Community turnover of wood-inhabiting fungi across hierarchical spatial scales.
For efficient use of conservation resources it is important to determine how species diversity changes across spatial scales. In many poorly known species groups little is known about at which spatial scales the conservation efforts should be focused. Here we examined how the community turnover of wood-inhabiting fungi is realised at three hierarchical levels, and how much of community variation is explained by variation in resource composition and spatial proximity. The hierarchical study design consisted of management type (fixed factor), forest site (random factor, nested within management type) and study plots (randomly placed plots within each study site). To examine how species richness varied across the three hierarchical scales, randomized species accumulation curves and additive partitioning of species richness were applied. To analyse variation in wood-inhabiting species and dead wood composition at each scale, linear and Permanova modelling approaches were used. Wood-inhabiting fungal communities were dominated by rare and infrequent species. The similarity of fungal communities was higher within sites and within management categories than among sites or between the two management categories, and it decreased with increasing distance among the sampling plots and with decreasing similarity of dead wood resources. However, only a small part of community variation could be explained by these factors. The species present in managed forests were in a large extent a subset of those species present in natural forests. Our results suggest that in particular the protection of rare species requires a large total area. As managed forests have only little additional value complementing the diversity of natural forests, the conservation of natural forests is the key to ecologically effective conservation. As the dissimilarity of fungal communities increases with distance, the conserved natural forest sites should be broadly distributed in space, yet the individual conserved areas should be large enough to ensure local persistence.


Like me, like you - relative importance of peers and siblings on children's fast food consumption and screen time but not sports club participation depends on age.
Lifestyle interventions to prevent paediatric obesity often target family and peer settings; their success is likely to depend on the influence that peers and families exert on children's lifestyle behaviors at different developmental stages. First, to determine whether children's lifestyle behavior more closely resembles their peers' or siblings' behaviors. Secondly, to investigate longitudinally whether children's behavioral change is predicted by that of their peers or their siblings as they grow older. The European prospective IDEFICS/I.Family cohort (baseline survey: 2007/2008, first follow-up: 2009/2010, and second follow-up: 2013/2014) aims at investigating risk factors for overweight and related behaviors during childhood and adolescence. The present investigation includes 2694 observations of children and their siblings aged 2 to 18 years. Peers were defined as same-sex, same-age children in the same community and identified from the full cohort. The longitudinal analysis (mean follow-up time: 3.7 years) includes 525 sibling pairs. Children's lifestyle behaviors including fast food consumption (frequency/week), screen time (hours/week) and sports club participation (hours/week) were assessed by questionnaire. Data were analyzed using multilevel linear models. Children's lifestyle behavior was associated with the respective behavior of their peers and sibling for all 3 behaviors. For fast food consumption, the peer resemblance was more than 6-fold higher than the sibling resemblance and the peer resemblance surpassed the sibling resemblance by the age of 9-10 years. The similarities with peers for fast food consumption and screen time steadily increased, while the similarities with siblings steadily decreased with increasing age of the children (Pinteraction < 0.001). In contrast, the relative importance of peers and siblings on sports club duration did not vary by the age of the children. Longitudinal results showed that children's changes in fast food consumption were more strongly associated with those in their peer group than their sibling, in particular if the age gap between siblings was large. In conclusion, our results support the implementation of multi-setting interventions for improving lifestyle behaviors in children. Our findings might also guide future intervention studies in the choice of timing and setting in which interventions are likely to be most effective. From the ages of 9-10 years onwards, family- or home-based interventions targeting children's fast food intake and screen time behavior may become less effective than school- or community-based interventions aimed at peer groups.


Water Solubilization and Guest Molecule-Induced Fluorescence Changes of Porous Host Molecules via Complexing with Polysaccharide or Polypeptide.
To detect small aromatic molecules in water, we prepared functional host molecules based on water-soluble N,N'-bis(2-aminobenzophenone)-1,4,5,8-naphthalenetetracarboxylic diimide (1) and a solubilizing agent using a high-speed vibration milling apparatus. The fluorescence response of host 1-solubilizing agent complexes before and after extraction of small aromatic guest molecules was large and the fluorescence maxima were dependent on the small aromatic guest molecules.


Cavernous angioma associated with venous angioma--two case reports.
A 17-year-old male and a 28-year-old female presented with cavernous angioma and venous angioma colocated in the same region, and manifesting as convulsions. The cavernous angiomas were extirpated without damage to the venous angioma. No postoperative neurological deficits were observed. The patients have been free of convulsions without anticonvulsant medication for 2 years. Early extirpation of cavernous angioma with presentation of venous angioma is recommended, preferably before bleeding occurs.


Characterization of calcium and magnesium binding domains of human 5-lipoxygenase.
Two calcium binding sites, separated by about 9.3A, present in the loops that connect the beta-sheets of N-terminal domain contain the ligating residues F14, A15, G16, D79, and D18, D19, L76, respectively. Magnesium is found to bind in regions, which are marginally different owing to the disparity in the ionic radii of Ca2+ and Mg2+. The entropy analysis on the loops of 5-lipoxygenase, implementing the wormlike chain model, explains that the N-terminal beta-barrel is well suited to accommodate calcium binding sites. The large buried side chain area of W102 (compared to W13 and W75) and comparatively smaller fraction of side chain exposed to polar atoms corroborate the calcium induced higher affinity to phosphatidylcholine (PC). However, W80 lying in close proximity of the calcium binding sites is expected to have considerable PC affinity but negligible calcium induced effect on PC binding.


Cytotoxicity of advanced glycation endproducts is mediated by oxidative stress.
Non-enzymatic glycation of proteins with reducing sugars and subsequent transition metal catalysed oxidations leads to the formation of protein bound "advanced glycation endproducts" (AGEs). They accumulate on long-lived proteins and are for example structural components of the beta-amyloid plaques in Alzheimer's disease. Since the oxidation of glycated proteins as well as the interaction of AGEs with cell surface receptors produces superoxide radicals, it was tested in BHK 21 hamster fibroblast cells and SH-SY5Y human neuroblastoma cells if AGEs can exert cytotoxic effects on cells. Cell viability was assessed with three independent tests: MTT-assay (activity of the mitochondrial respiratory chain), lactate dehydrogenase assay (release of cytoplasmatic enzymes, membrane integrity) and Neutral Red assay (active uptake of a hydrophilic dye). Two model AGEs, chicken egg albumin-AGE and BSA-AGE, both caused significant cell death in a dose-dependent manner. The cytotoxic effects of AGEs could be attenuated by alpha-ketoglutarate and pyruvate, by antioxidants such as thioctic acid and N-acetylcysteine, and by aminoguanidine, an inhibitor of nitric oxide synthase. This suggests that reactive oxygen species as well as reactive nitrogen species contribute to AGE mediated cytotoxicity. Since AGEs accumulate on beta-amyloid plaques in AD over time, they may additionally contribute to oxidative stress, cell damage, functional loss and even neuronal cell death in the Alzheimer's disease brain.


Erythropoietin as a performance-enhancing drug: Its mechanistic basis, detection, and potential adverse effects.
Erythropoietin (EPO) is the main hormone regulating red blood cell (RBC) production. The large-scale production of a recombinant human erythropoietin (rHuEPO) by biotechnological methods has made possible its widespread therapeutic use as well as its misuse in sports. Since the marketing of the first epoetin in 1989, the development has progressed to the third-generation analogs. However, the production of rHuEPO is costly, and the frequent administration of an injectable formula is not optimal for compliance of therapeutic patients. Hence, pharmaceutical industries are currently developing alternative approaches to stimulate erythropoiesis, which might offer new candidates for doping purposes. The hypoxia inducible factors (HIF) pathway is of particular interest. The introduction of new erythropoiesis-stimulating agents (ESAs) for clinical use requires subsequent development of anti-doping methods for detecting the abuse of these substances. The detection of ESAs is based on two different approaches, namely, the direct detection of exogenous substances and the indirect detection, for which the effects of the substances on specific biomarkers are monitored. Omics technologies, such as ironomics or transcriptomics, are useful for the development of new promising biomarkers for the detection of ESAs. Finally, the illicit use of ESAs associates with multiple health risks that can be irreversible, and an essential facet of anti-doping work is to educate athletes of these risks. The aim of this review is to provide an overview of the evolution of ESAs, the research and implementation of the available detection methods, and the side effects associated with the misuse of ESAs.


