window, and a paraboloidal window, were calculated by adopting the Reynolds-averaged Navier-Stokes equations with the Spalart-Allmaras model provided by FLUENT. The fourth-order Runge-Kutta algorithm based ray-tracing program was used to simulate the optical transmission through the aerodynamic flow field. Four kinds of imaging quality evaluation parameters were presented: wave aberration of the entrance pupil, point spread function, encircled energy, and modulation transfer function. The results show that the imaging quality of the airborne optical system was affected by the shape of the optical window and angle of attack of the aircraft.


Research on Vegetable Pest Warning System Based on Multidimensional Big Data.
Pest early warning technology is part of the prerequisite for the timely and effective control of pest outbreaks. Traditional pest warning system with artificial mathematical statistics, radar, and remote sensing has some deficiency in many aspects, such as higher cost, weakness of accuracy, low efficiency, and so on. In this study, Pest image data was collected and information about four major vegetable pests (Bemisia tabaci (Gennadius), Phyllotreta striolata (Fabricius), Plutella xylostella (Linnaeus), and Frankliniella occidentalis (Pergande) (Thysanoptera, Thripidae)) in southern China was extracted. A multi-sensor network system was constructed to collect small-scale environmental data on vegetable production sites. The key factors affecting the distribution of pests were discovered by multi-dimensional information, such as soil, environment, eco-climate, and meteorology of vegetable fields, and finally, the vegetable pest warning system that is based on multidimensional big data (VPWS-MBD) was implemented. Pest and environmental data from Guangzhou Dongsheng Bio-Park were collected from June 2017 to February 2018. The number of pests is classified as level I (0⁻56), level II (57⁻131), level III (132⁻299), and level IV (above 300) by K-Means algorithm. The Pearson correlation coefficient and the grey relational analysis algorithm were used to calculate the five key influence factors of rainfall, soil temperature, air temperature, leaf surface humidity, and soil moisture. Finally, Back Propagation (BP) Neural Network was used for classification prediction. The result shows: I-level warning accuracy was 96.14%, recall rate was 97.56%; II-level pest warning accuracy was 95.34%, the recall rate was 96.45%; III-level pest warning accuracy of 100%, the recall rate was 96.28%; IV-level pest warning accuracy of 100%, recall rate was 100%. It proves that the early warning system can effectively predict vegetable pests and achieve the early warning of vegetable pest's requirements, with high availability.


A standardization method to compare isotonic vs. isokinetic eccentric exercises.
The present study aimed to standardize isotonic (IT) and isokinetic (IK) eccentric exercises by equalizing the amount of work and the angular velocity at two intensity levels, to be able to compare specific effects of these exercise modes on the neuromuscular system. Fourteen subjects participated in three test sessions consisting of two IT and two IK sets on a customized isokinetic dynamometer. IT sets were comprised of 8 eccentric contractions of the knee extensors at 120% of the maximal repetition (1RM) in the first two sessions, and 100% in the third session. IK sets were performed at the same mean angular IT velocity and stopped when the amount of work performed corresponded to the IT set. External work, angular velocity and Root Mean Square (RMS) of electromyographic activity of three superficial muscles of the quadriceps femoris were calculated. Results showed concordance of work and angular velocity for each test session. Both modes involved the same number of repetitions at 120% and fewer repetitions in IK mode at 100% of 1RM. Work and RMS values remained steady in all sets. This study allowed the standardization of isotonic and isokinetic eccentric exercises, a first step before determining their specific effects on neuromuscular function.


The association of Streptococcus bovis/gallolyticus with colorectal tumors: the nature and the underlying mechanisms of its etiological role.
Streptococcus bovis (S. bovis) bacteria are associated with colorectal cancer and adenoma. S. bovis is currently named S. gallolyticus. 25 to 80% of patients with S. bovis/gallolyticus bacteremia have concomitant colorectal tumors. Colonic neoplasia may arise years after the presentation of bacteremia or infectious endocarditis of S. bovis/gallolyticus. The presence of S. bovis/gallolyticus bacteremia and/or endocarditis is also related to the presence of villous or tubular-villous adenomas in the large intestine. In addition, serological relationship of S. gallolyticus with colorectal tumors and direct colonization of S. gallolyticus in tissues of colorectal tumors were found. However, this association is still under controversy and has long been underestimated. Moreover, the etiological versus non-etiological nature of this associationis not settled yet. Therefore, by covering the most of up to date studies, this review attempts to clarify the nature and the core of S. bovis/gallolyicus association with colorectal tumors and analyze the possible underlying mechanisms.


