A method to facilitate and monitor expression of exogenous genes in the rat kidney using plasmid and viral vectors.
Gene therapy has been proposed as a novel alternative to treat kidney disease. This goal has been hindered by the inability to reliably deliver transgenes to target cells throughout the kidney, while minimizing injury. Since hydrodynamic forces have previously shown promising results, we optimized this approach and designed a method that utilizes retrograde renal vein injections to facilitate transgene expression in rat kidneys. We show, using intravital fluorescence two-photon microscopy, that fluorescent albumin and dextrans injected into the renal vein under defined conditions of hydrodynamic pressure distribute broadly throughout the kidney in live animals. We found injection parameters that result in no kidney injury as determined by intravital microscopy, histology, and serum creatinine measurements. Plasmids, baculovirus, and adenovirus vectors, designed to express EGFP, EGFP-actin, EGFP-occludin, EGFP-tubulin, tdTomato-H2B, or RFP-actin fusion proteins, were introduced into live kidneys in a similar fashion. Gene expression was then observed in live and ex vivo kidneys using two-photon imaging and confocal laser scanning microscopy. We recorded widespread fluorescent protein expression lasting more than 1 mo after introduction of transgenes. Plasmid and adenovirus vectors provided gene transfer efficiencies ranging from 50 to 90%, compared with 10-50% using baculovirus. Using plasmids and adenovirus, fluorescent protein expression was observed 1) in proximal and distal tubule epithelial cells; 2) within glomeruli; and 3) within the peritubular interstitium. In isolated kidneys, fluorescent protein expression was observed from the cortex to the papilla. These results provide a robust approach for gene delivery and the study of protein function in live mammal kidneys.


Epidemiology of tuberculosis by DNA fingerprinting.
In recent years, deoxyribonucleic acid (DNA) probes have become available to study the epidemiology of tuberculosis by DNA fingerprint techniques. These methods make it possible to distinguish different strains of Mycobacterium tuberculosis. Currently, DNA fingerprinting is integrated with conventional epidemiological approaches to improve understanding of the spread of tuberculosis. This molecular approach has led to the investigation of a wide variety of epidemiological issues, such as adequate identification of outbreaks, tracing of nosocomial infections in hospitals, and investigation of the relative contribution of newly acquired versus reactivated infections in different populations. This paper reviews the potentials of DNA fingerprinting for studying the epidemiology of tuberculosis, and the lessons learned from this strategy. In addition, future prospects for molecular epidemiology will be discussed.


[Psychoautonomic disorders in patients with hypertension, stage I and possibilities of their treatment].
The article is devoted to the study of psychoautonomic features of patients with essential hypertension stage 1. The authors present the results of the clinical and psychological examinations of 62 patients with this pathology. These results show an increase in the levels of state and trait anxiety, which confirms the leading role of the psycho-emotional sphere in the formation of hypertensive reactions. This study evaluated the efficacy of the drug adaptol usage during 8 weeks in daily dose 1500 mg divided into three parts in the treatment of this group of patients. The results clearly demonstrate the positive effect of adaptol on psychoautonomic adaptation of patients with essential hypertension stage 1.


Interaction of pseudomonic acid A with Escherichia coli B isoleucyl-tRNA synthetase.
Sodium pseudomonate was shown to be a powerful competitive inhibitor of Escherichia coli B isoleucyl-tRNA synthetase (Ile-tRNA synthetase). The antibiotic competitively inhibits (Ki 6 nM; cf. Km 6.3 microM), with respect top isoleucine, the formation of the enzyme . Ile approximately AMP complex as measured by the pyrophosphate-exchange reaction, and has no effect on the transfer of [14C]isoleucine from the enzyme . [14C]Ile approximately AMP complex to tRNAIle. The inhibitory constant for the pyrophosphate-exchange reaction was of the same order as that determined for the inhibition of the overall aminoacylation reaction (Ki 2.5 nM; cf. Km 11.1 microM). Sodium [9'-3H]pseudomonate forms a stable complex with Ile-tRNA synthetase. Gel-filtration and gel-electrophoresis studies showed that the antibiotic is only fully released from the complex by 5 M-urea treatment or boiling in 0.1% sodium dodecyl sulphate. The molar binding ratio of sodium [9'-3H]pseudomonate to Ile-tRNA synthetase was found to be 0.85:1 by equilibrium dialysis. Aminoacylation of yeast tRNAIle by rat liver Ile-tRNA synthetase was also competitively inhibited with respect to isoleucine, Ki 20 microM (cf. Km 5.4 microM). The Km values for the rat liver and E. coli B enzymes were of the same order, but the Ki for the rat liver enzyme was 8000 times the Ki for the E. coli B enzyme. This presumably explains the low toxicity of the antibiotic in mammals.


Incidence of capsular contracture in silicone versus saline cosmetic augmentation mammoplasty: A meta-analysis.
Capsular contracture after augmentation mammoplasty occurs at a rate of 15% to 45%. The purpose of the present study was to determine the effect of implant core type (silicone versus saline) on the rate of capsular contracture in augmentation mammoplasty. A systematic review was conducted through a search of three electronic databases. Two reviewers independently scanned titles yielded by the search and identified potentially relevant papers. Inter-reviewer variability and the scientific quality of the articles were assessed. Meta-analysis was performed. Eighty-eight titles of potential relevance were selected from the 393 articles yielded by the search. Inter-rater agreement for selection of potentially relevant articles was 84% (kappa=0.54). Four comparative studies were included in the analysis. Scientific quality scores of the included studies ranged from 5 of 14 to 9 of 14. Three of the four studies reported a higher rate of capsular contracture in patients with silicone implants. A combined odds ratio calculated on two of the studies found a 2.25-fold increased risk of capsular contracture in patients who received silicone implants. Eight series of patients who received cohesive gel silicone implants reported rates of capsular contracture from 0% to 13.6%. Higher rates of capsular contracture were found in patients who received silicone implants when compared with those who received saline implants. However, the scientific quality of the comparative studies to date on this subject is poor. Recent series evaluating cohesive gel implants report relatively low rates of capsular contracture. A randomized controlled trial comparing rates of capsular contracture in cohesive gel and saline implants is recommended.


Patient Organizations' Barriers in Pharmacovigilance and Strategies to Stimulate Their Participation.
European drug regulations aim for a patient-centered approach, including involving patients in the pharmacovigilance (PV) systems. However many patient organizations have little experience on how they can participate in PV activities. The aim of this study was to understand patient organizations' perceptions of PV, the barriers they face when implementing PV activities, and their interaction with other stakeholders and suggest methods for the stimulation of patient organizations as promoters of PV. A sequential qualitative method study was conducted and integrated with the quantitative study performed by Matos, Weits, and van Hunsel to complete a mixed method study. The qualitative phase expands the understanding of the quantitative results from a previous study by broadening the knowledge on external barriers and internal barriers that patient organizations face when implementing PV activities. The strategies to stimulate patient-organization participation are the creation of more awareness campaigns, more research that creates awareness, education for patient organizations, communication of real PV examples, creation of a targeted PV system, creation of a PV communication network that provides feedback to patients, improvement of understanding of all stakeholders, and a more proactive approach from national competent authorities. Both study phases show congruent results regarding patients' involvement and the activities patient organizations perform to promote drug safety. Patient organizations progressively position themselves as stakeholders in PV, carrying out many activities that stimulate awareness and participation of their members in drug safety, but still face internal and external barriers that can hamper their involvement.


Exendin-4 treatment expands graft beta-cell mass in diabetic mice transplanted with a marginal number of fresh islets.
Exendin-4 stimulates insulin secretion, suppresses glucagons secretion, increases beta-cell replication and neogenesis, and reduces beta-cell apoptosis. However, it has been shown that posttransplant exendin-4 treatment did not improve glucose homeostasis in diabetic mice transplanted with a large number of freshly isolated islets. The aim of this study was to test if exendin-4 is beneficial for hyperglycemic recipients with a marginal number of fresh islets. We transplanted 150 C57BL/6 mouse islets under the kidney capsule of inbred streptozotocin-diabetic mice, and then treated the recipients with and without exendin-4 for 6 weeks. Before and after transplantation, recipients' blood glucose, body weight, and intraperitoneal glucose tolerance test were measured. At 6 weeks, the grafts were removed to determine beta-cell mass. Blood glucose levels in both groups decreased progressively after transplantation, and the exendin-4-treated group had had lower blood glucose than controls since day 3. By 6 weeks, euglycemia was achieved more in mice treated with exendin-4 than in controls (100% vs. 62.5%, p = 0.018). The time to obtain normoglycemia was shorter in the exendin-4-treated group than in controls (12 +/- 8 vs. 29 +/- 13 days, p < 0.001). Blood glucose at 6 weeks was 123 +/- 18 and 170 +/- 62 mg/dl in the exendin-4-treated group and controls, respectively (p = 0.008). Additionally, the exendin-4-treated group had better glucose tolerance than controls at 2 and 4 weeks (p <0.02). However, both groups exhibited increased body weight over time, and weight changes did not significantly differ between the two groups throughout the study period. At 6 weeks after transplantation, grafts in the exendin-4-treated group were more prominent and contained more insulin-stained cells than those of controls. They had 2.3-fold beta-cell mass of the graft compared with controls (0.30 +/- 0.11 vs. 0.13 +/- 0.03 mg, p = 0.012). These results indicate posttransplant exendin-4 treatment in the diabetic recipient with a marginal number of fresh islets expands graft beta-cell mass and improves transplantation outcome.


The role of toothpaste in the aetiology and treatment of dentine hypersensitivity.
Dentine hypersensitivity (DH) is a common, painful dental condition with a multi-factorial aetiology. The hydrodynamic mechanism theory to explain dentine sensitivity also appears to fit DH: lesions exhibiting large numbers of open dentinal tubules at the surface and patent to the pulp. By definition, DH can only occur when dentine becomes exposed (lesion localisation) and tubules opened (lesion initiation), thus permitting increased fluid flow in tubules on stimulation. Erosion, particularly from dietary acids appears to play a dominant role in both processes. Toothbrushing with most toothpaste products alone cause clinically insignificant wear of enamel but are additive, even synergistic, to erosive enamel loss. Additionally, toothbrushing with toothpaste is implicated in 'healthy' gingival recession. Toothbrushing with most toothpastes removes the smear layer to expose tubules and again can exacerbate erosive loss of dentine. These findings thereby implicate toothbrushing with toothpaste in the aetiology of DH. Management of the condition should have secondary prevention at the core of treatment and therefore, must consider first and foremost the aetiology. Fluoride toothpaste at present appears to provide little primary or secondary preventive benefits to DH; additional ingredients can provide therapeutic benefits. Potassium-based products to block pulpal nerve response have caused much debate and are considered by many as unproven, which should not translate to ineffective. Several toothpaste technologies formulated to block tubules are from studies in vitro, in situ and controlled clinical trials considered proven for the treatment of DH.