Trio, a novel high fecundity allele: I. Transcriptome analysis of granulosa cells from carriers and noncarriers of a major gene for bovine ovulation rate.
A major gene for bovine ovulation rate has been mapped to a 1.2 Mb region of chromosome 10. Screening of coding regions of positional candidate genes within this region failed to reveal a causative polymorphism, leading to the hypothesis that the phenotype results from differences in candidate gene expression rather than alteration of gene structure. This study tested differences in expression of positional candidate genes in granulosa cells between carriers and noncarriers of the high fecundity allele, as well as characterizing differences in the transcriptomic profile between genotypes. Five carriers and five noncarriers, female descendants of "Trio," a carrier of the high fecundity allele were initially used in an RNA-seq analysis of gene expression. Four of ten samples were contaminated with theca cells, so that six samples were used in the final analysis (three of each genotype). Of 14 973 genes expressed, 143 were differentially expressed (false discovery rate P < 0.05) in carriers versus noncarriers. Among the positional candidate genes, SMAD6 was 6.6-fold overexpressed in the carriers compared to noncarriers (P < 5 × 10-5). This result was replicated in an independent group of 12 females (7 carriers and 5 noncarriers) using quantitative real-time PCR; SMAD6 was 9.3-fold overexpressed in carriers versus noncarriers (P = 1.17 × 10-6). Association of overexpression of SMAD6, an inhibitor of the BMP/SMAD signaling pathway, with high ovulation rate corresponds well with disabling mutations in ligands (BMP15 and GDF9) and a receptor (BMPR1B) of this pathway that cause increased ovulation rate in sheep.


Sirolimus (rapamycin) halts and reverses progression of allograft vascular disease in non-human primates.
Current immunosuppressive protocols fail to prevent chronic rejection often manifested as graft vascular disease (GVD) in solid organ transplant recipients. Several new immunosuppressants including sirolimus, a dual function growth factor antagonist, have been discovered, but studies of drug efficacy have been hampered by the lack of a model of GVD in primates, as a prelude to clinical trials. As described earlier, we have developed a novel non-human primate model of GVD where progression of GVD is quantified by intravascular ultrasound (IVUS). Twelve cynomolgus monkeys underwent aortic transplantation from blood group compatible but mixed lymphocyte reaction-mismatched donors. To allow the development of GVD in the allograft, no treatment was administered for the first 6 weeks. Six monkeys were treated orally with sirolimus from day 45 after transplantation to day 105. Progression of GVD measured as change in intimal area from day 42 to 105 was halted in sirolimus-treated monkeys compared to untreated monkeys (P<0.001, general linear model). On day 105, the intimal area +/- SEM was 3.7+/-1.0 and 6.4+/-0.5 mm2, respectively (P<0.05, t test). The magnitude of allograft intimal area on day 105 correlated inversely with sirolimus trough levels (R2=0.67, P<0.05). Regression of the intimal area was seen in four of six sirolimus-treated monkeys, which was significantly different from the untreated monkeys (P<0.05). Our results in the first non-human primate model of GVD showed that treatment with sirolimus not only halted the progression of preexisting GVD but also was associated with partial regression. Sirolimus trough blood levels were correlated with efficacy. Therefore, sirolimus has the potential to control clinical chronic allograft rejection.


Which Melbourne Metropolitan Areas Are Vulnerable to COVID-19 Based on Age, Disability, and Access to Health Services? Using Spatial Analysis to Identify Service Gaps and Inform Delivery.
Aging adults (65+) with disability are especially vulnerable to coronavirus disease 2019 (COVID-19), and on contracting, they are a cohort most likely to require palliative care. Therefore, it is very important that health services-particularly health services providing palliative care-are proximately available. Treating the Melbourne metropolitan area as a case study, a spatial analysis was conducted to clarify priority areas with a significantly high percentage and number of aging adults (65+) with disability and high barriers to accessing primary health services. Afterward, travel times from priority areas to palliative medicine and hospital services were calculated. The geographic dispersion of areas with people vulnerable to COVID-19 with poor access to palliative care and health services is clarified. Unique methods of health service delivery are required to ensure that vulnerable populations in underserviced metropolitan areas receive prompt and adequate care. The spatial methodology used can be implemented in different contexts to support evidence-based COVID-19 and pandemic palliative care service decisions.


Sex differences in cancer mechanisms.
We now know that cancer is many different diseases, with great variation even within a single histological subtype. With the current emphasis on developing personalized approaches to cancer treatment, it is astonishing that we have not yet systematically incorporated the biology of sex differences into our paradigms for laboratory and clinical cancer research. While some sex differences in cancer arise through the actions of circulating sex hormones, other sex differences are independent of estrogen, testosterone, or progesterone levels. Instead, these differences are the result of sexual differentiation, a process that involves genetic and epigenetic mechanisms, in addition to acute sex hormone actions. Sexual differentiation begins with fertilization and continues beyond menopause. It affects virtually every body system, resulting in marked sex differences in such areas as growth, lifespan, metabolism, and immunity, all of which can impact on cancer progression, treatment response, and survival. These organismal level differences have correlates at the cellular level, and thus, males and females can fundamentally differ in their protections and vulnerabilities to cancer, from cellular transformation through all stages of progression, spread, and response to treatment. Our goal in this review is to cover some of the robust sex differences that exist in core cancer pathways and to make the case for inclusion of sex as a biological variable in all laboratory and clinical cancer research. We finish with a discussion of lab- and clinic-based experimental design that should be used when testing whether sex matters and the appropriate statistical models to apply in data analysis for rigorous evaluations of potential sex effects. It is our goal to facilitate the evaluation of sex differences in cancer in order to improve outcomes for all patients.


The IDENTIFY study: the investigation and detection of urological neoplasia in patients referred with suspected urinary tract cancer - a multicentre observational study.
To evaluate the contemporary prevalence of urinary tract cancer (bladder cancer, upper tract urothelial cancer [UTUC] and renal cancer) in patients referred to secondary care with haematuria, adjusted for established patient risk markers and geographical variation. This was an international multicentre prospective observational study. We included patients aged ≥16 years, referred to secondary care with suspected urinary tract cancer. Patients with a known or previous urological malignancy were excluded. We estimated the prevalence of bladder cancer, UTUC, renal cancer and prostate cancer; stratified by age, type of haematuria, sex, and smoking. We used a multivariable mixed-effects logistic regression to adjust cancer prevalence for age, type of haematuria, sex, smoking, hospitals, and countries. Of the 11 059 patients assessed for eligibility, 10 896 were included from 110 hospitals across 26 countries. The overall adjusted cancer prevalence (n = 2257) was 28.2% (95% confidence interval [CI] 22.3-34.1), bladder cancer (n = 1951) 24.7% (95% CI 19.1-30.2), UTUC (n = 128) 1.14% (95% CI 0.77-1.52), renal cancer (n = 107) 1.05% (95% CI 0.80-1.29), and prostate cancer (n = 124) 1.75% (95% CI 1.32-2.18). The odds ratios for patient risk markers in the model for all cancers were: age 1.04 (95% CI 1.03-1.05; P < 0.001), visible haematuria 3.47 (95% CI 2.90-4.15; P < 0.001), male sex 1.30 (95% CI 1.14-1.50; P < 0.001), and smoking 2.70 (95% CI 2.30-3.18; P < 0.001). A better understanding of cancer prevalence across an international population is required to inform clinical guidelines. We are the first to report urinary tract cancer prevalence across an international population in patients referred to secondary care, adjusted for patient risk markers and geographical variation. Bladder cancer was the most prevalent disease. Visible haematuria was the strongest predictor for urinary tract cancer.


Perceived trustworthiness of faces drives trust behaviour in children.
Facial appearances can powerfully influence adults' trust behaviour, despite limited evidence that these cues constitute honest signals of trustworthiness. It is not clear, however, whether the same is also true for children. The current study investigated whether, like adults, 5-year-olds and 10-year-olds are more likely to place their trust in partners that look trustworthy than those that look untrustworthy. A second, closely related question was whether children also explicitly value the information from face cues when making trust decisions. We investigated these questions using Token Quest: an economic trust game that gave participants the opportunity to make investments with a series of partners who might (or might not) repay their trust with large returns. These interactions occurred under different conditions, including one in which participants were shown the face of each partner and another in which they could 'purchase' access to faces with a portion of their investment capital. Results indicated that, like adults, 10-year-old children selectively placed their trust in those partners they perceived as looking trustworthy and many were willing to 'pay' to purchase access to these face cues during the trust game. We observed a similar profile of trust behaviour in 5-year-olds, with no significant group difference in the impact of face cues on behaviour across the three age groups. Together, these findings indicate that the influence of face cues on trust behaviour emerges early, and highlight a capacity for sophisticated social cognition in young children.


