Morphometric, genetic diversity and phylogenetic analysis of Taenia hydatigena (Pallas, 1766) larval stage in Iranian livestock.
Cysticercus tenuicollis as metacestode of Taenia hydatigena is the most prevalent taeniid species in livestock. Eighty-eight C. tenuicollis samples were collected from sheep (n = 44) and goats (n = 44) of the northern Iran from 2015 to 2016. The isolated parasites were characterized by morphometric keys. The DNA of the larval stage was extracted, amplified and sequenced targeting mitochondrial 12S rRNA and Cox 1 markers. A significant difference in larval rostellar hook length was observed in 12S rRNA haplotypes. Analysis of molecular variance of 12S rRNA indicated a moderate genetic diversity in the C. tenuicollis isolates. The pairwise sequence distance of C. tenuicollis showed an intra-species diversity of 0.3-0.5% and identity of 99.5-100%. Using the 12S rRNA sequence data we found a moderate genetic difference (Fst; 0.05421) in C. tenucollis isolates collected from livestock of the northern and southeastern regions of Iran. We concluded that the genetic variants of C. tenuicollis are being undoubtedly distributing mostly in different parts of Iran. Further studies with a larger number of T. hydatigena isolates collected from various intermediate and definitive hosts are needed to study this evolutionary assumption and also to determine the apparent genetic differences observed in the studied regions.


COVID-19 crisis management in Luxembourg: Insights from an epidemionomic approach.
We develop an epidemionomic model that jointly analyzes the health and economic responses to the COVID-19 crisis and to the related containment and public health policy measures implemented in Luxembourg. The model has been used to produce nowcasts and forecasts at various stages of the crisis. We focus here on two key moments in time, namely the deconfinement period following the first lockdown, and the onset of the second wave. In May 2020, we predicted a high risk of a second wave that was mainly explained by the resumption of social life, low participation in large-scale testing, and reduction in teleworking practices. Simulations conducted 5 months later reveal that managing the second wave with moderately coercive measures has been epidemiologically and economically effective. Assuming a massive third (or fourth) wave will not materialize in 2021, the real GDP loss due to the second wave will be smaller than 0.4 percentage points in 2020 and 2021.


Expression of genes and proteins of multidrug resistance in gastric cancer cells treated with resveratrol.
Multidrug resistance (MDR) is a notable problem in the use of chemotherapy. Therefore, studies aimed at identifying substances capable of overcoming resistance of cancer cells are required. Examples of these compounds are polyphenols, including resveratrol, that exert a range of various biological activities. The aim of the present study was to demonstrate the effect of 3,5,4'-trihydroxy-trans-stilbene (resveratrol) on the expression of ATP binding cassette subfamily B member 1, Annexin A1 (ANXA1) and thioredoxin (TXN) genes, and the proteins encoded by these genes, which are associated with MDR. The experiments were performed in human gastric cancer cell lines EPG85-257RDB (RDB) and EPG85-257RNOV (RNOV), which are resistant to daunorubicin and mitoxantrone, respectively, in addition to EPG85-257P (control), which is sensitive to cytostatic drugs. Cells were treated with 30 or 50 µM resveratrol for 72 h and changes in the expression levels of the genes were analysed with the use of a reverse transcription-quantitative polymerase chain reaction. The cellular levels of P-glycoprotein (P-gp), ANXA1 and TXN were evaluated using immunofluorescence and western blot analysis. Resveratrol in both concentrations has been shown to have a statistically significant influence on expression of the mentioned genes, compared with untreated cells. In RDB cells, resveratrol reduced the expression level of all analyzed genes, compared with untreated cells. Similar results at the protein level were obtained for P-gp and TXN. In turn, in the RNOV cell line, resveratrol reduced TXN expression at mRNA and protein levels, compared with untreated cells. The results of the present study indicate that resveratrol may reduce the resistance of cancer cells by affecting the expression of a number of the genes and proteins associated with MDR.


IL-6 is a predictive biomarker for stroke associated infection and future mortality in the elderly after an ischemic stroke.
Stroke associated infection (within the first seven days) occurs in approximately half of stroke patients and is associated with a worse prognosis, especially in the elderly. It is uncertain what factors predict stroke associated infection, yet identification of a suitable biomarker for infection may allow early and appropriate intervention with antibiotics. The aims of this study were to: a) identify independent risk factors for stroke associated infection, and b) test relationships between these risk factors and mortality at 2 years. Eight-two elderly patients were assessed within 72 h of stroke. Data on stroke severity (Barthel Index), stroke associated infection and mortality at 2 years were collected. Inflammatory biomarkers at baseline and 6 months were measured by ELISA. Logistic regression was used to identify risk factors for stroke associated infection and death. Patients with stroke associated infection, especially pneumonia, had increased IL-6, more severe strokes, and higher mortality. IL-6 was independently associated with stroke associated infection (OR = 19.2, [95%CI 3.68, 100], p < 0.001), after adjustment for other risk factors and cytokines. IL-6 was also independently associated with 2 year mortality (OR = 9.2, [1.0, 85.1], p = 0.031). These data suggest that IL-6 may be a key biomarker for predicting stroke associated infection and mortality in the first two years post stroke.


Immunocytochemical demonstration of tissue transglutaminase indicative of programmed cell death (apoptosis) in hormone sensitive mammary tumours.
Tissue transglutaminase (TG) positive tumour cells in MXT mouse mammary cancer were localized by immunohistochemistry. The number of TG-positive tumour cells increased after treatment with analogs of luteinizing hormone-releasing hormone (LH-RH) and somatostatin, which inhibit tumour growth by the enhancement of apoptosis (programmed cell death). TG positivity has been previously reported to be closely linked with apoptosis. In this study, the increase in the number of TG positive tumour cells after hormonal treatment was well correlated with the augmented apoptotic index, i.e. the ratio of tumourous glandular structures which contained tumour cells showing the signs of programmed cell death. TG-positive tumour cells are, therefore, believed to be undergoing programmed cell death.


On circadian rhythms in human epidermal cell proliferation.
The ascertainment of consistent diurnal variations in human epidermal cell proliferation may have important implications for the treatment of many skin diseases. For the evaluation of diurnal rhythms in the growth of human epidermis, skin biopsies were taken every 4th hour for 48 h from each of two persons under synchronized living conditions. The epidermal cell proliferation was assessed by the fraction of cells in S and in G2-M phase as determined by measurements of the DNA content in the individual cells in single-cell suspensions. The existence of diurnally consistent body functions in the test persons was verified by monitoring the excretion of cortisol by urine. The fraction of cells in G2-M phase indicated circadian rhythmicity for the first 32 h of the test period. No regular variation according to time of day could be established in the fraction of cells in S-phase.


Allergic sensitisation in tuberculosis and leprosy patients.
A negative association has been observed between infections and allergy in several studies. The aim of the present study was to examine whether tuberculosis and leprosy patients have more or fewer allergies than healthy individuals. Sera from tuberculosis patients, leprosy patients and healthy controls were analysed by ELISA and Pharmacia Unicap for serological markers for allergy and mycobacterial infection. The serological markers for allergy were total IgE, specific IgE using Phadiatop and specific IgE to the dust mite allergen Dermatophagoides pteronyssinus 1 (Der p 1). Serological markers for mycobacterial infections included specific IgG to a mixture of bacille Calmette-Guérin culture filtrate antigens, to purified mannose-capped lipoarabinomannan (manLAM) and to purified secreted antigen 85B. Both tuberculosis and leprosy patients had significantly higher levels of total IgE than controls. Furthermore, a significantly higher level of specific IgE (Phadiatop) was also found in the tuberculosis patients compared with controls. A similar result, but not statistically significant, was observed for the leprosy group. Specific IgG to antigen 85B and to manLAM was found to be significantly higher in both tuberculosis and leprosy patients compared with controls. In addition, leprosy patients had significantly more IgG to the BCG culture filtrate antigen than controls. The results indicate that patients with mycobacterial infections have allergic sensitisation more frequently compared with healthy controls. This is seemingly in contrast with the notion that there is a negative association between allergy and infection ('hygiene hypothesis'). However, since only one in ten of those infected with Mycobacterium tuberculosis will develop the disease, patients with active mycobacterial disease represent a selected group. A similar relationship applies for leprosy. It is conceivable that those predisposed to allergy are less resistant to mycobacterial infections.


Community-associated methicillin-resistant Staphylococcus aureus infections.
Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has been recognized for over a decade, and usually refers to MRSA identified in previously healthy individuals with no recognized MRSA risk factors. Infections range from minor skin and soft tissue infections, through to severe pneumonia and necrotizing fasciitis. This review summarizes the current data on the epidemiology and molecular features of CA-MRSA, in addition to diagnosis and therapeutic measures. We also refer to current national guidelines for the management of these infections. Areas of agreement include the important genotypic and phenotypic differences of community MRSA strains compared with hospital strains. Areas of controversy include the precise epidemiological definition of community-acquired/associated MRSA. Fortunately, true CA-MRSA can be differentiated from hospital MRSA by molecular techniques, as discussed herein. Recent interest has focused on the changing epidemiology of CA-MRSA. Worldwide, CA-MRSA is now seen outside of the initial specific population groups, and in the USA, the successful USA300 community strain is beginning to spread back into hospitals. Reasons why USA300 remains relatively uncommon in Europe are unclear. Topics timely for research include the investigation of the epidemiology of infections and evolutionary genomics.


Effect of corneal thickness on intraocular pressure measurements with the pneumotonometer, Goldmann applanation tonometer, and Tono-Pen.
To compare intraocular pressure (IOP) measurements of the Ocular Blood Flow (OBF) pneumotonometer, Goldmann applanation tonometer, and Tono-Pen in eyes with normal corneas of various thicknesses. IOP was measured with an OBF pneumotonometer, Tono-Pen and Goldmann applanation tonometer in random order in 181 eyes with normal corneas. Central corneal thickness (CCT) was measured using an ultrasonic pachymeter after all IOP determinations had been made. Right and left eyes were analyzed separately for statistical purposes. With all instruments, IOP varied with CCT, even though the variation in IOP was large. Readings with the OBF pneumotonometer showed a mean increase in IOP with increasing CCT of 0.28 mm Hg/10 microm, an increase of 0.23 mm Hg/10 microm with the Goldmann tonometer, and of 0.10 mm Hg/10 microm with the Tono-Pen. The OBF pneumotonometer consistently recorded comparatively higher IOPs than the other two instruments. The Tono-Pen is least affected by CCT when used to measure IOP in eyes with normal corneas. The OBF pneumotonometer appears to be more affected by variation in CCT than the Goldmann tonometer. This is contrary to expectations, based on the mechanism of measurement of IOP of the OBF pneumotonometer.