Ventricular-arterial coupling as a potential therapeutic target in diabetes.
Patients with type 2 diabetes (T2D) are at high risk of cardiovascular complications. Novel anti-diabetic medications such as sodium-glucose cotransporter-2 inhibitors (SGLT-2i) and Glucagon-like peptide-1 receptor agonists (GLP-1RA) have been shown to possess cardiac and renal protective effects beyond their ability to lower plasma glucose. Use of SGLT-2i and GLP-1RA in patients with T2D and heart failure reduce cardiovascular risk and heart failure-related hospitalisations. SGLT-2i treatment has been shown to improve the long-term prognosis of patients with heart failure. Both drugs also have the potential to normalise ventricular-arterial coupling (VAC). VAC is the crosstalk between the left ventricular function and arterial system, and is an indicator of the global cardiovascular performance. In this overview, we will describe the concept of VAC and the features of diabetic cardiomyopathy, as well as VAC as a potential therapeutic target in diabetes by the use of novel anti-diabetic drugs, primarily SGLT-2i and GLP-1RA.


[Methods of medical and surgical correction of atherosclerotic aorto-arterial pathology in management of malignant neoplasms].
The article deals with 6 -year experience of the Centre of Oncoangiosurgery of the Chelyabinsk Regional Oncological Dispensary in treatment of patients with concomitant malignant neoplasms and atherosclerotic lesions to the aorta and arteries. The authors analyze therapeutic outcomes in a total of 158 patients. Of these, 81 patients were subjected to surgical correction of the blood flow. The presence of a clinically significant atherosclerotic lesion of the aorta and peripheral arteries in a patient should not be interpreted as an absolute or relative contraindication to contraindication combined treatment of the tumour concerned. In the majority of cases, atherosclerotic occlusion of femoral and tibial arteries was treated conservatively, using <<vasaprostan>>. In manifestation of an atherosclerotic lesion to the carotid arteries, aorta and iliac arteries, surgical correction of the blood flow was used more often. Usually, vascular reconstruction was carried out first. Simultaneous interventions were performed in 16 patients; three patients died. The technique of operations on the aorta and arteries was traditional, while for revascularization in peripheral lesions to the arteries of the limbs more frequently compromising methods were used (sympathectomy, profundoplasty, local endarterectomy) on the background of prostaglandin E1 infusions.


An In Vivo Lapine Model for Impact-Induced Injury and Osteoarthritic Degeneration of Articular Cartilage.
In this study, we applied a spring-loaded impactor to deliver traumatic forces to articular cartilage in vivo. Based on our recent finding that a 0.28-J impact induces maximal catabolic response in adult bovine articular cartilage in vitro using this device, we hypothesize that this impact will induce the formation of a focal osteoarthritic defect in vivo. The femoral condyle of New Zealand White rabbits was exposed and one of the following procedures performed: 0.28 J impact, anterior cruciate ligament transection, articular surface grooving, or no joint or cartilage destruction (control). After 24 hours, 4 weeks, or 12 weeks (n = 3 for each time point), wounds were localized with India ink, and tissue samples were collected and characterized histomorphometrically with Safranin O/Fast green staining and Hoechst 33342 nuclear staining for cell vitality. The spring-loaded device delivered reproducible impacts with the following characteristics: impact area of 1.39 ± 0.11 mm(2), calculated load of 326 ± 47.3 MPa, time-to-peak of 0.32 ± 0.03 ms, and an estimated maximal displacement of 25.1% ± 4.5% at the tip apex. The impact resulted in immediate cartilage fissuring and cell loss in the surface and intermediate zones, and it induced the formation of a focal lesion at 12 weeks. The degeneration was defined and appeared more slowly than after anterior cruciate ligament transection, and more pronounced and characteristic than after grooving. A single traumatic 0.28 J impact delivered with this spring-loaded impactor induces focal cartilage degeneration characteristic of osteoarthritis.


Toxicological assessment and food allergy of silk fibroin derived from Bombyx mori cocoons.
Recent studies have demonstrated silk fibroin protein's (SF) ability to extend the shelf life of foods by mitigating the hallmarks of spoilage, namely oxidation and dehydration. Due to the potential for this protein to become more widespread, its safety was evaluated comprehensively. First, a bacterial reverse mutation test (Ames test) was conducted in five bacterial strains. Second, an in vivo erythrocyte test was conducted with Sprague Dawley rats at doses up to 1,000mg/kg-bw/day. Third, a range-finder study was conducted with Sprague Dawley rats at the highest consumption amount given solubility and oral gavage volume constrains (500mg/kg-bw/day). Fourth, a 28-day sub-chronic study in Sprague Dawley rats was conducted with the high dose set at 500mg/kg-bw/day, as limited by solubility of the protein in a single-gavage per-day study. Fifth, an in vitro pepsin digestion assay was performed to assess the potential for protein allergenicity. Sixth, allergenic potential was further assessed using liquid chromatography-mass spectroscopy for detection of allergenic insect proteins. Seventh, the SF protein sequences were subjected to bioinformatic analyses. Together, these studies raise no mutagenic, genotoxic, toxicological, or allergenic concerns with the oral consumption of silk fibroin.


Elevating body termperature to reduce low-grade inflammation: a welcome strategy for those unable to exercise?
Chronic low-grade inflammation is increasingly recognized in the aetiology of a range of chronic diseases, including type 2 diabetes mellitus and cardiovascular disease, and may therefore serve as a promising target in their prevention or treatment. An acute inflammatory response can be induced by exercise; this is characterised by the acute increase in proinflammatory markers that subsequently stimulate the production of anti-inflammatory proteins. This may help explain the reduction in basal concentrations of pro-inflammatory markers following chronic exercise training. For sedentary populations, such as people with a disability, wheelchair users, or the elderly, the prevalence of chronic low-grade inflammation- related disease is further increased above that of individuals with a greater capacity to be physically active. Performing regular exercise with its proposed anti-inflammatory potential may not be feasible for these individuals due to a low physical capacity or other barriers to exercise. Therefore, alternatives to exercise that induce a transient acute inflammatory response may benefit their health. Manipulating body temperature may be such an alternative. Indeed, exercising in the heat results in a larger acute increase in inflammatory markers such as interleukin-6 and heat shock protein 72 when compared with exercising in thermoneutral conditions. Moreover, similar to exercise, passive elevation of body temperature can induce acute increases and chronic reductions in inflammatory markers and positively affect markers of glycaemic control. Here we discuss the potential benefits and mechanisms of active (i.e., exercise) and passive heating methods (e.g., hot water immersion, sauna therapy) to reduce chronic low-grade inflammation and improve metabolic health, with a focus on people who are restricted from being physically active.


Relationship between touch impairment and brain activation after lesions of subcortical and cortical somatosensory regions.
The neural basis underlying somatosensory impairment and recovery poststroke is virtually unexplored. To investigate the relationship between touch discrimination impairment and task-related brain activation in stroke survivors with somatosensory impairment following subcortical or cortical lesions. A total of 19 stroke survivors with touch impairment were investigated using fMRI and a touch discrimination paradigm 1-month poststroke; 11 had subcortical and 8 cortical sensory lesions; 12 age-matched healthy controls were also studied. Mean task-related contrast images were regressed with sensory impairment using random effects analysis for each subgroup and the total group. There was no significant difference in touch impairment between stroke subgroups. Touch discrimination of the affected hand correlated negatively with task-related activation in the ipsilesional primary somatosensory cortex (SI; adjacent to the SI hand area activated in healthy controls); ipsilesional secondary somatosensory cortex (SII); contralesional thalamus; and attention-related frontal and occipital regions in the subcortical group. In contrast, the cortical group did not show significant correlated activity. Yet there was no significant between-group difference in a priori somatosensory regions: only in the superior medial frontal gyrus. A negative correlation was observed in the contralesional thalamus for the total group, irrespective of lesion type. The findings provide novel evidence of neural correlates of poststroke touch impairment involving a distributed network of ipsilesional SI and SII, the contralesional thalamus, and frontal attention regions, particularly following subcortical lesions. Further systematic investigation of a modulatory role for ipsilesional SI, the thalamus, and frontal attention regions in sensory processing and recovery is warranted, particularly given implications for rehabilitation.


In vivo generation of C4d, Bb, iC3b, and SC5b-9 after OKT3 administration in kidney and lung transplant recipients.
OKT3 monoclonal antibody therapy results in an acute clinical syndrome (ACS) associated with the release of tumor necrosis factor and sequestration of neutrophils in the lungs. We have previously shown that inhibition of tumor necrosis factor does not completely eliminate OKT3-ACS, suggesting that other factors also contribute to the ACS. The current studies analyzed complement activation in vivo during the first hour after OKT3 administration. Renal (n=4) and lung (n=4) transplant recipients received OKT3 as treatment for rejection and induction therapy, respectively. Complement activation products C4d, Bb, iC3b, and SC5b-9 were measured by ELISA. Hemodynamic parameters were also monitored in the lung transplant recipients. Neutrophil expression of CD11a, CD11b, and CD18 was monitored by flow cytometry. Controls included patients receiving methylprednisolone for rejection (n=4), two adults with adult respiratory distress syndrome who received extracorporeal membrane oxygenation, and normal volunteers (n=5). P values less than 0.05 (*) were considered significant. Increases in the plasma levels of C4d, Bb, iC3b, and SC5b-9 were observed in seven of eight patients after OKT3 administration. Mean values (n=8) at 0, 15, and 60 min (in microg/ml) were as follows-C4d: 1.865, 2.644*, and 2.607*; Bb: 0.245, 0.411, and 0.385; iC3b: 10.881, 17.242*, and 15.145*; and SC5b-9: 0.232, 0.269, and 0.302*. An increase in CD11b and CD18 and a decrease of CD11a on neutrophils in parallel with complement activation was observed. In lung transplant recipients, C3 activation correlated with increases in mean pulmonary and central venous pressures (P<0.05). As compared with extracorporeal membrane oxygenation, which activated classical and alternative pathways, OKT3 predominantly activated complement by the classical pathway. Methylprednisolone pulses did not activate complement. Complement activation is an early event after OKT3 administration and is associated with the increased expression of adhesion molecules on neutrophils and with pulmonary hemodynamic changes. Effective therapeutic approaches to the control of early monoclonal antibody side effects may require measures that limit complement activation in addition to reducing cytokine activity.