In vivo brain delivery of v-myc overproduced human neural stem cells via the intranasal pathway: tumor characteristics in the lung of a nude mouse.
We aimed to monitor the successful brain delivery of stem cells via the intranasal route and to observe the long-term consequence of the immortalized human neural stem cells in the lungs of a nude mouse model. Stably immortalized HB1.F3 human neural stem cells with firefly luciferase gene (F3-effluc) were intranasally delivered to BALB/c nude mice. Bioluminescence images were serially acquired until 41 days in vivo and at 4 hours and 41 days ex vivo after intranasal delivery. Lungs were evaluated by histopathology. After intranasal delivery of F3-effluc cells, the intense in vivo signals were detected in the nasal area, migrated toward the brain areas at 4 hours (4 of 13, 30.8%), and gradually decreased for 2 days. The brain signals were confirmed by ex vivo imaging (2 of 4, 50%). In the mice with initial lung signals (4 of 9, 44.4%), the lung signals disappeared for 5 days but reappeared 2 weeks later. The intense lung signals were confirmed to originate from the tumors in the lungs formed by F3-effluc cells by ex vivo imaging and histopathology. We propose that intranasal delivery of immortalized stem cells should be monitored for their successful delivery to the brain and their tumorigenicity longitudinally.


[Treatment of basal cell carcinomas of the eyelid].
In the treatment of basal cell carcinomas of the eyelid, diagnostic and therapeutic problems are still existing. Primary wrong diagnosis and resulting inadequate treatment can lead to the loss of the bulbus. In the following study, 198 patients were operated on basal cell carcinoma between 1980 and 1988. In 155 patients a primary eyelid tumor was removed, and the recurrence rate was 4.52%. In 43 patients with secondary eyelid tumors (recurrent tumors), a recurrence rate of 18.6% was found. Most of the tumors were verified as solid, nodular or cystic types: 73.3%. The sclerodermiform type was found in 11.0%. The recurrence rate correlated clearly with horizontal extent and depth of the tumor, and with the appearance of the sclerodermiform type. Methods of examination, treatment and the results are presented.


Effect of acupuncture on regulating IL-17, TNF-ɑ and AQPs in Sjögren's syndrome.
The aim of the study was to observe the effect of acupuncture on regulating interleukin (IL)-17, tumor necrosis factor (TNF)-ɑ, and aquaporins (AQPs) in Sjögren's syndrome (SS) on patients and on non-obese diabetic (NOD) models. Levels of anti-AQP 1, 5, 8, and 9 antibodies, IL-17, and TNF-ɑ in the serum of SS patients were compared prior and following 20 acupuncture treatment visits during 8 weeks. While in murine model, five groups were divided to receive interventions for 4 weeks, including control, model, acupuncture, isoflurane, and hydroxychloroquine. The submaxillofacial gland index, histology, immunohistochemistry of AQP1, 5, salivary flow, together with IL-17, and TNF-ɑ expression in peripheral blood were compared among the groups. Acupuncture reduced IL-17, TNF-ɑ, and immunoglobin A levels, and numeric analog scale of dryness in 14 patients with SS (p < 0.05). The salivary flow was increased, and the water intake decreased in NOD mice receiving acupuncture treatments. IL-17 and TNF-ɑ levels in peripheral serum were down-regulated (p < 0.05) and AQP1, 5 expression in the submandibular glands up-regulated in mice. The effect on relieving xerostomia with acupuncture may be achieved by up-regulating the expression of AQP1. AQP5, down-regulating levels of IL-17 and TNF-ɑ, and a decrease in inflammation of glands.


Human IL-3 and GM-CSF act synergistically in stimulating hematopoiesis in primates.
Interleukin-3 (IL-3) is a member of a family of growth factors, each of which supports the proliferation and development of hematopoietic precursors in culture. Although the biologic effects of the different hematopoietic growth factors have been well documented in different culture systems, it has only recently become possible to study the activities of these molecules in vivo. In comparison with the later acting hematopoietic growth factors granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor, IL-3 elicited a delayed and relatively modest leukocytosis when continuously infused intravenously in primates. The IL-3 infusion, however, greatly potentiated the responsiveness of the animal to subsequent administration of a low dose of GM-CSF. These results suggest that IL-3 expands an early cell population in vivo that subsequently requires the action of a later acting factor such as GM-CSF to complete its development. Optimal stimulation of hematopoiesis may be achieved with combinations of hematopoietic growth factors.


Inhibition of the norepinephrine transporter improves behavioral flexibility in rats and monkeys.
Poor cognitive control, including reversal learning deficits, has been reported in children with attention deficit hyperactivity disorder, in stimulant-dependent humans, and in animal models of these disorders; these conditions have each been associated with abnormal catecholaminergic function within the prefrontal cortex. In the current studies, we sought to explore how elevations in extracellular catecholamine levels, produced by pharmacological inhibition of catecholamine reuptake proteins, affect behavioral flexibility in rats and monkeys. Adult male Long-Evans rats and vervet monkeys were trained, respectively, on a four-position discrimination task or a three-choice visual discrimination task. Following systemic administration of pharmacological inhibitors of the dopamine and/or norepinephrine membrane transporters, rats and monkeys were exposed to retention or reversal of acquired discriminations. In accordance with our a priori hypothesis, we found that drugs that inhibit norepinephrine transporters, such as methylphenidate, atomoxetine, and desipramine, improved reversal performance in rats and monkeys; this was mainly due to a decrease in the number of perseverative errors. Interestingly, the mixed dopamine and norepinephrine transporters inhibitor methylphenidate, if anything, impaired performance during retention in both rats and monkeys, while administration of the selective dopamine transporter inhibitor GBR-12909 increased premature responses but did not alter reversal learning performance. Our results suggest that pharmacological inhibition of the membrane norepinephrine, but not membrane dopamine, transporter is associated with enhanced behavioral flexibility. These data, combined with earlier reports, may indicate that enhanced extracellular catecholamine levels in cortical regions, secondary to norepinephrine reuptake inhibition, improves multiple aspects of inhibitory control over responding in rats and monkeys.


Uncoupling proteins-2 and 3 influence obesity and inflammation in transgenic mice.
To test the hypothesis that either uncoupling protein-2 UCP2 or UCP3 or both together influence obesity and inflammation in transgenic mice. We generated 12 lines of transgenic mice for both human UCP2 and 3 using native promoters from a human bacterial artificial chromosome (BAC) clone. The BAC expresses no genes other than UCP2 and 3. Mice used for experiments are N4 or higher of backcross to C57BL/6J (B6). Each experiment used transgenic mice and their nontransgenic littermates. Northern blots confirmed expression on human UCP2 in adipose and spleen, while human UCP3 expression was detectable in gastrocnemius muscle. Western blots demonstrated a four-fold increase of UCP2 protein in spleens of Line 32 transgenic animals. Heterozygous mice of four lines showing expression of human UCP2 in spleen were examined for obesity phenotypes. There were no significant differences between Lines 1 and 32, but female transgenics of both lines had significantly smaller femoral fat depots than the control (littermate) mice (P=0.015 and 0.005, respectively). In addition, total fat of transgenic females was significantly less in Line 1 (P=0.05) and almost significantly different in Line 32 (P=0.06). Male Line 1 mice were leaner (P=0.04) while male Line 32 mice were almost significantly leaner (P=0.06). Heterozygous mice of Lines 35 and 44 showed no significant differences from the nontransgenic littermate controls. Effects of the UCP2/UCP3 transgene on obesity in Line 32 mice were confirmed by crossing transgenic mice with the B6.Cg-Ay agouti obese mice. B6.Cg-Ay carrying the UCP2/UCP3 transgene from Line 32 were significantly leaner than nontransgenic B6.Cg-Ay mice. Line 32 UCP2/UCP3 transgenics showed increased hypothalamic Neuropeptide (NPY) levels and food intake, with reduced spontaneous physical activity. Transgenic baseline interleukin4 (IL-4) and interleukin6 (IL-6) levels were low with lower or later increases after endotoxin injection compared to wild-type littermates. Endotoxin-induced fever was also diminished in transgenic male animals. Low-density lipoprotein (LDL) cholesterol levels were significantly higher in both Line 1 and 32 transgenics (P=0.05 and 0.001, respectively) after they had been placed on a moderate fat-defined diet containing 32% of calories from fat for 5 weeks. Moderate overexpression of UCP2 and 3 reduced fat mass and increased LDL cholesterol in two independent lines of transgenic mice. Thus, the reduced fat mass cannot be due to insertional mutagenesis since virtually identical fat pad weights and masses were observed with the two independent lines. Line 32 mice also have altered inflammation and mitochondrial function. We conclude that UCP2 and/or 3 have small but significant effects on obesity in mice, and that their mechanism of action may include alterations of metabolic rate.


