Quality of life after I-125 seed implantation using computed tomography and three-dimensional-printed template guidance in patients with advanced malignant tumor.
I-125 seed implantation has been widely applied in the local treatment of advanced malignant tumor. It has the advantages of providing a high dose of treatment to the target sites and low dose to normal tissues. It has been mostly applied as palliative treatment for recurrences in advanced malignant tumor (except for prostate cancer), suppressing tumor development and improving the quality of life of patients. The objective of this study was to investigate changes in quality of life for patients with advanced malignant tumor after receiving I-125 seed implantation using a three-dimensional (3D)-printed individualized template and computed tomography (CT) guidance. In this prospective study, convenience sampling was applied for patients with advanced tumors attending a tertiary hospital. The European Organization for Research on Treatment of Cancer Quality of Life Questionnaire-C30 was involved to assess quality of life. Patients completed the questionnaire before and 24 h after seed implantation. The questionnaire of 1 and 3 months after seed implantation was completed by telephonic follow-up. A total of 42 patients were included (24 males and 18 females), with an average age of 58.86 ± 14.13 years (ranged 25-91 years). The average scale score after seed implantation was higher than that of before implantation. The order was the average scale score 1 month after seed implantation >3 months after seed implantation >24 h after seed implantation. The results suggested that the quality of life could be improved with I-125 seed implantation using a 3D-printed individualized template under CT guidance in patients with the advanced malignant tumor.


Lack of Influence of Calcium/Phosphorus Ratio on Hip and Lumbar Bone Mineral Density in Older Americans: NHANES 2005-2006 Cross-Sectional Data.
We sought to ascertain the effect of a low dietary calcium/phosphorus (Ca:P) ratio on the bone health of older adults in the United States. The present analysis assessed whether a high dietary consumption of P, which generally leads to a low dietary Ca:P ratio, has an unfavorable effect on the bone mineral density (BMD) of the hip and lumbar vertebrae in a representative sample of older US men and women. For the 1228 men and women aged 50 to 70 and ≥71 years included in the National Health and Nutrition Examination Survey (NHANES) 2005 to 2006 cycle, quintiles of the dietary Ca:P ratio were tested for their association with hip and lumbar BMD after adjusting for body mass index (BMI). All data in this observational study were cross-sectional. Women typically have higher dietary Ca:P ratios than men and lower BMDs. No trend emerged for any age or sex group when studying the relationship between the dietary Ca:P ratio and BMD with adjustment for BMI. A wide range of dietary Ca:P ratios in the diets of a cross-section of older adult men and women in the United States had little effect on the BMD of the hip (proximal femur) or the lumbar vertebrae (spine), even among those consuming large amounts of Ca supplements. Despite the lack of complete assessment of total P intake in the United States, these results suggest that high P consumption patterns and low dietary Ca:P ratios do not exert an adverse effect on BMD at major fracture sites in older adults.


Angiolymphoid hyperplasia with eosinophilia and entrapment of the ulnar nerve.
The angiolymphoid hyperplasia with eosinophilia (ALHE) is a sporadic vasoproliferative lesion of uncertain etiology involving the skin and the subcutaneous tissue. Occasionally, it involves also the large arteries compressing the near nerves. ALHE is commonly confused with Kimura's disease because of their clinical and histological similarities. We report a case of a 52-year-old female suffering from a 6-month pain and paresthesias in the fourth and fifth finger of the right hand. The angiography showed a pseudoaneurysm in the proximal third of the right ulnar artery. A complete surgical excision of the vascular lesion was undertaken. The lesion forced the right ulnar nerve. The histopathological diagnosis deposed for ALHE. Up to now, literature has described 8 cases of ALHE involving the arteries, and only one case originating from the ulnar nerve. The authors report a case of a female with ALHE involving the ulnar artery that compressed the ulnar nerve. Clinical aspects, radiological features, surgical treatment, and operative findings are discussed reviewing the pertinent literature.


A comparative analysis of the mutagenicity of platinum-containing chemotherapeutic agents reveals direct and indirect mutagenic mechanisms.
Platinum-based drugs are a mainstay of cancer chemotherapy. However, their mutagenic effect can increase tumour heterogeneity, contribute to the evolution of treatment resistance and also induce secondary malignancies. We coupled whole genome sequencing with phenotypic investigations on two cell line models to compare the magnitude and examine the mechanism of mutagenicity of cisplatin, carboplatin and oxaliplatin. Cisplatin induced significantly more base substitution mutations than carboplatin or oxaliplatin when used at equitoxic concentrations on human TK6 or chicken DT40 cells, and also induced the highest number of short insertions and deletions. The analysis of base substitution spectra revealed that all three tested platinum drugs elicit both a direct mutagenic effect at purine dinucleotides, and an indirect effect of accelerating endogenous mutagenic processes, whereas the direct mutagenic effect appeared to correlate with the level of DNA damage caused as assessed through histone H2AX phosphorylation and single-cell agarose gel electrophoresis, the indirect mutagenic effects were equal. The different mutagenicity and DNA-damaging effect of equitoxic platinum drug treatments suggest that DNA damage independent mechanisms significantly contribute to their cytotoxicity. Thus, the comparatively high mutagenicity of cisplatin should be taken into account in the design of chemotherapeutic regimens.


Pathophysiology of diabetic gastroparesis.
Our understanding of the nature of diabetic gastroparesis has advanced in the last decade due to new investigational procedures (electrogastrography, visceral evoked potential recording), and transferring these insights into clinical routine will be our task in the future. Meanwhile, the clinical relevance of gastroparesis--whether overt or silent--remains unquestioned: proper gastric emptying is a prerequisite for adequate metabolic control, and its disturbance may result not only in further progression of the chronic complications of the disease, but also in the false assumption that these patients are not compliant with their doctor's management--the patients just may have delayed emptying of their stomach without noticing it.


Influence of operator experience on non-carious cervical lesion restorations: Clinical evaluation with different adhesive systems.
To evaluate the influence of operator experience on adhesive technique and to assess the clinical performances of two different adhesive systems. 90 cervical lesions in 42 subjects, with a mean age of 52.4 years (range between 32-63) were selected. Lesions were divided into two groups according to operator experience (n = 42 restorations performed by a skilled operator, n = 48 restorations performed by five unexperienced operators) and further divided into two subgroups according to the adhesive system used: three-step etch-and-rinse (Optibond FL) or one step self-etch (G-Bond). Adhesives were applied according to manufacturers' instructions. Subjects underwent follow-up at 12, 24, and 36 months and restoration retention, enamel and dentin marginal integrity, marginal discoloration, caries occurrence, post-operative sensitivity, and preservation of tooth vitality were evaluated according to USPSH criteria. ANOVA test was performed to evaluate the influence of the adhesive system, the operator experience, and aging on restorations. Operator experience (expert vs. inexperienced), aging time (12, 24, or 36 months), and the adhesive system (self-etch vs. etch-and-rinse) all affected the results statistically, as did the interaction between the adhesive system and operator experience.


Increase in insulin antibodies during continuous subcutaneous insulin infusion and multiple-injection therapy in contrast to conventional treatment.
Forty-five insulin-dependent diabetics were randomized to 1 yr treatment with either continuous subcutaneous insulin infusion (CSII), multiple insulin injections (MI), or continued conventional treatment. The CSII group used regular insulin only, the MI group used 4-6 premeal injections of regular insulin and intermediate insulin at night, and the conventional group used two daily injections of combined regular and intermediate insulin. Only highly purified porcine insulin was used. Near normoglycemia was obtained during CSII and MI but not during conventional treatment. Antibodies against insulin were measured in serum samples by measuring the binding of iodinated porcine insulin to serum after removal of free and antibody-bound insulin from the samples by acid charcoal. The percent binding of 125I-labeled insulin increased significantly during MI and CSII, in contrast to conventional treatment. Nineteen patients had sufficient binding capacity for Scatchard analysis. In the CSII and MI groups, high- or low-affinity antibodies or both were induced. When insulin was administered subcutaneously during MI or CSII for 1 yr, the insulin antibody production increased, in contrast with conventional treatment.


Anisogomphus yanagisawai sp. nov., a new gomphid dragonfly from northern Thailand (Odonata: Anisoptera: Gomphidae). [Corrected].
A new species of Anisogomphus Selys, A. yanagisawai sp. nov. (holotype male and paratype specimens) from N. Thailand (Doi Inthanon, ca. 1,400 m a.s.l., Ban Luang, Chiang Mai Prov.), is described and illustrated. This species can apparently be distinguished from the other species of this genus by the morphology of the anal appendages, especially the straight cerci closely disposed to each other and bearing a very strong outer branch.


Clostridium algidixylanolyticum sp. nov., a psychrotolerant, xylan-degrading, spore-forming bacterium.
A psychrotolerant Clostridium species was isolated from vacuum-packed, temperature-abused raw lamb. Colonies of this micro-organism on sheep-blood agar were circular with an entire margin, grey-white, translucent and beta-haemolytic. Cells were single, tapered, motile rods. Elliptical subterminal spores were produced in the late stationary growth phase. Spores did not cause swelling of the maternal cells. The micro-organism was obligately anaerobic. In peptone yeast extract glucose starch (PYGS) broth at pH 7.0, the micro-organism grew optimally between 25.5 and 30.0 degrees C. The temperature range for growth was 2.5-32.2 degrees C. At 26 degrees C, the micro-organism grew optimally at pH 6.8 to 7.0. The pH range for anaerobic growth was 4.7-9.1. The micro-organism was saccharoclastic, hydrolysed starch and degraded xylan. The fermentation products formed in PYGS broth were acetate, formate, lactate, ethanol, butyrate, butanol, hydrogen and carbon dioxide. The G + C content of the DNA was 38.4 mol%. Phylogenetic analyses indicated that the strain belongs to cluster XIVa of the genus Clostridium (sensu Collins et al. 1994). The new strain differed from phylogenetically related clostridia in terms of cellular fatty acid composition, soluble protein profiles and phenotypic properties. On the basis of phenotypic and genotypic characterization data, the strain was assigned to a new species, namely Clostridium algidixylanolyticum. The type strain is strain SPL73T (= DSM 12273T).