Effect of charge on hepatic specific nature of vesicle encapsulated insulin in conscious dogs.
Positively [(+)-VEI] and negatively [(-)-VEI] charged formulations of vesicle encapsulated insulin (VEI) were compared with insulin on the basis of their ability to suppress hepatic glucose production (hepatic Ra) and stimulate glucose utilization (Rd) in conscious dogs. Our results indicate that (-)-VEI and insulin have an equivalent capacity to suppress hepatic glucose production at dose levels of 0.2, 1.2, and 2.4 mU X kg-1 X min-1. (+)-VEI is less effective at suppressing hepatic Ra than both insulin and (-)-VEI at dose levels of 1.2 and 2.4 mU X kg-1 X min-1. Both (+)-VEI and (-)-VEI induced significantly less glucose utilization than a comparable amount of insulin at dose levels of 1.2 and 2.4 mU X kg-1 X min-1. The amount of glucose utilization stimulated by (+)-VEI was significantly less than that induced by insulin at 0.6 mU X kg-1 X min-1. This difference was not evident with (-)-VEI. These results suggest that the insulin contained in (+)-VEI is less bioavailable than that contained in (-)-VEI. This difference in bioavailability is believed to be the result of greater serum stability of (+)-VEI vesicles when compared with that of (-)-VEI. In conclusion, both (+)-VEI and (-)-VEI have the capacity to shield encapsulated insulin from interacting with peripheral tissues and deliver insulin selectively to the liver. Both formulations afford one an opportunity to expand the therapeutic window for hepatically active compounds where their utility is limited by systemic toxicity.


Development of the Automated Primer Design Workflow Uniqprimer and Diagnostic Primers for the Broad-Host-Range Plant Pathogen Dickeya dianthicola.
Uniqprimer, a software pipeline developed in Python, was deployed as a user-friendly internet tool in Rice Galaxy for comparative genome analyses to design primer sets for PCRassays capable of detecting target bacterial taxa. The pipeline was trialed with Dickeya dianthicola, a destructive broad-host-range bacterial pathogen found in most potato-growing regions. Dickeya is a highly variable genus, and some primers available to detect this genus and species exhibit common diagnostic failures. Upon uploading a selection of target and nontarget genomes, six primer sets were rapidly identified with Uniqprimer, of which two were specific and sensitive when tested with D. dianthicola. The remaining four amplified a minority of the nontarget strains tested. The two promising candidate primer sets were trialed with DNA isolated from 116 field samples from across the United States that were previously submitted for testing. D. dianthicola was detected in 41 samples, demonstrating the applicability of our detection primers and suggesting widespread occurrence of D. dianthicola in North America.


Signal transduction and transcriptional regulation by glucocorticoid receptor-LexA fusion proteins.
The glucocorticoid receptor regulates transcriptional initiation upon binding to its cognate hormone. A series of fusion genes was constructed to examine the mechanism of hormone-regulated transcriptional enhancement. The DNA binding domain of the bacterial LexA repressor was fused to receptor derivatives lacking the region that is necessary and sufficient for specific DNA binding and transcriptional enhancement at glucocorticoid response elements (GRE's). The resultant hybrid proteins activated transcription from promoters linked to the lex operator. Enhancement still required hormone binding by the hybrid receptor regardless of the exact positioning of the LexA binding domain within the protein. Thus, the unliganded hormone binding domain of the receptor acts as a strong but reversible inhibitor of receptor activity in a manner that is independent of the means by which the receptor recognizes DNA. The results also show directly that the receptor contains at least one "enhancement domain" other than that overlapping the GRE binding region; the second domain, enh2, occupies a region near the receptor amino terminus.


Safety and feasibility of early hospital discharge in ST-segment elevation myocardial infarction--a prospective and randomized trial in low-risk primary percutaneous coronary intervention patients (the Safe-Depart Trial).
Patients with ST-segment elevation myocardial infarction (STEMI) have traditionally been hospitalized for 5 to 7 days to monitor for serious complications such as heart failure, arrhythmias, reinfarction, and death. The Zwolle Primary Percutaneous Coronary Intervention (PCI) Index is an externally validated risk score that has been used to identify low-risk STEMI patients who have undergone primary PCI and can safely be discharged from hospital within 72 hours. Previous studies have shown that many low-risk patients remain in hospital significantly longer. We randomly assigned 54 low-risk STEMI patients treated with primary or rescue PCI to 1 of 2 groups. Patients in the intervention group (n = 27) were actively targeted for early hospital discharge (48-72 hours) and received outpatient follow-up with an advanced practice nurse (APN). In the control group (n = 27), discharge planning and follow-up were left to the treating physician, and there was no added nursing intervention. The 2 primary outcomes of this pilot study were to demonstrate feasibility and safety. Secondary outcomes included compliance with medications, smoking cessation, attendance at cardiac rehabilitation, and quality of life, measured in both groups at 6 weeks time. In the intervention group, 74% of patients were discharged within 72 hours, 100% had follow-up with the APN within 3 days (74% in person, 26% by phone), and 100% had >/= 3 APN follow-ups in total, meeting our prespecified criteria for feasibility. The median length of stay was 55 hours in both groups. There were no deaths in either group, and there was no difference in rehospitalization between patients in the intervention and control groups (8% vs 4%, P = .56). There was no difference in rates of medication compliance, smoking cessation, attendance at cardiac rehabilitation, or quality of life between the 2 groups, although our small pilot study was not powered to detect a difference in these outcomes. In low-risk STEMI patients treated with primary or rescue PCI, a strategy of early hospital discharge facilitated by close nursing follow-up is feasible. Although our study did not identify differences in compliance or quality of life between the 2 groups, it did provide a functional study design for a larger trial powered to detect these important clinical end points.


Plasma homocysteine and liver tissue S-adenosylmethionine, S-adenosylhomocysteine status in vitamin B6-deficient rats.
The aim of this study was to evaluate plasma homocysteine (Hcy), malondialdehyde (MDA), glutathione (GSH) levels, glutathione peroxidase (GSH-Px) and glutathione-S-transferase (GST) activities and liver tissue S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) levels in control and vitamin B6-deficient rats. Thirty-two male rats with a weight of 65-75 g were used for the experiment. The rats were divided into control (n=16) and vitamin B6-deficient groups. At the end of the experiment, the animals were anesthetized with ketamine-HCl (Ketalar, 20 mg/kg, i.p.), and the blood was collected by cardiac puncture after thoracotomy. Plasma Hcy, pyridoxal phosphate (PLP), liver SAM, SAH levels measured by an isocratic system with high performance liquid chromatography. Plasma GSH-Px, GSH activities and GSH, MDA levels were carried out using a spectrophotometer. Plasma Hcy, MDA, liver tissue SAH levels were significantly increased, whereas plasma GSH, PLP, liver tissue SAM levels, plasma GST, GSH-Px activities and SAM/SAH ratio were decreased compared to those of control group. Vitamin B6 deficiency causes an increase in plasma homocysteine levels. Thus, we think that vitamin B6 supplementation could be used for therapeutic purposes in hyperhomocysteinemia condition.


Liquid chromatography with mass spectrometry method based two-step precursor ion scanning for the structural elucidation of flavonoids.
Plant flavonoids are very important secondary metabolites for insect and virus control of their host plant and are potent nutrients for humans. To be able to understand the bioavailability and functions of plant flavonoids, it is necessary to reveal their exact chemical structures. Liquid chromatography with tandem mass spectrometry is a powerful approach for structural elucidation of metabolites. In this report, a two-step precursor ion scanning based liquid chromatography with tandem mass spectrometry method was developed for the structural elucidation of plant flavonoids. The established method consists of the two-step precursor ions scanning for possible flavonoids extraction, MS(2) fragment spectra acquisition and comparison with an online database, liquid chromatography retention rules correction, and commercial standards verification. The developed method was used for the structure elucidation of flavonoids in flowers and leaves of tobacco (Nicotiana tabacum), and 17 flavonoids were identified in the tobacco variety Yunyan 97. Nine of the 17 identified flavonoids were considered to be found in tobacco flowers or/and leaves for the first time based on the available references. This method was proved to be very effective and can be used for the identification of flavonoids in other plants.


Twist viscosities and flow alignment of biaxial nematic liquid crystal phases of a soft ellipsoid-string fluid studied by molecular dynamics simulation.
We have calculated the twist viscosity and the alignment angle between the director and the stream lines in shear flow of a liquid crystal model system, which forms biaxial nematic liquid crystals, as functions of the density, from the Green-Kubo relations by equilibrium molecular dynamics simulation and by a nonequilibrium molecular dynamics algorithm, where a torque conjugate to the director angular velocity is applied to rotate the director. The model system consists of a soft ellipsoid-string fluid where the ellipsoids interact according a repulsive version of the Gay-Berne potential. Four different length-to-width-to-breadth ratios have been studied. On compression, this system forms discotic or calamitic uniaxial nematic phases depending on the dimensions of the molecules, and on further compression a biaxial nematic phase is formed. In the uniaxial nematic phase there is one twist viscosity and one alignment angle. In the biaxial nematic phase there are three twist viscosities and three alignment angles corresponding to the rotation around the various directors and the different alignments of the directors relative to the stream lines, respectively. It is found that the smallest twist viscosity arises by rotation around the director formed by the long axes, the second smallest one arises by rotation around the director formed by the normals of the broadsides, and the largest one by rotation around the remaining director. The first twist viscosity is rather independent of the density whereas the last two ones increase strongly with density. One finds that there is one stable director alignment relative to the streamlines, namely where the director formed by the long axes is almost parallel to the stream lines and where the director formed by the normals of the broadsides is almost parallel to the shear plane. The relative magnitudes of the components of the twist viscosities span a fairly wide interval so this model should be useful for parameterisation experimental data.