Effects of a 6-Week Faroese Chain Dance Programme on Postural Balance, Physical Function, and Health Profile in Elderly Subjects: A Pilot Study.
The present pilot study investigates the impact of a Faroese chain dance intervention on health profile, mobility, and postural balance in elderly subjects. Healthy elderly subjects (n=27; age 75 ± 5 yrs) were randomised into an intervention group (IG) and a control group (CG). IG performed twice-weekly sessions of Faroese chain dance over 6 weeks. Dancing sessions lasted 30 min in the initial 3 weeks and 45 min in the final 3 weeks. Health profile was determined before and after intervention by measuring blood pressure, resting heart rate, muscle mass, and body fat content. Postural balance was evaluated using the Berg Balance Scale (BBS) and Fullerton Advanced Balance Scale (FAB) tests, while mobility was assessed using the Short Physical Performance Battery (SPPB), the Timed Up & Go (TUG) test, the 6-min walk test, and the 30-s sit-to-stand test. Systolic and diastolic blood pressure were lowered (9 ± 6 and 6 ± 3 mmHg, respectively) in IG, with a tendency (P=0.07) for a greater change score than in CG. Mean arterial pressure declined (P<0.05) by 7 ± 3 mmHg in IG, which tended (P=0.09) to be greater than in CG. IG improved (P<0.05) on BBS and FAB scores by 3.6 ± 2.1% and 15.8 ± 8.3%, with the change score for FAB being greater (P<0.05) than in CG (0.3 ± 1.6). Moreover, the postintervention SPPB score was improved (P<0.05) more in IG (13.9 ± 7.4%) compared to CG, while performance in the 30-s sit-to-stand, 6-min walk, and TUG tests improved (4-15%; P<0.05) in IG only. Body fat content was reduced (P<0.05) from 36.3 ± 2.8% to 34.8 ± 2.8% in IG, with no between-group differences and no change in CG (34.1 ± 2.8% to 33.7 ± 3.1%). In conclusion, a 6-week Faroese chain dance programme lowers blood pressure and improves postural balance and physical function in elderly.


Testosterone induction of song in photosensitive and photorefractory male sparrows.
Song in male songbirds is activated by the sex steroid testosterone (T). Using male song sparrows (Melospiza melodia), we compared effects of T in the normal spring state of photosensitivity (i.e., when the pituitary-gonadal axis is sensitive to stimulation by increasing daylength) and in the late summer-early fall state of photorefractoriness (i.e., when they are insensitive to increasing daylength). Photosensitive males experienced short days for 8 weeks and then long days for another 22 weeks to induce photorefractoriness. T implants were given to the birds twice, first when on short days and photosensitive, and second when on long days and photorefractory. Song rates were compared among 5 conditions: (1) photosensitive, short days, low T titers; (2) photosensitive, short days, high T titers; (3) photosensitive, long days, high T titers; (4) photorefractory, long days, low T titers; and (5) photorefractory, long days, high T titers. Plasma levels of T were monitored throughout the experiment by radioimmunoassay. T was equally effective in inducing song in both the photosensitive and photorefractory conditions. Thus, no seasonal change was found in the sensitivity to hormone action of the neural target sites mediating this behavior in song sparrows. Photosensitive birds sang at a higher rate when on long days than when on short days, however, even though there was no concomitant increase in plasma levels of T. This finding suggests that environmental factors can alter the expression of song activated by similar levels of T.


Hormone replacement therapy reduces lipid oxidation directly at the arterial wall: A possible link to estrogens' cardioprotective effect through atherosclerosis prevention.
The first step in atherosclerosis formation is the ingurgitation of an oxidized low-density lipid (LDL) molecule by a macrophage which then turns into a foam cell within the vascular wall and initiates a cascade of inflammatory responses. Could it be that the potential cardioprotective effect observed in women receiving hormone replacement therapy (HRT) is modulated by estrogen's capacity to decrease LDL oxidation in the vascular wall and thus decrease atherosclerotic foam cells? Thirty-four adult female Wistar rats were divided into three groups. All were double oophorectomized. After recovery, Group 1 received Estradiol Valerate subcutaneous (SC) (2.5 mg/kg/week), Group 2 Estradiol Valerate SC (2.5 mg/kg/week) + Progesterone SC (10 mg/kg/48 h), and Group 3 Placebo SC. After 10 weeks, all rats were sacrificed and a vascular dissection performed. Malondialdehyde (MDA) was measured directly on the vascular extract to determine lipid oxidative levels and HRTs' effect. Renal and hepatic tissue was also studied. Total antioxidant status (TAS) was measured to determine overall oxidative behavior. Vascular MDA levels for Group 1 = 80.80 (±16.8) μmol/ml/g, Group 2 = 107.69 (±24.9) μmol/ml/g, and Group 3 = 140.96 (±32.4) μmol/ml/g. ANOVA (P < 0.05), with a post hoc Bonferroni corrective t-test, showed that both Group 1 and 2 have statistically significant lower levels of MDA than Group 3. Renal tissue showed less oxidative damage in the HRT groups, while hepatic tissue showed an inverse behavior with less lipid oxidation in the placebo group. TAS decreased with oophorectomy in all groups but decreased less in both groups that received HRT compared to placebo (P < 0.05). HRT significantly reduces lipid oxidation directly in the arterial wall.


Acoustic velocities of shear waves in La3Ga(5.5)Ta(0.5)O14 piezoelectric crystal using singular value decomposition.
La3Ga(5.5)Ta(0.5)O14 (LGT) is a new piezoelectric material with superior electromechanical properties and high Q.f product. Transverse-wave splitting behavior of LGT was observed in the acoustic velocity measurements. The velocities of the two shear waves propagating in Y-(010) direction are very close and they cannot be separated completely. This has a tremendous influence on determining the correct velocity of the shear modes. It is proposed that the singular value decomposition (SVD) technique can be used to determine the splitting time of two shear modes in Y-plane LGT crystal. Moreover, it also helps to correct the arrival time of the transverse waves. It is believed that the presented approach is efficient and accurate for determining the correct velocities of shear modes in anisotropic media.


Eosinophilia in Asthma Patients Is Protective Against Severe COVID-19 Illness.
There is a paucity of information on coronavirus disease 2019 (COVID-19) outcomes in asthmatics. To identify risk factors associated with admission and subsequent mortality among COVID-19-infected asthmatics. Adults at our institution with a positive polymerase chain reaction for COVID-19 between March 14 and April 27, 2020, were retrospectively identified. Comorbidities, laboratory results, and mortality rates during hospitalization were recorded. In total, 737 of 951 (77.5%) asthma patients with COVID-19 were seen in the emergency department (ED), and 78.8% of these ED patients (581 of 737) were admitted. Individuals with previously measured mean absolute eosinophil counts (AEC) ≥150 cells/μL were less likely to be admitted (odds ratio [OR] = 0.46, 95% confidence interval [CI]: 0.21-0.98, P = .04), whereas concomitant heart failure (CHF), chronic kidney disease (CKD), and chronic obstructive pulmonary disease (COPD) were risk factors for admission. Hospitalized patients with asthma with peak hospital-measured AEC ≥150 cells/μL (n = 104) were less likely to die compared with those whose AEC remained <150 cells/μL (n = 213) (mortality rate 9.6% vs 25.8%; OR = 0.006, 95% CI: 0.0001-0.64, P = .03). This group had also higher preadmission mean AEC (237 ± 181 vs 163 ± 147 cells/μL, P = .001, OR = 2012, 95% CI: 27.3-14,816). The mortality rate in patients with asthma alone (no associated CHF, CKD, COPD, diabetes, or hypertension) was similar to that of patients without asthma or any of these comorbidities. In asthmatics, pre-existing eosinophilia (AEC ≥150 cells/μL) was protective from COVID-19-associated admission, and development of eosinophilia (AEC ≥150 cells/μL) during hospitalization was associated with decreased mortality. Preadmission AEC influenced the AEC trend during hospitalization. Having a Th2-asthma phenotype might be an important predictor for reduced COVID-19 morbidity and mortality that should be further explored in prospective and mechanistic studies.