Three-dimensional culture environments enhance osteoblast differentiation.
In previous work from our laboratory, we demonstrated that the three-dimensional (3D) cell cultures developed in simulated microgravity environments enhanced osseous-like aggregate formation and accelerated preosteoblast cell differentiation. Thus, as described here, we hypothesize that aggregate formation and mineralization would occur with fewer than 10 x 10(6) cells as previously described. Human preosteoblastic cells were cultured at different concentrations in a rotary wall vessel to simulate microgravity for 7 days. Aggregate size was assessed, and mineralization and collagen expression detected using Von Kossa and Masson Trichrome staining. Scanning electron microscopy was used for structural and elemental analysis. Immunohistochemistry was used to detect expression of the osteogenic markers BSPII and osteopontin (OP). Size and calcium expression were dependent upon cultured cell number (p < 0.01). Calcium and collagen expression were detected throughout the aggregate, but organization was independent of cell number. Aggregates had similar microscopic structural patterns demonstrating organized development. Presence of BSPII and OP showed that the aggregates share common differentiation proteins with in vivo bone formation. These results may lead to novel bone engineering techniques associated with dental treatment.


Targeted strategies in the treatment of metastatic colon cancer.
Advances in the understanding of tumor biology have led to the identification of important cellular processes involved in the pathogenesis of colon cancer. Drugs that interfere with these critical pathways are known as targeted agents. The goal of these therapies is to selectively interrupt the signal transduction pathways responsible for tumor growth and survival. Some of these targeted agents have made important, albeit modest, contributions to the treatment of patients with metastatic colorectal cancer. However, the activity levels with the currently available targeted therapies are far lower than experts had hoped, and toxicities are often nontrivial. This article reviews the available therapies, the data that justify their use, and the challenges of optimizing targeted therapies through combinations with cytotoxic chemotherapies and other targeted agents. Finally, some newer drugs and strategies currently being tested in clinical trials are discussed.


From "Kickeando las malias" (kicking the withdrawals) to "Staying clean": The impact of cultural values on cessation of injection drug use in aging Mexican-American men.
Drug use among older adults is a growing concern, particularly for the burgeoning Hispanic population. Older adults seeking drug treatment will double over the next decade to almost 6 million. Cultural factors influence drug use, and more specifically, Hispanic cultural values influence heroin use. This study explored Mexican-American injection drug users' adherence to traditional Hispanic cultural values and their impact on cessation. Ethnographic interviews endorsed contextualized influences of values on heroin use. Cultural values functioned dichotomously, influencing both initiation and cessation. Understanding the impact of cultural values on substance abuse is critical given the changing demographics in American society.


Restoration of erect posture by deep brain stimulation of the globus pallidus in disabling dystonic spinal hyperextension.
Dystonia is a movement disorder notoriously difficult to treat. While primary dystonia is classically considered to respond well to deep brain stimulation (DBS), treatment of secondary dystonia yields variable results. Patient selection should be done on a case-by-case basis. Clearly, there is a need to accumulate additional information with regard to prognostic factors that may aid neurosurgeons in selecting those patients in whom the disorder is most likely to respond favorably to pallidal DBS. The authors report the case of a 29-year-old man with secondary dystonia due to perinatal hypoxia. The most prominent symptom was what we have termed ectatocormia-that is, severe, fixed truncal hyperextension and retrocollis, exacerbated by phasic, twisting movements of the trunk and head. This made it impossible for the patient to maintain a normal upright posture or to walk. The patient underwent bilateral DBS of the globus pallidus internus (GPi), and the authors observed impressive improvement in motor abilities and function. The patient's body adopted the normal upright posture and he became able to walk again, 4 months after the commencement of GPi stimulation. This report, along with others, emphasizes that the GPi as an ideal target for alleviating axial tonic symptoms. The presence of normal MR imaging findings, a phenotypical purity of predominantly dystonic symptoms, and a younger age seem to favor a positive outcome.


Effect of sealant viscosity on the penetration of resin into etched human enamel.
This study was designed to investigate the relationship between the etched depth, penetration of sealants and marginal seal. Sixty unerupted lower first premolars extracted from patients for orthodontic reasons were thoroughly cleaned and stored in an antiseptic Ringer's solution at 4 degrees C. A "window" on the occlusal of 15 of the premolars, including both mesial and distal pits, was developed by painting nail varnish around the border of the occlusal surfaces. Etching was then done with 35% phosphoric acid for 60 seconds to all 60 premolars. Five of the "window" teeth were evaluated by microradiography to determine the etched depth of the superficial and subsurface enamel. Five were prepared for SEM analysis to observe the change of the superficial etched enamel surface. Another five "window" teeth were embedded in epoxy resin and sectioned parallel to the long axis of the tooth through the fissures in order to observe the subsurface depth of the etch by SEM analysis. The other 45 teeth were divided into three groups of 15 teeth each. Fissures of each group of teeth were sealed using Prisma-Shield (LD Caulk), Concise White Sealant (3M Dental Products) or Teethmate A (Kuraray) sealants and stored in water (37 degrees C) for 24 hours. They were then sectioned and demineralized before being examined by a scanning electron microscope. Photographs of secondary electron image (SEI) were done to gradate the resin-infiltrated enamel and resin tags for these sealants. After SEM observation, the 15 samples of each applied sealant were polished to a high gloss again and placed in a silver nitrate solution for 24 hours before being examined under the SEM equipped with a back-scatter electron detector. Data were then analyzed using the Welch and Student t-tests. Results showed that fissured enamel of unerupted human lower first premolars became porous after etching with 35% phosphoric acid. The low viscosity sealant Teethmate A (approximately 260 mPa.s), penetrated fully and formed a resin-infiltrated layer in enamel beyond the etched depth. However, the high viscosity sealants (Prisma-Shield and Concise White Sealant) did not penetrate enough to ensure that the acid-etched enamel was infiltrated sufficiently by the sealant to insure good marginal seals.


Testing for polygenic effects in genome-wide association studies.
To confirm associations with a large number of single nucleotide polymorphisms (SNPs), each with only a small effect size, as hypothesized in the polygenic theory for schizophrenia, the International Schizophrenia Consortium (2009, Nature 460:748-752) proposed a polygenic risk score (PRS) test and demonstrated its effectiveness when applied to psychiatric disorders. The basic idea of the PRS test is to use a half of the sample to select and up-weight those more likely to be associated SNPs, and then use the other half of the sample to test for aggregated effects of the selected SNPs. Intrigued by the novelty and increasing use of the PRS test, we aimed to evaluate and improve its performance for GWAS data. First, by an analysis of the PRS test, we point out its connection with the Sum test [Chapman and Whittaker, Genet Epidemiol 32:560-566; Pan, Genet Epidemiol 33:497-507]; given the known advantages and disadvantages of the Sum test, this connection motivated the development of several other polygenic tests, some of which may be more powerful than the PRS test under certain situations. Second, more importantly, to overcome the low statistical efficiency of the data-splitting strategy as adopted in the PRS test, we reformulate and thus modify the PRS test, obtaining several adaptive tests, which are closely related to the adaptive sum of powered score (SPU) test studied in the context of rare variant analysis [Pan et al., 2014, Genetics 197:1081-1095]. We use both simulated data and a real GWAS dataset of alcohol dependence to show dramatically improved power of the new tests over the PRS test; due to its superior performance and simplicity, we recommend the whole sample-based adaptive SPU test for polygenic testing. We hope to raise the awareness of the limitations of the PRS test and potential power gain of the adaptive SPU test.


Intramedullary epidermoid associated with an intramedullary spinal abscess secondary to a dermal sinus.
Intramedullary spinal abscesses are rare and potentially devastating lesions. A heightened clinical awareness of patients at risk is essential for an early diagnosis and intervention to improve the prognosis of these patients. The first case of spinal abscess was described in 1830, and only 63 cases have been reported subsequently. Intraspinal epidermoids also are unusual lesions. These tumors were reported first in 1829 and represent less than 1% of all intraspinal tumors. We present a case of a 2-year-old girl who had rapidly progressive paraplegia with urinary and bowel retention and was found to have an intramedullary epidermoid and an intramedullary abscess as a result of a dermal sinus. The association of an intramedullary abscess and epidermoid tumor, two rare lesions, has not been reported previously. We also review the literature related to both intramedullary epidermoids and intramedullary abscesses of the spinal cord. Advances in sectional imaging as described should help lead to an early and safe diagnosis of these rare but often debilitating lesions. This case illustrates the importance of a complete neurological assessment of all patients with a congenital dermal sinus because of the potential for intradural extension and the frequent association with other dysraphic abnormalities. Prophylactic surgery is indicated in many cases to prevent dangerous and recurrent infections of the central nervous system.


Effects of olmesartan and enalapril at low or high doses on cardiac, renal and vascular interstitial matrix in spontaneously hypertensive rats.
We have evaluated the effects of different doses of an angiotensin-converting enzyme (ACE) inhibitor, enalapril (ENA) and of an angiotensin II type 1 receptor blocker olmesartan (OLM), on extracellular matrix of the heart, kidney, aorta and mesenteric artery of spontaneously hypertensive rats (SHR). Forty SHR and eight Wistar-Kyoto controls (WKY) were included in the study. Eight SHR were treated with high-dose OLM 15 mg/kg per day, eight with high-dose ENA 25 mg/kg per day, eight with low-dose OLM 1 mg/kg per day and eight with low-dose ENA (2 mg/kg per day). Eight SHR and eight WKY were kept untreated as controls. Treatment was from age 4 to 12 weeks. Systolic blood pressure (SBP) was measured non-invasively every week. The left ventricular weight to body weight (RLVM) was measured, and the cardiac, aortic and glomerular interstitial collagen content was evaluated using Sirius red staining and image analysis. Mesenteric small arteries were dissected and mounted on a micromyograph, and the media:lumen ratio (M/L) was calculated. Collagen subtypes were evaluated by polarized light microscopy. The SHR treated with high-dose OLM or ENA showed a normalization of SBP. The RLVM was significantly increased in untreated SHR compared with untreated WKY, whereas significantly lower values were observed in the groups of SHR treated with high-dose OLM or ENA. A significant increase in cardiac and glomerular collagen content was observed in untreated SHR. Both high- or low-dose OLM and ENA normalized collagen content in the heart and the kidney. Both high-dose OLM and high-dose ENA normalized M/L ratio; however, OLM proved to be more effective than ENA in normalizing collagen pattern. In fact, aortic collagen content was normalized by both high-dose and low-dose OLM, but only by high-dose ENA. In conclusion, both OLM and ENA were significantly and equally effective in the prevention of cardiac and renal damage in SHR, whereas OLM was more effective than ENA in terms of effects on vascular extracellular matrix.


