ICE COLD ERIC--International collaborative effort on chronic obstructive lung disease: exacerbation risk index cohorts--study protocol for an international COPD cohort study.
Chronic Obstructive Pulmonary Disease (COPD) is a systemic disease; morbidity and mortality due to COPD are on the increase, and it has great impact on patients' lives. Most COPD patients are managed by general practitioners (GP). Too often, GPs base their initial assessment of patient's disease severity mainly on lung function. However, lung function correlates poorly with COPD-specific health-related quality of life and exacerbation frequency. A validated COPD disease risk index that better represents the clinical manifestations of COPD and is feasible in primary care seems to be useful. The objective of this study is to develop and validate a practical COPD disease risk index that predicts the clinical course of COPD in primary care patients with GOLD stages 2-4. We will conduct 2 linked prospective cohort studies with COPD patients from GPs in Switzerland and the Netherlands. We will perform a baseline assessment including detailed patient history, questionnaires, lung function, history of exacerbations, measurement of exercise capacity and blood sampling. During the follow-up of at least 2 years, we will update the patients' profile by registering exacerbations, health-related quality of life and any changes in the use of medication. The primary outcome will be health-related quality of life. Secondary outcomes will be exacerbation frequency and mortality. Using multivariable regression analysis, we will identify the best combination of variables predicting these outcomes over one and two years and, depending on funding, even more years. Despite the diversity of clinical manifestations and available treatments, assessment and management today do not reflect the multifaceted character of the disease. This is in contrast to preventive cardiology where, nowadays, the treatment in primary care is based on patient-specific and fairly refined cardiovascular risk profile corresponding to differences in prognosis. After completion of this study, we will have a practical COPD-disease risk index that predicts the clinical course of COPD in primary care patients with GOLD stages 2-4. In a second step we will incorporate evidence-based treatment effects into this model, such that the instrument may guide physicians in selecting treatment based on the individual patients' prognosis. ClinicalTrials.gov Archive NCT00706602.


The generation of killer cells to trinitrophenyl-modified allogeneic targets by lymphocyte populations negatively selected to strong alloantigens.
Negatively selected mouse and rat lymphocyte populations, specifically deprived of alloreactivity to a particular major histocompatibility complex (MHC) haplotype, are nevertheless fully capable of responding to trinitrophenyl (TNP)-modified allogeneic stimulator cells and developing cytotoxic T-lymphocyte activity to TNP-altered allogeneic target cells. As for syngeneic systems, lytic expression of those responder killer cells also requires MHC identity between the target and stimulator cell populations. Such a finding argues strongly against two variations of the dual recognition hypothesis: like-like interactions and adaptive differentiation. Instead, these data favor either the altered self model or a third variation of the dual receptor model, where one of the relevent receptors is specific for the modifying antigen and the second is a low affinity receptor unable to be triggered in the absence of a modifying antigen.


Multiwavefront digital holographic television.
This paper presents the full technology chain supporting wide angle digital holographic television from holographic capture of real world objects/scenes to holographic display with an extended viewing angle. The data are captured with multiple CCD cameras located around an object. The display system is based on multiple tilted spatial light modulators (SLMs) arranged in a circular configuration. The capture-display system is linked by a holographic data processing module, which allows for significant decoupling of the capture and display systems. The presented experimental results, based on the reconstruction of real world, variable in time scenes, illustrates imaging dynamics, viewing angle and quality.


Raman Optical Activity Spectra from Density Functional Perturbation Theory and Density-Functional-Theory-Based Molecular Dynamics.
We describe the calculation of Raman optical activity (ROA) tensors from density functional perturbation theory, which has been implemented into the CP2K software package. Using the mixed Gaussian and plane waves method, ROA spectra are evaluated in the double-harmonic approximation. Moreover, an approach for the calculation of ROA spectra by means of density functional theory-based molecular dynamics is derived and used to obtain an ROA spectrum via time correlation functions, which paves the way for the calculation of ROA spectra taking into account anharmonicities and dynamic effects at ambient conditions.


M1 muscarinic agonists can modulate some of the hallmarks in Alzheimer's disease: implications in future therapy.
M1 muscarinic receptors (M1 mAChRs) play a role in an apparent linkage of three major hallmarks of Alzheimer's disease (AD): beta-amyloid (Abeta) peptide; tau hyperphosphorylation and paired helical filaments (PHFs); and loss of cholinergic function conducive to cognitive impairments. We evaluated the M1 muscarinic agonists AF102B (Cevimeline, EVOXAC trade mark : prescribed for Sjøgren's syndrome), AF150(S), and AF267B on some of these hallmarks of AD. Activation of M1 mAChRs with these agonists leads, inter alia, to enhanced secretion of amyloid precursor protein (alpha-APP), (via alpha-secretase activation), to decreased Abeta (via gamma-secretase inhibition), and to inhibition of Abeta- and/or oxidative stress-induced cell death. In several animal models mimicking different aspects of AD, these drugs restored cognitive impairments, and in select cases induced a decrease in brain Abeta elevation, with a high safety margin, following po administration. Notably, in mice with small hippocampi, unlike rivastigmine and nicotine, AF150(S) and AF267B restored cognitive impairments also on escape latency in a Morris water maze paradigm, in reversal learning. Studies from other labs showed that AF102B and talsaclidine (another M1 agonist) decreased cerbrospinal fluid (CSF) Abeta in AD patients following chronic treatment, being the first reported drugs with such a profile. The clinical significance of these studies remains to be elucidated, yet based on in vivo (rabbits) and in vitro studies (cell cultures), our M1 agonists can decrease brain Abeta, owing to a novel and dual complementary effect (e.g., inhibition of gamma-secretase and activation of alpha-secretase). Remarkably, although M1 agonists can decrease CSF Abeta in AD patients, an increased AD-type pathology in Parkinson's disease was recently been associated with chronic antimuscarinic treatment. In another aspect, these agonists decreased tau hyperphosphorylation in vitro and in vivo. Notably, nicotinic agonists or cholinesterase inhibitors increased tau hyperphosphorylation. In summary, the M1 agonists tested are effective on cognition and behavior and show unique disease-modifying properties owing to beneficial effects on major hallmarks of AD. This may place such drugs in the first line of modern AD therapies (e.g., beta- or gamma-secretase inhibitors, vaccines against Abeta, statins, and inhibitors of tau hyperphosphorylation).


Percutaneous transcatheter left atrial appendage exclusion in atrial fibrillation.
Stroke is one of the leading causes of mortality, morbidity and serious disability in the developed world. Atrial fibrillation (AF), one of the most common cardiac arrhythmias, is a well-known predisposing factor for stroke, raising the risk significantly. Oral anticoagulation with warfarin is currently the most effective therapy for stroke risk reduction; however, this therapy increases the risk of bleeding and is often underutilized, contraindicated, or when administered, often subtherapeutic. It has been documented that the left atrial appendage (LAA) is the main source of left atrial thrombus, especially in non-rheumatic AF. Therefore, LAA exclusion may reduce the risk of stroke in AF, and retrospective surgical data have demonstrated a reduced risk of embolic events if surgical LAA exclusion was also performed during mitral valve replacement. Recently, several less invasive percutaneous transcatheter techniques of LAA exclusion - the PLAATO device, the Watchman device, and the Amplatzer Septal Occluder - have been employed with initially encouraging results. There is currently an ongoing randomized trial comparing percutaneous LAA exclusion to long-term oral anticoagulation therapy. Until such data are available, however, oral anticoagulation should remain the standard of care for stroke prevention in patients with AF.


Secured medical imaging over the Internet.
The Internet has established itself as an affordable, extremely viable and ubiquitous communications network that can be easily accessed from virtually any point in the world. This makes it ideally suited for medical image communications. Issues regarding security and confidentiality of information on the Internet, however, need to be addressed for both occasional, individual users and consistent enterprise-wide users. In addition, the limited bandwidth of most Internet connections must be factored into the development of a realistic usermodel and resulting protocol. Open architecture issues must also be considered so that images can be communicated to recipients who do not have similar programs. Further, application-specific software is required to integrate image acquisition, encryption and transmission into a single, streamlined process. Using Photomailer software provided by PhysiTel Inc., the authors investigated the use of sending secured still images over the Internet. The scope of their investigation covered the use of the Internet for communicating images for consultation, referral, mentoring and education. Photomailer software was used at several local and remote sites. The program was used for both sending and receiving images. It was also used for sending images to recipients who did not have Photomailer, but instead relied on conventional email programs. The results of the investigation demonstrated that using products such as Photomailer, images could be quickly and easily communicated from one location to another via the Internet. In addition, the investigators were able to retrieve images off of their existing email accounts, thereby providing greater flexibility and convenience than other systems which require scheduled transmission of information on dedicated systems. We conclude that Photomailer and similar products may provide a significant benefit and improve communications among colleagues, providing an inexpensive means of sending secured images on the Internet.


Biological activities of Brucella abortus lipopolysaccharides.
Purified lipopolysaccharide (LPS) from smooth (s) and rough (R) strains of Brucella abortus and lipid A isolated from S-LPS by mild acid hydrolysis were examined in several assays of biological activity. Brucella S- and R-LPSs and Brucella lipid A activated the complement cascade. Previously reported mitogenic activation by Brucella LPSs of spleen cells from endotoxin-resistant C3H/HeJ mice was confirmed and also produced by isolated Brucella lipid A. Mitogenicity was not inhibited by polymyxin B, and amino acid analysis showed no binding of polymyxin B to Brucella LPS under conditions in which mitogenicity of phenol-water-extracted Escherichia coli LPS was inhibited. S and R Brucella LPSs and lipid A all produced equivalent polyclonal stimulation of C3H/HeJ and C3H/HeAU spleen cells. Crude and purified LPS from S but not from R B. abortus was toxic for outbred mice, with 50% lethal doses approximately six times greater than that for E. coli LPS. S- and R-LPSs were abortifacient in pregnant outbred mice. S Brucella LPS was lethal for carrageenen-pretreated C3H/HeJ and C3H/HeAU mice, whereas only C3H/HeAU mice were killed by E. coli LPS. The data are consistent with the hypothesis that the unique fatty acid composition of Brucella lipid A is responsible for its biological activity in endotoxin-resistant C3H/HeJ mice. The participation of the protein strongly bound to the lipid A cannot be excluded, but its mode of action, if any, is different from that of the lipid A-associated protein of enterobacterial LPS.