Effects of varying material properties on the load deformation characteristics of heel cushions.
Various insole materials were used in attenuation of heel-strike impact. This study presented a compression test to investigate the deformation characteristics of common heel cushions. There were two materials (thermoplastic elastomer "TPE" and silicone) with three hardness and six thickness being analyzed. They underwent consecutive loading-unloading cycles with a load control mode. The displacement of material thickness was recorded during cyclic compression being applied and released from 0 to 1050 N. The energy input, return and dissipation were evaluated based on the load deformation curves when new and after repeated compression. The TPE recovered more deformed energy and thickness than the silicone after the first loading cycle. The silicone would preserve more strain energy with increasing its hardness for the elastic recovery in the unloading process. The deformed energy was decreased as the original thickness did not completely recover under cyclic tests. The reduction in hysteresis area was gradually converged within 20 cycles. The silicone attenuated more impact energy in the initial cycles, but its energy dissipation was reduced after repeated loading. To increase hardness or thickness should be considered to improve resilience or accommodate persistent compression without flattening. The careful selection of cushion materials is imperative to meet individual functional demands.


Relationship Between Body Mass Index, ApoE4 Status, and PET-Based Amyloid and Neurodegeneration Markers in Amyloid-Positive Subjects with Normal Cognition or Mild Cognitive Impairment.
Body weight loss in late-life is known to occur at a very early stage of Alzheimer's disease (AD). Apolipoprotein E4 (ApoE4) represents a major genetic risk factor for AD and is linked to an increased cortical amyloid-β (Aβ) accumulation. Since the relationship between body weight, ApoE4, and AD pathology is poorly investigated, we aimed to evaluate whether ApoE4 allelic status modifies the association of body mass index (BMI) with markers of AD pathology. A total of 368 Aβ-positive cognitively healthy or mild cognitive impaired subjects had undergone [18F]-AV45-PET, [18F]-FDG-PET, and T1w-MRI examinations. Composite cortical [18F]-AV45 uptake and [18F]-FDG uptake in posterior cingulate cortex were calculated as surrogates of cortical Aβ load and glucose metabolism, respectively. Multiple linear regressions were performed to assess the relationships between these PET biomarkers with BMI, present cognitive performance, and cognitive changes over time. Multivariate analysis of covariance was conducted to test for statistical differences between ApoE4/BMI categories on the PET markers and cognitive scores. In carriers of the ApoE4 allele only, BMI was inversely associated with cortical amlyoid load (β= -0.193, p < 0.005) and recent cognitive decline (β= -0.209, p < 0.05), and positively associated with cortical glucose metabolism in an AD-vulnerable region (β= 0.145, p < 0.05). ApoE4/BMI category analyses demonstrated lower Aβ load, higher posterior cingulate glucose metabolism, improved cognitive performance, and lower progression of cognitive decline in obese ApoE4 carriers. The effect of ApoE4 in promoting the accumulation of cortical amyoid, which may itself be a driver for weight loss, may be moderated by altering leptin signaling in the hypothalamus.


Bone mineral density and risk factors for osteoporosis--a population-based study of 1600 perimenopausal women.
Population-based epidemiological studies on osteoporosis are few. Our study evaluated the effects of menopause and certain putative behavioural risk factors on bone mineral density (BMD). Spinal and femoral neck BMD were measured with dual X-ray absorptiometry (DXA) from 1600 perimenopausal women aged 48-59 years (mean 53.2 years) with no diseases or medications known to affect bone metabolism. These women were a selected sample of the Kuopio Osteoporosis Risk Factor and Prevention Study population (n = 14,220). There was a wide variation of BMD among perimenopausal women. Menopause had a major effect on BMD. Postmenopausal women had significantly lower BMD in both spine (-6.2%) and femoral neck (-3.9%) as compared with premenopausal women. Multiple regression analysis showed that weight, menopausal status, age, and grip strength were significant independent predictors of both spinal and femoral BMD. Additionally, physical activity was found to be a significant predictor of femoral BMD, and alcohol consumption was a significant predictor of spinal BMD. However, current anthropometric and lifestyle factors explained only 18.7-25.4% of the variability of BMD. Therefore, the estimation of the risk factor status at menopause is not an adequate substitute for bone densitometry. However, our results may in part help clinicians to identify the risk groups at which to direct bone density measurements.


Comparing medical code usage with the compression-based dissimilarity measure.
It is well known that medical coding practice is inconsistent and that differences in usage may exist even at the institutional level. In this paper we introduce a novel method for investigating code usage patterns in clinical documentation corpora. By applying the Compression-based Dissimilarity Measure to calculate similarities between encounter notes, we find that certain notes can be associated with a number of different classifications and that a given classification code can be documented in fundamentally different ways. The effect is that some notes need to be understood in the context of the classification code, a finding which has implications for data mining or information extraction tasks. In addition, the method opens for a number of interesting application areas that include highlighting code use anomalies, measuring how coding practice changes over time, comparing code usage across institutions, and, perhaps most importantly, provide valuable feedback to developers of classification coding systems.


Lumbosacral plexus injury following vaginal delivery with epidural analgesia -A case report-.
A 26 year old, healthy, 41 week primiparous woman received a patient-controlled epidural analgesia (PCEA) and experienced paraplegia 11 hours later after a vaginal delivery. This was thought to be the result of complications from PCEA but there was no specific abnormality on magnetic resonance imaging (MRI) of the lumbosacral spine. On an electromyography (EMG) study performed 15 days following delivery, signs of tibial neuropathy were present and peripheral nerve injury during vaginal delivery was suspected. Motor weakness and hypoesthesia of both lower extremities improved rapidly, but a decrease in the desire to urinate or defecate, followed by urinary incontinence and constipation persisted, We suspected the sacral plexus had been severely damaged during vaginal delivery. Seven months later, the patient's conditions improved but had not fully recovered.


Purinergic type 2 receptors at GABAergic synapses on ventral tegmental area dopamine neurons are targets for ethanol action.
The current study investigated whether ethanol alters ATP activation of purinergic type 2 receptors (P2Rs) in the ventral tegmental area (VTA). The VTA is a key region of the brain that has been implicated in the development of alcohol addiction. We investigated the effects of ATP and ethanol on spontaneous inhibitory postsynaptic currents (sIPSCs) and the spontaneous firings in the VTA dopaminergic neurons, obtained using an enzyme-free procedure. These neurons preserved some functional GABA-releasing terminals after isolation. We found that ATP (1-200 microM) either increased or decreased the frequency of sIPSCs and the activity of VTA dopaminergic neurons. The effects of ATP on sIPSC frequency inversely correlated with its effects on dopaminergic neuron activity. The ATP-induced changes in sIPSC frequency were blocked by tetrodotoxin (a sodium channel blocker) and by suramin (a nonselective P2R antagonist). Furthermore, alpha,beta-methylene ATP, a selective P2X(1) and P2X(3) receptor agonist, increased sIPSC frequency, whereas adenosine 5'-[beta-thio]diphosphate, a preferential agonist of P2Y receptors, decreased sIPSC frequency. In experiments testing the effects of ethanol (10 and 40 mM) on sIPSCs, we found that ethanol significantly attenuated ATP-induced increase and enhanced ATP-induced decrease in sIPSC frequency. Taken together, the results demonstrate that multiple subtypes of P2Rs exist on GABA-releasing terminals that make synapses on VTA dopaminergic neurons. It seems that ATP increases sIPSC frequency involving P2X(1) and/or P2X(3) receptors, and ATP decreases sIPSC frequency involving P2YRs. These findings are also consistent with the notion that P2Rs at GABA-releasing terminals on VTA dopaminergic neurons are important targets for ethanol action.


Tyramine studies and the safety of MAOI drugs.
A major source of concern in using MAOIs is the risk of hypertensive crises, often called the "cheese effect" and thought to represent a drug-induced enhancement of the tyramine pressor effect. Several approaches to reduction of this risk are being explored, including use of inhibitors specific for the B vs. the A type of the enzyme, coadministration of tricyclic with MAOI, and development of reversible inhibitors which might be competitively displaced by tyramine. Such approaches require assessment of sensitivity to tyramine, usually given by the intravenous route. The oral tyramine pressor test, while cumbersome and in need of technical refinement, represents a significantly closer laboratory analogue to the clinical situation.


Atmospheric Oxidation of Squalene: Molecular Study Using COBRA Modeling and High-Resolution Mass Spectrometry.
Squalene is a major component of skin and plant surface lipids and is known to be present at high concentrations in indoor dust. Its high reactivity toward ozone makes it an important ozone sink and a natural protectant against atmospheric oxidizing agents. While the volatile products of squalene ozonolysis are known, the condensed-phase products have not been characterized. We present an analysis of condensed-phase products resulting from an extensive oxidation of squalene by ozone probed by electrospray ionization (ESI) high-resolution mass spectrometry (HR-MS). A complex distribution of nearly 1300 peaks assignable to molecular formulas is observed in direct infusion positive ion mode ESI mass spectra. The distribution of peaks in the mass spectra suggests that there are extensive cross-coupling reactions between hydroxy-carbonyl products of squalene ozonolysis. To get additional insights into the mechanism, we apply a Computational Brewing Application (COBRA) to simulate the oxidation of squalene in the presence of ozone, and compare predicted results with those observed by the HR-MS experiments. The system predicts over one billion molecular structures between 0 and 1450 Da, which correspond to about 27 000 distinct elemental formulas. Over 83% of the squalene oxidation products inferred from the mass spectrometry data are matched by the simulation. The simulation indicates a prevalence of peroxy groups, with hydroxyl and ether groups being the second-most important O-containing functional groups formed during squalene oxidation. These highly oxidized products of squalene ozonolysis may accumulate on indoor dust and surfaces and contribute to their redox capacity.


Ethical issues in palliative care.
To review important issues that address respect for patient autonomy, beneficnce, non-maleficence, and justice, which are included in communication surrounding the determination of decision-making capacity, informed consent, breaking bad news, and creating shared goals of care. Review articles, and government and organizational reports. Palliative care and its proximity to end-of-life care issues frequently raises ethical issues for patients, their families, and the clinicians caring for them. Supporting the identification and honoring the patient's preferences for treatment are central components of ethical behavior. Advance care planning provides an important opportunity for respecting patient autonomy and may be helpful when discussing care options surrounding resuscitation, withholding or withdrawal of treatment, or the determination of medical futility.