The Gas-Phase Photophysics of Eosin Y and its Maleimide Conjugate.
The use of the xanthene family of dyes as fluorescent probes in a wide range of applications has provided impetus for the studying of their photophysical properties. In particular, recent advances in gas-phase techniques such as FRET that utilize such chromophores have placed a greater importance on the characterization of these properties in the gas phase. Additionally, the use of synthetic linker chains to graft the chromophores in a site-specific manner to their target system is ubiquitous. There is, however, often limited information on how the addition of such a linker chain may affect the photophysical properties of the chromophores, which is of fundamental importance for interpretation of experimental data reliant on grafted chromophores. Here, we present data on the optical spectroscopy of different protonation states of Eosin Y, a fluorescein derivative. We compare the photophysics of Eosin Y to its maleimide conjugate, and to the thioether product of the reaction of this conjugate with cysteamine. Comparison of the mass spectra following laser irradiation shows that very different relaxation takes place upon addition of the maleimide moiety but that the photophysics of the bare chromophore are restored upon addition of cysteamine. This radical change in the photophysics is interpreted in terms of charge-transfer states, whose energy relative to the S1 ← S0 transition of the chromophore is dependent on the conjugation of the maleimide moiety. We also show that the shape of the absorption band is unchanged in the gas-phase as compared to the solution-phase, showing a maximum with a shoulder toward the blue, and examination of isotope distributions of the isolated ions show that this shoulder cannot be due to the presence of dimers. Consideration of the fluorescence emission spectrum allows a tentative assignment of the shoulder to be due to a vibrational progression with a high Franck-Condon factor.


A multi-institutional analysis of prehospital tourniquet use.
Recent military studies demonstrated an association between prehospital tourniquet use and increased survival. The benefits of this prehospital intervention in a civilian population remain unclear. The aims of our study were to evaluate tourniquet use in the civilian population and to compare outcomes to previously published military experience. We hypothesized that incorporation of tourniquet use in the civilian population will result in an overall improvement in mortality. This is a preliminary multi-institutional retrospective analysis of prehospital tourniquet (MIA-T) use of patients admitted to nine urban Level 1 trauma centers from January 2010 to December 2013. Patient demographics and mortality from a previous military experience by Kragh et al. (Ann Surg. 2009;249:1-7) were used for comparison. Patients younger than 18 years or with nontraumatic bleeding requiring tourniquet application were excluded. Data were analyzed using a two-tailed unpaired Student's t test with p < 0.05 as significant. A total of 197 patients were included. Tourniquets were applied effectively in 175 (88.8%) of 197 patients. The average Injury Severity Score (ISS) for MIA-T versus military was 11 ± 12.5 versus 14 ± 10.5, respectively (p = 0.02). The overall mortality and limb amputation rates for the MIA-T group were significantly lower than previously seen in the military population at 6 (3.0%) of 197 versus 22 (11.3%) of 194 (p = 0.002) and 37 (18.8%) of 197 versus 97 (41.8%) of 232 (p = 0.0001), respectively. Our study is the largest evaluation of prehospital tourniquet use in a civilian population to date. We found that tourniquets were applied safely and effectively in the civilian population. Adaptation of this prehospital intervention may convey a survival benefit in the civilian population. Epidemiologic study, level V.


Standards of practice and uniformity in references style.
Authors are usually invited to follow the instructions to authors in scientific journals, because this section is likely to contain unique requirements, such as the references style requested. Authors not only must accurately check the instructions, but are also expected to verify references against the original documents before submitting the article, consuming a considerable amount of time. This aspect may contribute to the large number of references errors in scientific manuscripts, some of which would be prevented by introducing a more homogeneous and uniformed style. A variety of international, peer-reviewed journals currently requires a rather different style, especially for the numbers of authors to be cited, the placing of the year of publication, the style of the journal title. A broad implementation of reasonable standards of practice and consensus guidelines for the format of manuscript references should be promoted for an advantageous process for both the journals and the authors.


A Monte Carlo based analytic model of the in-room neutron ambient dose equivalent for a Mevion gantry-mounted passively scattered proton system.
The goal of this study was to develop a Monte Carlo (MC)-based analytical model that can predict the in-room ambient dose equivalent from a Mevion gantry-mounted passively scattered proton system. The Mevion S250 and treatment vault were simulated using the MCNPX MC code. The results of the in-room neutron dose measurements, using an FHT 762 WENDI-II detector, were employed to benchmark the MC-derived values. After tuning the MCNPX MC code, for the same beam delivery parameters, the code was used to calculate the neutron spectra and ambient dose equivalent in the vault and at varying angles from the isocenter. Then, based on the calculations, an analytical model was reconstructed and data were fitted to derive the model parameters at 95% confidence intervals (CI). The MCNPX codes were tuned to within about 19% of the measured values for most of the measurements in the vault. For the maze, up to 0.08 mSv Gy-1 discrepancies were found between the experimental measurements and MCNPX calculated results. The analytical model showed up to 18% discrepancy for distances between 100 and 600 cm from the isocenter compared to the MC calculations. The model may underestimate the neutron ambient dose equivalent up to 21% for distances less than 100 cm from the isocenter. The proposed analytical model can be used to estimate the contribution of the secondary neutron dose from the Mevion S250 for the design of local shielding inside the proton therapy treatment vault.


Elastic, piezoelectric, and dielectric properties of 0.955Pb(Zn(1/3)Nb(2/3))O(3)-0.45PbTiO(3 ) single crystal with designed multidomains.
The elastic, piezoelectric, and dielectric properties of a 0.955Pb(Zn(1/3)Nb(2/3))O(3)-0.045PbTiO(3 ) (PZN-4.5%PT) multi-domain single crystal, poled along [001] of the original cubic direction, have been determined experimentally using combined resonance and ultrasonic methods. At room temperature, the PZN-4.5%PT single crystal has rhombohedral symmetry. After being poled along [001], four degenerate states still remain. Statistically, such a domain-engineered crystal may be treated as having an average tetragonal symmetry, and its material constants were determined based on 4 mm symmetry. It was confirmed that the electromechanical coupling coefficient k(33) for the domain-engineered samples is >90%, and the piezoelectric constant d(33) is >2000 pC/N. A soft shear mode with a velocity of 700 m/s was found in the [110] direction. From the measured experimental data, the orientational dependence of phase velocities and electromechanical coupling coefficients was calculated. The results showed that the transverse and longitudinal coupling coefficients, k(31) and k(33), reach their maximum along [110] and [001], respectively.


Weight gain after treatment of Graves' disease in children.
The frequency of and risk factors for weight gain in children treated for Graves' disease have not been described. We evaluated change in BMI-Z score and predictors of weight gain in this population. Retrospective review of data from January 2000 to July 2011. Two hundred and twenty two children and adolescents with Graves' disease (ages 2-18 years) evaluated following radioactive iodine administration (RAI); (n = 101), thyroidectomy (n = 9) and initiation of medical therapy (n = 112). Changes in body mass index Z score over 12 months (ΔBMI-Z0-12 ). All treatment groups in each gender and race increased BMI-Z (median ΔBMI-Z0-12 was positive). T3 levels following RAI (P = .04) and weight lost at the time of administration (P = .02) in the RAI group and free T4 levels in the medical therapy group (P = .03) were positively correlated with ΔBMI-Z0-12 . Race was a significant predictor only in the medical therapy group (P = .01). Age negatively correlated with ΔBMI-Z0-12 in both the RAI (P < .001) and medical therapy groups (P = .003). Gender, maximum TSH in the 12 months after RAI and initial dose of LT4 replacement did not correlate with ΔBMI-Z0-12 . The prevalence of overweight and obesity in our cohort was similar to US children. Weight gain during treatment for Graves' disease is common in children, and many children become overweight or obese during treatment. Risk factors include greater degree of hyperthyroidism at presentation and time of RAI and younger age. Weight lost upon presentation may also predict greater weight gain. Control of subsequent hypothyroidism does not appear to affect weight gain.