Stochastic impact evaluation of an irrigation development intervention in Northern Ethiopia.
Irrigation plays a significant role in achieving food and nutrition security in dry regions. However, detailed ex-ante appraisals of irrigation development investments are required to efficiently allocate resources and optimize returns on investment. Due to the inherent system complexity and uncertain consequences of irrigation development interventions coupled with limited data availability, deterministic cost-benefit analysis can be ineffective in guiding formal decision-making. Stochastic Impact Evaluation (SIE) helps to overcome the challenges of evaluating investments in such contexts. In this paper, we applied SIE to assess the viability of an irrigation dam construction project in northern Ethiopia. We used expert knowledge elicitation to generate a causal model of the planned intervention's impact pathway, including all identified benefits, costs and risks. Estimates of the input variables were collected from ten subject matter experts. We then applied the SIE tools: Monte Carlo simulation, Partial Least Squares regression, and Value of Information analysis to project prospective impacts of the project and identify critical knowledge gaps. Model results indicate that the proposed irrigation dam project is highly likely to increase the overall benefits and improve food and nutrition status of local farmers. However, the overall value of these benefits is unlikely to exceed the sum of the investment costs and negative externalities involved in the intervention. Simulation results suggest that the planned irrigation dam may improve income, as well as food and nutrition security, but would generate negative environmental effects and high investment costs. The Stochastic Impact Evaluation approach proved effective in this study and is likely to have potential for evaluating other agricultural development interventions that face system complexity, data scarcity and uncertainty.


Tidally controlled stick-slip discharge of a West Antarctic ice.
A major West Antarctic ice stream discharges by sudden and brief periods of very rapid motion paced by oceanic tidal oscillations of about 1 meter. Acceleration to speeds greater than 1 meter per hour and deceleration back to a stationary state occur in minutes or less. Slip propagates at approximately 88 meters per second, suggestive of a shear wave traveling within the subglacial till. A model of an episodically slipping friction-locked fault reproduces the observed quasi-periodic event timing, demonstrating an ice stream's ability to change speed rapidly and its extreme sensitivity to subglacial conditions and variations in sea level.


A critical analysis of alcohol hangover research methodology for surveys or studies of effects on cognition.
Alcohol hangover may be defined as an adverse effect of heavy alcohol consumption present after sufficient time has elapsed for the alcohol to have been eliminated from the blood. Understanding how hangover may impair performance is important for public safety; yet, there is relatively little hangover research. This paper outlines good practice for future studies. This paper presents a critical analysis of hangover methodology for surveys or studies of effects on cognition with human subjects and provides suggestions for optimum research practice for laboratory-based and naturalistic alcohol hangover studies. Four hangover symptom scales have been developed and subjected to psychometric testing. For retrospective assessment, we recommend the Hangover Symptoms Scale (HSS) or the Alcohol Hangover Severity Scale (AHSS). For concurrent assessment of hangover symptoms, we recommend either the Acute Hangover Scale (AHS), the five-item version of the HSS, or the AHSS. In research aiming to assess the cognitive effects of alcohol hangover, we suggest focusing on the cognitive domains of attention, memory and executive function, and we specify a number of tests within these cognitive domains that are likely to be sensitive to any decrements due to hangover. Finally, we argue that naturalistic studies should assess biological markers to improve the accuracy of estimates of alcohol consumption. Specifically, we recommend the assessment of ethyl glucuronide (EtG) for this purpose. Recommendations are made with respect to assessing hangover symptoms, cognitive effects of hangover and biological markers of alcohol consumption.


A commitment contract to achieve virologic suppression in poorly adherent patients with HIV/AIDS.
Assess whether a commitment contract informed by behavioral economics leads to persistent virologic suppression among HIV-positive patients with poor antiretroviral therapy (ART) adherence. Single-center pilot randomized clinical trial and a nonrandomized control group. Publicly funded HIV clinic in Atlanta, Georgia, USA. The study involved three arms. First, participants in the provider visit incentive (PVI) arm received $30 after attending each scheduled provider visit. Second, participants in the incentive choice arm were given a choice between the above arrangement and a commitment contract that made the $30 payment conditional on both attending the provider visit and meeting an ART adherence threshold. Third, the passive control arm received routine care and no incentives. A total of 110 HIV-infected adults with a recent plasma HIV-1 viral load more than 200 copies/ml despite ART. The sample sizes of the three groups were as follows: PVI, n = 21; incentive choice, n = 19; and passive control, n = 70. Virologic suppression (plasma HIV-1 viral load ≤200 copies/ml) at the end of the incentive period and at an unanticipated postincentive study visit approximately 3 months later. The odds of suppression were higher in the incentive choice arm than in the passive control arm at the postincentive visit (adjusted odds ratio 3.93, 95% confidence interval 1.19-13.04, P = 0.025). The differences relative to the passive control arm at the end of the incentive period and relative to the PVI arm at both points in time were not statistically significant. Commitment contracts can improve ART adherence and virologic suppression. ClinicalTrials.gov identifier NCT01455740.


Jumping Joints: The Complex Relationship Between Osteoarthritis and Jumping Mechanography.
We investigated the relationship between lower limb osteoarthritis (OA) and muscle strength and power (assessed by jumping mechanography) in UK community-dwelling older adults. We recruited 249 older adults (144 males, 105 females). OA was assessed clinically at the knee according to ACR criteria and radiographically, at the knee and hip, using Kellgren and Lawrence grading. Two-footed jumping tests were performed using a Leonardo Mechanography Ground Reaction Force Platform to assess maximum muscle force, power and Esslinger Fitness Index. Linear regression was used to assess the relationship between OA and jumping outcomes. Results are presented as β (95% confidence interval). The mean age of participants was 75.2 years (SD 2.6). Males had a significantly higher maximum relative power during lift off (mean 25.7 W/kg vs. 19.9 W/kg) and maximum total force during lift off (mean 21.0 N/kg vs. 19.1 N/kg) than females. In adjusted models, we found significant associations in males between clinical knee OA and maximum relative power [- 6.00 (CI - 9.10, - 2.94)] and Esslinger Fitness Index [- 19.3 (- 29.0, - 9.7)]. In females, radiographic knee OA was associated with total maximum power [- 2.0 (- 3.9, - 0.1)] and Esslinger Fitness Index [- 8.2 (- 15.9, - 0.4)]. No significant associations were observed for maximum total force. We observed significant negative associations between maximum relative power and Esslinger Fitness Index and clinical knee OA in males and radiographic knee OA in females. We have used novel methodology to demonstrate relationships between muscle function and OA in older adults.


Pushing the Scalability of RDF Engines on IoT Edge Devices.
Semantic interoperability for the Internet of Things (IoT) is enabled by standards and technologies from the Semantic Web. As recent research suggests a move towards decentralised IoT architectures, we have investigated the scalability and robustness of RDF (Resource Description Framework)engines that can be embedded throughout the architecture, in particular at edge nodes. RDF processing at the edge facilitates the deployment of semantic integration gateways closer to low-level devices. Our focus is on how to enable scalable and robust RDF engines that can operate on lightweight devices. In this paper, we have first carried out an empirical study of the scalability and behaviour of solutions for RDF data management on standard computing hardware that have been ported to run on lightweight devices at the network edge. The findings of our study shows that these RDF store solutions have several shortcomings on commodity ARM (Advanced RISC Machine) boards that are representative of IoT edge node hardware. Consequently, this has inspired us to introduce a lightweight RDF engine, which comprises an RDF storage and a SPARQL processor for lightweight edge devices, called RDF4Led. RDF4Led follows the RISC-style (Reduce Instruction Set Computer) design philosophy. The design constitutes a flash-aware storage structure, an indexing scheme, an alternative buffer management technique and a low-memory-footprint join algorithm that demonstrates improved scalability and robustness over competing solutions. With a significantly smaller memory footprint, we show that RDF4Led can handle 2 to 5 times more data than popular RDF engines such as Jena TDB (Tuple Database) and RDF4J, while consuming the same amount of memory. In particular, RDF4Led requires 10%-30% memory of its competitors to operate on datasets of up to 50 million triples. On memory-constrained ARM boards, it can perform faster updates and can scale better than Jena TDB and Virtuoso. Furthermore, we demonstrate considerably faster query operations than Jena TDB and RDF4J.


Oral administration of geraniol ameliorates acute experimental murine colitis by inhibiting pro-inflammatory cytokines and NF-κB signaling.
Ulcerative colitis is associated with a considerable reduction in the quality of life of patients. The use of phyto-ingredients is becoming an increasingly attractive approach for the management of colitis. Geraniol is a monoterpene with anti-inflammatory and antioxidative properties. In this study, we investigated the therapeutic potential of geraniol as a complementary and alternative medicine against dextran sulphate sodium (DSS)-induced ulcerative colitis in mice. Disease activity indices (DAI) comprising body weight loss, presence of occult blood and stool consistency were assessed for evaluation of colitis symptoms. Intestinal damage was assessed by evaluating colon length and its histology. Pre-treatment with geraniol significantly reduced the DAI score, improved stool consistency (without occult blood) and increased the colon length. The amount of pro-inflammatory cytokines, specifically TNF-α, IL-1β and IL-6 and the activity of myeloperoxidase in colon tissue were significantly decreased in geraniol pre-treated mice. Western blot analyses revealed that geraniol interfered with NF-κB signaling by inhibiting NF-κB (p65)-DNA binding, and IκBα phosphorylation, degradation and subsequent increase in nuclear translocation. Moreover, the expressions of downstream target pro-inflammatory enzymes such as iNOS and COX-2 were significantly reduced by geraniol. Pre-treatment with geraniol also restored the DSS-induced decline in antioxidant parameters such as reduced glutathione and superoxide dismutase activity and attenuated the increase in lipid peroxidation marker, thiobarbituric acid reactive substances and nitrative stress marker, nitrites in colon tissue. Thus, our results suggest that geraniol is a potential therapeutic agent for inflammatory bowel disease.


Deliberate hypotensive anesthesia with the rapidly acting, vascular-selective, L-type calcium channel antagonist-clevidipine: a case report.
Deliberate hypotension is an important technique for use in select anesthetics for procedures such as orthognathic surgery, specifically LeFort I maxillary osteotomy. We present a case report of an anesthetic involving deliberate hypotension for a 17-year-old female patient who presented for a LeFort I osteotomy, bilateral sagittal split of the mandible, and a genioplasty in order to correct a skeletal class III malocclusion. After reaching a steady-state general anesthetic, deliberate hypotension was induced solely with a bolus and subsequent continuous infusion of the ultrashort acting calcium channel blocker, clevidipine. The preoperative, intraoperative, and postoperative course and anesthetic management are discussed.