Potential pathobionts in vaginal microbiota are affected by fish oil and/or probiotics intervention in overweight and obese pregnant women.
New means to stabilize the microbial balance during pregnancy could benefit maternal health. Our objectives were to investigate in overweight/obese pregnant women 1) the impact of long-chain polyunsaturated fatty acids (fish oil) and/or probiotics on the vaginal microbiota, 2) its relation to gestational diabetes mellitus (GDM) and 3) its interaction with vaginal active matrix metalloproteinase-8 (aMMP-8) and serum high sensitivity C-reactive protein (hsCRP) and phosphorylated insulin-like growth factor-binding protein-1 (phIGFBP-1), IGFBP-1 and aMMP-8. The women were allocated to fish oil + placebo, probiotics + placebo, fish oil + probiotics and placebo + placebo-groups, from early pregnancy onwards (fish oil: 1.9 g docosahexaenoic acid and 0.22 g eicosapentaenoic acid; probiotics: Lacticaseibacillus rhamnosus HN001 (formerly Lactobacillus rhamnosus HN001) and Bifidobacterium animalis ssp. lactis 420, 1010 colony-forming units each). Vaginal and serum samples (early pregnancy, n = 112; late pregnancy, n = 116), were analyzed for vaginal microbiota using 16S rRNA gene amplicon sequencing and vaginal aMMP-8 and serum hsCRP, aMMP-8, phIGFBP-1 and IGFBP-1 by immunoassays. GDM was diagnosed from a 2-h 75 g OGTT. ClinicalTrials.gov, NCT01922791. The intervention exerted effects on many low-abundant bacteria. Compared to the placebo-group, there was a lower abundance of potential pathobionts, namely Ureaplasma urealyticum in the fish oil-group, Ureaplasma, U. urealyticum and Prevotella disiens in the probiotics-group, Dialister invisus and Prevotella timonensis in the fish oil + probiotics-group. Moreover, probiotics decreased the abundance of a few potential pathobionts during pregnancy. Many bacteria were related to GDM. The vaginal aMMP-8 level correlated significantly with α-diversity and inversely with two Lactobacillus species. Dietary interventions, especially probiotics, may have beneficial effects on the vaginal microbiota during pregnancy.


Wnt signaling in age-related macular degeneration: human macular tissue and mouse model.
The wingless-type MMTV integration site (Wnt) signaling is a group of signal transduction pathways. In canonical Wnt pathway, Wnt ligands bind to low-density lipoprotein receptor-related protein 5 or 6 (LRP5 or LRP6), resulting in phosphorylation and activation of the receptor. We hypothesize that canonical Wnt pathway plays a role in the retinal lesion of age-related macular degeneration (AMD), a leading cause of irreversible central visual loss in elderly. We examined LRP6 phosphorylation and Wnt signaling cascade in human retinal sections and plasma kallistatin, an endogenous inhibitor of the Wnt pathway in AMD patients and non-AMD subjects. We also used the Ccl2 (-/-) /Cx3cr1 (-/-) /rd8 and Ccl2 (-/-) /Cx3cr1 (gfp/gfp) mouse models with AMD-like retinal degeneration to further explore the involvement of Wnt signaling activation in the retinal lesions in those models and to preclinically evaluate the role of Wnt signaling suppression as a potential therapeutic option for AMD. We found higher levels of LRP6 (a key Wnt signaling receptor) protein phosphorylation and transcripts of the Wnt pathway-targeted genes, as well as higher beta-catenin protein in AMD macula compared to controls. Kallistatin was decreased in the plasma of AMD patients. Retinal non-phosphorylated-β-catenin and phosphorylated-LRP6 were higher in Ccl2 (-/-) /Cx3cr1 (-/-) /rd8 mice than that in wild type. Intravitreal administration of an anti-LRP6 antibody slowed the progression of retinal lesions in Ccl2 (-/-) /Cx3cr1 (-/-) /rd8 and Ccl2 (-/-) /Cx3cr1 (gfp/gfp) mice. Electroretinography of treated eyes exhibited larger amplitudes compared to controls in both mouse models. A2E, a retinoid byproduct associated with AMD was lower in the treated eyes of Ccl2 (-/-) /Cx3cr1 (-/-) /rd8 mice. Anti-LRP6 also suppressed the expression of Tnf-α and Icam-1 in Ccl2 (-/-) /Cx3cr1 (-/-) /rd8 retinas. Wnt signaling may be disturbed in AMD patients, which could contribute to the retinal inflammation and increased A2E levels found in AMD. Aberrant activation of canonical Wnt signaling might also contribute to the focal retinal degenerative lesions of mouse models with Ccl2 and Cx3cr1 deficiency, and intravitreal administration of anti-LRP6 antibody could be beneficial by deactivating the canonical Wnt pathway.


Familial cases of pseudohypoaldosteronism type II harboring a novel mutation in the Cullin 3 gene.
Pseudohypoaldosteronism type II (PHA II) is inherited in an autosomal dominant manner and is characterized by hypertension, hyperkalemia, and hyperchloremic metabolic acidosis. The enhancement of with-no-lysine kinase (WNK) functions is correlated to the pathogenesis of the condition. Cullin 3 (CUL3) forms an E3 ubiquitin ligase complex, and it can ubiquitinate WNK. Most CUL3 gene mutations are distributed in sites, such as intron 8 splice acceptor, intron 9 splice donor, and putative intron 8 splice branch sites, which are involved in the splicing of exon 9. These mutations result in the deletion of exon 9, which reduces the activity of ubiquitination against WNK and inhibits the degradation of WNK. In this report, we identified a novel CUL3 c.1312A>G mutation in familial cases. A mutation prediction software showed that the significance of these mutations was not clear. However, using the Human Splicing Finder 3.1 software, in silico analyses revealed that these mutations induced splicing alterations, which affected the sites of exon 9, altered the balance between predicted exonic splicing enhancers and silencers, and led to the deletions of exon 9. This study presented a novel pathogenic splicing variant to the CUL3 mutation and provided a reference for further research about the mechanisms of splicing. Moreover, it showed that not only amino acid substitution caused by nonsynonymous mutations but also splicing motif changes due to base substitutions have important roles in the pathogenesis of PHA II.


Brain Network for the Core Deficits of Semantic Dementia: A Neural Network Connectivity-Behavior Mapping Study.
Individuals with semantic dementia (SD) typically suffer from selective semantic deficits due to degenerative brain atrophy. Although some brain regions have been found to be correlated with the semantic impairments of SD patients, it is unclear if the damage is actually responsible for SD patients' semantic disorders because these findings were primarily obtained by examining the roles of local individual regions themselves without considering the influence of other regions that are functionally or structurally connected to the local individual regions. To resolve this problem, we investigated, from the brain network perspective, the relationship between the brain-network measures of regions and connections with semantic performance in 17 SD patients. We found that the severity of semantic deficits of SD patients was significantly correlated with the degree centrality values of the left anterior hippocampus (aHIP). Moreover, the semantic performance of the patients was also significantly correlated with the strength of gray matter functional connectivity of this region and two other regions: the left temporal pole/insula (TP/INS) and the left middle temporal gyrus. We further observed that the strength of the white matter structural connectivity of the left aHIP-left TP/INS tract could effectively predict the semantic performance of SD patients. When we controlled for a wide range of potential confounding factors (e.g., total gray matter volume), the above effects still held well. These findings revealed the critical brain network with the left aHIP as the center that could be contributing to the semantic impairments of SD.


Orthotopic pleural mesothelioma in mice: SPECT/CT and MR imaging with HER1- and HER2-targeted radiolabeled antibodies.
To evaluate the potential of anti-human epidermal growth factor receptor (HER)1- and anti-HER2-targeted radiolabeled antibodies and magnetic resonance (MR) imaging for imaging of orthotopic malignant pleural mesothelioma (MPM) in mouse models. Animal studies with 165 mice were performed in accordance with National Institutes of Health guidelines for the humane use of animals, and all procedures were approved by the institutional Animal Care and Use Committee. Flow cytometry studies were performed to evaluate HER1 and HER2 expression in NCI-H226 and MSTO-211H mesothelioma cells. Biodistribution and single photon emission computed tomography (SPECT)/computed tomography (CT) imaging studies were performed in mice (four or five per group, depending on tumor growth) bearing subcutaneous and orthotopic MPM tumors by using HER1- and HER2-targeted indium 111 ((111)In)- and iodine 125 ((125)I)-labeled panitumumab and trastuzumab, respectively. Longitudinal MR imaging over 5 weeks was performed in three mice bearing orthotopic tumors to monitor tumor growth and metastases. SPECT/CT/MR imaging studies were performed at the final time point in the orthotopic models (n = 3). The standard unpaired Student t test was used to compare groups. Orthotopic tumors and pleural effusions were clearly visualized at MR imaging 3 weeks after tumor cell inoculation. At 2 days after injection, the mean (111)In-panitumumab uptake of 29.6% injected dose (ID) per gram ± 2.2 (standard error of the mean) was significantly greater than the (111)In-trastuzumab uptake of 13.6% ID/g ± 1.0 and the (125)I-panitumumab uptake of 7.4% ID/g ± 1.2 (P = .0006 and P = .0001, respectively). MR imaging fusion with SPECT/CT provided more accurate information about (111)In-panitumumab localization in the tumor, as the tumor was poorly visualized at CT alone. This study demonstrates the utility of radiolabeled anti-HER1 antibodies in the imaging of MPM in preclinical models. http://radiology.rsna.org/lookup/suppl/doi:10.1148/radiol.12121021/-/DC1.


[The use of drugs in migraine].
Drugs may be given either for treatment of the acute attack or as prophylaxis. Those most commonly used for the acute attack are analgesics, anti-emetics and ergotamine tartrate. A recent work (Volans, 1974) has shown that absorption may be impaired during a migraine attack. It is important therefore that not only is the analgesic given in an easily absorbed form but that a drug such as metaclopromide should be given to help restore the normal activity of the gastro-intestinal tract. Patients having one or more attacks of migraine a week may need prophylactic treatment. The drugs now used include: Methysergide, should only be used for severe cases when no other treatment has been found helpful. Dihydroergotamine, the vasoconstrictor activity is less than in ergotamine tartrate and can therefore be used prophylactically. Pizotifen, possesses powerful anti-serotonin properties. It also has marked antihistamine and antitryptamine properties as well as being a central sedative and anti-depressant. Clonidine, in doses of 1 mugm/Kg renders the blood vessels less sensitive to circulating amines and seems to be effective in about one third of patients with classical or common migraine. Sympathetic Blocking Agents: alpha-blockers: indoramine has recently given some good results; beta-blockers: such as propanolol and pindolol have also been used. Full trials of all the substances are now in progress. Tranquilisers and anti-depressants, two of those commonly used are diazepam and amitryptiline. In either cases a small dose only should be used. Anticonvulsants, phenytoin in doses of 50-100 mgs per day is sometimes helpful particularly in children or in those who have abnormal electroencephalograms.