Acid tolerance in Listeria monocytogenes: the adaptive acid tolerance response (ATR) and growth-phase-dependent acid resistance.
Listeria monocytogenes acquired increased acid tolerance during exponential growth upon exposure to sublethal acid stress, a response designated the acid tolerance response (ATR). Maximal acid resistance was seen when the organism was exposed to pH 5.0 for 1 h prior to challenge at pH 3.0, although intermediate levels of protection were afforded by exposure to pH values ranging from 4.0 to 6.0. A 60 min adaptive period was required for the development of maximal acid tolerance; during this period the level of acid tolerance increased gradually. Full expression of the ATR required de novo protein synthesis; chloramphenicol, a protein synthesis inhibitor, prevented full induction of acid tolerance. Analysis of protein expression during the adaptive period by two-dimensional gel electrophoresis revealed a change in the expression of at least 23 proteins compared to the non-adapted culture. Eleven proteins showed induced expression while 12 were repressed, implying that the ATR is a complex response involving a modulation in the expression of a large number of genes. In addition to the exponential phase ATR, L.monocytogenes also developed increased acid resistance upon entry into the stationary phase; this response appeared to be independent of the pH-dependent ATR seen during exponential growth.


Protein adsorption and transport in cation exchangers with a rigid backbone matrix with and without polymeric surface extenders.
We compare the properties and protein adsorption characteristics of two polymeric cation exchangers: UNOsphere S, which has an open macroporous architecture, and Nuvia S, which is based on a very similar backbone matrix but contains sulfonated polymeric surface extenders. A monoclonal IgG and lysozyme were used as model adsorbates. The characteristic pore sizes, determined by inverse size exclusion chromatography, were about 140 nm for UNOsphere S, and only about 10 nm for Nuvia S, indicating that the polymeric extenders occupy a substantial portion of the base matrix pores. Greater exclusion limits were found for Nuvia S in 1 M NaCl and for a similar matrix containing uncharged surface extenders, suggesting that the polymeric extenders collapse partially at high ionic strength or when they are uncharged. Large equilibrium binding capacities were obtained for Nuvia S, approaching 320 ± 10 mg/mL of particle volume for both proteins in comparison with the UNOsphere S values of 170 ± 10 and 120 ± 10 mg/mL for lysozyme and IgG, respectively. Much higher adsorption rates were also found for Nuvia S, and the rate was nearly independent of protein concentration in solution. Confocal laser scanning microscopy showed very sharp intraparticle protein concentration profiles for UNOsphere S, consistent with a pore diffusion mechanism but diffuse concentration profiles for Nuvia S, consistent with a solid diffusion mechanism. The improved capacity and transport afforded by the polymeric extenders provide substantial potential benefits for bioprocess applications without sacrificing the desirable flow properties of the backbone matrix.


Molecular systematics of the Eastern Fence Lizard (Sceloporus undulatus): a comparison of Parsimony, Likelihood, and Bayesian approaches.
Phylogenetic analysis of large datasets using complex nucleotide substitution models under a maximum likelihood framework can be computationally infeasible, especially when attempting to infer confidence values by way of nonparametric bootstrapping. Recent developments in phylogenetics suggest the computational burden can be reduced by using Bayesian methods of phylogenetic inference. However, few empirical phylogenetic studies exist that explore the efficiency of Bayesian analysis of large datasets. To this end, we conducted an extensive phylogenetic analysis of the wide-ranging and geographically variable Eastern Fence Lizard (Sceloporus undulatus). Maximum parsimony, maximum likelihood, and Bayesian phylogenetic analyses were performed on a combined mitochondrial DNA dataset (12S and 16S rRNA, ND1 protein-coding gene, and associated tRNA; 3,688 bp total) for 56 populations of S. undulatus (78 total terminals including other S. undulatus group species and outgroups). Maximum parsimony analysis resulted in numerous equally parsimonious trees (82,646 from equally weighted parsimony and 335 from weighted parsimony). The majority rule consensus tree derived from the Bayesian analysis was topologically identical to the single best phylogeny inferred from the maximum likelihood analysis, but required approximately 80% less computational time. The mtDNA data provide strong support for the monophyly of the S. undulatus group and the paraphyly of "S. undulatus" with respect to S. belli, S. cautus, and S. woodi. Parallel evolution of ecomorphs within "S. undulatus" has masked the actual number of species within this group. This evidence, along with convincing patterns of phylogeographic differentiation suggests "S. undulatus" represents at least four lineages that should be recognized as evolutionary species.


Protection against cold in prehospital care: evaporative heat loss reduction by wet clothing removal or the addition of a vapor barrier--a thermal manikin study.
In the prehospital care of a cold and wet person, early application of adequate insulation is of utmost importance to reduce cold stress, limit body core cooling, and prevent deterioration of the patient's condition. Most prehospital guidelines on protection against cold recommend the removal of wet clothing prior to insulation, and some also recommend the use of a waterproof vapor barrier to reduce evaporative heat loss. However, there is little scientific evidence of the effectiveness of these measures. Using a thermal manikin with wet clothing, this study was conducted to determine the effect of wet clothing removal or the addition of a vapor barrier on thermal insulation and evaporative heat loss using different amounts of insulation in both warm and cold ambient conditions. A thermal manikin dressed in wet clothing was set up in accordance with the European Standard for assessing requirements of sleeping bags, modified for wet heat loss determination, and the climatic chamber was set to -15 degrees Celsius (°C) for cold conditions and +10°C for warm conditions. Three different insulation ensembles, one, two or seven woollen blankets, were chosen to provide different levels of insulation. Five different test conditions were evaluated for all three levels of insulation ensembles: (1) dry underwear; (2) dry underwear with a vapor barrier; (3) wet underwear; (4) wet underwear with a vapor barrier; and (5) no underwear. Dry and wet heat loss and thermal resistance were determined from continuous monitoring of ambient air temperature, manikin surface temperature, heat flux and evaporative mass loss rate. Independent of insulation thickness or ambient temperature, the removal of wet clothing or the addition of a vapor barrier resulted in a reduction in total heat loss of 19-42%. The absolute heat loss reduction was greater, however, and thus clinically more important in cold environments when little insulation is available. A similar reduction in total heat loss was also achieved by increasing the insulation from one to two blankets or from two to seven blankets. Wet clothing removal or the addition of a vapor barrier effectively reduced evaporative heat loss and might thus be of great importance in prehospital rescue scenarios in cold environments with limited insulation available, such as in mass-casualty situations or during protracted evacuations in harsh conditions.


Cohnella algarum sp. nov., isolated from a freshwater green alga Paulinella chromatophora.
A Gram-stain-positive, facultatively aerobic and endospore-forming bacterium, designated strain Pch-40T, was isolated from a freshwater green alga, Paulinella chromatophora. Cells were motile rods with a monotrichous polar flagellum showing catalase- and oxidase-positive reactions. Strain Pch-40T grew at 20-50 °C (optimum, 37-40 °C), at pH 5.0-11.0 (optimum, pH 7.0) and in the presence of 0-4.0 % (w/v) NaCl (optimum, 0 %). Menaquinone-7 was detected as the sole isoprenoid quinone. The genomic DNA G+C content of strain Pch-40T was 55.6 mol%. The major cellular fatty acids of strain Pch-40T were C16 : 0, iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Pch-40T clearly belonged to the genus Cohnella of the family Paenibacillaceae. Strain Pch-40T was most closely related to Cohnella rhizosphaerae CSE-5610T with a 96.1 % 16S rRNA gene sequence similarity. The phenotypic and chemotaxonomic features and the phylogenetic inference clearly suggested that strain Pch-40T represents a novel species of the genus Cohnella, for which the name Cohnellaalgarum sp. nov. is proposed. The type strain is strain Pch-40T (=KACC 19279T=JCM 32033T).