TMEM16A confers receptor-activated calcium-dependent chloride conductance.
Calcium (Ca(2+))-activated chloride channels are fundamental mediators in numerous physiological processes including transepithelial secretion, cardiac and neuronal excitation, sensory transduction, smooth muscle contraction and fertilization. Despite their physiological importance, their molecular identity has remained largely unknown. Here we show that transmembrane protein 16A (TMEM16A, which we also call anoctamin 1 (ANO1)) is a bona fide Ca(2+)-activated chloride channel that is activated by intracellular Ca(2+) and Ca(2+)-mobilizing stimuli. With eight putative transmembrane domains and no apparent similarity to previously characterized channels, ANO1 defines a new family of ionic channels. The biophysical properties as well as the pharmacological profile of ANO1 are in full agreement with native Ca(2+)-activated chloride currents. ANO1 is expressed in various secretory epithelia, the retina and sensory neurons. Furthermore, knockdown of mouse Ano1 markedly reduced native Ca(2+)-activated chloride currents as well as saliva production in mice. We conclude that ANO1 is a candidate Ca(2+)-activated chloride channel that mediates receptor-activated chloride currents in diverse physiological processes.


MRI-detectable changes in mouse brain structure induced by voluntary exercise.
Physical exercise, besides improving cognitive and mental health, is known to cause structural changes in the brain. Understanding the structural changes that occur with exercise as well as the neuroanatomical correlates of a predisposition for exercise is important for understanding human health. This study used high-resolution 3D MR imaging, in combination with deformation-based morphometry, to investigate the macroscopic changes in brain structure that occur in healthy adult mice following four weeks of voluntary exercise. We found that exercise induced changes in multiple brain structures that are involved in motor function and learning and memory including the hippocampus, dentate gyrus, stratum granulosum of the dentate gyrus, cingulate cortex, olivary complex, inferior cerebellar peduncle and regions of the cerebellum. In addition, a number of brain structures, including the hippocampus, striatum and pons, when measured on MRI prior to the start of exercise were highly predictive of subsequent exercise activity. Exercise tended to normalize these pre-existing differences between mice.


Natural Selection Towards Wild-Type in Composite Cross Populations of Winter Wheat.
Most of our crops are grown in monoculture with single genotypes grown over wide acreage. An alternative approach, where segregating populations are used as crops, is an exciting possibility, but outcomes of natural selection upon this type of crop are not well understood. We tracked allelic frequency changes in evolving composite cross populations of wheat grown over 10 generations under organic and conventional farming. At three generations, each population was genotyped with 19 SSR and 8 SNP markers. The latter were diagnostic for major functional genes. Gene diversity was constant at SSR markers but decreased over time for SNP markers. Population differentiation between the four locations could not be detected, suggesting that organic vs. non-organic crop management did not drive allele frequency changes. However, we did see changes for genes controlling plant height and phenology in all populations independently and consistently. We interpret these changes as the result of a consistent natural selection towards wild-type. Independent selection for alleles that are associated with plant height suggests that competition for light was central, resulting in the predominance of stronger intraspecific competitors, and highlighting a potential trade-off between individual and population performance.


Apolipoprotein epsilon3 allele is associated with persistent hepatitis C virus infection.
Host genetic factors may significantly influence the ability to clear hepatitis C virus (HCV) following infection. HCV is associated with very low density lipoproteins (VLDL) and low density lipoproteins (LDL) in the host's circulation. Apolipoprotein E (APOE) is found in VLDL and binds to potential receptors involved in HCV entry into cells, the LDL receptor, and the scavenger receptor protein SR-B1. The APOE gene is polymorphic with three alleles coding for three isoforms: Apo-epsilon2, Apo-epsilon3, and Apo-epsilon4. The aim of this study was to assess if these functional polymorphisms determine disease outcome in HCV infected individuals. The APOE genotype was determined in 420 Northern European patients with evidence of exposure to HCV. Genotype and allele distribution were compared with those of 288 healthy controls and progression of liver disease and viral clearance were analysed according to APOE allele status. The APOE*E2 and APOE*E4 alleles were both associated with a reduced likelihood of chronic infection (odds ratio (OR) 0.39 (95% confidence interval (CI) 0.211-0.728), p = 0.003; and OR 0.6 (95% CI 0.38-0.96), p = 0.032) and there was a notable absence of the E2E2 genotype in the HCV antibody positive group compared with the control population (p = 0.0067). Overall the genotypes carrying the E2 allele (E2,E3 and E2,E4) were associated with the equivalent of a 3-5-fold reduction in the risk of chronic HCV infection (genotype relative risk 0.36 and 0.20, respectively). This study indicates that functional APOE gene polymorphisms may be a determinant of outcome in HCV infection. We hypothesise that the E2 allele may protect against viral persistence via defective binding of HCV lipoviral particles to the cellular receptors involved in entry of these infectious particles.


Smoking Behavior Changes in the Early Rheumatoid Arthritis Period and Risk of Mortality During Thirty-Six Years of Prospective Followup.
To investigate whether rheumatoid arthritis (RA) diagnosis influences smoking behavior changes and whether these changes were associated with mortality. We identified an incident RA cohort in the Nurses' Health Study (NHS; 1976-2012). Behavioral data were collected through biennial questionnaires. We created a comparison cohort, matching RA cases to women without RA by age and calendar year at the index date of RA diagnosis. To investigate smoking behavior changes in the early RA period, sustained cessation was defined as permanently quitting within 4 years of the RA/index date. We used Cox regression to obtain hazard ratios (HRs) for mortality, comparing sustained smoking cessation to continued smoking. Among 121,700 women in the NHS, we identified 938 with incident RA matched to 8,951 non-RA comparators. Among current smokers, 40.0% with RA permanently quit smoking in the early RA period, compared to 36.1% of comparators (odds ratio for sustained cessation 1.18 [95% confidence interval (95% CI) 0.88, 1.58]). There were 313 deaths (33.4%) in the RA cohort and 2,042 (22.8%) among comparators. Compared to continued smoking, sustained cessation was associated with similarly decreased mortality in both the RA (HR 0.58 [95% CI 0.33, 1.01]) and comparison (HR 0.47 [95% CI 0.39, 0.58]) cohorts. Women with RA had higher mortality for >5 post-RA pack-years (HR 3.67 [95% CI 2.80, 4.81]) than comparators with >5 post-index pack-years (HR 1.88 [95% CI 1.62, 2.17]; P < 0.001 for interaction; reference: ever-smoker non-RA women with 0 post-index pack-years). Sustained smoking cessation within 4 years of RA diagnosis reduced mortality risk, with a similar effect observed among non-RA comparators. Smoking >5 pack-years after RA diagnosis significantly increased mortality beyond the risk of non-RA comparators.


Ciliary body toxicities of systemic oxcarbazepine and valproic acid treatments: electron microscopic study.
Ciliary body is responsible for humour aqueous production in posterior chamber. Valproic acid (VPA) has been widely used for the treatment of epilepsy and other neuropsychiatric diseases such as bipolar disease and major depression. Oxcarbazepine (OXC) is a new anti-epileptic agent that has been used recently for childhood epilepsies such as VPA. In this study, we aimed to investigate the effects of VPA and OXC treatments used as antiepileptic in ciliary body by electron microscopy. In our study, 40 Wistar rats (21 days old) were divided equally into four groups which were applied saline (group 1), VPA (group 2), OXC (group 3) and VPA + OXC (group 4). The as-prepared ocular tissues were characterized by transmission electron microscopy (TEM) technique in scanning and transmission electron microscopy (SEM-TEM) (Carl Zeiss EVO LS10). The results confirmed that VPA caused dense ciliary body degeneration. Additionally, ciliary body degeneration in group 4 was supposed to be due to VPA treatment. Ciliary body damage and secondary outcomes should be considered in patients with long-term VPA therapy.


Proliferation of murine midbrain neural stem cells depends upon an endogenous sonic hedgehog (Shh) source.
The Sonic Hedgehog (Shh) pathway is responsible for critical patterning events early in development and for regulating the delicate balance between proliferation and differentiation in the developing and adult vertebrate brain. Currently, our knowledge of the potential role of Shh in regulating neural stem cells (NSC) is largely derived from analyses of the mammalian forebrain, but for dorsal midbrain development it is mostly unknown. For a detailed understanding of the role of Shh pathway for midbrain development in vivo, we took advantage of mouse embryos with cell autonomously activated Hedgehog (Hh) signaling in a conditional Patched 1 (Ptc1) mutant mouse model. This animal model shows an extensive embryonic tectal hypertrophy as a result of Hh pathway activation. In order to reveal the cellular and molecular origin of this in vivo phenotype, we established a novel culture system to evaluate neurospheres (nsps) viability, proliferation and differentiation. By recreating the three-dimensional (3-D) microenvironment we highlight the pivotal role of endogenous Shh in maintaining the stem cell potential of tectal radial glial cells (RGC) and progenitors by modulating their Ptc1 expression. We demonstrate that during late embryogenesis Shh enhances proliferation of NSC, whereas blockage of endogenous Shh signaling using cyclopamine, a potent Hh pathway inhibitor, produces the opposite effect. We propose that canonical Shh signaling plays a central role in the control of NSC behavior in the developing dorsal midbrain by acting as a niche factor by partially mediating the response of NSC to epidermal growth factor (EGF) and fibroblast growth factor (FGF) signaling. We conclude that endogenous Shh signaling is a critical mechanism regulating the proliferation of stem cell lineages in the embryonic dorsal tissue.