Efflux of radiolabeled polyethylene glycols and albumin from rat brain.
Experiments were carried out to evaluate the role of convection in the removal of large molecules from brain interstitial fluid. Radiolabeled test compounds were injected into the caudate nucleus of anesthetized rats through a guide cannula implanted 1 wk previously and the concentrations of isotope in brain and cerebrospinal fluid (CSF) determined at various times after injection. Control studies with 22Na indicate that the permeability of the blood-brain barrier is normal in tissue surrounding the intracerebral injection cannula. For 69,000 dalton serum albumin, 4,000 dalton polyethylene glycol, and 900 dalton polyethylene glycol, clearance from brain approximates a single exponential decay with half times of disappearance of 12.2, 12.6, and 14.4 h, respectively. Similarly in efflux rate, despite a fivefold difference in diffusion coefficient, is consistent with convective losses from brain, and the maximal rate of interstitial fluid removal estimated on the basis of these data is 0.11 microliter.g brain-1.min-1. Only 10-20% of total efflux is into bulk CSF withdrawn from the cisterna magna.


Tumor cell rejection through terminal cell differentiation.
Leukemic cells cultured in the presence of various conditioned media differentiate into macrophages. This finding suggested that the maintenance of undifferentiated state and self-renewal in vivo may be related to the inability of the host to generate an appropriate level of differentiation factor (DF). Evidence for this hypothesis was derived from experiments in vitro and in vivo with myeloid leukemia of rat. The following results were obtained: (i) in vitro, the percentage of cell differentiation at a fixed concentration of DF was inversely related to the concentration of cells; (ii) leukemic cell inoculates that were lethal to 7-day-old rats were rejected by 21-day-old rats; (iii) leukemic cells in diffusion chambers underwent differentiation in 21-day-old rats but not in 7-day-old rats; (iv) organs from 21-day-old rats contained more DF activity than those of 7-day-old rats; (v) treatment of rats with DF in diffusion chambers resulted in leukemic cell differentiation inside the chamber; and (vi) the development of leukemia in 7-day-old rats was aborted by treatment with DF. These results show that the differentiation of rat leukemia cells requires the appropriate level of DF. The proliferation of transplanted leukemia cells in 7-day-old rats goes unchecked because of inadequate generation of DF. Conversely, in the 21-day-old rats, rejection is accomplished by differentiation of the transplanted cells.


A fugacity model assessment of ibuprofen, diclofenac, carbamazepine, and their transformation product concentrations in an aquatic environment.
An updated version of FATEMOD, a multimedia fugacity model for environmental fate of organic chemicals, was set up to assess environmental behaviour of three pharmaceuticals in northern Lake Päijänne, Finland. Concentrations of ibuprofen, diclofenac, and carbamazepine were estimated at various depths at two sites: near a wastewater treatment plant and 3.5 km downstream the plant. When compared with environmental sampling data from corresponding depths and sites, the predicted concentrations, ranging from nanograms to hundreds of nanograms per litre, were found to be in good agreement. Weather data were utilised with the model to rationalise the effects of various environmental parameters on the sampling results, and, e.g. the roles of various properties of lake dynamics and photodegradation were identified. The new model also enables simultaneous assessment of transformation products. Environmentally formed transformation product concentrations were estimated to be at highest an order of magnitude lower than those of the parent compounds, and unlikely to reach a detectable level. However, a possibility that conjugates of ibuprofen are present at higher levels than the parent compound was identified. Simulation results suggest that environmental degradation half-lives of the inspected contaminants under stratified lake conditions are in the range of some weeks to months.


Effects of ethanol and ethanol withdrawal on nociception in rats.
The effect of acute and chronic administration of ethanol and ethanol withdrawal on a radiant heat tail-flick assay of nociception was examined in rats. Acute administration of ethanol (2.0 g/kg, i.p.) produced peak antinociception (68% of maximum) by 30 min, and effects were gone by 120 min. Cumulative doses of ethanol (0.5-2.0 g/kg, i.p.) produced dose-dependent increases in latencies to 49% of maximum. During chronic administration, a liquid diet containing ethanol (6.5%) was given for 10 days. Tail-flick latencies were measured on day 0 (baseline), day 2, 4, 6, 8, and 10 of chronic ethanol and at 3, 6, 12, and 36 hr after removal of ethanol. To test for behavioral tolerance, both between- and within-group designs were used. In both between- and within-group experiments, the antinociceptive effects of chronic ethanol peaked by day 4 of exposure to the liquid diet, and tolerance developed by day 10. When the liquid diet was removed, hyperalgesia was detected at 6 and 12 hr after withdrawal, and was gone by 36 hr after withdrawal. When cumulative doses of ethanol (0.5-2.0 g/kg) were administered starting 12 hr after withdrawal, ethanol (0.5 g/kg) fully reversed the hyperalgesia induced by ethanol withdrawal, even though this dose was without antinociceptive effect in the absence of withdrawal. Higher doses of ethanol during ethanol withdrawal did not increase tail-flick latencies over baseline. In summary: (1) ethanol produces antinociception when administered acutely or chronically; (2) tolerance to the antinociceptive effects develops during chronic administration; (3) ethanol withdrawal induced hyperalgesia, which was reversed by ethanol; and (4) repeated testing did not produce behavioral tolerance.


Early identification and delay to treatment in myocardial infarction and stroke: differences and similarities.
The two major complications of atherosclerosis are acute myocardial infarction (AMI) and acute ischemic stroke. Both are life-threatening conditions characterised by the abrupt cessation of blood flow to respective organs, resulting in an infarction. Depending on the extent of the infarction, loss of organ function varies considerably. In both conditions, it is possible to limit the extent of infarction with early intervention. In both conditions, minutes count. This article aims to describe differences and similarities with regard to the way patients, bystanders and health care providers act in the acute phase of the two diseases with the emphasis on the pre-hospital phase. A literature search was performed on the PubMed, Embase (Ovid SP) and Cochrane Library databases. In both conditions, symptoms vary considerably. Patients appear to suspect AMI more frequently than stroke and, in the former, there is a gender gap (men suspect AMI more frequently than women).With regard to detection of AMI and stroke at dispatch centre and in Emergency Medical Service (EMS) there is room for improvement in both conditions. The use of EMS appears to be higher in stroke but the overall delay to hospital admission is shorter in AMI. In both conditions, the fast track concept has been shown to influence the delay to treatment considerably. In terms of diagnostic evaluation by the EMS, more supported instruments are available in AMI than in stroke. Knowledge of the importance of early treatment has been reported to influence delays in both AMI and stroke. Both in AMI and stroke minutes count and therefore the fast track concept has been introduced. Time to treatment still appears to be longer in stroke than in AMI. In the future improvement in the early detection as well as further shortening to start of treatment will be in focus in both conditions. A collaboration between cardiologists and neurologists and also between pre-hospital and in-hospital care might be fruitful.


Lycopene for the prevention of prostate cancer.
Prostate cancer is a common cause of death in developed countries, yet the benefits of screening for prostate cancer still remain controversial. A prostate-specific antigen (PSA) test result greater than 4 ng/mL (nanograms/millilitre) has commonly been used as the cut-off level for seeking further tests to diagnose the presence (or absence) of prostate cancer. An increase in PSA levels may not necessarily be associated with an increased risk of prostate cancer, as PSA levels may also be increased in men with benign prostatic hyperplasia and prostatitis. Despite the uncertainty of the net benefit of early detection and treatment, safe and effective methods to prevent prostate cancer are of value. Consumers, seeking greater involvement in their healthcare, are increasingly turning to lifestyle modification and complementary and alternative medicines (CAMs) to maintain their health and prevent disease. Lycopene is a member of the carotenoid family, which is found abundantly in tomatoes, tomato-based products, strawberries, and watermelon. It has been hypothesised that lycopene is a strong antioxidant, which may lower the risk of cancer (including prostate cancer) in people who have diets rich in lycopene. To determine whether lycopene reduces the incidence of prostate cancer and prostate cancer-specific mortality. Secondary objectives include changes in PSA levels, prostate symptoms and the nature of adverse events associated with lycopene use. Electronic searches were conducted across MEDLINE, EMBASE and the Cochrane Central Register of Controlled Trials (CENTRAL) databases. No language or other limitations were imposed. Randomised controlled trials (RCTs) that investigated the use of lycopene for the prevention of prostate cancer were eligible for inclusion in this review. A search of electronic databases, performed in August 2011, identified 64 citations. All articles were selected for full-text review. From these citations, three studies were identified as meeting the inclusion criteria. Handsearching did not provide any additional studies. Three RCTs, with a total of 154 participants were included in this review. None of the studies reported data on prostate cancer mortality. All of the included studies differed with respect to design, participants included and allocation of lycopene. This clinical heterogeneity limits the value on the pooled estimated of the meta-analyses. The methodological quality of two of the three included studies was assessed as posing a 'high' risk of bias. Meta-analysis indicated no statistical difference in PSA levels between men randomised to receive lycopene and the comparison group (MD (mean difference) -0.34, 95% CI (confidence interval) -2.01, 1.32). Only one study reported incidence of prostate cancer (10% in the lycopene group versus 30% in control group). The level of lycopene was also not statistically different in men randomised to receive lycopene and the comparison group (MD 0.39 µg/mL (micrograms/millilitre), 95% CI -0.19, 0.98). No other meta-analyses were possible since other outcomes assessed only had one study contributing data. Given that only three RCTs were included in this systematic review, and the high risk of bias in two of the three studies, there is insufficient evidence to either support, or refute, the use of lycopene for the prevention of prostate cancer. Similarly, there is no robust evidence from RCTs to identify the impact of lycopene consumption upon the incidence of prostate cancer, prostate symptoms, PSA levels or adverse events.