Breakthrough Virus Neutralization Resistance as a Correlate of Protection in a Nonhuman Primate Heterologous Simian Immunodeficiency Virus Vaccine Challenge Study.
Comprehensive assessments of immune correlates of protection in human immunodeficiency virus (HIV) vaccine trials are essential to vaccine design. Neutralization sieve analysis compares the neutralization sensitivity of the breakthrough transmitted/founder (TF) viruses from vaccinated and control animals to infer the molecular mechanisms of vaccine protection. Here, we report a robust neutralization sieve effect in a nonhuman primate simian immunodeficiency virus (SIV) vaccine trial (DNA prime/recombinant adenovirus type 5 [rAd5] boost) (VRC-10-332) that demonstrated substantial protective efficacy and revealed a genetic signature of neutralization resistance in the C1 region of env. We found significant enrichment for neutralization resistance in the vaccine compared to control breakthrough TF viruses when tested with plasma from vaccinated study animals, plasma from chronically SIV-infected animals, and a panel of SIV-specific monoclonal antibodies targeting six discrete Env epitopes (P < 0.008 for all comparisons). Neutralization resistance was significantly associated with the previously identified genetic signature of resistance (P < 0.0001), and together, the results identify virus neutralization as a correlate of protection. These findings further demonstrate the in vivo relevance of our previous in vitro analyses of the SIVsmE660 challenge stock, which revealed a broad range of neutralization sensitivities of its component viruses. In sum, this report demonstrates proof-of-concept that phenotypic sieve analyses can elucidate mechanistic correlates of immune protection following vaccination and raises a cautionary note for SIV and SHIV (simian-human immunodeficiency virus) vaccine studies that employ challenge strains with envelope glycoproteins that fail to exhibit neutralization resistance profiles typical of TF viruses. With more than 2 million new infections annually, the development of an effective vaccine against HIV-1 is a global health priority. Understanding immunologic correlates of protection generated in vaccine trials is critical to advance vaccine development. Here, we assessed the role of vaccine-elicited neutralizing antibodies in a recent nonhuman primate study of a vaccine that showed significant protection against simian immunodeficiency virus (SIV) challenge and suggested a genetic signature of neutralization sensitivity. We found that breakthrough viruses able to establish infection in vaccinated animals were substantially more resistant to antibody-mediated neutralization than were viruses from controls. These findings suggest that vaccine-elicited neutralizing antibodies selectively blocked the transmission of more sensitive challenge viruses. Sieve analysis also corroborated a genetic signature of neutralization sensitivity and highlighted the impact of challenge swarm diversity. Our findings suggest an important role for neutralization sieve analyses as an informative component of comprehensive immune-correlates analyses.


Distinct activation of Cu-MOR for direct oxidation of methane to methanol.
N2O activation of Cu-MOR enhanced methanol production at elevated temperatures, to a greater extent than activation by O2 under optimal conditions. The increase in methanol production by N2O activation was attributed to the facile formation of an active copper center with no formation of inactive intermediate species.


Sun Exposure - Hazards and Benefits.
There are carcinogenic effects of sun exposure that increase the risk for skin cancer, especially for fair-skinned individuals. Therefore, there are recommendations to avoid sun exposure and to apply sun blockers. A more nuanced and balanced message for sun safety guidelines is now advocated. Despite an increased risk of death due to skin cancer, fair skinned women seem to have an overall survival advantage. In addition, an inverse association between sun exposure and hypertension, thromboembolism, and type 2 diabetes mellitus has been shown. Furthermore, low sun exposure habits result in increased risk of cardiovascular disease (CVD), and non-CVD/non-cancer mortality among women. There are also data supporting that the prognosis of cancer is improved with increasing levels of vitamin D/sun exposure. In this narrative review we will provide a brief update of hazards and benefits of sun exposure focused on an updated, balanced, and evidence-based view.


Circulatory support with larger volume intra-aortic balloon pump vs. standard volume or no-balloon pump during high-risk percutaneous coronary interventions. A randomised study.
Percutaneous coronary intervention in high-risk patients (HRPCI) is associated with increased risk of periprocedural complications such as hypotension and shock. Mechanical circulatory support devices may the bridge patient safely throughout the procedure and are often used in this setting. We assessed the outcomes of patients subjected to HRPCI and supported with intra-aortic balloon pump (IABP) of larger volume (MEGA) compared to standard volume (STRD) or no balloon support at all (CTRL). In this single-centre, open-label, randomised, controlled trial, HRPCI patients were randomly assigned to three groups: MEGA, STRD, and CTRL in a 1 : 1 : 1 scheme. Screening failure patients were assigned to the registry (REG). Composite haemodynamic endpoint (CHEP) was assessed during the procedure and major adverse cardiac even (MACE)/safety endpoints up to 1-year follow-up (FU). A total of 36 patients were randomised (13 MEGA, 14 STRD, and 9 CTRL). The incidence of in-hospital MACE was observed in 23.1% of MEGA, 7.1% of STRD and 33.3% of CTRL (p = 0.25) patients; MACE at FU in 50.0%, 35.7%, and 55.6% (p = 0.61); major bleeding in 46.2%, 28.6%, and 22.2%, (p = 0.45); and CHEP in 15.4%, 50.0%, and 44.4%, respectively (p = 0.13). On per-treatment (PT) analysis (16 MEGA, 10 STRD, and 21 CTRL), including 11 patients from REG, in-hospital MACE was observed in 18.8% of MEGA, 10.0% of STRD, and 23.8% of CTRL (p = 0.64) patients; MACE at FU in 53.3%, 20.0%, and 57.1% (p = 0.12); major bleeding in 37.5%, 20.0%, and 33.3% (p = 0.62); and CHEP in 15.5%, 50.0%, and 52.4%, respectively (p = 0.023). Larger volume intra-aortic balloon pump might be effective at reducing haemodynamic instability during HRPCI without a statistically significant effect on safety endpoints or MACE.


Fetal thigh circumference: a critical evaluation of its relationship to menstrual age.
To provide standards for evaluating fetal thigh circumference measurements obtained with ultrasound, a study of the growth of this parameter during pregnancy was made. Because of possible sampling problems before 22 weeks (menstrual age), this investigation was limited to the 22-40 week time period. Regression analysis indicated that the optimal model (R2 = 88.9%) for describing thigh circumference (ThC) growth was a power function. ThC variability increased with menstrual age, from 1.3 cm at 22 weeks to 3.1 cm at 40 weeks. These data were used to construct a standard growth curve for the ThC.


Permissive hypercapnia: what to remember.
Hypercapnia is a central component of diverse respiratory disorders, while 'permissive hypercapnia' is frequently used in ventilatory strategies for patients with severe respiratory failure. This review will present data from recent studies relating to hypercapnia, focusing on issues that are of importance to anesthesiologists caring for the surgical and/or critically ill patient. Protective ventilatory strategies involving permissive hypercapnia are widely used in patients with severe respiratory failure, particularly in acute respiratory distress syndrome, status asthmaticus, chronic obstructive pulmonary disease and neonatal respiratory failure. The physiologic effects of hypercapnia are increasingly well understood, and important recent insights have emerged regarding the cellular and molecular mechanisms of action of hypercapnia and acidosis. Acute hypercapnic acidosis is protective in multiple models of nonseptic lung injury. These effects are mediated in part through inhibition of the NF-κB pathway. Hypercapnia-mediated NF-κB inhibition may also explain several deleterious effects, including delayed epithelial wound healing and decreased bacterial killing, which has been demonstrated to cause worse lung injury in prolonged untreated pneumonia models. The mechanisms of action of hypercapnia and acidosis continue to be elucidated, and this knowledge is central to determining the safety and therapeutic utility of hypercapnia in protective lung ventilatory strategies.


[A method for extracting landmark geometric data from X-ray images].
Extracting landmark geometric data in C-arm X-ray images is a key technique of camera calibration process in a fluoroscopic-image-based surgical navigation system. The geometric data is the basis of X-ray image distortion correction and C-arm camera calibration. In this paper we present an integrated clustering algorithm for landmark geometric data extraction. This proposed alorithm, with the advantages of an adaptive thresholding method as well as of a fast quasi connected-component labeling method, needs only one pass to extract the needed geometric data in a gray scale image. It is of high executive efficiency and good robustness as well.


Inverse Design of Light Manipulating Structural Phase Transition in Solids.
This Perspective focuses on recent advances in understanding ultrafast processes involved in photoinduced structural phase transitions and proposes a strategy for precise manipulation of such transitions. It has been demonstrated that photoexcited carriers occupying empty antibonding or bonding states generate atomic driving forces that lead to either stretching or shortening of associated bonds, which in turn induce collective and coherent motions of atoms and yield structural transitions. For instance, phase transitions in IrTe2 and VO2, and nonthermal melting in Si, can be explained by the occupation of specific local bonding or antibonding states during laser excitation. These cases reveal the electronic-orbital-selective nature of laser-induced structural transitions. Based on this understanding, we propose an inverse design protocol for achieving or preventing a target structural transition by controlling the related electron occupations with orbital-selective photoexcitation. Overall, this Perspective provides a comprehensive overview of recent advancements in dynamical structural control in solid materials.


Foreign medical graduates in the 1980s: trends in specialization.
Secondary analysis of data collected by the American Medical Association and the Graduate Medical Education National Advisory Committee (GMENAC) suggests that measures to diminish the flow of alien Foreign Medical Graduates (FMGs) into the United States have been less effective than planned. Declining trends in the proportion of FMG house officers in the mid- to late-1970s have recently stabilized around 19 per cent. There has also been a dramatic increase in the number of US citizen Foreign Medical Graduates ( USFMGs ) in house officer positions. A pattern of alien FMG and USFMG house officer specialization correlates with specialties designated by the GMENAC as shortage areas by 1990 (r = -.49, p less than .05). Despite the GMENAC prediction of a surplus of physicians by 1990, differential selection of alien FMGs and USFMGs into shortage specialties may assure their substantial future presence in the US health care system.