The presence of hrp genes on the pathogenicity-associated plasmid of the tumorigenic bacterium Erwinia herbicola pv. gypsophilae.
The pathogenicity-associated plasmid (pPATH) of Erwinia herbicola pv. gypsophilae (Ehg), which is present only in pathogenic strains, contains a gene cluster encoding indole-3-acetic acid and cytokinin biosynthesis. The transposon-reporter Tn3-Spice was used to generate nonpathogenic mutants on two overlapping cosmids, pLA150 and pLA352, of the pPATH. A cluster of such mutations, which spanned 16 kb, mapped approximately 15 kb from the gene cluster involved in phytohormone biosynthesis. Non-pathogenic mutants also failed to elicit the hypersensitive reaction (HR) on tobacco. Pathogenicity and HR were restored concomitantly to these mutants by in trans complementation with wild-type Ehg DNA. A 3.8-kb HindIII DNA fragment that complemented the hrp mutants was sequenced and six complete and two partial open reading frames (ORFs) were identified. Comparison of the deduced amino acid sequences of the eight ORFs showed striking homology and co-linearity with hrp genes of E. amylovora as well as with other plant and mammalian pathogenic bacterial genes encoding proteins of the type III secretion system. Limited DNA sequencing at various sites on the remaining 11-kb region of pLA352 also showed high identity to Hrp proteins of E. amylovora, E. stewartii, and Pseudomonas syringae. These results suggest that hrp genes are mandatory for gall formation by E. herbicola pv. gypsophilae.


beta-cell function in obesity: effects of weight loss.
In nondiabetic subjects, obesity is associated with a modest expansion of beta-cell mass, possibly amounting-according to the best available estimates-to 10-30% for each 10 kg of weight excess. Whether age of onset and duration of obesity, recent changes in body weight, and body fat distribution have any effect on beta-cell mass in humans is unknown. Both fasting insulin secretion and the total insulin response to oral glucose have the following characteristics: 1) they increase with BMI in an approximately linear fashion, 2) both fat-free and fat mass are significant positive correlates, and 3) BMI exerts a positive effect separate from that of insulin resistance (i.e., obesity may be a state of primary insulin hypersecretion). The mechanisms are currently unknown, though chronic small increments in plasma glucose may play a role. In contrast, dynamic properties of beta-cell function, such as glucose sensitivity (i.e., dose-response function), rate sensitivity, and potentiation, do not appear to be substantially altered by the presence of obesity, body fat distribution, or insulin resistance as long as glucose tolerance is maintained. Weight loss, by diet or restrictive bariatric surgery, is associated with consensual decrements in insulin resistance and insulin hypersecretion. The latter, however, seems to be more persistent, suggesting that the postobese state may reproduce the primary insulin hypersecretion of the obese state. Malabsorptive bariatric surgery, in contrast, normalizes insulin sensitivity and abolishes insulin hypersecretion even before achievement of ideal body weight. Lipid-triggered messages from the gastrointestinal tract to the insulin target tissues and endocrine pancreas are the subject of intense investigation.


S-nitrosoglutathione reductase affords protection against pathogens in Arabidopsis, both locally and systemically.
Nitric oxide and S-nitrosothiols (SNOs) are widespread signaling molecules that regulate immunity in animals and plants. Levels of SNOs in vivo are controlled by nitric oxide synthesis (which in plants is achieved by different routes) and by S-nitrosoglutathione turnover, which is mainly performed by the S-nitrosoglutathione reductase (GSNOR). GSNOR is encoded by a single-copy gene in Arabidopsis (Arabidopsis thaliana; Martínez et al., 1996; Sakamoto et al., 2002). We report here that transgenic plants with decreased amounts of GSNOR (using antisense strategy) show enhanced basal resistance against Peronospora parasitica Noco2 (oomycete), which correlates with higher levels of intracellular SNOs and constitutive activation of the pathogenesis-related gene, PR-1. Moreover, systemic acquired resistance is impaired in plants overexpressing GSNOR and enhanced in the antisense plants, and this correlates with changes in the SNO content both in local and systemic leaves. We also show that GSNOR is localized in the phloem and, thus, could regulate systemic acquired resistance signal transport through the vascular system. Our data corroborate the data from other authors that GSNOR controls SNO in vivo levels, and shows that SNO content positively influences plant basal resistance and resistance-gene-mediated resistance as well. These data highlight GSNOR as an important and widely utilized component of resistance protein signaling networks conserved in animals and plants.


Deep anterior lamellar keratoplasty using precut anterior lamellar cap for herpes simplex keratitis: a long-term follow-up study.
To evaluate the feasibility, safety and efficacy of deep anterior lamellar keratoplasty (DALK) using precut anterior lamellar cap (ALC) for patients with herpes simplex keratitis. In this single-centre retrospective study, 48 consecutive patients with herpes simplex keratitis and having undergone DALK using either big bubble technique or manual lamellar dissection, between February 2009 and March 2010, were included. Each patient received DALK using either a precut ALC or a full-thickness stroma (FTS), and completed a minimum follow-up time of 36 months. There were no significant differences in the age, sex, eyes and sizes of trephination between two groups. Postoperative best spectacle-corrected visual acuity of 0.3 logarithm of the minimum angle of resolution or better at the last follow-up visit was achieved in 74.1% of eyes in the FTS group and 71.4% in the ALC group (p=0.84). The mean central corneal thickness and corneal volume were significantly higher in the FTS group (550.7 ± 34.3 µm and 61.21 ± 4.12 mm(3)) than the ALC group (393 ± 45.7 µm and 54.68 ± 4.55 mm(3); p<0.0001). The mean simulated keratometry value was lower in the ALC group as compared with the FTS group (p=0.03; 44.51 ± 2.60 vs 46.06 ± 2.31). There was no difference in biomechanical behaviour of two types of corneal tissues after DALK. Corneal curvature remained stable in the ALC group, and no signs of corneal ectasia were observed during follow-ups. DALK using precut ALC should be considered as a selective surgical approach. A postoperative subnormal central corneal thickness of about 400 µm would not influence corneal curvature and refractive status.


The Epstein-Barr virus (EBV) BZLF2 gene product associates with the gH and gL homologs of EBV and carries an epitope critical to infection of B cells but not of epithelial cells.
Glycoprotein gp85, the product of the BXLF2 open reading frame (ORF), is the gH homolog of Epstein-Barr virus (EBV) and has been implicated in penetration of virus into B cells. Like its counterparts in other herpesviruses, it associates with a gL homolog, gp25, which is the product of the BKRF2 ORF. Unlike the gH homologs of other herpesviruses, however, gp85 also complexes with two additional glycoproteins of 42 and 38 kDa. Glycoproteins gp42 and gp38 were determined to be alternatively processed forms of the BZLF2 gene product. Coexpression of EBV gH and gL facilitated transport of gH to the cell surface and resulted in formation of a stable complex of gH and gL. It also restored expression of an epitope recognized by monoclonal antibody E1D1, which immunoprecipitates the native gH complex but not recombinant gH expressed in isolation. Coexpression of gH, gL, and the BZLF2 ORF restored expression of an epitope recognized by a second monoclonal antibody, F-2-1, which immunoprecipitates the native gH-gL-gp42/38 complex but not the complex of recombinant gH and gL alone. The epitope recognized by antibody F-2-1 was mapped to the BZLF2 gene product itself. Antibody F-2-1 inhibited the ability of EBV to infect B lymphocytes but had no effect on the ability of the virus to infect the epithelial cell line SVK-CR2. In contrast, antibody E1D1 had no effect on infection of the B-cell line but inhibited infection of the epithelial cell line. These results indicate that penetration of the two cell types by EBV involves differential use of the gH-gL-gp42/38 complex and suggest the hypothesis that the BZLF2 gene product has evolved as a unique adaptation to infection of B lymphocytes by EBV.


Anterior face height values in a nigerian population.
Withan increasing demand in the number of patients seeking orthodontictreatment at the Orthodontic unit of University of Benin teaching hospital it becomes imperative to provide normative values for anterior face heights. These values will then form a basis for clinical diagnosis, treatment planning and evaluation of facial proportions following orthodontic treatments. The aim of this study was to measure anterior face height proportions of school children in Benin City. One hundred Lateral cephalometric radiographs of 12-15 years old school children, in Benin City were taken to establish anterior face height values. Manual tracings of the cephalograms were carried out and the linear measurements were recorded, summarized, and statistically analyzed using the Microsoft Excel Programme 2000. The comparative test was conducted using the Student's t-test to demonstrate any statistically significant difference between the values for the males and females, at 95% confidence level P < 0.05, was regarded as significant. Out of the 100 subjects who participated in the study, there were 40 males 40% (40/100) and 60 females 60% (60/100), with a mean age of 12.2 years. Following a combined data analysis for both males and females, the mean biological values obtained for the parameters investigated are; Anterior Lower Face Height (ALFH) 60.9 (5) mm, Anterior Upper Face Height 47.7 (4) mm, Anterior Total Face Height (ATFH) 108.5 (5) mm, ratio of ALFH to ATFH ALFH: ATFH 56 (4)%. This study provides anterior face height measurements, which will be of great significance in evaluating facial proportions andesthetics in orthodontics, orthognathic surgery, and prosthetic dentistry.


Growth modes and chiral selectivity of single-walled carbon nanotubes.
Chemical vapor deposition synthesis of single-walled carbon nanotubes, using an Fe catalyst, and alternating methane and carbon monoxide as carbon feedstocks, leads to the reversible formation of junctions between tubes of different diameters. Combined with an atomistic modeling of the tube/catalyst interface, this shows that the ratio of diameters of the tube and its seeding particle, denoting the growth mode, depends on the carbon fraction inside the catalyst. With carbon monoxide, nanoparticles are strongly carbon enriched, and tend to dewet the tube, in a perpendicular growth mode. Cross-checking our results with the available reports from the literature of the last decade strongly suggests that these latter conditions should favor the near armchair chiral selectivity observed empirically.


Cytoimmunotherapy for persistent virus infection reveals a unique clearance pattern from the central nervous system.
The mechanism(s) by which infectious or malignant material is cleared by the host has long been an area of intensive study. We have used the murine model of infection with lymphocytic choriomeningitis virus (LCMV) to look at immune clearance during persistent infection. LCMV was selected because the mouse is its natural host, it easily induces acute or persistent infection in vivo, and the mechanism by which it is cleared in vivo during acute infection is now well understood. Clearance, although associated with several antiviral immune effector mechanisms, is primarily dependent on the activity of virus-specific cytotoxic T lymphocytes (CTL) restricted by H-2 molecules of the mouse major histocompatibility complex (MHC). If these cells fail to generate or are depleted, progression from acute to persistent infection occurs. Here, using molecular probes, we show that viral nucleic acid sequences, viral proteins and infectious materials can be efficiently and effectively cleared by adoptive transfer of antiviral H-2-restricted lymphocytes bearing the Lyt 2+ phenotype. Viral materials are cleared from a wide variety of tissues and organs where they normally lodge during persistent infection. Unexpectedly, the mode by which viral materials are removed from the central nervous system (CNS) differed markedly from the mechanism of clearance occurring at other sites. These observations indicate the possible use of adoptive lymphocyte therapy for treatment of persistent infections and suggest that immune clearance of products from the CNS probably occurs by a process distinct from those in other organs.