Depression and coronary heart disease: recommendations for screening, referral, and treatment: a science advisory from the American Heart Association Prevention Committee of the Council on Cardiovascular Nursing, Council on Clinical Cardiology, Council on Epidemiology and Prevention, and Interdisciplinary Council on Quality of Care and Outcomes Research: endorsed by the American Psychiatric Association.
Depression is commonly present in patients with coronary heart disease (CHD) and is independently associated with increased cardiovascular morbidity and mortality. Screening tests for depressive symptoms should be applied to identify patients who may require further assessment and treatment. This multispecialty consensus document reviews the evidence linking depression with CHD and provides recommendations for healthcare providers for the assessment, referral, and treatment of depression.


Connecting functional and statistical definitions of genotype by genotype interactions in coevolutionary studies.
Predicting how species interactions evolve requires that we understand the mechanistic basis of coevolution, and thus the functional genotype-by-genotype interactions (G × G) that drive reciprocal natural selection. Theory on host-parasite coevolution provides testable hypotheses for empiricists, but depends upon models of functional G × G that remain loosely tethered to the molecular details of any particular system. In practice, reciprocal cross-infection studies are often used to partition the variation in infection or fitness in a population that is attributable to G × G (statistical G × G). Here we use simulations to demonstrate that within-population statistical G × G likely tells us little about the existence of coevolution, its strength, or the genetic basis of functional G × G. Combined with studies of multiple populations or points in time, mapping and molecular techniques can bridge the gap between natural variation and mechanistic models of coevolution, while model-based statistics can formally confront coevolutionary models with cross-infection data. Together these approaches provide a robust framework for inferring the infection genetics underlying statistical G × G, helping unravel the genetic basis of coevolution.


Comparing patient and healthcare worker experiences during a dengue outbreak in Singapore: understanding the patient journey and the introduction of a point-of-care test (POCT) toward better care delivery.
In the aftermath of an upsurge in the number of dengue cases in 2013 and 2014, the SD BIOLINE Dengue Duo rapid diagnostic Point-of-Care Test (POCT) kit was introduced in Tan Tock Seng Hospital, Singapore in June 2013. It is known that the success of POCT usage is contingent on its implementation within the health system. We evaluated health services delivery and the Dengue Duo rapid diagnostic test kit application in Singapore from healthcare workers' perspectives and patient experiences of dengue at surge times. Focus group discussions were conducted with dengue patients, from before and after the POCT implementation period. In-depth interviews with semi-structured components with healthcare workers were carried out. A patient centred process mapping technique was used for evaluation, which mapped the patient's journey and was mirrored from the healthcare worker's perspective. Patients and healthcare workers confirmed a wide range of symptoms in adults, making it challenging to determine diagnosis. There were multiple routes to help seeking, and no 'typical patient journey', with patients either presenting directly to the hospital emergency department, or being referred there by a primary care provider. Patients groups diagnosed before and after POCT implementation expressed some differences between speed of diagnoses and attitudes of doctors, yet shared negative feelings about waiting times and a lack of communication and poor information delivery. However, the POCT did not in its current implementation do much to help waiting times. Healthcare workers expressed that public perceptions of dengue in recent years was a major factor in changing patient management, and that the POCT kit was helpful in improving the speed and accuracy of diagnoses. Health service delivery for dengue patients in Singapore was overall perceived to be of an acceptable clinical standard, which was enhanced by the introduction of the POCT. However, improvements can be focused on Adapting to outbreaks by reducing and rendering Waiting experiences more comfortable; Advancing education about symptom recognition, while also Recognising better communication strategies; and Expanding follow-up care options. This is presented as the Dengue AWARE model of care delivery.


Effects of a 1 per cent hydrocortisone conditioner on the prevention of immediate and late-phase reactions in canine skin.
Ten laboratory beagles were used to determine whether a 1 per cent hydrocortisone conditioner applied topically for three consecutive days would inhibit IgE-mediated immediate and late-phase reactions induced in their skin. The trial was blinded, controlled with the product's vehicle and designed with a crossover. It consisted of three phases: one period without treatment (control phase) and two periods of treatment with either the active ingredient or the vehicle. Immediate and late-phase reactions were induced by the intradermal injection of rabbit anti-canine IgE polyclonal antibodies. Twenty minutes after the intradermal challenge, the diameter of the wheal, but not the erythematous flares, were significantly reduced after the application of the active product. In contrast, IgE-mediated cutaneous late-phase reactions, evaluated by measurements of dermal thickness and eosinophil counts six hours after challenge and the numbers of dermal CD3-positive T lymphocytes after 24 hours, were not reduced by its application.


Activation of Bacteroides fragilis toxin by a novel bacterial protease contributes to anaerobic sepsis in mice.
Bacteroides fragilis is the leading cause of anaerobic bacteremia and sepsis. Enterotoxigenic strains that produce B. fragilis toxin (BFT, fragilysin) contribute to colitis and intestinal malignancy, yet are also isolated in bloodstream infection. It is not known whether these strains harbor unique genetic determinants that confer virulence in extra-intestinal disease. We demonstrate that BFT contributes to sepsis in mice, and we identify a B. fragilis protease called fragipain (Fpn) that is required for the endogenous activation of BFT through the removal of its auto-inhibitory prodomain. Structural analysis of Fpn reveals a His-Cys catalytic dyad that is characteristic of C11-family cysteine proteases that are conserved in multiple pathogenic Bacteroides spp. and Clostridium spp. Fpn-deficient, enterotoxigenic B. fragilis has an attenuated ability to induce sepsis in mice; however, Fpn is dispensable in B. fragilis colitis, wherein host proteases mediate BFT activation. Our findings define a role for B. fragilis enterotoxin and its activating protease in the pathogenesis of bloodstream infection, which indicates a greater complexity of cellular targeting and activity of BFT than previously recognized. The expression of fpn by both toxigenic and nontoxigenic strains suggests that this protease may contribute to anaerobic sepsis in ways that extend beyond its role in toxin activation. It could thus potentially serve as a target for disease modification.


Upregulation of imidazoline receptors in the medulla oblongata accounts for the enhanced hypotensive effect of clonidine in aortic barodenervated rats.
The present study tested the hypothesis that an upregulation of the imidazoline receptor in the rostral ventrolateral medulla (RVLM) of aortic barodenervated (ABD) rats may account for the enhanced hypotensive effect of clonidine. In vitro autoradiographic radioligand binding studies were utilized to investigate the binding characteristics of imidazoline receptors in the RVLM and nucleus tractus solitarius (NTS), areas that play critical roles in cardiovascular regulation and elicitation of clonidine responses. ABD but not sham operation (SO) caused immediate and significant (P < 0.05) increases in mean arterial pressure (MAP) and heart rate (HR) and an impairment of the baroreflex-mediated HR response (baroreflex sensitivity, BRS). Two days after ABD, these parameters, except BRS, subsided to near-control (SO) levels. Intracisternal (i.c.) administration of clonidine (0.1 micrograms) elicited a 3-fold greater decrease in BP of conscious ABD compared with SO rats (-20.3 +/- 2.6 vs. -7.4 +/- 0.9 mmHg) thus demonstrating the ability of ABD to enhance centrally-mediated hypotensive responses. Autoradiographic visualization of brain sections obtained from separate groups of ABD and SO rats 48 h after surgery preincubated with [3H]idazoxan (2.5-3.5 nM) showed that [3H]idazoxan binding in RVLM, middle NTM (mNTS) and rostral NTS (rNTS) was saturable and of high affinity. Uneven distribution of imidazoline binding sites was evident since in control (SO) rats, Scatchard analysis of binding data revealed similar densities (Bmax) of [3H]idazoxan binding sites in the RVLM and mNTS versus significantly higher density in the rNTS. In ABD rats, the binding dissociation constant (Kd) was significantly decreased in both the RVLM (8.1 +/- 3.1 vs. 21.4 +/- 5.0 nM) and rNTS (12.3 +/- 1.3 vs. 18.6 +/- 3.1 nM) compared with SO rats while the Bmax was not affected. This finding suggests an increased receptor affinity in the RVLM and rNTS of barodenervated rats. The mNTS of ABD rats exhibited significant increases in the Bmax (861 +/- 96 vs. 570 +/- 87 fmol/mg protein) compared with values of SO rats but the receptor affinity was not affected. It is concluded that: (i) aortic baroreceptors exert a tonic inhibitory influence on central imidazoline receptor function; and (ii) the enhanced hypotensive effect of clonidine in conscious ABD rats may be accounted for by the increased affinity of the medullary imidazoline receptors particularly in the RVLM.


Surgical Approaches to the Proximal Interphalangeal Joint.
The proximal interphalangeal (PIP) joint may be affected by many conditions such as arthropathy, fractures, dislocations, and malunions. Whereas some of these conditions may be treated nonsurgically, many require open surgical intervention. Open interventions include implant arthroplasty or arthrodesis for arthropathy, open reduction internal fixation, or hemi-hamate arthroplasty for dorsal fracture-dislocations. Volar plate arthroplasty and corrective osteotomy for malunion about the PIP joint are also surgeries that may be required. The traditional approach to the PIP joint has been dorsal, which damages the delicate extensor apparatus with subsequent development of an extensor lag. This has led surgeons to explore volar and lateral approaches to the PIP joint. In this article, we describe each of these surgical approaches, discuss their advantages and disadvantages, and provide some guidance on which approach to choose based on the surgery that is to be performed.


Persistently high levels of intersexuality in male-biased amphipod populations.
In a study conducted over 30 years ago, a remarkable population of amphipod crustaceans (Gammarus minus) was discovered in which 100% of the females displayed intersex characteristics. Such high levels of intersexuality are atypical amongst gonochoristic crustaceans and current theories relating to sex allocation suggest intermediate (intersex) forms should be competitively selected against. This study set out to confirm the existence of this extraordinary population and to compare it with other G. minus populations in various spring and cave systems across Pennsylvania, Virginia and West Virginia (USA), with varying environmental characteristics in an attempt to elucidate the prevalence and cause of intersexuality in this species. Results from this study verify the continued existence of these highly abundant, intersex female populations of G. minus, with proportions ranging between 73% and 100%. These populations were also associated with highly male-biased sex ratios, which is equally unusual amongst gonochoristic crustaceans. Intersex female specimens had a reduced fecundity compared to normal females, although this result was not statistically significant. Although no feminising microsporidian parasites were observed through histological examination, the presence of a novel feminising parasitic factor cannot be ruled out. The continued vigour of these populations over three decades suggests that anthropomorphic factors are not causing the high levels of intersexuality observed. Intersexuality was absent or very low in frequency (<1%) in most other springs that we investigated. Therefore, G. minus appears to represent an interesting and novel model system for studying sex determination.