Fostering a culture of engagement: a pilot study of the outcomes of training mental health nurses working in two UK acute admission units in brief solution-focused therapy techniques.
It is widely acknowledged that there are major concerns about quality of care, ward atmosphere, the nature of nurse-patient interactions and patient outcomes in UK psychiatric acute admission units. Brief solution-focused therapy (SFT) is an approach which aims to shift the focus of interactions in professional care away from the traditional concentration on an individual's problems and weaknesses towards a more proactive identification of their strengths and positive coping mechanisms. This approach relies on a collaborative engagement with patients, in which the nurse or therapist using simple language aims to help the patient construct a plan to ensure their immediate safety while working to identify, focus on and reinforce their strengths and coping mechanisms in the achievement of identified future goals. This paper reports on a pilot study whose principal objective was to determine whether a short training in brief SFT for psychiatric nurses can produce measurable improvements in nurse-patient interactions in two psychiatric acute admission wards. In this study, 36 nurses undertook a 2-day training course in SFT and were followed up 3 months after training. Positive results were obtained on a number of measures indicating that nurses had acquired knowledge and skills and were applying SFT techniques in their clinical work.


Distribution of immobilized enzymes on porous membranes.
In this article, the results from a theoretical and experimental investigation of enzyme immobilization in porous membranes are reported. A theoretical model of the immobilization process, which accounts for restricted diffusion of enzyme in the pores of the membrane, has been developed. The model predicts the effect of immobilization kinetics and time of immobilization on the enzyme distribution in the pores of the membrane. The immobilization of glucose oxidase and glucose oxidase-biotin conjugate on porous alumina membranes was experimentally investigated. Enzyme uptake data was correlated to the theory to determine the rate constant of immobilization and the distribution of the enzyme in the pore. Immobilization studies were carried out for enzyme adsorption and for enzyme attachment by covalent coupling. The distribution of enzyme was experimentally studied by assembling five membranes in the diffusion cell. Following immobilization, the membranes were separated and each was assayed for activity. The amount of active enzyme present in each membrane yielded a discrete distribution that compared well with that predicted by theory.


[Quadricuspid Aortic Valve with Atrial Septal Defect;Report of a Case].
The patient was a 69-year-old man who had 1st experienced exertional dyspnea approximately 1 month earlier. He visited our hospital for treatment because his symptoms had gradually worsened and he had developed orthopnea. Echocardiography revealed an atrial septal defect(ASD) and a quadricuspid aortic valve with moderate regurgitation. As patch closure of the ASD alone could pose a risk of exacerbating the untreated aortic regurgitation, we performed both procedures patch closure and aortic valve replacement through median sternotomy concomitantly. The postoperative course was favorable. We report our experience with a rare case of quadricuspid aortic valve with ASD and a review the relevant literature.


Converting a Geriatrics Clinic to Virtual Visits during COVID-19: A Case Study.
To characterize the experience of converting a geriatrics clinic to telehealth visits in early stages of a pandemic. An organizational case study with mixed methods evaluation from the first 8 weeks of converting a geriatrics clinic from in-person visits to video and telephone visits. Veteran's Health Administration in Northern California Participants Community-dwelling older Veterans receiving care at VA Palo Alto Geriatrics clinic. Veterans had a mean age of 85.7 (SD = 6.8) and 72.1% had cognitive impairment. Veterans with face-to-face appointments were converted to video or telephone visits to mitigate exposure to community spread of COVID-19. Thirty-two patient evaluations and 80 clinician feedback evaluations were completed. This provided information on satisfaction, care access during pandemic, and travel and time savings. Of the 62 scheduled appointments, 43 virtual visits (69.4%) were conducted. Twenty-six (60.5%) visits were conducted via video, 17 (39.5%) by telephone. Virtual visits saved patients an average of 118.6 minutes each. Patients and providers had similar, positive perceptions about telehealth to in-person visit comparison, limiting exposure, and visit satisfaction. After the telehealth appointment, patients indicated greater comfort with using virtual visits in the future. Thirty-one evaluations included comments for qualitative analysis. We identified 3 main themes of technology set-up and usability, satisfaction with visit, and clinical assessment and communication. During a pandemic that has limited the ability to safely conduct inperson services, virtual formats offer a feasible and acceptable alternative for clinically-complex older patients. Despite potential barriers and additional effort required for telehealth visits, patients expressed willingness to utilize this format. Patients and providers reported high satisfaction, particularly with the ability to access care similar to in-person while staying safe. Investing in telehealth services during a pandemic ensures that vulnerable older patients can access care while maintaining social distancing, an important safety measure.


Production of omega-3 eicosapentaenoic acid by metabolic engineering of Yarrowia lipolytica.
The availability of the omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) is currently limited because they are produced mainly by marine fisheries that cannot keep pace with the demands of the growing market for these products. A sustainable non-animal source of EPA and DHA is needed. Metabolic engineering of the oleaginous yeast Yarrowia lipolytica resulted in a strain that produced EPA at 15% of dry cell weight. The engineered yeast lipid comprises EPA at 56.6% and saturated fatty acids at less than 5% by weight, which are the highest and the lowest percentages, respectively, among known EPA sources. Inactivation of the peroxisome biogenesis gene PEX10 was crucial in obtaining high EPA yields and may increase the yields of other commercially desirable lipid-related products. This technology platform enables the production of lipids with tailored fatty acid compositions and provides a sustainable source of EPA.


Ultrasonically guided percutaneous aspiration biopsy of the pancreas.
One hundred and twenty-six patients had fine needle aspiration of the pancreas under ultrasonic guidance. The aspirates of 66 patients contained malignant cells. Two of 38 patients who had negative cytology subsequently were diagnosed at laparotomy as having carcinoma of the pancreas. The results of this study show that the sensitivity of the cytologic method was 97%, and the specificity and predictive value were 100%.


AN INTERACTION BETWEEN ENVIRONMENTAL TEMPERATURE AND GENETIC VARIATION FOR BODY SIZE FOR THE FITNESS OF ADULT FEMALE DROSOPHILA MELANOGASTER.
- Drosophila and other ectotherms show geographic genetic variation in body size, with larger individuals at higher latitudes and altitudes. Temperature is implicated as an important selective agent because long-term laboratory culture of Drosophila leads to the evolution of larger body size at lower temperatures. In this paper, we tested the hypothesis that, in Drosophila melanogaster, larger size is favored at lower temperatures in part because of selection on adult females. We used replicated lines of D. melanogaster artificially selected for increased and decreased wing area with constant cell area. The resulting size differences between the selected lines were due solely to differences in cell number, and thereby were similar to the cellular basis of clinal variation in body size in nature. We examined life-history traits of adult females at 18 and 25°C. Rearing for two generations at the two temperatures did not affect the extent of the size differences between lines from the different selection regimes. There was a strong interaction between temperature and size selection for both survival and lifetime reproductive success, with larger females living significantly longer and producing more offspring over their lifetime only when reared and tested in the colder environment. There was also an increase in average daily progeny production in large-line females relative to the control and small lines again, only in the colder environment. Thus, the females from the large selection lines were relatively fitter at the colder temperature. At both experimental temperatures, especially the lower one, the small- line females rescheduled their progeny production to later ages. Larger body size may have evolved at higher latitudes and altitudes because of the advantages to the adult female of being larger at lower temperatures.


Urea Cycle Defects: Early-Onset Disease Associated with A208T Mutation in OTC Gene-Expanding the Clinical Phenotype.
Ornithine transcarbamylase deficiency (OMIM: 311250) is the most common disorder of urea cycle disorders, accounting for nearly 50% of all cases. We report a case of a two-month- old male patient, who attends our medical genetics consultation because of low citrulline levels and elevated glutamine to citrulline ratio detected by expanded newborn screening with tandem mass spectrometry. He is an asymptomatic male with a normal physical examination and appropriate neurodevelopmental milestones. The patient has a family history of one older brother who died at 18 months old from severe and sudden hyperammonemia and a maternal aunt who suddenly died at two years old. He had high plasma ammonium concentration and a confirmed OTC mutation (p.A208T). Usually, this mutation causes OTC deficiency of late onset in adult males. However, this report raises awareness about mutations previously described as a late-onset causing disease, which can cause severe hyperammonemia and high risk of dying at an early age.


Oxidative stress, redox, and the tumor microenvironment.
Cellular metabolism is critical for the generation of energy in biological systems; however, as a result of electron transfer reactions, reactive oxygen species (ROS) are generated in aerobic cells. Although low amounts of ROS are easily tolerated by the cell, abnormally high levels of ROS induce oxidative stress. ROS are also produced after exposure to ionizing radiation, selected chemotherapeutic agents, hyperthermia, inhibition of antioxidant enzymes, or depletion of cellular reductants such as NADPH and glutathione. Oxidative stress such as ionizing radiation produces a variety of highly reactive free radicals that damage cells, initiate signal transduction pathways, and alter gene expression. Cells are capable of countering the effects of oxidative stress by virtue of a complex redox buffering system. With respect to the radiation treatment of cancer, components of the cellular redox armamentarium may be targeted to enhance cell killing in the case of tumors and/or protection in the case of normal tissues.


Supraventricular tachycardia as a complication of severe diabetic ketoacidosis in an adolescent with new-onset type 1 diabetes.
Diabetic ketoacidosis (DKA) is one of the most common causes of morbidity and mortality in new-onset type 1 diabetes (T1D). Supraventricular tachycardia (SVT), however, is a very rare complication of DKA. We present the case of a patient with new-onset T1D who presented with DKA. He received intravenous fluid resuscitation, insulin and potassium supplementation and subsequently developed SVT, confirmed on a 12-lead electrocardiograph despite a structurally normal heart. Vagal manoeuvres and adenosine failed to restore sinus rhythm, but flecainide was successful. We conclude that SVT can occur as a complication of DKA, including in new-onset T1D. Our case is the first of this phenomenon occurring in new-onset childhood diabetes, as the few prior documented cases had established diabetes. Furthermore, a combination of potassium derangement, hypophosphataemia and falling magnesium levels may have precipitated the event.