Therapeutic significance of penicillin tolerance in experimental streptococcal endocarditis.
Tolerance to penicillin exists among the viridans group of streptococci, but its therapeutic significance is unknown. We studied the effect of penicillin alone and in combination with streptomycin, in vivo and in vitro, on three strains of dextran-producing Streptococcus sanguis serotype II which possess widely various degrees of penicillin tolerance. In rabbits with experimental endocarditis, treatment with procaine penicillin (250 mg/kg intramuscularly twice daily for 5 days) decreased the number of viable organisms in valvular vegetations from 8.82 log10 +/- 0.98 CFU/g in untreated controls to 5.31 +/- 1.19 for a highly tolerant strain, 4.22 +/- 1.05 for a less tolerant strain, and 1.79 +/- 1.72 for a nontolerant strain (P less than or equal to 0.01 for comparison between any of the four groups). None of 36 rabbits infected with tolerant strains were cured by 5 days of treatment with penicillin, but 10 of 23 animals infected with the nontolerant strain were cured (P = 0.00002). When streptomycin was given in combination with penicillin, rabbits infected with the nontolerant strain were cured within 3 days, and rabbits infected with the tolerant strain were cured within 5 days. These findings indicate that tolerance can exert a critical influence on the response of S. sanguis to penicillin therapy in vivo and that the combination of penicillin plus streptomycin exerts a synergistic effect against tolerant as well as nontolerant organisms.


Losartan hydroxylation phenotype in an Ecuadorian population: influence of CYP2C9 genetic polymorphism, habits and gender.
To describe for the first time CYP2C9 hydroxylation phenotype with CYP2C9 genotypes in a Hispanic (Ecuadorian) population using losartan; and the relevance of gender, tobacco, ethanol and caffeine consumption on the enzyme hydroxylation capacity. Ecuadorian healthy volunteers (n = 194) received a single oral dose of 25 mg losartan. Losartan metabolic ratio was defined as losartan:E3174 concentration. CYP2C9 alleles *2, *3, *4, *5 and *6 were analyzed. No phenotypically poor metabolizers were found. The metabolic ratio (mean ± standard deviation) was higher (p < 0.05) in CYP2C9*1/*3 carriers (12.4 ± 13.8; n = 6) versus CYP2C9*1/*1 (4.9 ± 7.0; n = 167), as well as in females versus males (6.72 ± 9.72 and 3.76 ± 4.48, respectively; p < 0.05). Only the following genotypes, CYP2C9*1/*1, CYP2C9*1/*2 and CYP2C9*1/*3, were found with a frequency of 86.1%, 10.8% and 3.1%, respectively. Despite the mean metabolic ratio being higher in this population than in others previously studied across genotypes, no poor metabolizers, either phenotypically or genotypically, were found.


Multiplexed short tandem repeat polymorphisms of the Weber 8A set of markers using tailed primers and infrared fluorescence detection.
Short tandem repeat polymorphism (STRP) markers have become important reagents for mapping genetic diseases. These markers are available as screening sets, which are located in all chromosomes at discrete intervals, allowing the entire genome to be analyzed. Mapping studies that include many individuals in the analysis necessitate the production of large numbers of genotypes. In an effort to increase the efficiency and lower the cost of using these STRP screening sets, we have divided the amplification primers of the Weber 8A screening set into groups that can be amplified in single polymerase chain reaction (PCR) amplification reactions, resulting in a reduction of both time and cost. Fluorescently-labeled amplification products were produced using a three primer reaction. The forward STRP amplification primer for each marker contained a 19 bp sequence at the 5' end. A fluorescently-labeled primer, with a sequence identical to the 19 bp tail, was added to the amplification reaction as the sole source of fluorescent label. The STRP banding pattern is detected using an automated fluorescent DNA sequencer. Use of this multiplexed genomic screening set should greatly enhance the mapping of human disease loci.


The burden of disease associated with being African-American in the United States and the contribution of socio-economic status.
The burden of disease associated with being African-American in the US, and the contribution of socio-economic status (SES) to that burden have not been quantified. We derived burden of disease estimates for African-Americans and whites by age group, with and without adjustment for SES (income and education). We used (1) EQ-5D Index scores from the 2000 US Medical Expenditure Panel Survey to derive quality-adjusted life year (QALY) compatible estimates of health-related quality of life (HRQL); (2) 1990-1992 US National Health Interview Survey data linked to National Death Index data through 1995 to derive mortality risks; and (3) 2000 US mortality data from the National Center for Health Statistics to derive current mortality estimates for the US population. We found that relative to whites, African-Americans suffer 67,000 more deaths annually, resulting in 2.2 million years of life lost, and 1.1 million years after SES adjustment. Total QALYs lost (HRQL and mortality) dropped from 2.3 million to 902,000 after SES adjustment. SES differences between African-Americans and whites appear to explain all the HRQL disparity but only half the mortality disparity. Better understanding of the disparate effects of SES may inform interventions to address health disparities adversely affecting African-Americans.


Structures of the Streptococcus sanguinis SrpA Binding Region with Human Sialoglycans Suggest Features of the Physiological Ligand.
Streptococcus sanguinis is a leading cause of bacterial infective endocarditis, a life-threatening infection of heart valves. S. sanguinis binds to human platelets with high avidity, and this adherence is likely to enhance virulence. Previous studies suggest that a serine-rich repeat adhesin termed SrpA mediates the binding of S. sanguinis to human platelets via its interaction with sialoglycans on the receptor GPIbα. However, in vitro binding assays with SrpA and defined sialoglycans failed to identify specific high-affinity ligands. To improve our understanding of the interaction between SrpA and human platelets, we determined cocrystal structures of the SrpA sialoglycan binding region (SrpABR) with five low-affinity ligands: three sialylated trisaccharides (sialyl-T antigen, 3'-sialyllactose, and 3'-sialyl-N-acetyllactosamine), a sialylated tetrasaccharide (sialyl-LewisX), and a sialyl galactose disaccharide component common to these sialoglyans. We then combined structural analysis with mutagenesis to further determine whether our observed interactions between SrpABR and glycans are important for binding to platelets and to better map the binding site for the physiological receptor. We found that the sialoglycan binding site of SrpABR is significantly larger than the sialoglycans cocrystallized in this study, which suggests that binding of SrpA to platelets either is multivalent or occurs via a larger, disialylated glycan.


Dealing with the health state 'dead' when using discrete choice experiments to obtain values for EQ-5D-5L heath states.
To evaluate two different methods to obtain a dead (0)--full health (1) scale for EQ-5D-5L valuation studies when using discrete choice (DC) modeling. The study was carried out among 400 respondents from Barcelona who were representative of the Spanish population in terms of age, sex, and level of education. The DC design included 50 pairs of health states in five blocks. Participants were forced to choose between two EQ-5D-5L states (A and B). Two extra questions concerned whether A and B were considered worse than dead. Each participant performed ten choice exercises. In addition, values were collected using lead-time trade-off (lead-time TTO), for which 100 states in ten blocks were selected. Each participant performed five lead-time TTO exercises. These consisted of DC models offering the health state 'dead' as one of the choices--for which all participants' responses were used (DCdead)--and a model that included only the responses of participants who chose at least one state as worse than dead (WTD) (DCWTD). The study also estimated DC models rescaled with lead-time TTO data and a lead-time TTO linear model. The DC(dead) and DCWTD models produced relatively similar results, although the coefficients in the DCdead model were slightly lower. The DC model rescaled with lead-time TTO data produced higher utility decrements. Lead-time TTO produced the highest utility decrements. The incorporation of the state 'dead' in the DC models produces results in concordance with DC models that do not include 'dead'.


Syphilis increases HIV viral load and decreases CD4 cell counts in HIV-infected patients with new syphilis infections.
Syphilitic ulcers are known to facilitate the transmission of HIV infection, but the effect of syphilis infection on HIV viral loads and CD4 cell counts is poorly understood. We abstracted medical records for HIV-infected male syphilis patients seen at three clinics in San Francisco and Los Angeles from January 2001 to April 2003. We compared plasma HIV-RNA levels and CD4 cell counts during syphilis infection with those before syphilis infection and after syphilis treatment, using the Wilcoxon signed rank test. Fifty-two HIV-infected men with primary or secondary syphilis had HIV viral load and CD4 cell count data available for analysis; 30 (58%) were receiving antiretroviral therapy. Viral loads were higher during syphilis compared with pre-syphilis levels by a mean of 0.22 RNA log10 copies/ml (P = 0.02) and were lower by a mean of -0.10 RNA log10 copies/ml (P = 0.52) after syphilis treatment. CD4 cell counts were lower during syphilis infection than before by a mean of -62 cells/mm3 (P = 0.04), and were higher by a mean of 33 cells/mm3 (P = 0.23) after syphilis treatment. Increases in the HIV viral load and reductions in the CD4 cell count were most substantial in men with secondary syphilis and those not receiving antiretroviral therapy. Syphilis infection was associated with significant increases in the HIV viral load and significant decreases in the CD4 cell count. The findings underscore the importance of preventing and promptly treating syphilis in HIV-infected individuals.


Whole-genome deep-learning analysis identifies contribution of noncoding mutations to autism risk.
We address the challenge of detecting the contribution of noncoding mutations to disease with a deep-learning-based framework that predicts the specific regulatory effects and the deleterious impact of genetic variants. Applying this framework to 1,790 autism spectrum disorder (ASD) simplex families reveals a role in disease for noncoding mutations-ASD probands harbor both transcriptional- and post-transcriptional-regulation-disrupting de novo mutations of significantly higher functional impact than those in unaffected siblings. Further analysis suggests involvement of noncoding mutations in synaptic transmission and neuronal development and, taken together with previous studies, reveals a convergent genetic landscape of coding and noncoding mutations in ASD. We demonstrate that sequences carrying prioritized mutations identified in probands possess allele-specific regulatory activity, and we highlight a link between noncoding mutations and heterogeneity in the IQ of ASD probands. Our predictive genomics framework illuminates the role of noncoding mutations in ASD and prioritizes mutations with high impact for further study, and is broadly applicable to complex human diseases.