Droplet digital PCR measurement of HER2 copy number alteration in formalin-fixed paraffin-embedded breast carcinoma tissue.
Human epidermal growth factor receptor 2 (HER2) testing is routinely performed by immunohistochemistry (IHC) and/or fluorescence in situ hybridization (FISH) analyses for all new cases of invasive breast carcinoma. IHC is easier to perform, but analysis can be subjective and variable. FISH offers better diagnostic accuracy and added confidence, particularly when it is used to supplement weak IHC signals, but it is more labor intensive and costly than IHC. We examined the performance of droplet digital PCR (ddPCR) as a more precise and less subjective alternative for quantifying HER2 DNA amplification. Thirty-nine cases of invasive breast carcinoma containing ≥30% tumor were classified as positive or negative for HER2 by IHC, FISH, or both. DNA templates for these cases were prepared from formalin-fixed paraffin-embedded (FFPE) tissues to determine the HER2 copy number by ddPCR. ddPCR involved emulsifying hydrolysis probe-based PCR reaction mixtures containing the ERBB2 [v-erb-b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian); also known as HER2] gene and chromosome 17 centromere assays into nanoliter-sized droplets for thermal cycling and analysis. ddPCR distinguished, through differences in the level of HER2 amplification, the 10 HER2-positive samples from the 29 HER2-negative samples with 100% concordance to HER2 status obtained by FISH and IHC analysis. ddPCR results agreed with the FISH results for the 6 cases that were equivocal by IHC analyses, confirming 2 of these samples as positive for HER2 and the other 4 as negative. ddPCR can be used as a molecular-analysis tool to precisely measure copy number alterations in FFPE samples of heterogeneous breast tumor tissue.


Case series: Combined spinal epidural anesthesia for Cesarean delivery and ex utero intrapartum treatment procedure.
To report the use of regional anesthesia and iv nitroglycerin to provide anesthesia and uterine relaxation for three Cesarean deliveries (CD) involving ex utero intrapartum treatment (EXIT) of potentially life-threatening airway obstruction in the newborn. Case 1--a 36-yr-old woman at 38 weeks' gestation was scheduled for an elective CD for fetal skeletal dysplasia and micrognathia. Case 2--a 34-yr-old woman at 35 weeks gestation had a fetal ultrasound revealing fixed neck flexion and micrognathia consistent with fetal arthrogryposis. Case 3--a 27-yr-old woman presented at 38 weeks gestation for CD for severe fetal micrognathia, with mandibular growth below the fifth percentile. For each case, a combined spinal epidural anesthetic was performed with 0.75% bupivacaine, fentanyl and morphine intrathecally followed by placement of a multiorifice epidural catheter. Prior to uterine incision patients received a loading dose followed by an iv infusion of nitroglycerin. Uterine relaxation was sufficient in all cases for delivery of the fetus, and allowed for evaluation by direct laryngoscopy and intubation while maintaining fetal-placental circulation. The surgical procedures were completed without incident. Anesthesia and uterine relaxation for CD and EXIT procedures can be safely provided with regional anesthesia and iv nitroglycerin.


Hypoxia-inducible factor-2alpha is a catabolic regulator of osteoarthritic cartilage destruction.
Osteoarthritic cartilage destruction is caused by an imbalance between anabolic and catabolic factors. Here, we show that hypoxia-inducible factor-2alpha (HIF-2alpha, encoded by EPAS1) is a catabolic transcription factor in the osteoarthritic process. HIF-2alpha directly induces the expression in chondrocytes of genes encoding catabolic factors, including matrix metalloproteinases (MMP1, MMP3, MMP9, MMP12 and MMP13), aggrecanase-1 (ADAMTS4), nitric oxide synthase-2 (NOS2) and prostaglandin-endoperoxide synthase-2 (PTGS2). HIF-2alpha expression was markedly increased in human and mouse osteoarthritic cartilage, and its ectopic expression triggered articular cartilage destruction in mice and rabbits. Moreover, mice transgenic for Epas1 only in chondrocytes showed spontaneous cartilage destruction, whereas heterozygous genetic deletion of Epas1 in mice suppressed cartilage destruction caused by destabilization of the medial meniscus (DMM) or collagenase injection, with concomitant modulation of catabolic factors. Our results collectively demonstrate that HIF-2alpha causes cartilage destruction by regulating crucial catabolic genes.


Urokinase-mediated recruitment of myeloid-derived suppressor cells and their suppressive mechanisms are blocked by MUC1/sec.
The transmembrane isoform of mucin 1 (MUC1/TM) is a well-recognized tumor antigen, contributing to tumorigenesis and immune evasion. Although MUC1/TM has been correlated with malignancy, we have previously reported on antitumor properties and prevention of tumor development by a secreted splice variant of MUC1 (MUC1/sec). Because myeloid-derived suppressor cells (MDSCs) play a critical role in tumor-induced immunosuppression, we investigated their recruitment by tumor cells expressing either MUC1/TM or MUC1/sec. DA-3 tumor cells expressing MUC1/sec recruit dramatically lower levels of MDSCs, relative to MUC1/TM-expressing DA-3 cells. Because MUC1/sec was previously shown to down-regulate tumor expression of urokinase plasminogen activator (uPA), a protease linked to tumor aggressiveness and metastasis, the potential role of uPA in MDSC recruitment was investigated. Tumor-derived uPA is capable of recruiting MDSCs, and correlates with tumor development. In addition to diminishing recruitment of MDSCs, the effect of MUC1/sec on MDSC-suppressive mechanisms was investigated. MUC1/sec, or its unique immunoenhancing peptide, is capable of blocking expression of arginase 1 and production of reactive oxygen species in MDSCs, implicated in the suppression of T cells. These findings demonstrate a new mechanism of MDSC recruitment, and provide evidence that MUC1/sec has antitumor properties affecting MDSCs.


Surveillance for Ixodes scapularis (Acari Ixodidae) and Borrelia burgdorferi in Eastern South Dakota State Parks and Nature Areas.
Ixodes scapularis (Acari: Ixodidae) is the principal vector of Borrelia burgdorferi (the etiologic agent of Lyme disease) in the eastern and midwestern United States. Recent efforts have documented the first established population of I. scapularis in South Dakota, representing a western expansion of the known species distribution. Our goal was to describe the current distribution of I. scapularis in eastern South Dakota and to survey for the presence of B. burgdorferi in questing I. scapularis. We surveyed for the presence of adult and nymphal I. scapularis in seven counties within South Dakota, including 13 locales from 2016 to 2017. We then tested all I. scapularis, including those collected in 2015 from a previous study, for the presence of B. burgdorferi sensu stricto, via quantitative and conventional polymerase chain reaction. Here, we document the presence of I. scapularis in four new counties in South Dakota, and report the first instance of B. burgdorferi in a questing tick in South Dakota. Coupled with data from the Centers for Disease Control and Prevention and Companion Animal Parasite Council our data show that the risk of contracting Lyme disease in South Dakota is low, but existent and should be an important consideration with regard to public health, pets, and wildlife.


Molecular mechanisms of cellular mechanics.
Mechanical forces play an essential role in cellular processes as input, output, and signals. Various protein complexes in the cell are designed to handle, transform and use such forces. For instance, proteins of muscle and the extracellular matrix can withstand considerable stretching forces, hearing-related and mechanosensory proteins can transform weak mechanical stimuli into electrical signals, and regulatory proteins are suited to forcing DNA into loops to control gene expression. Here we review the structure-function relationship of four protein complexes with well defined and representative mechanical functions. The first example is titin, a protein that confers passive elasticity on muscle. The second system is the elastic extracellular matrix protein, fibronectin, and its cellular receptor integrin. The third protein system is the transduction apparatus in hearing and other mechanical senses, likely containing cadherin and ankyrin repeats. The last system is the lac repressor protein, which regulates gene expression by looping DNA. This review focuses on atomic level descriptions of the physical mechanisms underlying the various mechanical functions of the stated proteins.


Health insurance and other factors associated with mammography surveillance among breast cancer survivors: results from a national survey.
Whether disparities in mammography surveillance among breast cancer survivors exist by health insurance type is unclear. To determine the association of surveillance mammography with insurance and other factors among breast cancer survivors. DESIGN/PARTICIPANTS/MEASURES: We examined mammography within the prior year among 1511 breast cancer survivors aged 30 or older and ≥1 year after diagnosis from the 2000, 2003, 2005, and 2008 National Health Interview Surveys. Insurance included private (health maintenance organization/independent practice association, other, unspecified), Medicare+private, public only (Medicare+Medicaid, other Medicare, other public), and uninsured. Multivariable logistic regression was used to determine factors associated with mammography, including age, race/ethnicity, education, time since diagnosis, health status, insurance, income to poverty threshold ratios, having a usual provider, and survey year. Results are presented as predictive margins. For insurance, pairwise comparisons were used to compare Medicare+private versus other groups. Overall 75% reported a mammogram. Mammography reporting was lower for uninsured survivors and those with only public insurance versus Medicare+private or private (50% and 68% vs. 76% and 80%, respectively; P=0.001). In each insurance group, 20% to 50% reported not receiving a mammogram. After adjustment, use remained lower for women with only public insurance versus Medicare+private (71% vs. 78%, P=0.029). Age of at least 75 years, greater time since diagnosis, and no usual provider were associated with not reporting a mammogram. One in 4 breast cancer survivors did not report guideline-concordant mammography surveillance. Women with only public insurance were less likely than those with Medicare+private coverage to report a mammogram. Efforts to understand barriers and promote mammography among survivors are needed.