Highly efficient uptake of phosphorus in epiphytic bromeliads.
Vascular epiphytes which can be abundant in tree crowns of tropical forests have to cope with low and highly intermittent water and nutrient supply from rainwater, throughfall and stem flow. Phosphorus rather than nitrogen has been suggested as the most limiting nutrient element, but, unlike nitrogen, this element has received little attention in physiological studies. This motivated the present report, in which phosphate uptake kinetics by leaves and roots, the subsequent distribution within plants and the metabolic fate of phosphate were studied as a step towards an improved understanding of physiological adaptations to the conditions of tree canopies. Radioactively labelled [(32)P]phosphate was used to study uptake kinetics and plant distribution of phosphorus absorbed from bromeliad tanks. The metabolism of low molecular phosphorus metabolites was analysed by thin-layer chromatography followed by autoradiography. Uptake of phosphate from tanks is an ATP-dependent process. The kinetics of phosphorus uptake suggest that epiphytes possess effective phosphate transporters. The K(m) value of 1.05 microm determined for leaves of the bromeliad Aechmea fasciata is comparable with values obtained for the high affinity phosphate transporters in roots of terrestrial plants. In this species, young leaves are the main sink for phosphate absorbed from tank water. Within these leaves, phosphate is then allocated from the basal uptake zone into distal sections of the leaves. More than 80 % of the phosphate incorporated into leaves is not used in metabolism but stored as phytin. Tank epiphytes are adapted to low and intermittent nutrient supply by different mechanisms. They possess an effective mechanism to take up phosphate, minimizing dilution and loss of phosphorus captured in the tank. Available phosphorus is taken up from the tank solution almost quantitatively, and the surplus not needed for current metabolism is accumulated in reserves, i.e. plants show luxury consumption. Young, developing leaves are preferentially supplied with this nutrient element. Taken together, these features allow epiphytes the efficient use of scarce and variable nutrient supplies.


Development of sensitivity to orthographic errors in children: An event-related potential study.
To study the development of orthographic sensitivity during elementary school, we recorded event-related brain potentials (ERPs) from 2nd and 4th grade children who were exposed to line drawing of object or animals upon which the correctly or incorrectly spelled name was superimposed. Stimulus-locked ERPs showed a modulation of a frontocentral negativity between 200 and 500ms which was larger for the 4th grade children but did not show an effect of correctness of spelling. This effect was followed by a pronounced positive shift which was only seen in the 4th grade children and which showed a modulation of spelling correctness. This effect can be seen as an electrophysiological correlate of orthographic sensitivity and replicates earlier findings in adults. Moreover, response-locked ERPs triggered to the children's button presses indicating orthographic (in)-correctness showed a succession of waves including the frontocentral error-related negativity and a subsequent negativity with a more posterior distribution. This latter negativity was generally larger for the 4th grade children. Only for the 4th grade children, this negativity was smaller for the false alarm trials suggesting a conscious registration of the error in these children.


Classification of pancreatitis.
An international group of doctors interested in pancreatic disease met in Cambridge in March 1983, under the auspices of the Pancreatic Society of Great Britain and Ireland, to discuss the classification of pancreatitis in the light of developments that have taken place in the 20 years since the crucial conference in Marseille.


yucca6, a dominant mutation in Arabidopsis, affects auxin accumulation and auxin-related phenotypes.
Auxin plays critical roles in many aspects of plant growth and development. Although a number of auxin biosynthetic pathways have been identified, their overlapping nature has prevented a clear elucidation of auxin biosynthesis. Recently, Arabidopsis (Arabidopsis thaliana) mutants with supernormal auxin phenotypes have been reported. These mutants exhibit hyperactivation of genes belonging to the YUCCA family, encoding putative flavin monooxygenase enzymes that result in increased endogenous auxin levels. Here, we report the discovery of fertile dominant Arabidopsis hypertall1-1D and hypertall1-2D (yucca6-1D, -2D) mutants that exhibit typical auxin overproduction phenotypic alterations, such as epinastic cotyledons, increased apical dominance, and curled leaves. However, unlike other auxin overproduction mutants, yucca6 plants do not display short or hairy root phenotypes and lack morphological changes under dark conditions. In addition, yucca6-1D and yucca6-2D have extremely tall (>1 m) inflorescences with extreme apical dominance and twisted cauline leaves. Microarray analyses revealed that expression of several indole-3-acetic acid-inducible genes, including Aux/IAA, SMALL AUXIN-UP RNA, and GH3, is severalfold higher in yucca6 mutants than in the wild type. Tryptophan (Trp) analog feeding experiments and catalytic activity assays with recombinant YUCCA6 indicate that YUCCA6 is involved in a Trp-dependent auxin biosynthesis pathway. YUCCA6:GREEN FLUORESCENT PROTEIN fusion protein indicates YUCCA6 protein exhibits a nonplastidial subcellular localization in an unidentified intracellular compartment. Taken together, our results identify YUCCA6 as a functional member of the YUCCA family with unique roles in growth and development.


Membrane surface properties of normal and malignant cells: partition in aqueous two-polymer phase systems.
The partition of normal and malignantly transformed fibroblast lines and cell lines initiated from malignant human astrocytomas and a benign ganglioneuroma has been examined in aqueous dextran-polyethylene glycol phase system containing phosphate buffer with a low phosphate/sodium chloride ratio. The malignant astrocytomas showed a significantly lower partition coefficient as compared with the benign ganglioneuroma. Treatment of astrocytoma cells with dexamethasone caused an increase in the partitioning of the cell population. No differences were found in the partition behaviour of normal BHK-21 cells and their malignant transformants, the TRES fibrosarcoma cells. Polyoma and simian virus-transformed 3T3 fibroblasts showed partition ratios similar to the untransformed cells. Dexamethasone pre-treatment had no effect on the partition behaviour of these cells. The significance of these observations has been discussed in relation to the surface hydrophobicity and the neoplastic state.


Zinc deficiency in sheep and goats: three field cases.
Zinc deficiency was diagnosed in 2 flocks of sheep and a herd of goats. Alopecia and eating of wool were observed. Skin lesions were hyperkeratosis and parakeratosis. Abnormal hoof growth and a closed stance with arched back and bowed hindlimbs were seen. Anorexia, depression, and foot soreness diminished quickly after zinc supplementation. Skin lesions healed and hair and wool growth resumed. A cause for the deficiency could not be established.


Surface modification of cellulose fiber via supramolecular assembly of biodegradable polyesters by the aid of host-guest inclusion complexation.
In this article, we report a novel surface modification method for cellulose fiber that is based on supramolecular assembly. Beta-cyclodextrin (beta-CD) was first covalently grafted onto the fiber surface. Then poly(epsilon-caprolactone) (PCL) oligomers having both ends capped with adamantane motifs (i.e., PCL-AD) were immobilized to the cellulose fiber surface through the host-guest inclusion complexation between beta-CD and AD motif. FTIR-ATR and XPS analyses confirmed the successful assembly of PCL-ADs, which was further supported by the increasing trend of weight gain with the concentration of CDs on the fiber surface. Contact angle and TGA measurements reflect the enhanced hydrophobicity and thermal stability of the cellulose fiber as a consequence of this modification. The morphologies of the cellulose fiber before and after the assembly process have also been compared by SEM.


Evaluation of the spectral accuracy of mass spectrometers using compounds containing Cl or Br atoms.
Current procedures for the evaluation of spectral accuracy of mass spectrometers are limited by the lack of certified isotopic reference materials and the high uncertainty in the isotopic composition of natural abundance molecules. The calculated uncertainties in the ratio M + 1/M for natural abundance molecules containing any number of C, H, N and/or O atoms are close to 5% relative because of the natural variability of the isotopic composition of carbon. So, we have developed two alternative measurement procedures with much lower theoretical uncertainties for a better evaluation of spectral accuracy in both single and triple quadrupole analysers. The first method is based on the measurement of the M + 2/M, M + 4/M + 2, etc. ratios for halogenated organic compounds containing either Cl or Br. The theoretical uncertainties for these ratios because of natural variability are in the order of 0.3 to 1.0% making them suitable for the evaluation of spectral accuracy with the additional advantage that there is no need to take into account other limitations such as cluster purity or poor mass resolution. This procedure was applied to the evaluation of a single quadrupole GC-MS instruments using natural abundance PCB and PBDE standards with satisfactory results. The second method can be applied to tandem instruments and takes advantage of the loss of two halogen atoms when PCB and PBDE standards are fragmented by Collision Induced Dissociation. Theoretical SRM transition ratios can be calculated as a pure combinatorial probability with theoretical uncertainties lower than 0.1%. By combining PCBs and PBDEs with different number of halogen atoms, a mass range from 100 to 700 u and abundance ratios from 0.1 to 10 can be evaluated. The use of penta-chlorinated PCBs and/or penta-brominated PBDEs is finally recommended for the evaluation of spectral accuracy of mass spectrometers with the EI source. Copyright © 2016 John Wiley & Sons, Ltd.


High-resolution infrared spectroscopy of the charge-transfer complex [Ar-N2]+: a combined experimental/theoretical study.
A combined experimental and theoretical study of the charge-transfer complex [Ar-N(2)](+) is presented. Nearly 50 transitions split by spin-rotation interaction have been observed by means of infrared diode laser absorption spectroscopy in a supersonic planar plasma expansion. The band origin is at 2272.2563(18) cm(-1) and rotational constants in the ground and vibrationally (NN-stretch) excited state amount to 0.128701(8) cm(-1) and 0.128203(8) cm(-1), respectively. The interpretation of the data in terms of a charge switch upon complexation is supported by new ab initio calculations. The best estimate for a linear equilibrium structure yields R(e)(NN)=1.102 A and R(e)(Ar-N)=2.190 A. Predictions for molecular parameters not directly available from the experimental results are presented as well. Furthermore, the electronic structure and Ar-N bonding mechanism of [Ar-N(2)](+) have been analyzed in detail. The Ar-N bond is a textbook example of a classical 2-center-3-electron bond.


A new gene involved in stationary-phase survival located at 59 minutes on the Escherichia coli chromosome.
We determined the DNA sequence of a 2,232-bp region immediately upstream of the pcm gene at 59 min on the Escherichia coli chromosome that encodes an L-isoaspartyl protein methyltransferase with an important role in stationary-phase survival. Two open reading frames of 477 and 1,524 bp were found oriented in the same direction as that of the pcm gene. The latter open reading frame overlapped the 5' end of the pcm gene by 4 bp. Coupled in vitro transcription-translation analysis of DNA containing the 1,524-bp open reading frame directly demonstrated the production of a 37,000-Da polypeptide corresponding to a RNA species generated from a promoter within the open reading frame. The deduced amino acid sequence showed no similarity to known protein sequences. To test the function of this gene product, we constructed a mutant strain in which a kanamycin resistance element was inserted at a BstEII site in the middle of its coding region in an orientation that does not result in reduction of Pcm methyltransferase activity. These cells were found to survive poorly in stationary phase, at elevated temperatures, and in high-salt media compared with parent cells containing the intact gene, and we thus designate this gene surE (survival). surE appears to be the first gene of a bicistronic operon also containing the pcm gene. The phenotypes of mutations in either gene are very similar and indicate that both gene products are important for the viability of E. coli cells under stressful conditions.