Differential expression of GHRH receptor and its splice variant 1 in human normal and malignant mucosa of the oesophagus and colon.
Recent evidence indicates that growth hormone-releasing hormone (GHRH) functions as a growth factor for gastrointestinal (GI) tumours. The tumourigenic effects of GHRH appear to be mediated by the splice variant 1 (SV-1) of GHRH receptor as well as the full length pituitary type receptor for GHRH (GHRH-R). We examined the protein and mRNA expression of GHRH-R and SV-1 in normal human tissues and tumours of the gastrointestinal (GI-) tract by immunohistochemical staining and reverse transcriptase (RT)-PCR. Squamous cells and squamous cell carcinoma of the oesophagus were negative for GHRH-R and SV-1, while Barrett's mucosa and adenocarcinomas of the oesophagus showed a strong expression of both receptors. The expression of GHRH-R was absent in normal colonic mucosa other than neuroendocrine cells (NE) and lining epithelium (LE) but strong in tubular adenomas of the colon, while the staining for SV-1 was absent in cells other than NE. However, the expression of both receptors was significantly increased in tubulovillous adenomas and colorectal cancers. No differences were seen in protein levels for both receptors between normal and neoplastic tissues of the stomach, pancreas and liver. Because of low mRNA levels for both receptors in all samples tested, only a qualitative assessment could be made. However, mRNA for GHRH-R and SV-1 showed a near-perfect correlation with the assessment of receptor proteins by immunostaining. Our study shows that in contrast to normal mucosa, transformed mucosa of the oesophagus and the colon expresses GHRH-R and SV-1. This aberrant expression of GHRH-R and SV-1 in oesophageal and colorectal malignancies may provide a molecular target for a therapeutic approach based on GHRH antagonists.


Epidermal Lamellar Granules.
In the mid-1950s and 1960s, transmission electron microscopes became widely available, leading to many studies of the ultrastructure of various tissues including the epidermis. Most of these studies involved tissue fixation with formaldehyde and postfixation with osmium tetroxide. A few studies employed freeze-fracture electron microscopy. One set of these studies identified a small organelle variously called lamellar granules (LGs), lamellar bodies, membrane-coating granules, cementsomes, and Odland bodies. LGs are round to ovoid in shape, with a diameter of about 200 nm. They have a bounding membrane surrounding a stack of internal lipid lamellae. These small organelles are first seen in the spinous layer and accumulate with differentiation in the granular layer. In the uppermost granular cells, the bounding membrane of the LG fuses into the cell plasma membrane, and the internal contents are extruded into the intercellular space. The initially extruded contents of the LG then rearrange to form the intercellular lamellae of the stratum corneum. In this context, LGs serve as the precursor to the permeability barrier of the skin. Various studies have provided evidence that they are derived from the Golgi apparatus, specifically the trans-Golgi. Isolated LGs contain phosphoglycerides, sphingomyelin, and glucosylceramides. The most unusual lipid component is a linoleate-containing glucosylceramide comprising 30- to 34-carbon ω-hydroxy-acids. Isolated granules also contain acid hydrolases including glucocerebrosidase, sphingomyelinase, and phospholipase A. They also contain proteases and antimicrobial peptides. Defective LGs have been associated with a number of skin diseases including ichthyotic conditions and defective barrier function. Recently, studies employing cryo-transmission electron microscopy have called into question the validity of observations on LGs with more conventional electron microscopic techniques. These studies suggest a continuity of the membrane structure from the Golgi through the intercellular lamellae of the stratum corneum.


A secreted 80 x 10(3) Mr protein mediates sensing of cell density and the onset of development in Dictyostelium.
In submerged monolayer culture, Dictyostelium cells can differentiate into prespore and prestalk cells at high cell densities in response to cAMP but not at low cell densities. However, cells at low densities will differentiate in medium taken from developing cells starved at a high density. The putative factor in the medium was designated CMF for conditioned medium factor (Mehdy and Firtel, Molec. cell. Biology 5, 705-713, 1985). In this report, we size-fractionate conditioned medium and show that the activity that allows low density cells to differentiate can be separated into high and low Mr (relative molecular mass) fractions. Interestingly, the two fractions both have the same activity and do not need to be combined to allow differentiation. The large conditioned medium factor is a protein, as determined by trypsin sensitivity, that can be purified to a single 80 x 10(3) Mr band on a silver-stained SDS-polyacrylamide gel, and has CMF activity at a concentration of approximately 4 pM (0.3 ng ml-1). Our results suggest that CMF is a secreted factor that functions in vivo as an indicator of cell density in starved cells. At high cell densities, the concentration of CMF is sufficient to enable cells to enter the multicellular stage of the developmental cycle. When present below a threshold concentration, cells do not initiate the expression of genes required for early development. This factor plays an essential role in the regulatory pathway necessary for cells to obtain the developmental competence to induce prestalk and prespore gene expression in response to cAMP.


Cosi2: an efficient simulator of exact and approximate coalescent with selection.
Efficient simulation of population genetic samples under a given demographic model is a prerequisite for many analyses. Coalescent theory provides an efficient framework for such simulations, but simulating longer regions and higher recombination rates remains challenging. Simulators based on a Markovian approximation to the coalescent scale well, but do not support simulation of selection. Gene conversion is not supported by any published coalescent simulators that support selection. We describe cosi2, an efficient simulator that supports both exact and approximate coalescent simulation with positive selection. cosi2 improves on the speed of existing exact simulators, and permits further speedup in approximate mode while retaining support for selection. cosi2 supports a wide range of demographic scenarios, including recombination hot spots, gene conversion, population size changes, population structure and migration. cosi2 implements coalescent machinery efficiently by tracking only a small subset of the Ancestral Recombination Graph, sampling only relevant recombination events, and using augmented skip lists to represent tracked genetic segments. To preserve support for selection in approximate mode, the Markov approximation is implemented not by moving along the chromosome but by performing a standard backwards-in-time coalescent simulation while restricting coalescence to node pairs with overlapping or near-overlapping genetic material. We describe the algorithms used by cosi2 and present comparisons with existing selection simulators. A free C++ implementation of cosi2 is available at http://broadinstitute.org/mpg/cosi2.


Distinct functions of condensin I and II in mitotic chromosome assembly.
Condensin is a protein complex associated with mitotic chromosomes that has been implicated in chromosome condensation. In vertebrates, two types of condensin complexes have recently been identified, called condensin I and II. Here, we show that in mammalian cells condensin II associates with chromatin in prophase, in contrast to condensin I which is cytoplasmic and can thus interact with chromosomes only after nuclear envelope breakdown. RNA interference experiments in conjunction with imaging of live and fixed cells revealed that condensin II is required for chromosome condensation in early prophase, whereas condensin I appears to be dispensable at this stage. By contrast, condensin I is required for the complete dissociation of cohesin from chromosome arms, for chromosome shortening and for normal timing of progression through prometaphase and metaphase, whereas normal condensin II levels are dispensable for these processes. After depletion of both condensin complexes, the onset of chromosome condensation is delayed until the end of prophase, but is then initiated rapidly before nuclear envelope breakdown. These results reveal that condensin II and I associate with chromosomes sequentially and have distinct functions in mitotic chromosome assembly.


Nuclear transport factor GmNTF2B-1 enhances soybean drought tolerance by interacting with oxidoreductase GmOXR17 to reduce reactive oxygen species content.
Drought is a critical abiotic stressor that modulates soybean yield. Drought stress drastically enhances reactive oxygen species (ROS) formation, and maintaining ROS content above a cytostatic level but below a cytotoxic level is essential for normal biology processes in plants. At present, most of the known ROS-scavenging systems are in the cytoplasm, and the mechanism of ROS regulation in the nucleus remains unclear. GmNTF2B-1 is a member of the IV subgroup in the nucleus transporter family. Its expression is localized to the roots and is stimulated by drought stress. In this study, the overexpression of GmNTF2B-1 was found to improve the drought tolerance of transgenic soybean by influencing the ROS content in plants. An oxidoreductase, GmOXR17, was identified to interact with GmNTF2B-1 in the nucleus through the yeast two-hybrid, coimmunoprecipitation and bimolecular fluorescence complementation assays. The drought tolerance of GmOXR17 transgenic soybean was similar to that of GmNTF2B-1. GmNTF2B-1 was expressed in both cytoplasm and nucleus, and GmOXR17 transferred from the cytoplasm to the nucleus under drought stress. The overexpression of GmNTF2B-1 enhanced the nuclear entry of GmOXR17, and the overexpression of GmNTF2B-1 or GmOXR17 could decrease the H2 O2 content and oxidation level in the nucleus. In conclusion, the interaction between GmNTF2B-1 and GmOXR17 may enhance the nuclear entry of GmOXR17, thereby enhancing nuclear ROS scavenging to improve the drought resistance of soybean.


Indices of Diet Quality and Risk of Lung Cancer in the Women's Health Initiative Observational Study.
Prospective evidence on associations between diet quality indices and lung cancer risk is limited, particularly among older women. We investigated associations between 4 diet quality indices [Healthy Eating Index-2015 (HEI-2015), Alternate Healthy Eating Index 2010 (AHEI-2010), alternate Mediterranean Diet (aMED), and Dietary Approaches to Stop Hypertension (DASH)] and lung cancer incidence and mortality in the Women's Health Initiative Observational Study. Postmenopausal women aged 50-79 y at baseline (1993-1998) self reported their diet intake and information on relevant covariates. We used Cox proportional hazards regression models to estimate HRs and 95% CIs after controlling for age, smoking, and other relevant covariates. During ∼17 y of follow-up among 86,090 participants, 1491 lung cancer cases and 1393 lung cancer deaths were documented. Dietary indices were not associated with overall lung cancer incidence but were protective against squamous cell carcinoma (12.8% of total lung cancer) cases (HEI-2015: HR: 0.85; 95% CI: 0.76, 0.96; AHEI-2010: HR: 0.87; 95% CI: 0.78, 0.98; aMED: HR: 0.90; 95% CI: 0.81, 0.99; DASH: HR: 0.87; 95% CI: 0.77, 0.98). Among the indices, only HEI-2015 showed an inverse trend (P-trend = 0.02) with overall lung cancer mortality. Smoking status or participant age at baseline did not modify the association between dietary indices and lung cancer incidence or mortality. After comprehensive control of smoking exposure, we found that diet quality was not associated with overall lung cancer among postmenopausal women. However, a high-quality diet was inversely related to incident lung cancer of the squamous cell subtype. Future studies in populations with diverse age, smoking history, and dietary intake may further elucidate the relation between diet quality indices and lung cancer, especially by histological subtype.


Assessing the needs of breast cancer patients and their families.
Patient needs have long been elicited only through the use of customer satisfaction surveys, which unfortunately are not designed to obtain the in-depth information needed to guide the improvement of health care services. Through the use of the integrative group process and the help of many patients and hospital staffs, we developed a needs assessment survey for women with breast cancer. This article outlines this process and the results and discusses how we used these results to create a computer program designed to help women with breast cancer and their families.