Detection of a geographical and endemic cluster of hyper-invasive meningococcal strains.
From 2006 to December 2009, 45 out of the 513 strains isolated from patients with invasive meningococcal disease in Belgium, were identified as Neisseria meningitidis serogroup B, non-serotypeable, subtype P1.14 (B:NT:P1.14). Most cases were geographically clustered in the northern part of the country. Multilocus Sequence Typing and antigen gene sequencing combined with Pulsed-Field Gel electrophoresis were used to investigate this cluster. Molecular typing showed that 39 out of these 45 N. meningitidis strains belonged to the clonal complex cc-269. The presence of the same PorA Variable Regions (VR1-VR2: 22, 14), the FetA allele (F5-1) and the highly similar Pulsed-Field Gel Electrophoresis profiles, supported genetic relatedness for 38 out of these 39 isolates. Retrospective analysis of B:NT:P1.22,14 isolates from 1999 onwards suggested that these strains belonging to the cc-269 complex, first emerged in the Belgian province of West-Flanders in 2004. This study showed that the combination of molecular tools with classical methods enabled reliable outbreak detection as well as a cluster identification.


Identification of tryptic peptides unique to a 110,000-molecular weight polyprotein encoded by the T-8 isolate of murine leukemia virus.
Mink cells nonproductively-infected with the weakly-transforming T-8 isolate of murine leukemia virus (MuLV) express a 110,000 mol wt polyprotein designated T-8 P110. By immunoprecipitation analysis, T-8 P110 is shown to contain AKR-MuLV amino terminal gag gene-specific components (p15, p12) but to lack p30, p10, gp70, and p15(E) antigenic determinants. These observations are further substantiated by tryptic peptide analysis indicating T-8 P110 to share approximately six lysine-containing tryptic peptides with AKR-MuLV Pr65gag, and none with AKr-MuLV Pr82env. Furthermore, of seven methionine-containing T-8 P110 tryptic peptides, at least four can be conclusively shown not to be present in either AKr-MuLV Pr180gag/pol or Pr82env. A clonal mink cell line nonproductively infected by T-8, and expressing high levels of P110, although not morphologically transformed, is shown to lack elevated levels of tyrosine-specific protein kinase activity and reduction of epidermal growth factor binding sites characteristic of cells transformed by many other RNA-transforming viruses. These findings argue either that the T-8 viral genome contains acquired cellular sequences encoding a portion of P110, or that T-8 P110 represents an inphase deletion of AKR-MuLV Pr180gag/pol with extensive posttranlational modification and that an as yet unidentified protein is responsible for T-8 associated transformation.


Electrical burns; pathophysiology and complications.
The authors review the pathophysiology and complications of electrical injuries. Such injuries can be categorized into direct electrical injuries, arc burns and flame burns. Each has a distinct appearance and prognosis. Nine patients with electrical injuries were admitted to the Toronto General Hospital over the 5-year period 1973 to 1978. All were men and the average age was 33 years. Five patients required amputation. In one patient a convulsive disorder developed 3 weeks after injury. One patient demonstrated peripheral nerve injury with complete axonal lesions of the right median and ulnar nerves; there was progressive full return of nerve function over 12 months. Direct electrical injury to nerves is usually transient and return of function can be expected unless there has been a concomitant thermal or mechanical injury.


New challenges in gene expression data analysis and the extended GEPAS.
Since the first papers published in the late nineties, including, for the first time, a comprehensive analysis of microarray data, the number of questions that have been addressed through this technique have both increased and diversified. Initially, interest focussed on genes coexpressing across sets of experimental conditions, implying, essentially, the use of clustering techniques. Recently, however, interest has focussed more on finding genes differentially expressed among distinct classes of experiments, or correlated to diverse clinical outcomes, as well as in building predictors. In addition to this, the availability of accurate genomic data and the recent implementation of CGH arrays has made mapping expression and genomic data on the chromosomes possible. There is also a clear demand for methods that allow the automatic transfer of biological information to the results of microarray experiments. Different initiatives, such as the Gene Ontology (GO) consortium, pathways databases, protein functional motifs, etc., provide curated annotations for genes. Whereas many resources on the web focus mainly on clustering methods, GEPAS has evolved to cope with the aforementioned new challenges that have recently arisen in the field of microarray data analysis. The web-based pipeline for microarray gene expression data, GEPAS, is available at http://gepas.bioinfo.cnio.es.


Imaging real-time gene expression in Mammalian cells with single-transcript resolution.
INTRODUCTIONThe MS2 system provides optimal sensitivity for single-molecule detection in cells. It requires two genetically encoded moieties: a reporter mRNA that contains MS2 binding site (MBS) stem loops and a fluorescent MS2 coat protein (MCP-xFP) that binds to the stem loops with high affinity, thus tagging the mRNA within the cell. This protocol describes transfection of COS-7 cells with reporter RNA (e.g., pRSV-Z-24 MBS-β-actin) and MCP-xFP (e.g., pPolII-MCP-GFP-NLS) plasmids using calcium phosphate precipitation. The reporter mRNA plasmid must be co-transfected with the MCP-xFP-NLS plasmid for simultaneous expression in a cell. The unbound MCP-xFP-NLS is sequestered in the nucleus, leaving only the MCP-xFP-NLS that is bound to the reporter mRNA in the cytoplasm. This provides a high signal-to-noise ratio (SNR) that permits detection of single mRNA molecules. The Delta T Imaging System is used for image acquisition of fluorescent particles in the cells.


[Homeostatic changes in burn patients treated by a laminar flow of sterile heated air].
Under observation were 102 burned patients with different degrees of injuries treated under conditions of the laminar flow of sterile air without dressing. The results obtained were compared with those obtained in the similar group of patients treated by a traditional dressing method. The abacterial medium with controlled humidity and temperature was established to exercise a favourable effect on homeostasis and such stages of the burn disease as shock and toxemia.


Image detection under low-level illumination.
Image detection under low-light-level conditions is treated as a hypothesis-testing problem in which the observations are modeled as a shot-noise process. Since computing the likelihood ratio for shot-noise processes is not feasible, the use of a one-dimensional test statistic obtained by filtering and sampling the observations is proposed. The filter is chosen to maximize a generalized signal-to-noise ratio. The likelihood ratio for the one-dimensional test statistic is evaluated numerically by inverting the corresponding characteristic function under each hypothesis.


Evaluation of Standard and Proposed Reference Values for Entheseal Thickening by Using Musculoskeletal Ultrasound.
Ultrasound (US) is increasingly used to evaluate enthesitis. One of the US features of enthesitis is thickening. However, there is no consensus on how the entheseal thickening needs to be defined, and existing cut-off levels have been criticized for being frequently positive in healthy controls (HCs). Our objective was to determine the frequency of thickening of entheses on US using the existing cut-off values in HCs and in patients with axial spondyloarthritis (axSpA) and propose new values to improve discriminative value. Eighty HCs and 100 patients with axSpA had US scans of 2160 entheses. Sensitivity, specificity, odds ratio (OR), and accuracy were calculated according to accepted cut-off levels from the literature and proposed cut-offs were calculated as the mean ± 2 SD. Thickening according to current cut-off levels was found in 20.4% (196/960) of healthy participants' entheses and 33% (396/1200) of entheses of patients with axSpA. Thickening according to proposed cut-off levels decreased frequency of thickening in both groups, and therefore increased specificity at the cost of decreasing sensitivity. The only anatomical site where the thickness had a value to discriminate disease from health was seen at the triceps tendon enthesis with an OR of 13.4 (95% CI 4.0-44.8) according to the current cut-offs compared to 10.3 (95% CI 4.0-26.6) with the proposed cut-off levels. Although using cut-offs appears to be an appealing method to evaluate entheseal thickness, the measurements may be affected by several confounding factors, leading to a low discriminative value, except for at the triceps tendon enthesis.


Metal-containing polymers: building blocks for functional (nano)materials.
The incorporation of metallic units into polymer chains has emerged as a promising route towards functional metal-containing (nano)materials. The resulting polymers possess rich functions derived from their metallic elements, such as redox, optical, catalytic and magnetic properties. In addition, the directional and dynamic nature of metal coordination interactions provides further variables for the exploration of novel materials with designed nanostructures. These types of polymers can be synthesized through direct metal-ligand coordination or chain polymerization of metal containing monomers. Depending on the polymerization techniques and starting components, the resulting polymers, akin to their organic counterparts, can be produced in the form of insoluble networks, processible chain structures, gels or colloids. Research into this rising multidisciplinary subject has benefited from recent progress in several related areas such as supramolecular chemistry, colloidal chemistry etc., with the combination of the relative merits of each ensuring further developments in each individual discipline. For example, as a result of studies into organometallic block copolymers self-assembly behavior, living supramolecular polymerization has been unprecedentedly realized for the architectural design of micelles (see image on the right). Nevertheless, the field is still in a developmental stage and offers ample opportunities for fundamental research, as well as material exploration. In this Feature Article, we intend to overview the field with a brief survey of recent literature.


Fine-Tuning ER Stress Signal Transducers to Treat Amyotrophic Lateral Sclerosis.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive loss of motoneurons and paralysis. The mechanisms underlying neuronal degeneration in ALS are starting to be elucidated, highlighting disturbances in motoneuron proteostasis. Endoplasmic reticulum (ER) stress has emerged as an early pathogenic event underlying motoneuron vulnerability and denervation in ALS. Maintenance of ER proteostasis is controlled by a dynamic signaling network known as the unfolded protein response (UPR). Inositol-requiring enzyme 1 (IRE1) is an ER-located kinase and endoribonuclease that operates as a major ER stress transducer, mediating the establishment of adaptive and pro-apoptotic programs. Here we discuss current evidence supporting the role of ER stress in motoneuron demise in ALS and build the rational to target IRE1 to ameliorate neurodegeneration.


Binding and inhibition in working memory: individual and age differences in short-term recognition.
Two studies investigated the relationship between working memory capacity (WMC), adult age, and the resolution of conflict between familiarity and recollection in short-term recognition tasks. Experiment 1 showed a specific deficit of young adults with low WMC in rejecting intrusion probes (i.e., highly familiar probes) in a modified Sternberg task, which was similar to the deficit found in old adults in a parallel experiment (K. Oberauer, 2001). Experiment 2 generalized these results to 3 recognition paradigms (modified Sternberg, local recognition, and n back tasks). Old adults showed disproportional performance deficits on intrusion probes only in terms of reaction times, whereas young adults with low WMC showed them only in terms of errors. The generality of the effect across paradigms is more compatible with a deficit in content-context bindings subserving recollection than with a deficit in inhibition of irrelevant information in working memory. Structural equation models showed that WMC is related to the efficiency of recollection but not of familiarity.