Enzyme inactivation through sulfhydryl oxidation by physiologic NO-carriers.
Nitric oxide (NO) is a pluripotent regulatory molecule, yet the molecular mechanisms by which it exerts its effects are largely unknown. Few physiologic target molecules of NO have been identified, and even for these, the modifications caused by NO remain uncharacterized. Human glutathione reductase (hGR), a central enzyme of cellular antioxidant defense, is inhibited by S-nitrosoglutathione (GSNO) and by diglutathionyl-dinitroso-iron (DNIC-[GSH]2), two in vivo transport forms of NO. Here, crystal structures of hGR inactivated by GSNO and DNIC-[GSH]2 at 1.7 A resolution provide the first picture of enzyme inactivation by NO-carriers: in GSNO-modified hGR, the active site residue Cys 63 is oxidized to an unusually stable cysteine sulfenic acid (R-SOH), whereas modification with DNIC-[GSH]2 oxidizes Cys 63 to a cysteine sulfinic acid (R-SO2H). Our results illustrate that various forms of NO can mediate distinct chemistry, and that sulfhydryl oxidation must be considered as a major mechanism of NO action.


An evaluation of inexpensive methods for root image acquisition when using rhizotrons.
Belowground processes play an essential role in ecosystem nutrient cycling and the global carbon budget cycle. Quantifying fine root growth is crucial to the understanding of ecosystem structure and function and in predicting how ecosystems respond to climate variability. A better understanding of root system growth is necessary, but choosing the best method of observation is complex, especially in the natural soil environment. Here, we compare five methods of root image acquisition using inexpensive technology that is currently available on the market: flatbed scanner, handheld scanner, manual tracing, a smartphone application scanner and a time-lapse camera. Using the five methods, root elongation rate (RER) was measured for three months, on roots of hybrid walnut (Juglans nigra × Juglans regia L.) in rhizotrons installed in agroforests. When all methods were compared together, there were no significant differences in relative cumulative root length. However, the time-lapse camera and the manual tracing method significantly overestimated the relative mean diameter of roots compared to the three scanning methods. The smartphone scanning application was found to perform best overall when considering image quality and ease of use in the field. The automatic time-lapse camera was useful for measuring RER over several months without any human intervention. Our results show that inexpensive scanning and automated methods provide correct measurements of root elongation and length (but not diameter when using the time-lapse camera). These methods are capable of detecting fine roots to a diameter of 0.1 mm and can therefore be selected by the user depending on the data required.


Potassium 3-cyano-4-(dicyanomethylene)-5-oxo-4,5-dihydro-1H-pyrrol-2-olate.
The crystal structure of the title potassium salt, K(+).C(8)HN(4)O(2)(-), of the organic anion 3-cyano-4-(dicyanomethylene)-5-oxo-4,5-dihydro-1H-pyrrol-2-olate shows that the dicyanomethylene moiety is able to accept an electron in the same way as does tetracyanoethylene, to yield the novel product. The organic anion is nearly planar, with deviations caused by steric crowding among the exocyclic cyano groups. The K(+) cations lie within tricapped trigonal prisms that stack to form channels. The three-dimensional structure is completed by the formation of hydrogen-bonded chains by the anions.


Underdiagnosed Roifman syndrome manifested as non-ischaemic cardiomyopathy: a case report.
Roifman syndrome is a rare congenital disorder characterized by growth retardation, cognitive delay, spondyloepiphyseal dysplasia, immunodeficiency, and retinal dystrophy. However, very rarely, with only one case reported to date, a patient with Roifman syndrome may develop cardiomyopathy in their lifetime. We reported a case with underdiagnosed Roifman syndrome confirmed by whole genome sequencing, manifested as non-ischaemic cardiomyopathy, which has broadened the association between non-ischaemic cardiomyopathy and the genetic disorder Roifman syndrome. We also underscored that cardiomyopathy might be part of the clinical manifestations of Roifman syndrome and the importance of whole genome sequencing for diagnosis, as RNU4ATAC is not targeted by many commercially available exome capture kits.


Congenital hypogonadotrophic hypogonadism, induction of minipuberty, and future fertility.
A 33-year-old man with Kallmann syndrome had received pulsatile GnRH as an infant for the treatment of cryptorchidism. As an adult, his treatment for fertility with gonadotrophins was unusually rapid compared with expectations, with a total sperm count of 25 million after only 12 months of gonadotrophin therapy. We propose that pulsatile GnRH treatment as an infant induced minipuberty and facilitated his successful, rapid response to therapy. We also propose that identification of the absence of minipuberty in infants with clinical signs suggesting congenital hypogonadotrophic hypogonadism (CHH) is an opportunity for intervention with pulsatile GnRH yielding benefits for fertility decades later. Absence of minipuberty in males with CHH results in low Sertoli cell numbers and delayed response to induction of spermatogenesis in adulthood. Presentation with 'red flags' for androgen deficiency including cryptorchidism at birth, with or without micropenis, should prompt screening for CHH and minipuberty by measurement of gonadotrophins and testosterone in the first 2 months after birth. Pulsatile GnRH therapy in patients with CHH, given prior to age of attainment of Sertoli cell maturation, can replicate the normal physiology of minipuberty, thereby priming the testis for future fertility.


Functioning insulinoma--incidence, recurrence, and long-term survival of patients: a 60-year study.
For the 60-year period from 1927 through 1986, we assessed the incidence, recurrence, and long-term survival among all Mayo Clinic patients with histologically confirmed functioning insulinoma. With use of the complete medical record system at Mayo and the comprehensive epidemiologic data base of residents of Olmsted County, Minnesota, we found 224 patients in whom an initial pancreatic exploration at Mayo had confirmed the presence of insulinoma. The median age (and range) of these patients at surgical diagnosis was 47 (8 to 82) years, and 59% were female patients. During the study period, eight cases of insulinoma occurred among residents of Olmsted County; their age and gender distributions were similar to those of the total cohort. The incidence of insulinoma among residents of Olmsted County increased during the study period to a stable level during the last 2 decades of 4 cases per 1 million person-years. For the total cohort, 7.6% had multiple endocrine neoplasia type I (MEN I), and 5.8% had malignant insulinoma. The risk of recurrence was greater among patients with MEN I (21% at 10 and 20 years) than in those without MEN I (5% at 10 years and 7% at 20 years). Although survival of the total cohort was not significantly impaired, it was significantly worse than expected for patients with malignant insulinoma (29% versus 88% expected at 10 years postoperatively). We conclude that insulinoma is less rare than previously suspected. After successful surgical removal, the long-term risk of recurrent insulinoma is relatively high in patients with MEN I; for patients with benign disease, the long-term survival is normal.


Role for platelet von Willebrand factor in supporting platelet-vessel wall interactions in von Willebrand disease.
Twelve infusions of plasma concentrates of von Willebrand factor (vWF) were given to four patients with severe (type III) von Willebrand disease (vWD). Their prolonged bleeding times were either completely or partially corrected after five infusions and had not changed after the remaining seven. In contrast, the low platelet coverage of the subendothelial surface of rabbit aorta perfused with normal washed platelets and red cells resuspended in preinfusion patient plasma was completely or partially corrected in ten instances by replacing preinfusion plasma with postinfusion plasma and remained unchanged in two. Postinfusion improvement in surface coverage was greater than that in bleeding time, suggesting that vWF from normal platelets is needed to support optimal platelet-vessel wall interactions in vWD. This possibility was further explored through other perfusion experiments. The subendothelial surface covered by platelets from an untreated patient with type III vWD (containing no measurable vWF) or from a type IIA vWD patient (containing dysfunctional vWF) resuspended in normal plasma was much smaller than that covered by normal platelets resuspended in normal plasma. These results establish that platelet vWF is important in supporting platelet-vessel wall interactions in vWD and also provide experimental support in favour of the therapeutic transfusion of normal platelets in addition to vWF concentrates to correct the bleeding time in vWD patients.


Social Workers' Experiences of Support in the Workplace during the COVID-19 Pandemic.
This qualitative study included in-depth semistructured interviews with 17 social workers from across the state of Texas that took place during the summer of 2021. The purpose of this study was to describe social workers' experiences of workplace support during the COVID-19 pandemic and provide implications for how to improve support in the workplace. The interviews were analyzed using a phenomenological approach to describe the main themes of the participants' experiences. The findings combined with current literature suggest that following the pandemic and in the case of similar prolonged disasters organizational leaders should consider supporting their social work staff by clearly communicating protocol changes, providing more autonomy and flexible work arrangements, increasing time off and financial support, and providing tangible assistance such as technological support. Leaders should also consider their social work staff's emotional well-being following the pandemic by providing therapeutic resources and bonding opportunities for staff to promote a more resilient team-like culture in the workplace.


Long-Term Aberrations To Cerebellar Endocannabinoids Induced By Early-Life Stress.
Emerging evidence points to the role of the endocannabinoid system in long-term stress-induced neural remodeling with studies on stress-induced endocannabinoid dysregulation focusing on cerebral changes that are temporally proximal to stressors. Little is known about temporally distal and sex-specific effects, especially in cerebellum, which is vulnerable to early developmental stress and is dense with cannabinoid receptors. Following limited bedding at postnatal days 2-9, adult (postnatal day 70) cerebellar and hippocampal endocannabinoids, related lipids, and mRNA were assessed, and behavioral performance evaluated. Regional and sex-specific effects were present at baseline and following early-life stress. Limited bedding impaired peripherally-measured basal corticosterone in adult males only. In the CNS, early-life stress (1) decreased 2-arachidonoyl glycerol and arachidonic acid in the cerebellar interpositus nucleus in males only; (2) decreased 2-arachidonoyl glycerol in females only in cerebellar Crus I; and (3) increased dorsal hippocampus prostaglandins in males only. Cerebellar interpositus transcriptomics revealed substantial sex effects, with minimal stress effects. Stress did impair novel object recognition in both sexes and social preference in females. Accordingly, the cerebellar endocannabinoid system exhibits robust sex-specific differences, malleable through early-life stress, suggesting the role of endocannabinoids and stress to sexual differentiation of the brain and cerebellar-related dysfunctions.