DNA recognition by the homing endonuclease PI-SceI involves a divalent metal ion cofactor-induced conformational change.
PI-SceI, a homing endonuclease of the LAGLIDADG family, consists of two domains involved in DNA cleavage and protein splicing, respectively. Both domains cooperate in binding the recognition sequence. Comparison of the structures of PI-SceI in the absence and presence of substrate reveals major conformational changes in both the protein and DNA. Notably, in the protein-splicing domain the loop comprising residues 53-70 and adopts a "closed" conformation, thus enabling it to interact with the DNA. We have studied the dynamics of DNA binding and subsequent loop movement by fluorescence techniques. Six amino acids in loop53-70 were individually replaced by cysteine and modified by fluorescein. The interaction of the modified PI-SceI variants with the substrate, unlabeled or labeled with tetramethylrhodamine, was analyzed in equilibrium and stopped-flow experiments. A kinetic scheme was established describing the interaction between PI-SceI and DNA. It is noteworthy that the apparent hinge-flap motion of loop53-70 is only observed in the presence of a divalent metal ion cofactor. Substitution of the major Mg2+-binding ligands in PI-SceI, Asp-218 and Asp-326, by Asn or "nicking" PI-SceI with trypsin at Arg-277, which interferes with formation of an active enzyme.substrate complex, both prevent the conformational change of loop53-70. Deletion of the loop inactivates the enzyme. We conclude that loop53-70 is an important structural element that couples DNA recognition by the splicing domain with DNA cleavage by the catalytic domain and as such "communicates" with the Mg2+ binding sites at the catalytic centers.


[Special anesthesiological aspects in patients with Niikawa-Kuroki ("Kabuki Make-Up") syndrome].
The Niikawa-Kuroki ("Kabuki Make-up") syndrome (NKS) is a rare disease with a characteristic array of multiple congenital anomalies. The syndrome is characterised by distinct craniofacial dysmorphias, moderate mental retardation, skeletal abnormalities, cardiovascular and urogenital disorders, growth retardation and skeletal muscle abnormalities. The inheritance of NKS is not completely clear. However, the available data are compatible with an autosomal dominant mutation with variable expressivity. There is no specific treatment for NKS. The patients require intensive support for the development of their mental and physical capabilities. Furthermore, therapy includes operative correction of the multiple anomalies. The risk of anaesthesia is increased in these patients because of a possible difficult airway, cardiovascular and urogenital diseases, and abnormalities of the skeletal muscles. Special problems associated with the anaesthetic management in patients with NKS are discussed in this case report.


Muscle repositioning: combining subjective and objective feedbacks in the teaching and practice of a reflex-based myofascial release technique.
Muscle Repositioning (MR) is a new style of myofascial release that elicits involuntary motor reactions detectable by electromyography. This article describes the principal theoretical and practical concepts of MR, and summarizes a workshop presented October 31, 2009, after the Second International Fascia Research Congress, held at Vrije Universitiet, Amsterdam.The manual mechanical input of MR integrates the client's body segments into a block, which is evident as a result of the diagnostic manual oscillations the practitioner imparts to the client's body. Segmental integration is achieved when the client's body responds as a unit to the oscillatory assessment. It appears that manually sustaining the condition of segmental integration evokes involuntary muscle reactions, which reactions might correspond to mechanisms that maintain homeostasis, such as pandiculation. It might be that these reactions are part of the MR mechanism of action and underlie its clinically observed efficacy in the treatment of musculoskeletal disorders.For the practitioner and the client alike, segmental integration provides unique sensations. In teaching MR, these paired sensations can be used as kinesthetic feedback resources, because quality of touch can be guided by the client's reported sensations, which should match the practitioner's sensations. Another form of feedback with respect to quality of touch is the visually discernable degree of segmental integration. Finally, because the involuntary motor activity elicited by the MR touch can be objectively monitored through electromyography and possibly other instrumented measurements, the MR approach might yield objectivity, precision, and reproducibility-features seldom found in manual therapies.


Clinical significance of glomerular C3 deposition in primary membranous nephropathy.
We aimed to investigate the effects of glomerular C3 deposition on clinical, histopathological features, and outcomes of patients with primary membranous nephropathy (MN). A total of 261 patients with biopsy-proven primary MN, who were on follow up for at least 6 months, were included in the study. The patients were grouped according to their C3 immunostaining in kidney biopsy samples at the time of diagnosis: Low intensity [LI; (C3 1 +)] and high intensity [HI; (C3 2 + or C3 3 +)]. The primary outcome was the development of kidney failure. Complete (CR) or partial remission (PR) was defined as secondary outcome. Sixteen patients reached the primary outcome after a median follow-up of 33.8 months. Patients in the high intensity group (119 cases) had lower eGFR and higher proteinuria at admission and last follow-up compared to patients in the low intensity group (142 cases). Also, more patients in the high intensity group reached the primary outcome compared to patients in the low intensity group: twelve patients (10.1%) in the high intensity group and four patients (2.8%) in the low intensity group reached the primary outcome (p = 0.015). Kaplan-Meier analysis demonstrated that patients in the high intensity group had a higher risk for kidney failure (p = 0.02). In multivariate logistic regression analysis, high intensity C3 deposition and initial estimated glomerular filtration rate (eGFR) indepenently predicted primary outcome. Extensive glomerular C3 deposition is a predictor of kidney failure in patients with MN.


Time-trends in rates of hospital admission of adolescents for violent, self-inflicted or drug/alcohol-related injury in England and Scotland, 2005-11: population-based analysis.
Incidence of emergency admissions for violent injury in 10- to 18-year olds decreased in England and Scotland between 2005 and 2011, but more steeply in Scotland. To generate hypotheses about causes of these differences, we determined whether trends were consistent across admissions for three common types of adversity-related injury (violent, self-inflicted and drug/alcohol-related). Emergency admissions to NHS hospitals were captured using Hospital Episode Statistics and Scottish Morbidity Records. Adversity-related injury was defined using ICD-10 codes. Analyses were stratified by sex/age groups (10-12, 13-15 and 16-18 years) and adjusted for background trends in admissions for injury. During 2005-11, rates declined in all sex/age groups in Scotland (reductions adjusted for background trends ranged from -22.0 to -103.7/100 000) and in girls and boys aged <16 years in England (adjusted reductions -12.0 to -49.9/100 000). However, these rates increased in England for both sexes aged 16-18 years (adjusted increases, girls 71.8/100 000; boys 28.0/100 000). However, throughout 2005-11 overall rates remained relatively similar in England and Scotland for both sexes aged <16 years, and remained higher in Scotland for both sexes aged 16-18 years. A greater decline in the rates of emergency admissions for adversity-related injury for adolescents in Scotland compared with England could signal more effective policies in Scotland for reducing violence, self-harm, or drug/alcohol misuse, particularly for 16 to 18-year olds.


The hormonal response to intravenous 5-hydroxytryptophan in bulimia nervosa.
In recent years evidence has accumulated to implicate a disturbance in serotonin function in the eating disorder bulimia nervosa. This study employs a neuroendocrine technique to assess the effect of intravenous 5-hydroxytryptophan, the immediate precursor to serotonin, on peripheral hormones in bulimia nervosa subjects and controls. Blunted prolactin and growth hormone responses were observed among bulimia nervosa subjects; the possible pathophysiology and implications of the findings are discussed.


Peripartum cardiomyopathy: the need for a national database.
Peripartum cardiomyopathy (PPCM) is a rare and life-threatening disease that affects young women in the last month of pregnancy or within 5 months of delivery. It is a form of dilated cardiomyopathy with left-sided systolic dysfunction. The incidence rate in the Western world is estimated to be 1:3000. Symptoms of PPCM vary greatly and may be obscured by common physiological aspects of pregnancy. Therefore, the incidence rate might be higher. Echocardiography or MRI can confirm or rule out PPCM. Unfortunately, there is no specific risk factor profile available. The clinical course varies from complete recovery to deterioration of cardiac function. Patients with PPCM, especially those whose ventricular function has not returned to normal, are advised against further pregnancy. Recently, more disease-specific therapeutic strategies have been developed with promising results for prolactin blockade by bromocriptine. Increasing awareness for PPCM among general practitioners, gynaecologists and cardiologists may help to diagnose patients efficiently in order to start adequate treatment. A national registry is warranted to identify risk factor profiles and to optimise treatment strategies.