Medication calculation skills of practicing paramedics.
To assess the medication calculation skills among a group of practicing paramedics, the types of computations they find most difficult, and the relationship between drug calculation skills and various demographic characteristics. A demographic survey and a ten-item drug calculation examination were administered to a convenience sample of 109 practicing paramedics representing a cross-section of emergency medical services (EMS) system characteristics in North Carolina. Examinations were scored independently by two graders and error types were assigned to incorrect responses. Examination results were then correlated with demographic and EMS system characteristics. Overall performance on the drug calculation examination was poor. The mean score was 51.4% (SD 27.4). Intravenous flow rate problems and medication bolus problems were calculated correctly in 68.8% of the cases, followed by non-weight-based medication infusions (33.9%), weight-based medication infusions (32.5%), and percentage-based medication infusions (4.5%). Examination scores were higher among paramedics with college level education, but scores were lower among paramedics with more years of EMS experience. Conceptual errors (i.e., errors in setting up the problem) were more prevalent than mathematical errors, errors in weight conversion, or errors in unit conversion (e.g., grams to milligrams). The participants reported that drug calculations were infrequently performed in daily practice and were rarely a topic of continuing education programs. Similar to findings among other allied health professions, medication calculation skills were found to be lacking among a group of practicing paramedics. In addition, the paramedics reported infrequent opportunities to perform this skill in the clinical setting and that medication calculations were not a routine part of EMS continuing education programs.


Dosimetry and techniques for simultaneous hyperthermia and external beam radiation therapy.
An increased biological effect is realized when hyperthermia and radiation therapy are combined simultaneously. To take advantage of this effect, techniques have been developed that combine existing hyperthermia devices with a linear accelerator. This allows concomitant delivery of either ultrasound or microwave hyperthermia with photon radiation therapy. Two techniques have been used clinically: the orthogonal technique, in which the microwave or ultrasound beam and the radiation beam are orthogonal to one another, and the en face technique, in which the ultrasound or microwave beam and the radiation beam travel into the tumour through the same treatment window. The en face technique has necessitated the development of special attachments so that the hyperthermia device can be mounted to the linear accelerator and so that non-uniform portions of the hyperthermia device can be removed from the radiation beam. For microwave therapy, applicators are mounted onto the linear accelerator using the compensating filter tray holder. For ultrasound, special reflector devices are mounted to a frame that is mounted onto the compensating filter tray holder of the linear accelerator. Because the linear accelerator is an isocentric device, the height of the radiation source is fixed, and this has necessitated specially designed devices so that the ultrasound support system is compatible with the linear accelerator. The treatment setups for both the en face technique and the orthogonal technique require the interaction of both hyperthermia and radiation therapy personnel and equipment. The dosimetry and day-to-day operations for each technique are unique. The simulation for the en face technique is much different from the simulation of a normal radiation treatment and requires the presence of a hyperthermia physicist. Also, for the en face technique, the attenuation of the microwave applicator and the thickness and attenuation of the ultrasound reflector system are taken into account for radiation dosimetry. This paper presents details of the dosimetry and logistics of the techniques for simultaneous thermoradiotherapy based on 7 years of experience treating more than 50 patients.


Validation of a Clinical Scoring System for Outcome Prediction in Dogs with Acute Kidney Injury Managed by Hemodialysis.
A scoring system for outcome prediction in dogs with acute kidney injury (AKI) recently has been developed but has not been validated. The scoring system previously developed for outcome prediction will accurately predict outcome in a validation cohort of dogs with AKI managed with hemodialysis. One hundred fifteen client-owned dogs with AKI. Medical records of dogs with AKI treated by hemodialysis between 2011 and 2015 were reviewed. Dogs were included only if all variables required to calculate the final predictive score were available, and the 30-day outcome was known. A predictive score for 3 models was calculated for each dog. Logistic regression was used to evaluate the association of the final predictive score with each model's outcome. Receiver operating curve (ROC) analyses were performed to determine sensitivity and specificity for each model based on previously established cut-off values. Higher scores for each model were associated with decreased survival probability (P < .001). Based on previously established cut-off values, 3 models (models A, B, C) were associated with sensitivities/specificities of 73/75%, 71/80%, and 75/86%, respectively, and correctly classified 74-80% of the dogs. All models were simple to apply and allowed outcome prediction that closely corresponded with actual outcome in an independent cohort. As expected, accuracies were slightly lower compared with those from the previously reported cohort used initially to develop the models.


Perspectives of health professionals towards deprescribing practice in Asian nursing homes: a qualitative interview study.
To examine the determinants of deprescribing among health professionals in nursing homes, focusing on knowledge, practice and attitude. This was a qualitative study comprising semi-structured face-to-face interviews guided by 10 open-ended questions. Interviews were conducted until data saturation was achieved and no new ideas were formed. The interviews were audio-recorded, transcribed verbatim and analysed for themes. To derive themes, we employed directed content analysis of transcript data. Coding was completed using a combination of open, axial and selective coding. Four nursing homes in Singapore. The study involved 17 participants (comprising 4 doctors, 4 pharmacists and 9 nurses). Two key themes were identified, enablers and challenges. These were enablers and challenges faced by doctors, pharmacists and nurses towards deprescribing. The identified subthemes for enablers of deprescribing were: (1) awareness of medications that are unnecessary or could be targeted for deprescribing; (2) improving quality of life for patients with limited life expectancy; (3) improving communication between doctors, pharmacists and nurses; (4) systematic deprescribing practice and educational tools and (5) acknowledgement of possible benefits of deprescribing. The identified subthemes for challenges of deprescribing were: (1) symptoms not acknowledged as possibly drug-related; (2) lack of knowledge in patient's and family members' preferences; (3) lack of coordination between health professionals in hospitals and nursing homes and (4) limited tools for deprescribing. The development of a local guideline, mentoring nurses, case discussions, better shared decision-making and improving multidisciplinary communication, may help to support the process of deprescribing. In conclusion, this study highlighted that deprescribing in the nursing homes is perceived by health professionals to be challenging and future research could assess how routine case studies, mentoring and better multidisciplinary communication could improve deprescribing knowledge and process in the nursing homes.


Uses and Misuses of Recorded Mental Health Lived Experience Narratives in Healthcare and Community Settings: Systematic Review.
Mental health lived experience narratives are first-person accounts of people with experience of mental health problems. They have been published in journals, books and online, and used in healthcare interventions and anti-stigma campaigns. There are concerns about their potential misuse. A four-language systematic review was conducted of published literature characterizing uses and misuses of mental health lived experience narratives within healthcare and community settings. 6531 documents in four languages (English, Danish, Swedish, Norwegian) were screened and 78 documents from 11 countries were included. Twenty-seven uses were identified in five categories: political, societal, community, service level and individual. Eleven misuses were found, categorized as relating to the narrative (narratives may be co-opted, narratives may be used against the author, narratives may be used for different purpose than authorial intent, narratives may be reinterpreted by others, narratives may become patient porn, narratives may lack diversity), relating to the narrator (narrator may be subject to unethical editing practises, narrator may be subject to coercion, narrator may be harmed) and relating to the audience (audience may be triggered, audience may misunderstand). Four open questions were identified: does including a researcher's personal mental health narrative reduce the credibility of their research?: should the confidentiality of narrators be protected?; who should profit from narratives?; how reliable are narratives as evidence?).


Nonlinear screening in multilayer graphene systems.
Electrostatic screening in multilayer graphene is highly nonlinear due to the vanishing density of states at the Fermi level. Using a discrete model we study the charge screening normal to the layers. Our model shows a strong charge and temperature dependence and has a simple continuum limit at T=0 for undoped systems. Doped systems can exhibit more complex behavior due to minority-carrier screening. Most importantly we find that the screening length can vary more than an order of magnitude depending on the experimental conditions, reconciling the large range of screening lengths reported in previous experiments. This has important consequences for technological applications of multilayer graphene used in electrodes or transistor channels.


Localization during development of alternatively spliced forms of cytotactin mRNA by in situ hybridization.
Cytotactin, an extracellular glycoprotein found in neural and nonneural tissues, influences a variety of cellular phenomena, particularly cell adhesion and cell migration. Northern and Western blot analysis and in situ hybridization were used to determine localization of alternatively spliced forms of cytotactin in neural and nonneural tissues using a probe (CT) that detected all forms of cytotactin mRNA, and one (VbVc) that detected two of the differentially spliced repeats homologous to the type III repeats of fibronectin. In the brain, the levels of mRNA and protein increased from E8 through E15 and then gradually decreased until they were barely detectable by P3. Among the three cytotactin mRNAs (7.2, 6.6, and 6.4 kb) detected in the brain, the VbVc probe hybridized only to the 7.2-kb message. In isolated cerebella, the 220-kD polypeptide and 7.2-kb mRNA were the only cytotactin species present at hatching, indicating that the 220-kD polypeptide is encoded by the 7.2-kb message that contains the VbVc alternatively spliced insert. In situ hybridization showed cytotactin mRNA in glia and glial precursors in the ventricular zone throughout the central nervous system. In all regions of the nervous system, cytotactin mRNAs were more transient and more localized than the polypeptides. For example, in the radial glia, cytotactin mRNA was observed in the soma whereas the protein was present externally along the glial fibers. In the telencephalon, cytotactin mRNAs were found in a narrow band at the edge of a larger region in which the protein was wide-spread. Hybridization with the VbVc probe generally overlapped that of the CT probe in the spinal cord and cerebellum, consistent with the results of Northern blot analysis. In contrast, in the outermost tectal layers, differential hybridization was observed with the two probes. In nonneural tissues, hybridization with the CT probe, but not the VbVc probe, was detected in chondroblasts, tendinous tissues, and certain mesenchymal cells in the lung. In contrast, hybridization with both probes was observed in smooth muscle and lung epithelium. Both epithelium and mesenchyme expressed cytotactin mRNA in varying combinations: in the choroid plexus, only epithelial cells expressed cytotactin mRNA; in kidney, only mesenchymal cells; and in the lung, both of these cell types contained cytotactin mRNA. These spatiotemporal changes during development suggest that the synthesis of the various alternatively spliced cytotactin mRNAs is responsive to tissue-specific local signals and prompt a search for functional differences in the various molecular forms of the protein.