A simple spatial typology for assessment of complex coastal ecosystem services across multiple scales.
This paper aims to present and demonstrate the applicability of a methodology to characterise supply and demand for ecosystem services (ES) on the basis of spatial properties and interdependence, and economic properties of ES. The typology is demonstrated to support inclusion of ES flow in social-ecological systems management. Ria de Aveiro coastal lagoon, Portugal, is used as a showcase for the proposed spatial typology for ecosystem services assessment across multiple scales. To address the proposed objective a four-step methodology was developed: i) choice of the management area and definition of the boundaries of the system to be governed; ii) assessment and classification of ES produced or consumed within the management area, where supply or demand of an ES may be endogenous, exogenous or partially exogenous/endogenous; iii) mapping of production and consumption areas covering the major categories of ES (biotic and abiotic provisioning services, regulating and maintenance services, and cultural services) and major types of spatial relationships, which may be independent; interdependent, dependent or controlling; and iv) spatial characterisation of the ES production and benefits based on excludability and rivalness of ES. In Ria de Aveiro from the variety of ES and their spatial relationships we highlight interdependencies between the Lagoon and other locations outside the management area. This approach shows that the proposed methodology can support the understanding of these interrelationships and improve these connections by, identifying inherent transactions and key-actors.


Lethal mutagenesis of an RNA plant virus via lethal defection.
Lethal mutagenesis is an antiviral therapy that relies on increasing the viral mutation rate with mutagenic nucleoside or base analogues. Currently, the molecular mechanisms that lead to virus extinction through enhanced mutagenesis are not fully understood. Increasing experimental evidence supports the lethal defection model of lethal mutagenesis of RNA viruses, where replication-competent-defectors drive infective virus towards extinction. Here, we address lethal mutagenesis in vivo using 5-fluorouracil (5-FU) during the establishment of tobacco mosaic virus (TMV) systemic infections in N. tabacum. The results show that 5-FU decreased the infectivity of TMV without affecting its viral load. Analysis of molecular clones spanning two genomic regions showed an increase of the FU-related base transitions A → G and U → C. Although the mutation frequency or the number of mutations per molecule did not increase, the complexity of the mutant spectra and the distribution of the mutations were altered. Overall, our results suggest that 5-FU antiviral effect on TMV is associated with the perturbation of the mutation-selection balance in the genomic region of the RNA-dependent RNA polymerase (RdRp). Our work supports the lethal defection model for lethal mutagenesis in vivo in a plant RNA virus and opens the way to study lethal mutagens in plant-virus systems.


Effect of adenosine triphosphate on the accessory pathways.
To determine the site of the anterograde and retrograde conduction in the Wolff-Parkinson-White syndrome (WPW), 40 mg of adenosine triphosphate (ATP) was injected during electrophysiological studies in 53 patients with ventricular preexcitation. In 40 cases, the accessory pathway was evident (group 1) and in 13 cases it was concealed (group 2). In 10 cases in group 1, anterograde conduction was abolished with disappearance of the features of preexcitation. In 7 patients of group 1 and in 4 patients of group 2, retrograde conduction in the accessory pathway was prolonged or abolished. These effects were unexpected because ATP is a parasympathomimetic drug. There was a correlation between the Kent effective refractory period (ERP) and the action of ATP. When the drug did not change the anterograde and retrograde conduction in the Kent bundle, the anterograde accessory pathway ERP was always less than 230 ms. When ATP only decreased retrograde conduction in the Kent bundle, anterograde accessory pathway ERP was always more than 280 ms.


Bone marrow stromal cells express neural phenotypes in vitro and migrate in brain after transplantation in vivo.
To investigate the differentiation of bone marrow stromal cells (BMSC) into neuron-like cells and to explore their potential use for neural transplantation. BMSC from rats and adult humans were cultured in serum-containing media. Salvia miltiorrhiza was used to induce human BMSC (hBMSC) to differentiate. BMSC were identified with immunocytochemistry. Semi-quantitative RT-PCR was used to examine mRNA expression of neurofilamentl (NF1), nestin and neuron-specific enolase (NSE) in rat BMSC (rBMSC). Rat BMSC labelled by Hoschst33258 were transplanted into striatum of rats to trace migration and distribution. rBMSC expressed NSE, NF1 and nestin mRNA, and NF1 mRNA and expression was increased with induction of Salvia miltiorrhiza. A small number of hBMSC were stained by anti-nestin, anti-GFAP and anti-S100. Salvia miltiorrhiza could induce hBMSC to differentiate into neuron-like cells. Some differentiated neuron-like cells, that expressed NSE, beta-tubulin and NF-200, showed typical neuron morphology, but some neuron-like cells also expressed alpha smooth muscle protein, making their neuron identification complicated. rBMSC could migrate and adapted in the host brains after being transplanted. Bone marrow stromal cells could express phenotypes of neurons, and Salvia miltiorrhiza could induce hBMSC to differentiate into neuron-like cells. If BMSC could be converted into neurons instead of mesenchymal derivatives, they would be an abundant and accessible cellular source to treat a variety of neurological diseases.


T-type calcium channels drive migration/invasion in BRAFV600E melanoma cells through Snail1.
Melanoma is a malignant tumor derived from melanocytes. Once disseminated, it is usually highly resistant to chemotherapy and is associated with poor prognosis. We have recently reported that T-type calcium channels (TTCCs) are overexpressed in melanoma cells and play an important role in melanoma progression. Importantly, TTCC pharmacological blockers reduce proliferation and deregulate autophagy leading to apoptosis. Here, we analyze the role of autophagy during migration/invasion of melanoma cells. TTCC Cav3.1 and LC3-II proteins are highly expressed in BRAFV600E compared with NRAS mutant melanomas, both in cell lines and biopsies. Chloroquine, pharmacological blockade, or gene silencing of TTCCs inhibit the autophagic flux and impair the migration and invasion capabilities, specifically in BRAFV600E melanoma cells. Snail1 plays an important role in motility and invasion of melanoma cells. We show that Snail1 is strongly expressed in BRAFV600E melanoma cells and patient biopsies, and its expression decreases when autophagy is blocked. These results demonstrate a role of Snail1 during BRAFV600E melanoma progression and strongly suggest that targeting macroautophagy and, particularly TTCCs, might be a good therapeutic strategy to inhibit metastasis of the most common melanoma type (BRAFV600E).


Modulation of KC/gro protein (interleukin-8 related protein in rodents) release from hepatocytes by biologically active mediators.
Modulation of the release of KC/gro protein (a chemoattractant for neutrophils; IL-8 related protein in rodents) from isolated hepatocytes after stimulation with biologically active mediators was investigated. The release of KC/gro protein from hepatocytes of control rats was enhanced by stimulation with lipopolysaccharide, IL-1 beta or TNF-alpha in a dose-dependent manner, but was not enhanced by IL-6. In contrast, although spontaneous release of KC/gro protein from the hepatocytes of chronically ethanol-fed rats was markedly enhanced as compared with control rats, the relative increase by stimulation with lipopolysaccharide, IL-1 beta or TNF-alpha was significantly smaller than in controls. These findings suggest that the regulation of hepatocyte KC/gro protein production might be disturbed in chronically ethanol-fed rats.


The localization of myocardial ischaemia with technetium-99m methoxy isobutyl isonitrile and single photon emission computed tomography.
Thirty-two patients with suspected coronary artery disease were studied by single photon emission computed tomography (SPECT) imaging with oblique reconstructions of the myocardium following the intravenous administration of technetium-99m methoxy isobutyl isonitrile at peak exercise. All patients also underwent three-vessel coronary angiography. The SPECT technique produced very detailed images allowing easy delineation of localized myocardial defects. Segmental myocardial uptake defects were compared with diseased vessels as shown at angiography. A good correlation was shown between right coronary artery (RCA) disease and mid and proximal inferior segments and between left circumflex (LCx) artery disease and mid and proximal lateral segments, allowing accurate localization of a defect to one of these two vessels' territories. Sensitivity and specificity of detection of disease of the RCA and LCx artery were high. Defects associated with a lesion of the left anterior descending vessel were more variable.


Rates and predictors of 5-year survival in a national cohort of asymptomatic elderly patients undergoing carotid revascularization.
Current American Heart Association guidelines recommend carotid revascularization for asymptomatic patients on the basis of life expectancy. To determine the rates and predictors of 5-year survival in elderly patients with asymptomatic carotid artery stenosis who underwent either carotid artery stent placement (CAS) or carotid endarterectomy (CEA). The rates of 5-year survival were determined by use of Kaplan-Meier survival methods in a representative sample of fee-for-service Medicare beneficiaries ≥65 years of age who underwent CAS or CEA for asymptomatic carotid artery stenosis with postprocedural follow-up of 3.4 ± 1.7 years. Cox proportional hazards analysis was used to assess the relative risk of all-cause mortality for patients in the presence of selected comorbidities, including ischemic heart disease, chronic renal failure, and atrial fibrillation, after adjustment for potential confounders such as age, sex, race/ethnicity, and procedure type. A total of 22,177 patients with asymptomatic carotid artery stenosis were treated with either CAS (n = 2144) or CEA (n = 20,033). The overall estimated 5-year survival rate (±SE) was 95.3 ± 0.00149; it was 95.5% and 93.8% in patients treated with CEA and CAS, respectively. After adjustment for potential confounders, relative risk of all-cause 5-year mortality was significantly higher among patients with atrial fibrillation (relative risk, 1.8; 95% confidence interval, 1.5-2.1) and those with chronic renal failure (relative risk, 2.1; 95% confidence interval, 1.7-2.6). Risks and benefits must be carefully weighed before carotid revascularization in elderly patients with asymptomatic carotid artery stenosis who have concurrent atrial fibrillation or chronic renal failure.


4,6-Diaminopyrimidines as Highly Preferred Troponin I-Interacting Kinase (TNNI3K) Inhibitors.
Structure-guided progression of a purine-derived series of TNNI3K inhibitors directed design efforts that produced a novel series of 4,6-diaminopyrimidine inhibitors, an emerging kinase binding motif. Herein, we report a detailed understanding of the intrinsic conformational preferences of the scaffold, which impart high specificity for TNNI3K. Further manipulation of the template based on the conformational analysis and additional structure-activity relationship studies provided enhancements in kinase selectivity and pharmacokinetics that furnished an advanced series of potent inhibitors. The optimized compounds (e.g., GSK854) are suitable leads for identifying new cardiac medicines and have been employed as in vivo tools in investigational studies aimed at defining the role of TNNI3K within heart failure.