Retinoic acid-induced pancreatic stellate cell quiescence reduces paracrine Wnt-β-catenin signaling to slow tumor progression.
Patients with pancreatic ductal adenocarcinoma are deficient in vitamin A, resulting in activation of pancreatic stellate cells (PSCs). We investigated whether restoration of retinol to PSCs restores their quiescence and affects adjacent cancer cells. PSCs and cancer cell lines (AsPc1 and Capan1) were exposed to doses and isoforms of retinoic acid (RA) in 2-dimensional and 3-dimensional culture conditions (physiomimetic organotypic culture). The effects of all-trans retinoic acid (ATRA) were studied in LSL-KrasG12D/+;LSL-Trp53R172H/+;Pdx-1-Cre mice, a model of human pancreatic ductal adenocarcinoma. After incubation with ATRA, PSCs were quiescent and had altered expression of genes that regulate proliferation, morphology, and motility; genes that encode cytoskeletal proteins and cytokines; and genes that control other functions, irrespective of culture conditions or dosage. In the organotypic model, and in mice, ATRA induced quiescence of PSCs and thereby reduced cancer cell proliferation and translocation of β-catenin to the nucleus, increased cancer cell apoptosis, and altered tumor morphology. ATRA reduced the motility of PSCs, so these cells created a "wall" at the junction between the tumor and the matrix that prevented cancer cell invasion. Restoring secreted frizzled-related protein 4 (sFRP4) secretion to quiescent PSCs reduced Wnt-β-catenin signaling in cancer cells and their invasive ability. Human primary and metastatic pancreatic tumor tissues stained strongly for cancer cell nuclear β-catenin but had low levels of sFRP4 (in cancer cells and PSCs). RA induces quiescence and reduces motility of PSCs, leading to reduced proliferation and increased apoptosis of surrounding pancreatic cancer cells. RA isoforms might be developed as therapeutic reagents for pancreatic cancer.


A comparison of the effect of hyperthermia on DNA polymerase in hamster and human glioma cells.
The hyperthermia response of two human glioma cells lines (87MG and 373MG) was compared to the CHO cell line for cell killing and DNA polymerase inactivation. Glioma cells were found to be more thermally resistant than CHO cells over a temperature range of 41-46 degrees C. Inactivation of polymerase alpha and beta by hyperthermia was also more resistant in glioma cells than in CHO cells. The relative order of resistance for both killing and polymerase inactivation was 373MG > 87MG > CHO. While polymerase inactivation correlated with cell killing at high thermal doses, such correlation at low doses was absent; i.e. thermal killing was characterized by survival curves with shoulders while polymerase inactivation was not. Thus at low thermal doses the mechanism of thermal cell killing is probably not related to the degree of polymerase inactivation. Arrhenius analysis of the survival data showed that the inactivation energy for the glioma cells was 133-135 kcal/mol. The inactivation energies of alpha and beta polymerase were also evaluated and were 102-104 and 140-146 kcal/mol, respectively. Further analysis of the temperature-time relationship of hyperthermia treatment resulting in 50% cell kill showed the degree of polymerase beta inactivation to be a good indicator of thermal dose.


Collagen cross-linking: current status and future directions.
Collagen cross-linking (CXL) using UVA light and riboflavin (vitamin B2) was introduced as a clinical application to stabilize the cornea by inducing cross-links within and between collagen fibers. CXL has been investigated extensively and has been shown clinically to arrest the progression of keratoconic or post-LASIK ectasia. With its minimal cost, simplicity, and proven positive clinical outcome, CXL can be regarded as a useful approach to reduce the number of penetrating keratoplasties performed. Small case series have also indicated that CXL is beneficial in corneal edema by reducing stromal swelling behavior and in keratitis by inhibiting pathogen growth. Despite these encouraging results, CXL remains a relatively new method that is potentially associated with complications. Aspects such as side effects and recurrence rates have still to be elucidated. In light of the growing interest in CXL, our paper summarizes present knowledge about this promising approach. We have intentionally endeavored to include the more relevant studies from the recent literature to provide an overview of the current status of CXL.


Pesticides and their degradates in groundwater reflect past use and current management strategies, Long Island, New York, USA.
Long Island, New York, has a mix of urban/suburban to agricultural/horticultural land use and nearly 3 million residents that rely on a sole-source aquifer for drinking water. The analysis of shallow groundwater (<40 m below land surface) collected from 54 monitoring wells across Long Island detected 53 pesticides or pesticide degradates. Maximum concentrations for individual pesticides or pesticide degradates ranged from 3 to 368,000 ng/L. The highest concentrations and most frequent pesticide detections occurred in samples collected from the pesticide management (PM) network, set in an agricultural/horticultural area in eastern Long Island with coordinated pesticide management by state and local agencies. The other two networks (Suffolk and Nassau/Queens) were set in suburban and urban areas, respectively, and had less frequent detections and lower pesticide concentrations than the PM network. Pesticide detections and concentration patterns (herbicide, insecticide, or fungicide) differed among the three networks revealing broad differences in land use. The predominance of fungicides metalaxyl, 1H-1,2,4-triazole (propiconazole/myclobutanil degradate), and 4-hydroxychlorothalonil (HCTL, chlorothalonil degradate) in samples from the PM network reflects their intensive use in agricultural settings. Total fungicide concentrations in the PM network ranged from <10 to >300,000 ng/L. The widespread detection of imidacloprid and triazine herbicides, simazine and atrazine, reveal a mixture of current and past use pesticides across the Long Island region. Low concentrations (<200 ng/L) of the triazines in the Suffolk and Nassau/Queens networks may reflect a change in land use and application. Acetanilide herbicides and aldicarb have been discontinued for 20 and 40 years, respectively, yet the concentrations of their degradates were among the highest observed in this study. Acetanilide (total concentrations up to 10,000 ng/L) and aldicarb degradates (up to 270 ng/L) are present in the PM network at much lower concentrations than previous Long Island studies and reflect changes in agricultural practices and pesticide management.