New vessel formation after surgical brain injury in the rat's cerebral cortex II. Formation of the blood vessels distal to the surgical injury.
We investigated the features of newly formed blood vessels after surgical brain injury of the rat's cerebral cortex distal to the operated region. We document the process of split mature blood vessels by an endothelial bridge and morphological features of newly formed vessels. We did not observe a disruption of brain parenchyma. The endothelial lining in vessels was complete. The morphological features of the endothelial cells and basement membrane show that non-sprouting angiogenesis takes place distally to the surgical injury.


[Pathogenicity of hemosporidians (Sporozoa, Haemosporidia) of birds].
Ultrastructure of the cyst's wall was studied in monocercus type larvae of 5 species of the family Dilepididae: Dichoanotaenia tundra, Particterotaenia porosa, Platyscolex ciliata, Sacciuterina stellifera and Trichocephaloides megalocephala. In all examined cysticercoids the cyst's wall is of identical structure. It differs from the cyst's wall of larvae of the family Hymenolepididae by the character of material filling tegument, by fibrous and muscular, elements forming independent layers, by the absence of myelin-like layer limiting the cyst cavity. In addition to cytons and myofibroblasts in the cyst parenchima there are secretory cells producing electronic dense granules similar to those produced by neurosecretory cells of tape forms of cestodes.


Nondestructive production of exosomes loaded with ultrathin palladium nanosheets for targeted bio-orthogonal catalysis.
The use of exosomes as selective delivery vehicles of therapeutic agents, such as drugs or hyperthermia-capable nanoparticles, is being intensely investigated on account of their preferential tropism toward their parental cells. However, the methods used to introduce a therapeutic load inside exosomes often involve disruption of their membrane, which may jeopardize their targeting capabilities, attributed to their surface integrins. On the other hand, in recent years bio-orthogonal catalysis has emerged as a new tool with a myriad of potential applications in medicine. These bio-orthogonal processes, often based on Pd-catalyzed chemistry, would benefit from systems capable of delivering the catalyst to target cells. It is therefore highly attractive to combine the targeting capabilities of exosomes and the bio-orthogonal potential of Pd nanoparticles to create new therapeutic vectors. In this protocol, we provide detailed information on an efficient procedure to achieve a high load of catalytically active Pd nanosheets inside exosomes, without disrupting their membranes. The protocol involves a multistage process in which exosomes are first harvested, subjected to impregnation with a Pd salt precursor followed by a mild reduction process using gas-phase CO, which acts as both a reducing and growth-directing agent to produce the desired nanosheets. The technology is scalable, and the protocol can be conducted by any researcher having basic biology and chemistry skills in ~3 d.


Emerging dilemmas in the diagnosis and management of gastroesophageal reflux disease.
Gastroesophageal reflux disease (GERD) is common, but less so than widely reported because of inconsistencies in definition. In clinical practice, the diagnosis is usually based on a symptom assessment without testing, and the extent of diagnostic testing pursued should be limited to that which guides management or which protects the patient from the risks of a potentially morbid treatment or an undetected early (or imminent) esophageal adenocarcinoma or which does both. When testing is pursued, upper gastrointestinal endoscopy is the most useful initial diagnostic test because it evaluates for the major potential morbidities (Barrett's, stricture, and cancer) associated with GERD and facilitates the identification of some alternative diagnostic possibilities such as eosinophilic esophagitis. However, endoscopy is insensitive for diagnosing GERD because most patients with GERD have non-erosive reflux disease, a persistent diagnostic dilemma. Although many studies have tried to objectify the diagnosis of GERD with improved technology, this is ultimately a pragmatic diagnosis based on response to proton pump inhibitor (PPI) therapy, and, in the end, response to PPI therapy becomes the major indication for continued PPI therapy. Conversely, in the absence of objective criteria for GERD and the absence of apparent clinical benefit, PPI therapy is not indicated and should be discontinued. PPIs are well tolerated and safe, but nothing is perfectly safe, and in the absence of measurable benefit, even a miniscule risk dominates the risk-benefit assessment.


Hodgkin's disease and transplantation: a room with a (nontransplanter's) view.
Hodgkin's disease is a malignancy curable by combination chemotherapy in newly diagnosed patients. Management of refractory disease or relapse has been unsatisfactory with salvage chemotherapy alone, although some patients with long initial remission can be cured with this approach. Because correlations between dose intensity and response to treatment in animal models and in clinical studies have been positive, high-dose chemotherapy (HDT) with bone marrow (BM) or peripheral blood stem-cell (PBSC) transplantation support has been used in an attempt to improve disease-free and overall survival in patients with refractory disease or relapse. Controversy exists over who and when to transplant. Results are difficult to interpret because of the heterogeneous nature of the patients under study and short follow-up times. In general, patient disease-free and overall survival appear to be improved with HDT and PBSC support when compared with historical controls. Allografting has also been tried in Hodgkin's disease as well, but is not recommended due to high associated mortality. Improvements in supportive care for transplantation in general, and antiviral and graft-versus-host disease prophylaxis in particular, have decreased early mortality associated with allografting in other malignancies, but not yet in Hodgkin's disease. The fact that allografted patients who survive the initial transplant procedure have an impressively lower relapse rate makes Hodgkin's disease patients potential candidates for future studies of allografting under more modern circumstances. Some suggestions are made about the introduction of new approaches to treatment.


Risk assessment of SWEN21 a suggested new dive table for the Swedish armed forces: bubble grades by ultrasonography.
To develop the diving capacity in the Swedish armed forces the current air decompression tables are under revision. A new decompression table named SWEN21 has been created to have a projected risk level of 1% for decompression sickness (DCS) at the no stop limits. The aim of this study was to evaluate the safety of SWEN21 through the measurement of venous gas emboli (VGE) in a dive series. A total 154 dives were conducted by 47 divers in a hyperbaric wet chamber. As a proxy for DCS risk serial VGE measurements by echocardiography were conducted and graded according to the Eftedal-Brubakk scale. Measurements were done every 15 minutes for approximately 2 hours after each dive. Peak VGE grades for the different dive profiles were used in a Bayesian approach correlating VGE grade and risk of DCS. Symptoms of DCS were continually monitored. The median (interquartile range) peak VGE grade after limb flexion for a majority of the time-depth combinations, and of SWEN21 as a whole, was 3 (3-4) with the exception of two decompression profiles which resulted in a grade of 3.5 (3-4) and 4 (4-4) respectively. The estimated risk of DCS in the Bayesian model varied between 4.7-11.1%. Three dives (2%) resulted in DCS. All symptoms resolved with hyperbaric oxygen treatment. This evaluation of the SWEN21 decompression table, using bubble formation measured with echocardiography, suggests that the risk of DCS may be higher than the projected 1%.


A comparison of disease burden in rheumatoid arthritis, psoriatic arthritis and axial spondyloarthritis.
The main objective of this study was to compare disease burden in rheumatoid arthritis (RA), psoriatic arthritis (PsA) and axial spondyloarthritis (ax-SpA). In this cross-sectional study, all the RA (1093), PsA (365) and ax-SpA (333) patients who visited the out-patient clinic of the Hospital of Southern Norway Trust during the year 2013 were included; the RA patients all had a RA diagnosis verified by the treating rheumatologist, the PsA patients all fulfilled the ClASsification for Psoriatic ARthritis (CASPAR) criteria and the ax-SpA patients all fulfilled the Assessment of SpondyloArthritis international Society (ASAS) classification criteria for ax-SpA. Patient-reported health status, demographic variables, medications, and composite scores of disease activity were assessed. The main analyses were performed using General Linear Models adjusted for age, sex and multiple comparisons. Correlation analyses were performed using Spearman's rho. The reported pain, joint pain, patient's global assessment and fatigue were similar in PsA and ax-SpA, but significantly lower in RA. The 28-joint Disease Activity Score (DAS28) (0.3±0.1, p = 0.003), Clinical Disease Activity Index (CDAI) (1.0±0.4, p = 0.028) and Routine Assessment of Patient Index Data 3 (RAPID3) (0.4±0.1, p = 0.004) were all significantly higher in PsA vs. RA. RAPID3 showed moderate to high correlation with DAS28 (rho = 0.521, p<0.001) and CDAI (rho = 0.768, p<0.001) in RA and PsA, and with Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) (rho = 0.902, p<0.001) and Bath Ankylosing Spondylitis Functional Index (BASFI) (0.865, p<0.001) in ax-SpA and PsA. In conclusion, patient- reported outcome measures were similar in our population of PsA and ax-SpA patients, but significantly lower for the RA patients. Composite disease activity measures were lower in RA than in PsA and ax-SpA, but the magnitude of these differences was small and probably not of clinical significance. Our study indicates that disease burden in RA, PsA and ax-SpA may be more similar than previously demonstrated.


Simulation of the single-vibronic-level emission spectrum of HPS.
We have computed the potential energy surfaces of the X¹A' and ùA" states of HPS using the explicitly correlated multi-reference configuration interaction (MRCI-F12) method, and Franck-Condon factors between the two states, which include anharmonicity and Duschinsky rotation, with the aim of testing the assignment of the recently reported single-vibronic-level (SVL) emission spectrum of HPS [R. Grimminger, D. J. Clouthier, R. Tarroni, Z. Wang, and T. J. Sears, J. Chem. Phys. 139, 174306 (2013)]. These are the highest level calculations on these states yet reported. It is concluded that our spectral simulation supports the assignments of the molecular carrier, the electronic states involved and the vibrational structure of the experimental laser induced fluorescence, and SVL emission spectra proposed by Grimminger et al. [J. Chem. Phys. 139, 174306 (2013)]. However, there remain questions unanswered regarding the relative electronic energies of the two states and the geometry of the excited state of HPS.


Encoding generalized phase functions on Dammann gratings.
We extend the Dammann encoding concept to more general phase functions, i.e., to encode a phase pattern so that a number of diffracted orders have the same intensity. We demonstrate a Dammann vortex grating that generates different vortex patterns in the different diffracted orders. We also show a Fresnel lens that creates seven transverse focal spots in each of seven axial planes.


Vitamin B12 requirement of a marine blue-green alga.
A species of Synechocystis isolated from a marine mud has an absolute requirement for vitamin B(12). Analogues of B(12), including cobinamide and alpha-(adenyl) cobamide cyanide, satisfactorily substitute for vitamin B(12). Methionine alone or in combination with other compounds supports only a low level of growth.