How Shall I Count the Ways? A Method for Quantifying the Qualitative Aspects of Unscripted Movement With Laban Movement Analysis.
There is significant clinical evidence showing that creative and expressive movement processes involved in dance/movement therapy (DMT) enhance psycho-social well-being. Yet, because movement is a complex phenomenon, statistically validating which aspects of movement change during interventions or lead to significant positive therapeutic outcomes is challenging because movement has multiple, overlapping variables appearing in unique patterns in different individuals and situations. One factor contributing to the therapeutic effects of DMT is movement's effect on clients' emotional states. Our previous study identified sets of movement variables which, when executed, enhanced specific emotions. In this paper, we describe how we selected movement variables for statistical analysis in that study, using a multi-stage methodology to identify, reduce, code, and quantify the multitude of variables present in unscripted movement. We suggest a set of procedures for using Laban Movement Analysis (LMA)-described movement variables as research data. Our study used LMA, an internationally accepted comprehensive system for movement analysis, and a primary DMT clinical assessment tool for describing movement. We began with Davis's (1970) three-stepped protocol for analyzing movement patterns and identifying the most important variables: (1) We repeatedly observed video samples of validated (Atkinson et al., 2004) emotional expressions to identify prevalent movement variables, eliminating variables appearing minimally or absent. (2) We use the criteria repetition, frequency, duration and emphasis to eliminate additional variables. (3) For each emotion, we analyzed motor expression variations to discover how variables cluster: first, by observing ten movement samples of each emotion to identify variables common to all samples; second, by qualitative analysis of the two best-recognized samples to determine if phrasing, duration or relationship among variables was significant. We added three new steps to this protocol: (4) we created Motifs (LMA symbols) combining movement variables extracted in steps 1-3; (5) we asked participants in the pilot study to move these combinations and quantify their emotional experience. Based on the results of the pilot study, we eliminated more variables; (6) we quantified the remaining variables' prevalence in each Motif for statistical analysis that examined which variables enhanced each emotion. We posit that our method successfully quantified unscripted movement data for statistical analysis.


The Surgical Management of Small Bowel Neuroendocrine Tumors: Consensus Guidelines of the North American Neuroendocrine Tumor Society.
Small bowel neuroendocrine tumors (SBNETs) have been increasing in frequency over the past decades, and are now the most common type of small bowel tumor. Consequently, general surgeons and surgical oncologists are seeing more patients with SBNETs in their practices than ever before. The management of these patients is often complex, owing to their secretion of hormones, frequent presentation with advanced disease, and difficulties with making the diagnosis of SBNETs. Despite these issues, even patients with advanced disease can have long-term survival. There are a number of scenarios which commonly arise in SBNET patients where it is difficult to determine the optimal management from the published data. To address these challenges for clinicians, a consensus conference was held assembling experts in the field to review and discuss the available literature and patterns of practice pertaining to specific management issues. This paper summarizes the important elements from these studies and the recommendations of the group for these questions regarding the management of SBNET patients.


Application of statistical experimental design for optimisation of bioinsecticides production by sporeless Bacillus thuringiensis strain on cheap medium.
In order to overproduce bioinsecticides production by a sporeless Bacillus thuringiensis strain, an optimal composition of a cheap medium was defined using a response surface methodology. In a first step, a Plackett-Burman design used to evaluate the effects of eight medium components on delta-endotoxin production showed that starch, soya bean and sodium chloride exhibited significant effects on bioinsecticides production. In a second step, these parameters were selected for further optimisation by central composite design. The obtained results revealed that the optimum culture medium for delta-endotoxin production consists of 30 g L(-1) starch, 30 g L(-1) soya bean and 9 g L(-1) sodium chloride. When compared to the basal production medium, an improvement in delta-endotoxin production up to 50% was noted. Moreover, relative toxin yield of sporeless Bacillus thuringiensis S22 was improved markedly by using optimised cheap medium (148.5 mg delta-endotoxins per g starch) when compared to the yield obtained in the basal medium (94.46 mg delta-endotoxins per g starch). Therefore, the use of optimised culture cheap medium appeared to be a good alternative for a low cost production of sporeless Bacillus thuringiensis bioinsecticides at industrial scale which is of great importance in practical point of view.


Hyperbaric oxygen therapy to treat diabetes impaired wound healing in rats.
Wound healing in diabetes is frequently impaired and its treatment remains a challenge. Hyperbaric oxygen therapy (HBOT) receives a wide attendance and is often used as a last resort treatment option, however, its effectiveness for many conditions is unproven. We tested the effect of HBOT on healing of diabetic ulcers in an animal experimental setting. Experimental diabetes was induced by intraperitoneal injection of streptozotocin. Four weeks after diabetes induction, rats were ulcerated by clamping a pair of magnet disks on the dorsal skin for 16 h. After magnet removal, the animals received HBOT, daily on weekdays, for 4 weeks. To examine the effect of HBOT on diabetes impaired wound healing, the degree of wound tissue perfusion, inflammation, angiogenesis, and tissue breaking strength were evaluated. HBOT effects on the degree of inflammation and number of blood vessels could not be observed. HBOT improved the tissue breaking strength of the wound, however, this did not reach statistical significance. Twenty hours after ending the HBOT, a significantly improved oxygen saturation of the hemoglobin at the venous end of the capillaries and the quantity of hemoglobin in the micro-blood vessels was measured.


Genes necessary for C. elegans cell and growth cone migrations.
The migrations of cells and growth cones contribute to form and pattern during metazoan development. To study the mechanisms that regulate cell motility, we have screened for C. elegans mutants defective in the posteriorly directed migrations of the canal-associated neurons (CANs). Here we describe 14 genes necessary for CAN cell migration. Our characterization of the mutants has led to three conclusions. First, the mutations define three gene classes: genes necessary for cell fate specification, genes necessary for multiple cell migrations and a single gene necessary for final positioning of migrating cells. Second, cell interactions between the CAN and HSN, a neuron that migrates anteriorly to a position adjacent to the CAN, control the final destination of the HSN cell body. Third, C. elegans larval development requires the CANs. In the absence of CAN function, larvae arrest development, with excess fluid accumulating in their pseudocoeloms. This phenotype may reflect a role of the CANs in osmoregulation.


Water aging reverses residual stresses in hydrophilic dental composites.
Dental composites develop residual stresses during polymerization due to shrinkage. These stresses may change with time because of relaxation and water sorption in the oral environment. This phenomenon is likely dependent on the composition of the materials, specifically their hydrophilic characteristics, and could result in deleterious stresses on restorative materials and tooth structure. The purpose of this experiment was to use the thin ring-slitting method to compare the residual stress generated within composite materials of varying hydrophilicity when aged in wet and dry conditions after polymerization. Water sorption, solubility, elastic modulus, and residual stresses were measured in 6 commercial composites/cements aged in water and dry conditions. The self-adhesive resin cement showed the highest water sorption and solubility. All composites showed initial residual contraction stresses, which were maintained when aged dry. Residual stresses in 2 of the self-adhesive cements and the polyacid-modified composite aged in wet conditions resulted in a net expansion. This experiment verified that residual shrinkage stresses in dental composites can be reversed during aging in water, resulting in a net expansion, with the effect directly related to their hydrophilic properties.


Presence of plicidentine in the oral teeth of the coelacanth Latimeria chalumnae Smith 1939 (Sarcopterygii; Actinistia).
The extant coelacanth Latimeria is a sarcopterygian predatory fish with caniniform teeth on its upper and lower jaws. The teeth are constituted of a cone of dentine with an apical cap of enamel, and they are fixed to the osseous component of the jaws by an attachment bone. Internal walls of the tooth base show folds that have been firstly interpreted in the past as radial vascular canals. Three-dimensional visualisation of these foldings using X-ray tomographic techniques and new histological interpretation lead to reconsider these structures as true plicidentine. The folds of the dentine do not invade the whole pulp cavity of the tooth contrary to the plicated condition of most fossil sarcopterygian fishes (e.g., Eusthenopteron, Porolepis, Megalichthys) certain fossil marine reptiles (ichthyosaurs) and extant varanids; in Latimeria they are limited to the lower third to the half of the pulp cavity. The presence of plicidentine in Latimeria's teeth is proposed to be a plesiomorphic character for sarcopterygians.


Mating-type switching by chromosomal inversion in methylotrophic yeasts suggests an origin for the three-locus Saccharomyces cerevisiae system.
Saccharomyces cerevisiae has a complex system for switching the mating type of haploid cells, requiring the genome to have three mating-type (MAT)-like loci and a mechanism for silencing two of them. How this system originated is unknown, because the three-locus system is present throughout the family Saccharomycetaceae, whereas species in the sister Candida clade have only one locus and do not switch. Here we show that yeasts in a third clade, the methylotrophs, have a simpler two-locus switching system based on reversible inversion of a section of chromosome with MATa genes at one end and MATalpha genes at the other end. In Hansenula polymorpha the 19-kb invertible region lies beside a centromere so that, depending on the orientation, either MATa or MATalpha is silenced by centromeric chromatin. In Pichia pastoris, the orientation of a 138-kb invertible region puts either MATa or MATalpha beside a telomere and represses transcription of MATa2 or MATalpha2. Both species are homothallic, and inversion of their MAT regions can be induced by crossing two strains of the same mating type. The three-locus system of S. cerevisiae, which uses a nonconservative mechanism to replace DNA at MAT, likely evolved from a conservative two-locus system that swapped genes between expression and nonexpression sites by inversion. The increasing complexity of the switching apparatus, with three loci, donor bias, and cell lineage tracking, can be explained by continuous selection to increase sporulation ability in young colonies. Our results provide an evolutionary context for the diversity of switching and silencing mechanisms.


SPT4, SPT5 and SPT6 interactions: effects on transcription and viability in Saccharomyces cerevisiae.
The SPT4, SPT5 and SPT6 genes of Saccharomyces cerevisiae were identified originally by mutations that suppress delta insertion mutations at HIS4 and LYS2. Subsequent analysis has demonstrated that spt4, spt5 and spt6 mutations confer similar pleiotropic phenotypes. They suppress delta insertion mutations by altering transcription and are believed to be required for normal transcription of several other loci. We have now analyzed interactions between SPT4, SPT5 and SPT6. First, the combination of mutations in any two of these three genes causes lethality in haploids. Second, some recessive mutations in different members of this set fail to complement each other. Third, mutations in all three genes alter transcription in similar ways. Finally, the results of coimmunoprecipitation experiments demonstrate that at least the SPT5 and SPT6 proteins interact physically. Taken together, these genetic and biochemical results indicate that SPT4, SPT5 and SPT6 function together in a transcriptional process that is essential for viability in yeast.