Efficacy of soluble glycoprotein fraction from Allium sativum purified by size exclusion chromatography on murine Schistosomiasis mansoni.
In this work, the efficiency of crude MeOH extracts and soluble glycoprotein fraction of Allium sativum purified by size-exclusion chromatography (SEC) on parasitological, histopathological and some biochemical parameters in Schistosoma mansoni infected mice were investigated. Animals were infected by tail immersion with 100 cercariae/each mouse and divided into five groups in addition to the normal control. The results revealed a significant decrease in mean worm burden in all treated mice especially in the group treated with soluble glycoprotein fraction of A. sativum as compared to infected non-treated control with the disappearance of female worms. Administration of the studied extracts revealed remarkable amelioration in the levels of all the measured parameters in S. mansoni infected mice. In addition, treatment of mice with crude A. sativum MeOH extract and soluble glycoprotein fraction of A. sativum decreased significantly the activities of studied enzymes as compared to the infected untreated group. The highest degrees of enhancement in pathological changes was observed in the treated one with soluble glycoprotein fraction of A. sativum compared to the infected group represented by small sized, late fibro-cellular granuloma, the decrease in cellular constituents and degenerative changes in eggs. In conclusion, A. sativum treatment had effective schistosomicidal activities, through reduction of worm burden and tissue eggs, especially when it was given in purified glycoprotein fraction. Moreover, the soluble glycoprotein fraction of A. sativum largely modulates both the size and the number of granulomas.


Presentation of atrial fibrillation and its management by cardiologists in the ambulatory and hospital setting: MOVE cross-sectional study.
The aim of the study was to collect comprehensive data on atrial fibrillation (AF) in ambulatory and hospital-based management in Germany. Consecutive patients with ECG-confirmed AF in the previous 12 months were documented in a non-interventional study in 638 physician offices (78.0%) or hospitals (12.7%). Of the 3354 patients (mean age 68.9 ± 10.1 years; CHADS(2) score 1.9 ± 1.3), a total of 1136 (33.9%) had paroxysmal, 899 (26.8%) persistent, 1295 (38.6%) permanent and 24 (0.7%) unspecified AF. In the 12 months prior to documentation, pharmacological conversion was attempted in 18.2%, electric cardioversion in 17.5%, the combination of both in 31.2%, and catheter ablation of AF in 5.5%. Only 41.4% of patients met the definition of stable disease (having neither AF related intervention nor change in antiarrhythmic therapy in the previous 12 months). As treatment strategy, physicians stated rate control in 64%, rhythm control in 8%, and both in 19% (not reported: 8%). Patients received antiarrhythmic drugs of class IA in 1.3%, IC in 13.8%, II in 78.1%, III in 17.9%, IV in 9.7% and digitalis in 26.7%. Drugs for thromboembolic prevention (oral anticoagulants and/or antithrombotics) were administered in 81.5%. Hospitalisations for AF or associated diseases in the previous 12 months were reported in 34.2%. Possible limitations include the open, observational design, selection of physicians with particular interest in the field and selection of patients (i.e. underrepresentation of critically ill individuals). While treatment rates with regards to the prevention of thromboembolic events were among the highest reported to date, the low proportion of stable patients and in particular, the high hospitalisation rate hint at difficulties in the management of patients with AF in clinical practice.


Tangram of Sodium and Fluid Balance: Salt Series.
Homeostasis of fluid and electrolytes is a tightly controlled physiological process. Failure of this process is a hallmark of hypertension, chronic kidney disease, heart failure, and other acute and chronic diseases. While the kidney remains the major player in the control of whole-body fluid and electrolyte homeostasis, recent discoveries point toward more peripheral mechanisms leading to sodium storage in tissues, such as skin and muscle, and a link between this sodium and a range of diseases, including the conditions above. In this review, we describe multiple facets of sodium and fluid balance from traditional concepts to novel discoveries. We examine the differences between acute disruption of sodium balance and the longer term adaptation in chronic disease, highlighting areas that cannot be explained by a kidney-centric model alone. The theoretical and methodological challenges of more recently proposed models are discussed. We acknowledge the different roles of extracellular and intracellular spaces and propose an integrated model that maintains fluid and electrolyte homeostasis and can be distilled into a few elemental players: the microvasculature, the interstitium, and tissue cells. Understanding their interplay will guide a more precise treatment of conditions characterized by sodium excess, for which primary aldosteronism is presented as a prototype.


Interobserver agreement of Medical Research Council sum-score and handgrip strength in the intensive care unit.
Muscle weakness often complicates critical illness and is associated with devastating short- and long-term consequences. For interventional studies, reliable measurements of muscle force in the intensive care unit (ICU) are needed. To examine interobserver agreement, two observers independently measured Medical Research Council (MRC) sum-score (n = 75) and handgrip strength (n = 46) in a cross-sectional ICU sample. The intraclass correlation coefficient (ICC) for MRC sum-score was 0.95 (0.92-0.97). The kappa coefficient for identifying "significant weakness" (MRC sum-score <48, MRC subtotal upper limbs <24) and "severe weakness" (MRC sum-score <36) was 0.68 ± 0.09, 0.88 ± 0.07, and 0.93 ± 0.07, respectively. The ICC for left and right handgrip strength was 0.97 (0.94-0.98) and 0.93 (0.86-0.97), respectively. Interobserver agreement on MRC sum-score and handgrip strength in the ICU was very good. Agreement on "severe weakness" (MRC sum-score <36) was excellent and supports its use in interventional studies. Agreement on "significant weakness" (MRC sum-score <48) was good, but even better using the equivalent cut-off in the upper limbs. It remains to be determined whether this may serve as a substitute.


Interaction energies for the purine inhibitor roscovitine with cyclin-dependent kinase 2: correlated ab initio quantum-chemical, DFT and empirical calculations.
The interaction between roscovitine and cyclin-dependent kinase 2 (cdk2) was investigated by performing correlated ab initio quantum-chemical calculations. The whole protein was fragmented into smaller systems consisting of one or a few amino acids, and the interaction energies of these fragments with roscovitine were determined by using the MP2 method with the extended aug-cc-pVDZ basis set. For selected complexes, the complete basis set limit MP2 interaction energies, as well as the coupled-cluster corrections with inclusion of single, double and noninteractive triples contributions [CCSD(T)], were also evaluated. The energies of interaction between roscovitine and small fragments and between roscovitine and substantial sections of protein (722 atoms) were also computed by using density-functional tight-binding methods covering dispersion energy (DFTB-D) and the Cornell empirical potential. Total stabilisation energy originates predominantly from dispersion energy and methods that do not account for the dispersion energy cannot, therefore, be recommended for the study of protein-inhibitor interactions. The Cornell empirical potential describes reasonably well the interaction between roscovitine and protein; therefore, this method can be applied in future thermodynamic calculations. A limited number of amino acid residues contribute significantly to the binding of roscovitine and cdk2, whereas a rather large number of amino acids make a negligible contribution.


Quantification of underivatized fatty acids from vegetable oils by HPLC with UV detection.
We propose a chromatographic method for the separation of saturated and unsaturated fatty acids by a high-performance liquid chromatography system, equipped with a photo diode array detector. Central to the method is the use of an appropriate mobile phase composed of acetonitrile, methanol, and n-hexane in ratio 90:8:2 acidified with 0.2% acetic acid, which allows the detection of fatty acids without a preliminary derivatization with chromophores or fluorescent dyes. Calibration on solutions of standards mixtures gives a quantification limit (at a wavelength of 208 nm) of 0.232, 0.093, 0.039, 0.056, 0.068, 0.004, 0.0005, 0.067 mg/mL for the myristic, palmitic, palmitoleic, stearic, oleic, linoleic, linolenic, and erucic acids, respectively. The method, applied to different vegetable oils (olive, sunflower, soybean, and palm) was able to distinguish the main fatty acids and quantify their amount. Data reliability was tested by comparing our results (on the relative percentages of some fatty acids in the olive oil) with those obtained by gas chromatographic analysis. Differences of the order of 0.3%, 0.6%, 2%, and 6% were observed for the oleic, linoleic, palmitic, and linolenic acids. Although less accurate, our method proved to be a simple alternative to standard gas chromatographic technique, as it can be applied even using a simple UV detector.


A Comprehensive Development Agenda on Tenofovir Alafenamide in Clinical Practice.
The introduction of tenofovir (TFV) alafenamide (TAF) into clinical practice will be a further revolution in antiretroviral therapy. Currently available HIV-1 regimens are wide enough to allow diversified usage in different settings. Despite the fact that TAF is not capillary accessible, even in industrialized countries, ultimate International Guidelines have already included TAF in backbone or in single-tablet regimens. Due to a better safety profile, TAF will progressively replace TFV disoproxil fumarate, both in naïve and experienced patients. However, therapeutic innovations have to deal with budget constraints and different global spending-review patterns. The aim of this article is to give a comprehensive agenda of TAF use in naïve and experienced HIV-1 infected patients, providing a full review of the studies present in the literature and contextualizing these findings into daily clinical practice.


Transactivation of human immunodeficiency virus type 1 long terminal repeats by cell surface tumor necrosis factor alpha.
Tumor necrosis factor alpha (TNF-alpha) is expressed in secreted and cell surface (csTNF-alpha) forms by activated monocytic and T cells. In this report, we demonstrate that csTNF-alpha may predominantly regulate the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) activation in the promonocytic cell line U937 and in the Epstein-Barr virus-transformed B-cell line BH1. Anti-TNF-alpha antibody suppressed both the constitutive expression of the HIV-1 LTR in BH1 cells and the expression induced by phorbol 12-myristate 13-acetate in U937 cells. This suppression was found to be mediated via csTNF-alpha. No correlation between the HIV-1 LTR activation and the secretion of TNF-alpha was evident in these cell lines. Suppression of TNF-alpha secretion by cyclosporin A or by a serine protease inhibitor did not suppress the HIV-1 LTR activation. These observations suggest a novel biological role for csTNF-alpha in the immunopathogenesis of AIDS.