Physiological and pathological left ventricular hypertrophy of comparable degree is associated with characteristic differences of in vivo hemodynamics.
Left ventricular (LV) hypertrophy is a physiological or pathological response of LV myocardium to increased cardiac load. We aimed at investigating and comparing hemodynamic alterations in well-established rat models of physiological hypertrophy (PhyH) and pathological hypertrophy (PaH) by using LV pressure-volume (P-V) analysis. PhyH and PaH were induced in rats by swim training and by abdominal aortic banding, respectively. Morphology of the heart was investigated by echocardiography. Characterization of cardiac function was completed by LV P-V analysis. In addition, histological and molecular biological measurements were performed. Echocardiography revealed myocardial hypertrophy of similar degree in both models, which was confirmed by post-mortem heart weight data. In aortic-banded rats we detected subendocardial fibrosis. Reactivation of fetal gene program could be observed only in the PaH model. PhyH was associated with increased stroke volume, whereas unaltered stroke volume was detected in PaH along with markedly elevated end-systolic pressure values. Sensitive indexes of LV contractility were increased in both models, in parallel with the degree of hypertrophy. Active relaxation was ameliorated in athlete's heart, whereas it showed marked impairment in PaH. Mechanical efficiency and ventriculo-arterial coupling were improved in PhyH, whereas they remained unchanged in PaH. Myocardial gene expression of mitochondrial regulators showed marked differences between PaH and PhyH. We provided the first comparative hemodynamic characterization of PhyH and PaH in relevant rodent models. Increased LV contractility could be observed in both types of LV hypertrophy; characteristic distinction was detected in diastolic function (active relaxation) and mechanoenergetics (mechanical efficiency), which might be explained by mitochondrial differences.


[The Mais Médicos program - a conceptual mistake].
The Mais Médicos program (involving the influx of Cuban physicians) has been a topic of heated debate in the media, mainly due to the difference of opinion between the government and the medical profession. The population in general is left in a climate of anxiety and uncertainty. This paper discusses the misconceptions that have been misleading and confusing the actors involved in the conflict surrounding the program in question.


Extra-pituitary expressed follicle-stimulating hormone: Is it physiologically important?
Pituitary gonadotropes synthesize and secrete follicle-stimulating hormone (FSH). FSH is a heterodimer that consists of an α- and β-subunit. The α-subunit is common to other pituitary and placental glycoprotein hormones, and the β-subunit is the hormone/receptor-specific subunit. Although the pituitary is the main tissue that accounts for circulating hormone, previous and recent reports indicate extra-pituitary sources of FSH production including mouse gonads, human stomach, prostate, umbilical cord vein endothelial cells, uterine myometrium, placenta, and chicken abdominal adipose tissue. Whether extra-pituitary derived FSH exerts any physiologically significant actions is not known. In this review, we have comprehensively analyzed the expression of mRNAs that encode mouse and human FSH subunits and also their corresponding expressed sequence tags in normal tissues, cancer cell lines, and primary tumors by public database mining. We propose criteria to assess the significance of individual FSH subunit or FSH dimer expression as well as genetic approaches to unambiguously define the physiological relevance of extra-pituitary FSH expression.


Longitudinal coherence measurements of a transient collisional x-ray laser.
We present what is to our knowledge the first longitudinal coherence measurement of a transient inversion collisional x-ray laser. We investigated the picosecond output of a Ni-like Pd x-ray laser at 14.68 nm generated by the COMET laser facility at the Lawrence Livermore National Laboratory. Interference fringes were generated with a Michelson interferometer setup in which a thin multilayer membrane was used as a beam splitter. We determined the longitudinal coherence for the 4d1S0 --> 4p1P1 lasing transition to be approximately 400 microm (1/e half-width) by changing the length of one interferometer arm and measuring the resultant variation in fringe visibility. The inferred gain-narrowed linewidth of approximately 0.29 pm is a factor of 4 less than previously measured in quasi-steady-state x-ray laser schemes.


Fingerprinting and characterization of the polysaccharides from Polygonatum odoratum and the in vitro fermented effects on Lactobacillus johnsonii.
Polygonatum odoratum (Yu-Zhu) can be utilized to treat the digestive and respiratory illness. Previous studies have revealed that the underlying therapeutic mechanism of P. odoratum polysaccharides (POPs) is associated with remodeling the gut microbiota. However, POPs in terms of the chemical composition and fermentation activities have been understudied. Here we developed the three-level fingerprinting approaches to characterize the structures of POPs and probed into the beneficial effects on promoting the growth and fermentation of Lactobacillus johnsonii. POPs were prepared by water decoction followed by alcohol sedimentation, while trifluoroacetic acid under different conditions to prepare the hydrolyzed oligosaccharides and monosaccharides. POPs exhibited three main molecular distribution of 601-620 kDa, 4.12-6.09 kDa, and 3.57-6.02 kDa. Hydrolyzed oligosaccharides with degree of polymerization (DP) 2-13 got primarily characterized by analyzing the rich fragmentation information obtained by hydrophilic interaction chromatography/ion mobility-quadrupole time-of-flight mass spectrometry (HILIC/IM-QTOF-MS). Amongst them, the DP5 oligosaccharide was characterized as 1,6,6-kestopentaose. The molecular ratio of Fru: Ara: Glc: Gal: Xyl was 87.72: 0.30: 11.56: 0.19: 0.23. In vitro fermentation demonstrated that 4.5 mg/mL of POPs could significantly promote the growth of L. johnsonii. Co-cultivated with 4.5 mg/mL of POPs, L. johnsonii exhibited stronger antimicrobial activity against Klebsiella pneumoniae. The concentrations of short-chain fatty acids in the POPs-lactobacilli fermented products, including acetic acid, isobutyric acid, and isovaleric acid, were increased. Conclusively, POPs represent the promising prebiotic candidate to facilitate lactobacilli, which is associated with exerting the health benefits.


Role of mast cells in inflammatory bowel disease and inflammation-associated colorectal neoplasia in IL-10-deficient mice.
Inflammatory bowel disease (IBD) is hypothesized to result from stimulation of immune responses against resident intestinal bacteria within a genetically susceptible host. Mast cells may play a critical role in IBD pathogenesis, since they are typically located just beneath the intestinal mucosal barrier and can be activated by bacterial antigens. This study investigated effects of mast cells on inflammation and associated neoplasia in IBD-susceptible interleukin (IL)-10-deficient mice with and without mast cells. IL-10-deficient mast cells produced more pro-inflammatory cytokines in vitro both constitutively and when triggered, compared with wild type mast cells. However despite this enhanced in vitro response, mast cell-sufficient Il10(-/-) mice actually had decreased cecal expression of tumor necrosis factor (TNF) and interferon (IFN)-gamma mRNA, suggesting that mast cells regulate inflammation in vivo. Mast cell deficiency predisposed Il10(-/-) mice to the development of spontaneous colitis and resulted in increased intestinal permeability in vivo that preceded the development of colon inflammation. However, mast cell deficiency did not affect the severity of IBD triggered by non-steroidal anti-inflammatory agents (NSAID) exposure or helicobacter infection that also affect intestinal permeability. Mast cells thus appear to have a primarily protective role within the colonic microenvironment by enhancing the efficacy of the mucosal barrier. In addition, although mast cells were previously implicated in progression of sporadic colon cancers, mast cells did not affect the incidence or severity of colonic neoplasia in this inflammation-associated model.


Prevalent, protective, and convergent IgG recognition of SARS-CoV-2 non-RBD spike epitopes in COVID-19 convalescent plasma.
Although humoral immunity is essential for control of SARS-CoV-2, the molecular composition, binding epitopes and effector functions of the immunoglobulin G (IgG) antibodies that circulate in blood plasma following infection are unknown. Proteomic deconvolution of the circulating IgG repertoire (Ig-Seq 1 ) to the spike ectodomain (S-ECD 2 ) in four convalescent study subjects revealed that the plasma response is oligoclonal and directed predominantly (>80%) to S-ECD epitopes that lie outside the receptor binding domain (RBD). When comparing antibodies directed to either the RBD, the N-terminal domain (NTD) or the S2 subunit (S2) in one subject, just four IgG lineages (1 anti-S2, 2 anti-NTD and 1 anti-RBD) accounted for 93.5% of the repertoire. Although the anti-RBD and one of the anti-NTD antibodies were equally potently neutralizing in vitro , we nonetheless found that the anti-NTD antibody was sufficient for protection to lethal viral challenge, either alone or in combination as a cocktail where it dominated the effect of the other plasma antibodies. We identified in vivo protective plasma anti-NTD antibodies in 3/4 subjects analyzed and discovered a shared class of antibodies targeting the NTD that utilize unmutated or near-germline IGHV1-24, the most electronegative IGHV gene in the human genome. Structural analysis revealed that binding to NTD is dominated by interactions with the heavy chain, accounting for 89% of the entire interfacial area, with germline residues uniquely encoded by IGHV1-24 contributing 20% (149 Å 2 ). Together with recent reports of germline IGHV1-24 antibodies isolated by B-cell cloning 3,4 our data reveal a class of shared IgG antibodies that are readily observed in convalescent plasma and underscore the role of NTD-directed antibodies in protection against SARS-CoV-2 infection.


Pattern of cerebral metabolic interactions in a subject with isolated amnesia at risk for Alzheimer's disease: a longitudinal evaluation.
A pattern of reduced cerebral metabolic rate for glucose (rCMRglc) has been shown by positron emission tomography (PET) in patients with dementia of the Alzheimer type. To verify if a similar rCMRglc pattern is present in subjects 'at risk' for Alzheimer's disease (AD), we used high-resolution PET to longitudinally study a subject with isolated memory impairment and a family history for autosomal dominant AD. Initial rCMRglc data did not reveal any consistent abnormality as compared to a group of sex- and age-matched healthy controls. However, 1 year later, a follow-up evaluation did reveal reduced parietal rCMRglc values coinciding with a worsening of cognitive impairment, which suggested that standard analyses of resting rCMRglc data may not be useful in the early diagnosis of AD. In contrast, when a previously determined discriminant function for distinguishing controls from AD patients was applied, the subject was correctly identified as an AD patient on both PET scans.