Novel DNA methylation sites associated with cigarette smoking among African Americans.
Cigarette smoking has been associated with adverse health outcomes for mothers and children and is a major contributor to heart disease. Although cigarette smoking is known to affect the epigenome, few studies have been done in African American populations. In this study, we investigated the association between cigarette smoking and DNA methylation (DNAm) among African Americans from the Intergenerational Impact of Genetic and Psychological Factors on Blood Pressure Study (InterGEN), and the Genetic Epidemiology Network of Arteriopathy (GENOA). The InterGEN study aims to examine the effects of genetic and psychological factors on blood pressure among African American women and their children. Current cigarette smoking was assessed at baseline. DNAm of saliva was assessed using the 850K EPIC Illumina BeadChip for Epigenome-Wide Association analyses. A replication study was conducted among 1100 participants in the GENOA study using the same BeadChip. After controlling for age, body mass index, population structure and cell composition, 26 epigenome-wide significant sites (FDR q < 0.05) were identified, including the AHRR and PHF14 genes associated with atherosclerosis and lung disease, respectively. Six novel CpG sites were discovered in the InterGEN sample and replicated in the GENOA sample. Genes mapped include RARA, FSIP1, ALPP, PIK3R5, KIAA0087, and MGAT3, which were largely associated with cancer development. We observed significant epigenetic associations between smoking and disease-associated genes (e.g., cardiovascular disease, lung cancer). Six novel CpG sites were identified and replicated across saliva and blood samples.


Reflections on a new medical cosmology.
Since the nineteenth century the theory and practice of mainstream Western medicine has been grounded in the biomedical model. In the later years of the twentieth century, however, it has faced a range of serious problems, which when viewed collectively, remain unresolved despite a variety of responses. The question we now face is whether these problems can be dealt with by modifying and extending the principles underlying the biomedical model, or whether a more radical solution is required. Recent critiques of Western medicine have focused mainly on the biopsychosocial model in relation to the former approach, but it will be contended that this cannot deal adequately with the challenges that medicine currently faces, because although it addresses both the scientific and humanistic aspects of medicine it fails to harmonise them. I shall therefore argue for the necessity of a more radical approach, and suggest that what is required to accomplish this is the development of a new medical cosmology, rooted in an older and more global framework. Such a fundamental change would inevitably involve a long term process which it is not yet possible to fully comprehend let alone specify in detail. Some of the necessary features of such a new medical cosmology can, however, already be distinguished and the outline of these is described.


Yeast genome sequencing: the power of comparative genomics.
For decades, unicellular yeasts have been general models to help understand the eukaryotic cell and also our own biology. Recently, over a dozen yeast genomes have been sequenced, providing the basis to resolve several complex biological questions. Analysis of the novel sequence data has shown that the minimum number of genes from each species that need to be compared to produce a reliable phylogeny is about 20. Yeast has also become an attractive model to study speciation in eukaryotes, especially to understand molecular mechanisms behind the establishment of reproductive isolation. Comparison of closely related species helps in gene annotation and to answer how many genes there really are within the genomes. Analysis of non-coding regions among closely related species has provided an example of how to determine novel gene regulatory sequences, which were previously difficult to analyse because they are short and degenerate and occupy different positions. Comparative genomics helps to understand the origin of yeasts and points out crucial molecular events in yeast evolutionary history, such as whole-genome duplication and horizontal gene transfer(s). In addition, the accumulating sequence data provide the background to use more yeast species in model studies, to combat pathogens and for efficient manipulation of industrial strains.


Inhibition of herpes simplex virus replication by purine deoxyribonucleosides.
The inhibition of herpes simplex virus (HSV) replication by 2'-deoxyadenosine (dAdo) is greatly potentiated by the presence of the inhibitor of adenosine deaminase, erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA). HSV replication is inhibited by dAdo [in the presence of EHNA] or by 2'-deoxyguanosine (dGuo) at concentrations slightly lower than are necessary to inhibit growth of uninfected HeLa cells. Under conditions where virus replication is inhibited by greater than 99% with dAdo and EHNA, the level of dATP increases 50-fold or more, and synthesis of HSV DNA is inhibited. However, there is no depletion of any other DNA precursor, and HSV multiplication is not restored by simultaneous provision of dGuo, deoxythymidine, deoxycytidine, or a combination of all three of these nucleosides. Thus, the inhibition of HSV replication by dAdo cannot be explained as a block of precursor provision through inhibition of ribonucleotide reductase. In contrast, dGuo treatment of HSV-infected cells leads to depletion of dCTP, and virus multiplication is partially restored by provision of deoxycytidine. HSV-infected cells may serve as a useful system for study of the toxic effects of dAdo that are unrelated to inhibition of ribonucleotide reductase by dATP.


Neurological perspectives on voltage-gated sodium channels.
The activity of voltage-gated sodium channels has long been linked to disorders of neuronal excitability such as epilepsy and chronic pain. Recent genetic studies have now expanded the role of sodium channels in health and disease, to include autism, migraine, multiple sclerosis, cancer as well as muscle and immune system disorders. Transgenic mouse models have proved useful in understanding the physiological role of individual sodium channels, and there has been significant progress in the development of subtype selective inhibitors of sodium channels. This review will outline the functions and roles of specific sodium channels in electrical signalling and disease, focusing on neurological aspects. We also discuss recent advances in the development of selective sodium channel inhibitors.


Leukoaraiosis and ventricular enlargement in patients with ischemic stroke.
We studied the relationship between ventricular size and nonspecific periventricular lucency on computed tomograms (leukoaraiosis) in 192 patients with ischemic stroke. Leukoaraiosis did not occur in 21 patients less than 50 years of age; ventricular size could not be measured in an additional 29. Leukoaraiosis was graded from 0 to 4 on a semiquantitative scale; bicaudate, frontal horn, and posterior horn indices were used as measures of ventricular size. Patients with leukoaraiosis were older (difference between means 7 years, t = 5.3, df = 140, p less than 0.0001) and had larger bicaudate indices (difference between means 0.023, t = 3.54, df = 140, p = 0.0007) than patients without leukoaraiosis. Multiple regression analysis demonstrated that the effects of age and leukoaraiosis were independent. No effect of lesion type (cortical or lacunar infarct, or both) on bicaudate index could be demonstrated. Larger values for the bicaudate index were associated with a predominantly anterior location of leukoaraiosis. The frontal horn and occipital horn indices increased with age, but we could not find an effect of leukoaraiosis on these indices.


Hydrogen sulfide is a partially redox-independent activator of the human jejunum Na+ channel, Nav1.5.
Hydrogen sulfide (H(2)S) is produced endogenously by L-cysteine metabolism. H(2)S modulates several ion channels with an unclear mechanism of action. A possible mechanism is through reduction-oxidation reactions attributable to the redox potential of the sulfur moiety. The aims of this study were to determine the effects of the H(2)S donor NaHS on Na(V)1.5, a voltage-dependent sodium channel expressed in the gastrointestinal tract in human jejunum smooth muscle cells and interstitial cells of Cajal, and to elucidate whether H(2)S acts on Na(V)1.5 by redox reactions. Whole cell Na(+) currents were recorded in freshly dissociated human jejunum circular myocytes and Na(V)1.5-transfected human embryonic kidney-293 cells. RT-PCR amplified mRNA for H(2)S enzymes cystathionine β-synthase and cystathionine γ-lyase from the human jejunum. NaHS increased native Na(+) peak currents and shifted the half-point (V(1/2)) of steady-state activation and inactivation by +21 ± 2 mV and +15 ± 3 mV, respectively. Similar effects were seen on the heterologously expressed Na(V)1.5 α subunit with EC(50)s in the 10(-4) to 10(-3) M range. The reducing agent dithiothreitol (DTT) mimicked in part the effects of NaHS by increasing peak current and positively shifting steady-state activation. DTT together with NaHS had an additive effect on steady-state activation but not on peak current, suggesting that the latter may be altered via reduction. Pretreatment with the Hg(2+)-conjugated oxidizer thimerosal or the alkylating agent N-ethylmaleimide inhibited or decreased NaHS induction of Na(V)1.5 peak current. These studies show that H(2)S activates the gastrointestinal Na(+) channel, and the mechanism of action of H(2)S is partially redox independent.


Usefulness of comprehensive cardiothoracic computed tomography in the evaluation of acute undifferentiated chest discomfort in the emergency department (CAPTURE).
Newer cardiac computed tomographic (CT) technology has permitted comprehensive cardiothoracic evaluations for coronary artery disease, pulmonary embolism, and aortic dissection within a single breath hold, independent of the heart rate. We conducted a randomized diagnostic trial to compare the efficiency of a comprehensive cardiothoracic CT examination in the evaluation of patients presenting to the emergency department with undifferentiated acute chest discomfort or dyspnea. We randomized the emergency department patients clinically scheduled to undergo a dedicated CT protocol to assess coronary artery disease, pulmonary embolism, or aortic dissection to either the planned dedicated CT protocol or a comprehensive cardiothoracic CT protocol. All CT examinations were performed using a 64-slice dual source CT scanner. The CT results were immediately communicated to the emergency department providers, who directed further management at their discretion. The subjects were then followed for the remainder of their hospitalization and for 30 days after hospitalization. Overall, 59 patients (mean age 51.2 ± 11.4 years, 72.9% men) were randomized to either dedicated (n = 30) or comprehensive (n = 29) CT scanning. No significant difference was found in the median length of stay (7.6 vs 8.2 hours, p = 0.79), rate of hospital discharge without additional imaging (70% vs 69%, p = 0.99), median interval to exclusion of an acute event (5.2 vs 6.5 hours, p = 0.64), costs of care (p = 0.16), or the number of revisits (p = 0.13) between the dedicated and comprehensive arms, respectively. In addition, radiation exposure (11.3 mSv vs 12.8 mSv, p = 0.16) and the frequency of incidental findings requiring follow-up (24.1% vs 33.3%, p = 0.57) were similar between the 2 arms. Comprehensive cardiothoracic CT scanning was feasible, with a similar diagnostic yield to dedicated protocols. However, it did not reduce the length of stay, rate of subsequent testing, or costs. In conclusion, although this "triple rule out" protocol might be helpful in the evaluation of select patients, these findings suggest that it should not be used routinely with the expectation that it will improve efficiency or reduce resource use.


Anti-neuropathic effects of Rosmarinus officinalis L. terpenoid fraction: relevance of nicotinic receptors.
Traditional uses and current results highlight the neuroprotective properties of Rosmarinus officinalis L. The compelling need for novel strategies able to relieve neuropathic pain encouraged us to analyze different rosemary leaf extracts in rats following chronic constriction injury (CCI) of sciatic nerve. Ethanol, acetone, and the innovative ultrasound-hexane extractive methods were used to obtain: EE, AE, and for hexane extracts UREprel and URE. Extracts were characterized in terms of typical constituents and repeatedly administered to CCI-rats (13-days treatment, from the day of surgery). URE showed the best efficacy and potency in reducing hypersensitivity to noxious- and non-noxious stimuli and spontaneous pain. URE contained the higher quantity of the terpenoid carnosic acid (CA) and its efficacy was compared to pure CA. Histological analysis of the sciatic nerve revealed that URE prevented axon and myelin derangement, edema and inflammatory infiltrate. In the dorsal horn of the spinal cord, URE did not reduce astrocyte activation. Both the pain reliever and the neuroconservative effects of URE were significantly prevented by the nicotinic receptor (nAChR) antagonist mecamylamine. In conclusion, the hexane-ultrasound rosemary extract is able to reduce neuropathic hypersensitivity and protect nervous tissues. Effectiveness is mainly related to the terpenoid fraction by mechanisms involving nAChRs.


Activation of the coagulation cascade after infusion of a factor XI concentrate in congenitally deficient patients.
Virally inactivated, high-purity factor XI concentrates are available for treatment of patients with factor XI deficiency. However, preliminary experience indicates that some preparations may be thrombogenic. We evaluated whether a highly purified concentrate produced signs of activation of the coagulation cascade in two patients with severe factor XI deficiency infused before and after surgery. Signs of heightened enzymatic activity of the common pathway of coagulation (elevated plasma levels of prothrombin fragment 1 + 2 and fibrinopeptide A) developed in the early post-infusion period, accompanied by more delayed signs of fibrin formation with secondary hyperfibrinolysis (elevated D-dimer and plasmin-antiplasmin complex). These changes occurred in both patients, but were more severe in the older patient with breast cancer when she underwent surgery, being accompanied by fibrinogen and platelet consumption. There were no concomitant signs of heightened activity of the factor VII-tissue factor mechanism on the factor Xase complex (plasma levels of activated factor VII and of factor IX and X activation peptides did not increase). The observed changes in biochemical markers of coagulation activation indicate that concentrate infusions increased thrombin generation and activity and that such changes were magnified by malignancy and surgery. Because some factor XI concentrates may be thrombogenic, they should be used with caution, especially in patients with other risk factors for thrombosis.


Murine amebiasis: the role of the macrophage in host defense.
The congenitally athymic nude (nu/nu) mouse was studied as a model for amebic liver disease, using Entamoeba histolytica. Despite intrahepatic inoculation of massive numbers of amebae, we could not produce sustained infections in nu/nu mice. We also failed to induce hepatic amebiasis in thymus-intact nu/+ and +/+ littermates, or in nu/+ mice pretreated with rabbit antimouse thymocyte globulin. Humoral response was measured in both the nu/nu and nu/+ mice. Nu/+ but not nu/nu animals developed a specific IgG response after challenge with E. histolytica. There were no significant IgM responses. Pretreatment with silica increased susceptibility of both nu/nu and nu/+ mice to development of liver abscesses. These studies suggest that host resistance in murine amebiasis is critically dependent upon the macrophage, but not upon T cell-mediated defenses.


Photoperiod affects daily torpor and tissue fatty acid composition in deer mice.
Photoperiod and dietary lipids both influence thermal physiology and the pattern of torpor of heterothermic mammals. The aim of the present study was to test the hypothesis that photoperiod-induced physiological changes are linked to differences in tissue fatty acid composition of deer mice, Peromyscus maniculatus ( approximately 18-g body mass). Deer mice were acclimated for >8 weeks to one of three photoperiods (LD, light/dark): LD 8:16 (short photoperiod), LD 12:12 (equinox photoperiod), and LD 16:8 (long photoperiod). Deer mice under short and equinox photoperiods showed a greater occurrence of torpor than those under long photoperiods (71, 70, and 14%, respectively). The duration of torpor bouts was longest in deer mice under short photoperiod (9.3 +/- 2.6 h), intermediate under equinox photoperiod (5.1 +/- 0.3 h), and shortest under long photoperiod (3.7 +/- 0.6 h). Physiological differences in torpor use were associated with significant alterations of fatty acid composition in approximately 50% of the major fatty acids from leg muscle total lipids, whereas white adipose tissue fatty acid composition showed fewer changes. Our results provide the first evidence that physiological changes due to photoperiod exposure do result in changes in lipid composition in the muscle tissue of deer mice and suggest that these may play a role in survival of low body temperature and metabolic rate during torpor, thus, enhancing favourable energy balance over the course of the winter.


[The effect of rate of administration of isradipine on the hemodynamic response].
As the magnitude of drug response may depend on its input rate, we investigated the rate of administration-effect relationship of the new dihydropyridine isradipine. Ten healthy volunteers received (double-blind and cross-over) isradipine 1 mg i.v. (5 min), 5 mg solution, 5 mg tablet, 10 mg sustained release, and placebo. Blood pressure and heart rate were recorded for 24 h. A single dose of slow-release formulation isradipine does not induce significant hemodynamic effects in healthy subjects. The maximal BP fall was comparable with either i.v., solution, and conventional tablet administration. This is due to a stronger heart rate counter-regulation, linked to rapid isradipine administration. These results imply that slow input rates of isradipine are more effective in lowering blood pressure.


Experimental comparison of point-of-use filters for drinking water ultrafiltration.
Waterborne pathogens such as Pseudomonas spp. and Legionella spp. may persist in hospital water networks despite chemical disinfection. Point-of-use filtration represents a physical control measure that can be applied in high-risk areas to contain the exposure to such pathogens. New technologies have enabled an extension of filters' lifetimes and have made available faucet hollow-fibre filters for water ultrafiltration. To compare point-of-use filters applied to cold water within their period of validity. Faucet hollow-fibre filters (filter A), shower hollow-fibre filters (filter B) and faucet membrane filters (filter C) were contaminated in two different sets of tests with standard bacterial strains (Pseudomonas aeruginosa DSM 939 and Brevundimonas diminuta ATCC 19146) and installed at points-of-use. Every day, from each faucet, 100 L of water was flushed. Before and after flushing, 250 mL of water was collected and analysed for microbiology. There was a high capacity of microbial retention from filter C; filter B released only low Brevundimonas spp. counts; filter A showed poor retention of both micro-organisms. Hollow-fibre filters did not show good micro-organism retention. All point-of-use filters require an appropriate maintenance of structural parameters to ensure their efficiency.


Human T-cell lymphotropic virus type I and severe neoplasia of the cervix in Jamaica.
Human T-cell lymphotropic virus type I (HTLV-I) was associated with carcinoma of the cervix in Japan in a recent study that compared hospital cases with healthy population-based controls. To test this relationship in women more alike for cervical neoplasia risk factors (including sexual behavior and human papilloma virus; HPV), we enrolled consecutive patients from a colposcopy clinic in Kingston, Jamaica (an HTLV-I endemic area). Patients underwent Pap smear, colposcopy, biopsy and cervical swab for detection of HPV by polymerase chain reaction. Cases were defined as women with CIN-3 or invasive cancer (CIN-3/CA). Controls included all patients with either CIN-I or koilocytotic atypia, atypical squamous cells of undetermined significance or benign cervical pathology (all but one had at least inflammatory changes). Patients with CIN-2 were excluded to minimize risk of case-control misclassification. Cases were much more likely to be HTLV-I seropositive than controls. Although mean age differed significantly between cases (mean age = 39 years) and controls (mean age = 33 years), control for age did not explain the relation of CIN-3/CA with HTLV-I. Among HPV DNA positive subjects the age-adjusted association was not diminished but lost statistical significance. HTLV-I seroprevalence may be independently associated with progression to severe neoplasia of the cervix.


Induction of inflammatory cytokines and alteration of urothelial ATP, acetylcholine and prostaglandin E2 release by doxorubicin.
Intravesical treatment with cytotoxic drugs such as doxorubicin is associated with local adverse effects in bladder cancer patients. Here we investigate the effects of doxorubicin on urothelial release of ATP, acetylcholine and prostaglandin E(2), and production of inflammatory cytokines. Urothelial cells were treated with doxorubicin for 1h at 37 °C. Immediately or 24 h following treatment the level of ATP, acetylcholine and prostaglandin E(2) released under basal and stimulated conditions was measured and compared to release from vehicle treated control cultures. The presence of inflammatory cytokines, in culture medium was also assessed 24 h after doxorubicin pre-treatment. Immediately following treatment, stimulated ATP release was inhibited at doxorubicin concentrations ≥1 μg/ml and showed partial recovery at 24 h. Immediately following treatment, basal acetylcholine release was increased by doxorubicin at its clinical concentration (1 mg/ml), while a concentration-dependent decrease in stimulated acetylcholine release was observed. Twenty four hour after treatment, basal acetylcholine release was increased in culture treated with 0.01 mg/ml doxorubicin while stimulated acetylcholine release remained depressed. A significant increase in prostaglandin E(2) release was observed in cells immediately and 24 h after treatment with doxorubicin. A 5.5- and 2-fold increase in interleukin -8 and -1β secretion, respectively was detected 24 h following doxorubicin treatment. These findings indicate that inflammatory cytokines interleukin-8 and -1β are induced and urothelial mediator release is affected by treatment with doxorubicin at clinically relevant concentrations and durations of treatment. These changes may play a role in the adverse effects associated with intravesical doxorubicin treatment.


Adaptive response to oxidative stress in the filamentous fungus Aspergillus niger B1-D.
In the present study, we used a recombinant filamentous fungus strain, Aspergillus niger B1-D, as a model system, and investigated the antioxidant defences in this organism. Our findings indicate that pretreatment with low concentrations of H(2)O(2) completely prevents killing by this oxidant at high concentrations. It shows that A. niger adapts to exposure to H(2)O(2) by reducing growth and inducing a number of antioxidant enzyme activities, including superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, of which the induction of catalase is the most pronounced. Moreover the decline of these antioxidant enzymes activities after H(2)O(2) detoxification, coincides with recommencement of growth. Results from monitoring the extracellular H(2)O(2) concentration clearly indicate a very rapid detoxification rate for H(2)O(2) in adapted A. niger cultures. A mathematical model predicts only very low concentrations of intracellular H(2)O(2) accumulating in such cultures. Our results also show that glutathione plays a role in the oxidative defence against H(2)O(2) in A. niger. On addition of H(2)O(2), the intracellular pool of glutathione increases while the redox state of glutathione becomes more oxidized.