Design and Selection of a Synthetic Feedback Loop for Optimizing Biofuel Tolerance.
Feedback control allows cells to dynamically sense and respond to environmental changes. However, synthetic controller designs can be challenging because of implementation issues, such as determining optimal expression levels for circuit components within a feedback loop. Here, we addressed this by coupling rational design with selection to engineer a synthetic feedback circuit to optimize tolerance of Escherichia coli to the biojet fuel pinene. E. coli can be engineered to produce pinene, but it is toxic to cells. Efflux pumps, such as the AcrAB-TolC pump, can improve tolerance, but pump expression impacts growth. To address this, we used feedback to dynamically regulate pump expression in response to stress. We developed a library with thousands of synthetic circuit variants and subjected it to three types of pinene treatment (none, constant, and varying pinene). We were able to select for strains that were biofuel tolerant without a significant growth cost in the absence of biofuel. Using next-generation sequencing, we found common characteristics in the designs and identified controllers that dramatically improved biofuel tolerance.


Healthcare Providers' Experiences with Hiccups in Patients with Cancer: Report of a United States National Survey.
Hiccups occur in 15-40% of cancer patients, but previous research has not sought the perspectives of cancer healthcare providers. The objective of this research is to report on United States cancer healthcare providers' awareness of their patients' hiccups and these healthcare providers' perceived need for further palliation options. A survey was developed and then distributed throughout the United States via email to cancer healthcare providers; results are reported descriptively. Six hundred eighty-four cancer healthcare providers completed 2 eligibility screening questions which required them to have cared for an adequate number of patients (> 10 in the past 6 months) with "clinically significant" hiccups (defined as hiccups that persisted for >48 hours or occurred from cancer or from cancer care). Of 113 eligible healthcare providers, 90 completed the survey. Healthcare providers described hiccups as associated with stress/anxiety, fatigue, sleep problems, and decreased work/school productivity. In 49% of patients, healthcare providers initially prescribed medications (commonly chlorpromazine or baclofen); 18% expressed dissatisfaction with current palliation. Proffered comments included, "When current therapies do not work, it can be very demoralizing to our patients; " and "…my biggest complaint is that current treatments also come with their own side effects which can be quite severe." Discordance appears to exist between the percentage of cancer patients with hiccups and the percentage of cancer healthcare providers with awareness of their patients' hiccups. Nonetheless, healthcare providers described notable hiccup-associated symptoms in their patients and a need for more palliative options.


Amino terminus of reovirus nonstructural protein sigma NS is important for ssRNA binding and nucleoprotein complex formation.
Reovirus nonstructural protein sigma NS exhibits a ssRNA-binding activity thought to be involved in assembling the reovirus mRNAs for genome replication and virion morphogenesis. To extend analysis of this activity, recombinant sigma NS (r sigma NS) was expressed in insect cells using a recombinant baculovirus. In infected-cell extracts, r sigma NS was found in large complexes (> or = 30 S) that were disassembled into smaller, 13-19 S complexes upon treatment with RNase A. R sigma NS also bound to poly(A)-Sepharose beads both before and after purification. Treatment with high salt during purification caused r sigma NS to sediment in even smaller, 7-9 S complexes, consistent with more complete loss of RNA. To localize the RNA-binding site, limited proteolysis was used to fragment the r sigma NS protein. Upon mild treatment with thermolysin, 11 amino acids were removed from the amino terminus of r sigma NS, and the resulting protein no longer bound to poly(A). In addition, when r sigma NS in cell extracts was treated with thermolysin to generate the amino-terminally truncated from, it sedimented at 7-9 S, also consistent with the loss of RNA-binding capacity. To confirm these findings, a deletion mutant lacking amino acids 2-11 was constructed and expressed in insect cells from a recombinant baculovirus. The mutant protein in cell extracts showed greatly reduced poly(A)-binding activity and sedimented as 7-9 S complexes. These data suggest that the first 11 amino acids of sigma NS, which are predicted to form an amphipathic alpha-helix, are important for both ssRNA binding and formation of complexes larger than 7-9 S.


Internal circuit noise of hearing aids and speech intelligibility.
The circuit noise generated internally by various components of a hearing aid is influenced mostly by circuit gain and frequency response. The measurement of this variable and its effect, if any, on the listener's ability to unders-and monosyllabic words through a hearing-aid system was investigated. The overall circuit noise of 10 behind-the-ear hearing aids of different brands with comparable gain and frequency response was measured. Based on these measurements, a low-noise and a high-noise hearing aid were selected as the test aids. The internal circuit noise of each test aid was then measured automatically by utilizing a 1/3-octave-band tracking filter. Monosyllabic words were recorded through the test aids at two gain settings. The prerecorded test tapes were presented to 32 normal-hearing listeners. Findings indicate that different amounts of circuit noise generated internally by a hearing aid do exist among various aids. However, the noise generated by the hearing aids used in this study does not affect significantly the intelligibility of monosyllabic words.


Cumulative glycemic exposure and microvascular complications in insulin-dependent diabetes mellitus. The glycemic threshold revisited.
The development of microvascular insulin-dependent diabetes mellitus (IDDM) complications has been shown to be related to both duration of diabetes and the degree of glycemic exposure. However, controversy exists as to whether there is a threshold of glycemic exposure, below which there is minimal risk. Furthermore, there are few data describing the relationship of total glycemic exposure (duration x degree) to complications rates-a potentially useful research and clinical tool. To determine a cumulative glycemic exposure variable that combines the effect of both degree and duration of hyperglycemia and to evaluate this variable in terms of its relation to microvascular complications. The association between cumulative glycemic exposure and complication risk was also examined to evaluate whether there was a threshold effect. A total of 353 patients with IDDM who had completed the first 6 years of follow-up in the Pittsburgh Epidemiology of Diabetes Complications Study were included in this analysis. These subjects had a mean age of 27.9 years, and the mean duration of the disease was 19.4 years. Subjects were examined at baseline (cycle 1) and then biennially (cycle 2, cycle 3, and cycle 4) for diabetes complications. Total glycosylated hemoglobin (HbA1) was measured at each cycle. A cumulative glycemic exposure variable, named A1months, was calculated by multiplying the number of HbA1 units above normal at each cycle by the number of months between the midpoints of the preceding and succeeding cycle intervals. The mean number of A1months experienced at the time of diagnosis of proliferative retinopathy (914), microalbuminuria (952), overt nephropathy (1043), and distal symmetrical polyneuropathy (1043) did not vary by duration of diabetes. Thus, approximately 1000 A1months were needed (on average) for the advanced complications to develop. Although the risk for developing proliferative retinopathy rose gradually as A1months increased, a more abrupt increase in the risk was seen (again at approximately 1000 A1months) for microalbuminuria (odds ratio, 6.9; 95% confidence interval, 2.5-19.1), overt nephropathy (odds ratio, 6.5; 95% confidence interval, 2.0-21.7), and distal symmetrical polyneuropathy (odds ratio, 6.5; 95% confidence interval, 2.4-17.8). Nonetheless, complications developed in the majority of cases at glycemic exposures below 1000 A1months. The cumulative glycemic exposure variable A1months does not predict complications any better than its component variables (duration and HbA1). Furthermore, formal statistical testing failed to show a definitive threshold for any complication. Although A1months does not enhance prediction of complications, it may be a useful summary measure of glycemic exposure for both patients and physicians. However, although subjects with 1000 A1months or more appear to be at increased risk of developing most microvascular complications, because the majority of complications arise in subjects with less than this exposure, this threshold value should only be considered a minimal goal. For example, our data suggest that for most microvascular complications to develop, it would take, on average, 83 years with an HbA1 unit at 1% above normal, 42 years at 2% above normal, 28 years at 3% above normal. 21 years at 4% above normal, and 18 years at 5% above normal.


Serial casting of the ankles in Duchenne muscular dystrophy: can it be an alternative to surgery?
The aim of this non-randomized pilot study was to assess whether serial casting can be used in Duchenne muscular dystrophy (DMD) patients to reduce tendo-achilles (TA) contractures and to allow the fitting of the knee-ankle-foot orthoses (KAFOs) at the end of the functional walking. TA range and age at loss of standing with KAFOs were prospectively assessed in nine DMD patients who underwent serial casting prior to rehabilitation into KAFOs and compared to a group of 20 DMD patients who had TA (group II) or TA and iliotibial band (ITB) surgical resection (group III). TA range after casting was reduced at least to plantargrade/neutral in 8 of the 9 patients studied although patients who had surgery had better correction of TA angles. The mean interval between rehabilitation and loss of standing ability with KAFOs was only minimally different among the three groups 1.8 years (9 months-2.8 years) in group I, 2 years (1 month-4.3 years) in group II, 1.9 years (6 months-4.7 years) in group III. The results of this pilot study suggest that serial casting of the TAs can be considered in DMD patients with moderate TA contractures and in whom there are no significant iliotibial band (ITB) tightness, although this intervention is probably not as effective as surgery in maintaining long-term TA range. This procedure may therefore be considered both in patients with medical contraindication to surgery, and also in patients or families who are not prepared to cope with the stress and discomfort of TA surgery.


Perceived Impact of an Online Community Care Platform for Dutch Older Adults on Local Participation, Informal Caregiving, and Feelings of Connectedness: Pretest-Posttest Observational Study.
In a changing ageing society wherein older adults are increasingly expected to take care of themselves instead of relying on health care services, online community care platforms can help older adults to meet these expectations. A considerable number of these online community care platforms have been introduced in several European countries based on their potential. However, their actual impact is unclear. The aim of this study was to investigate the self-reported use, expectations, and perceived impact of a Dutch online community care platform called Grubbenvorst-Online among Dutch older adults. The following 2 questions were studied: (1) What is the self-reported use of Grubbenvorst-Online among older adults? (2) What are their expectations and perceived impact of Grubbenvorst-Online regarding local participation, their social network, mutual informal caregiving, and feelings of connectedness? An observational pretest-posttest study was conducted. Participants were recruited via a web-based message on the Grubbenvorst-Online platform and data were collected via postal questionnaires among older users at the start of the study and 4 months later. Data regarding the expectations and the perceived impact of Grubbenvorst-Online were compared and tested. Forty-seven Grubbenvorst-Online users with an average age of 74 years participated in this study. They were healthy, predominantly "internet-skilled," and they found the internet important for maintaining social contacts. In general, the use of the online community care platform decreased during the 4-month follow-up period. The perceived impact of Grubbenvorst-Online was significantly lower than that expected regarding information provision (P=.003), seeking help from fellow villagers (P<.001), giving help to fellow villagers (P<.001), and consulting care or welfare services (P<.001). The findings of this study indicate that online community care platforms perhaps do not provide enough "added value" in their current form. We suggest a new direction in which online community care platforms primarily support existing offline initiatives aimed at stimulating local participation, informal caregiving, and feelings of connectedness.


[Morphology of the optic papilla in glaucoma. II. Secondary chronic open angle glaucoma].
Previous studies have shown that the chronic open-angle glaucomas form a heterogeneous spectrum of diseases which have in common an open anterior chamber angle and glaucomatous optic nerve damage. Purpose of this study was to evaluate whether the appearance of the optic disc shows specific features among various types of secondary chronic open-angle glaucoma. Clinical data and color-stereo optic disc photographs of 126 patients with pseudoexfoliative glaucoma and 47 patients with pigmentary glaucoma were compared with those of 501 patients with primary open-angle glaucoma (POAG) and of 481 normal subjects. The glaucoma groups did not differ in neuroretinal rim nor in perimetric mean defect. Mean optic disc area was significantly smaller in the pseudoexfoliative glaucoma eyes (2.54 +/- 0.51 mm2 vs. 2.71 +/- 0.63 mm2, p = 0.03) than in the primary open-angle glaucoma eyes. The pigmentary glaucoma group did not vary significantly from the primary open-angle glaucoma group in size of the optic disc. No significant differences were found for neuroretinal rim area, configuration of neuroretinal rim, depth of optic cup and diameters of the retinal arterioles and venules at the disc border between the secondary glaucoma groups and the POAG group respectively. Size of zone beta of the parapapillary atrophy was slightly, but not significantly smaller in the secondary glaucoma groups than in POAG. In the secondary glaucoma groups, the maximal intraocular pressure measurements were significantly (p < 0.001) higher than in the group with POAG. All glaucoma groups had a significantly smaller neuroretinal rim, significantly smaller retinal arterioles, and significantly larger parapapillary atrophy compared to the normal group. Except of a slightly smaller optic disc in eyes with pseudoexfoliative glaucoma, eyes with secondary glaucoma due to pseudoexfoliation or due to pigmentary dispersion do not vary significantly in their optic disc morphology compared to POAG and do not show pathognomonic features of the optic disc despite marked changes in the anterior segment of the eye.


Effect of pathogen concentrations on removal of Cryptosporidium and Giardia by conventional drinking water treatment.
The presence of waterborne enteric pathogens in municipal water supplies contributes risk to public health. To evaluate the removal of these pathogens in drinking water treatment processes, previous researchers have spiked raw waters with up to 10(6) pathogens/L in order to reliably detect the pathogens in treated water. These spike doses are 6-8 orders of magnitude higher than pathogen concentrations routinely observed in practice. In the present study, experiments were conducted with different sampling methods (i.e., grab versus continuous sampling) and initial pathogen concentrations ranging from 10(1) to 10(6) pathogens/L. Results showed that Cryptosporidium oocyst and Giardia cyst removal across conventional treatment were dependent on initial pathogen concentrations, with lower pathogen removals observed when lower initial pathogen spike doses were used. In addition, higher raw water turbidity appeared to result in higher log removal for both Cryptosporidium oocysts and Giardia cysts.


Monoaminergic control of spinal locomotor networks in SOD1G93A newborn mice.
Mutations in the gene that encodes Cu/Zn-superoxide dismutase (SOD1) are the cause of approximately 20% of familial forms of amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disease characterized by the progressive loss of motor neurons. While ALS symptoms appear in adulthood, spinal motoneurons exhibit functional alterations as early as the embryonic and postnatal stages in the murine model of ALS, the SOD1 mice. Monoaminergic - i.e., dopaminergic (DA), serotoninergic (5-HT), and noradrenergic (NA) - pathways powerfully control spinal networks and contribute significantly to their embryonic and postnatal maturation. Alterations in monoaminergic neuromodulation during development could therefore lead to impairments in the motoneuronal physiology. In this study, we sought to determine whether the monoaminergic spinal systems are modified in the early stages of development in SOD1 mice. Using a post-mortem analysis by high performance liquid chromatography (HPLC), monoaminergic neuromodulators and their metabolites were quantified in the lumbar spinal cord of SOD1 and wild-type (WT) mice aged one postnatal day (P1) and P10. This analysis underscores an increased content of DA in the SOD1 lumbar spinal cord compared to that of WT mice but failed to reveal any modification of the other monoaminergic contents. In a next step, we compared the efficiency of the monoaminergic compounds in triggering and modulating fictive locomotion in WT and SOD1 mice. This study was performed in P1-P3 SOD1 mice and age-matched control littermates using extracellular recordings from the lumbar ventral roots in the in vitro isolated spinal cord preparation. This analysis revealed that the spinal networks of SOD1(G93A) mice could generate normal locomotor activity in the presence of NMA-5-HT. Interestingly, we also observed that SOD1 spinal networks have an increased sensitivity to NA compared to WT spinal circuits but exhibited similar DA responses.


Morpho-molecular characterization of Discosia ravennica sp. nov. and a new host record for Sporocadus rosigena.
Collections of fungal samples from two dead leaf specimens from Italy were subjected to morphological examination and phylogenetic analyses. Two coelomycetous taxa belonging to two different genera in Xylariomycetidae, Sordariomycetes, namely Discosia and Sporocadus, were identified. The Discosia taxon is revealed as a new species and is herein introduced as Discosia ravennica sp. nov. while the Sporocadus taxon is identified as Sporocadus rosigena. Multi-locus phylogeny based on DNA sequence data of the large subunit (LSU) and internal transcribed spacer (ITS) of nuclear ribosomal genes, β-tubulin (β-tub) and RNA polymerase II second largest subunit (rpb2) showed that D. ravennica is related to D. neofraxinea but it forms an independent lineage that supports its new species status. The new taxon also differs from other Discosia species by its unilocular to bilocular, superficial and applanate conidiomata with basal stroma composed of cells of textura angularis, elongate-ampulliform conidiogenous cells and conidia smaller in size. Sporocadus rosigena is here reported as a new host record from Quercus ilex from Italy. Descriptions, illustrations and molecular data for both species are provided in this paper.


[Risk factors for sexual behaviors in girl's high school students].
The purpose of this study was to examine risk factors for sexual behaviors in Korean female high school students. Data was collected by a written questionnaire from June 22 to July 18, 2002 from 522 girls, who were stratified samples from a target population of 63,375 11th grade students from 200 regular high schools and 70 vocational high schools in Seoul, Korea. We conducted multiple regression analysis using the SAS pc+ program. Risk factors for intimate behaviors were association with boy friends, differential association, family attachment, and family abuse experience. These variables cause 68% of intimate behaviors. Risk factors for sexual experiences were differential association, association with boy friend, and family attachment. These variables cause 14% of sexual experiences. Efforts to reduce sexual behaviors in girls should include the possible role of peers and develop peer leader programs.


A delayed role for nitric oxide-sensitive guanylate cyclases in a migratory population of embryonic neurons.
Neuronal differentiation requires a coordinated intracellular response to diverse extracellular stimuli, but the role of specific signaling mechanisms in regulating this process is still poorly understood. Soluble guanylate cyclases (sGCs), which can be stimulated by diffusible free radical gasses such as nitric oxide (NO) and carbon monoxide (CO) to produce the intracellular messenger cGMP, have recently been found to be expressed within a variety of embryonic neurons and implicated in the control of both neuronal motility and differentiation. Using the enteric nervous system (ENS) of the moth, Manduca sexta, we examined the role of NO and NO-sensitive sGCs during the migration and differentiation of an identified set of migratory neurons (the EP cells). Shortly after the onset of their migration, a subset of EP cells began to express NO-sensitive sGC activity (visualized with an anti-cGMP antiserum). Unlike many neurons in the central nervous system, the expression of sGC activity in the EP cells was not transient but persisted throughout subsequent periods of axon elongation and terminal branch formation on the gut musculature. In contrast, nitric oxide synthase activity (visualized using NADPH-diaphorase histochemistry) was undetectable in the vicinity of the EP cells until the period of synapse formation. Manipulations designed to alter sGC and NOS activity in an in vivo embryonic culture preparation had no discernible effect on either the migration or axonal outgrowth of the EP cells. In contrast, inhibition of both of these enzymes resulted in a significant reduction in terminal synaptic branch formation within the postmigratory neurons. These results indicate that while NO-sensitive sGC activity is expressed precociously within the EP cells during their initial migratory dispersal, a role for this signaling pathway can only be demonstrated well after migration is complete, coincident with the formation of mature synaptic connections.


Population fluctuation of Anopheles (Diptera: Culicidae) in forest and forest edge habitats in Tucumán province, Argentina.
The aim of this work was to study the possible effects of forest and forest edge habitats on the population fluctuation of the Anopheles species in northwestern Argentina, taking into consideration the relationship between this fluctuation and climatic variables. This study is one of the first that involves the Anopheles fauna in the country and its dynamics in two different habitats. Sampling was carried out from October, 2002 to October, 2003, in the forest and on the forest edge. Both habitats were compared for species diversity and abundance, and multiple regression analyses were performed to analyze the effects of environmental variables on the population dynamics. Five hundred and sixteen adult specimens of Anopheles species were collected, the most numerous group being Arribalzaga (52.1%), followed by Anopheles (Nyssorhynchus) strodei (20.5%) and Anopheles (Nyssorhynchus) evansae (6.4%). Mosquito abundance was greatest in the forest, the most productive habitat. Samples were collected throughout the sampling period, with a smaller peak in summer. Small numbers of Anopheles (Anopheles) pseudopunctipennis were found throughout the year. Relative humidity, with a 15-day delay, was the factor that most strongly contributed to the temporal sample fluctuation. We conclude that the best season for anopheline development in the study area is from spring to fall, although the period with the greatest transmission risk is the fall, with the greatest An. pseudopunctipennis abundance.


Evidence of Chlamydophila abortus vaccine strain 1B as a possible cause of ovine enzootic abortion.
Chlamydophila abortus, the agent of ovine enzootic abortion (OEA), is a major cause of lamb mortality worldwide. Disease can be controlled through the use of vaccines based on the 1B temperature-sensitive mutant strain of C. abortus. This study investigated suspected OEA cases across Scotland for the presence of the 1B strain by analysis of recently identified unique point mutations (9). Thirty-five cases were C. abortus-positive and 14 came from vaccinated flocks. Analysis of single nucleotide polymorphisms by PCR-RFLP and sequence analysis revealed the presence of point mutations consistent with the presence of the 1B vaccine strain in 5 of these 14 samples. Quantitative real-time PCR revealed comparable numbers of genome copies of the 1B strain in infected placentas to those present following wild-type infection. This study is the first to demonstrate the presence of the 1B vaccine strain in the placentas of OEA cases and suggests a probable causal role in the disease.