The alveolin IMC1h is required for normal ookinete and sporozoite motility behaviour and host colonisation in Plasmodium berghei.
Alveolins, or inner membrane complex (IMC) proteins, are components of the subpellicular network that forms a structural part of the pellicle of malaria parasites. In Plasmodium berghei, deletions of three alveolins, IMC1a, b, and h, each resulted in reduced mechanical strength and gliding velocity of ookinetes or sporozoites. Using time lapse imaging, we show here that deletion of IMC1h (PBANKA_143660) also has an impact on the directionality and motility behaviour of both ookinetes and sporozoites. Despite their marked motility defects, sporozoites lacking IMC1h were able to invade mosquito salivary glands, allowing us to investigate the role of IMC1h in colonisation of the mammalian host. We show that IMC1h is essential for sporozoites to progress through the dermis in vivo but does not play a significant role in hepatoma cell transmigration and invasion in vitro. Colocalisation of IMC1h with the residual IMC in liver stages was detected up to 30 hours after infection and parasites lacking IMC1h showed developmental defects in vitro and a delayed onset of blood stage infection in vivo. Together, these results suggest that IMC1h is involved in maintaining the cellular architecture which supports normal motility behaviour, access of the sporozoites to the blood stream, and further colonisation of the mammalian host.


[Postneonatal infant mortality and quality of medical care: a case-control study]
OBJECTIVE: To identify the risk factors for postneonatal infant mortality caused by diarrhea and pneumonia in relation to the quality of medical assistance. METHODS: Population-based case-control study of 277 postneonatal infant deaths caused by diarrhea and pneumonia occurring in the metropolitan area of Belo Horizonte, Brazil, between May/1991 and April/1992. The cases were compared with hospitalized controls, and matched by pathology, age and hospital. Information on cases and controls were collected from medical records and through home interviews. Some variables related to the quality of medical care were analyzed. McNemar test and conditional logistic regression were used to define the risk factors for postneonatal deaths. RESULTS: Multiple logistic regression analysis showed the following factors independently associated with increased risk of postneonatal death induced by diarrhea and pneumonia: delayed immunization (OR = 2.48; 95%CI=1.17-5.23), general status (serious) on hospital admission (OR=10.94; 95%CI=4.91-24.34), unaccomplished hospital procedures (OR=10.08; 95%CI = 3.55-20.59) and malnutrition on hospital admission (OR=3.58; 95%CI=1.42-9.07). CONCLUSIONS: The results indicate the low quality of medical assistance as an important risk factor for avoidable causes of postneonatal deaths. The authors highlight the lack of integration between the outpatient clinic and hospital activities as an important determinant of low quality. It is necessary that the performance of health services and their effect on avoidable infant mortality be widely discussed, also taking into consideration the preponderant role of socioeconomic variables


Use of image analysis for sludge characterisation: studying the relation between floc shape and sludge settleability.
This paper starts by presenting a fully automatic image analysis procedure for characterisation of flocs and filaments in activated sludge images. Thereafter the attention is directed towards the results of four lab-scale experiments, in which image information is related to sludge settleability in terms of sludge volume index. This relation is statistically confirmed by applying a principal component analysis to the data. In addition, the redundancy in the data sets is studied with regard to floc shape descriptors and the monitoring potential of image analysis is demonstrated by means of a multiple linear regression exercise.


Bioactivation of the nasal toxicant 2,6-dichlorobenzonitrile: an assessment of metabolic activity in human nasal mucosa and identification of indicators of exposure and potential toxicity.
The herbicide 2,6-dichlorobenzonitrile (DCBN) is a potent nasal toxicant in rodents; however, it is not known whether DCBN causes similar nasal toxicity in humans. The tissue-selective toxicity of DCBN in mouse nasal mucosa is largely dependent on target tissue bioactivation by CYP2A5. The human orthologues of CYP2A5, CYP2A6 and CYP2A13, are both expressed in nasal mucosa and are capable of activating DCBN. In this study, we directly determined the ability of human nasal mucosa to bioactivate DCBN. We also tested the suitability of a glutathione conjugate of DCBN (GS-DCBN) or its derivatives as biomarkers of DCBN exposure and nasal toxicity in mouse models. We found that human fetal nasal mucosa microsomes catalyze the formation of GS-DCBN, with a Km value comparable to that of adult mouse nasal mucosa microsomes. The activity of the human nasal mucosa microsomes was inhibited by 8-methoxypsoralen, a known CYP2A inhibitor. GS-DCBN and its metabolites were detected in the nasal mucosa and nasal-wash fluid obtained from DCBN-treated mice, in amounts that increased with escalations in DCBN dose, and they were all still detectable at 24 h after a DCBN treatment (at 10 mg/kg). Further studies in Cyp2a5-null mice indicated that GS-DCBN and its metabolites in nasal-wash fluid were generated in the nasal mucosa, rather than in other organs. Thus, our data indicate for the first time that the human nasal mucosa is capable of bioactivating DCBN and that GS-DCBN and its metabolites in nasal-wash fluid may collectively serve as indicators of DCBN exposure and potential nasal toxicity in humans.


Ensemble Inversion for Brain Tumor Growth Models With Mass Effect.
We propose a method for extracting physics-based biomarkers from a single multiparametric Magnetic Resonance Imaging (mpMRI) scan bearing a glioma tumor. We account for mass effect, the deformation of brain parenchyma due to the growing tumor, which on its own is an important radiographic feature but its automatic quantification remains an open problem. In particular, we calibrate a partial differential equation (PDE) tumor growth model that captures mass effect, parameterized by a single scalar parameter, tumor proliferation, migration, while localizing the tumor initiation site. The single-scan calibration problem is severely ill-posed because the precancerous, healthy, brain anatomy is unknown. To address the ill-posedness, we introduce an ensemble inversion scheme that uses a number of normal subject brain templates as proxies for the healthy precancer subject anatomy. We verify our solver on a synthetic dataset and perform a retrospective analysis on a clinical dataset of 216 glioblastoma (GBM) patients. We analyze the reconstructions using our calibrated biophysical model and demonstrate that our solver provides both global and local quantitative measures of tumor biophysics and mass effect. We further highlight the improved performance in model calibration through the inclusion of mass effect in tumor growth models-including mass effect in the model leads to 10% increase in average dice coefficients for patients with significant mass effect. We further evaluate our model by introducing novel biophysics-based features and using them for survival analysis. Our preliminary analysis suggests that including such features can improve patient stratification and survival prediction.


DUSP1 mRNA modulation during porcine circovirus type 2 and porcine reproductive and respiratory syndrome virus co-infection regulates viruses replication.
The effects of porcine circovirus type 2b (PCV2b) and porcine reproductive and respiratory syndrome virus (PRRSV) co-infection in epithelial cells of the swine respiratory tract is unknown. In the present study, the newborn pig trachea cell line NPTr-CD163, which is permissive to both viruses, was persistently infected with PCV2b and then with PRRSV. Viral replication, cell viability, cytokines' mRNA expression, and modulation of cellular genes expression were evaluated in infected cells. In NPTr-CD163 co-infection model, PCV2b replication was enhanced while PRRSV replication was suppressed. Cell viability was significantly decreased during PCV2b single infection and co-infection compared to mock-infected and PRRSV single infected cells. However, no difference was observed in cell viability between PCV2b and PCV2b/PRRSV infected cells. The IL6, IL8 and IL10 mRNA expression was significantly higher in co-infected cells compared to PCV2b and PRRSV single infected cells. Moreover, the IFN-α/β expression was significantly reduced in co-infected cells compared to PCV2b infected cells whereas it remained higher compared to PRRSV infected cells. The differential gene expression analysis revealed that the mRNA expression level of the cellular gene DUSP1 was significantly higher in all PRRSV infection models compared to PCV2b single infected cells. Knockdown of DUSP1 expression in co-infected cells significantly reduced PCV2b replication, suggesting a role for DUSP1 in PCV2b/PRRSV pathogenesis.


Prevalence and Associations, by Age Group, of IPV Among AGYW in Rural South Africa.
The prevalence of intimate partner violence (IPV) is alarmingly high among South African adolescent girls and young women (AGYW). Limited data exist exploring how IPV prevalence and its risk factors differ by age. Study data were from the baseline visit of HPTN 068, a randomized controlled trial (RCT) conducted from 2011 to 2015 in Mpumalanga, South Africa. A cohort of 2,533 AGYW, aged 13 years to 20 years, answered survey questions on demographics and behaviors, including their experiences of physical and sexual violence ever and in the past 12 months. We calculated the prevalence of IPV and related risk factors, as well as prevalence ratios with 95% confidence intervals, stratified by age. Nearly one quarter (19.5%, 95% CI = [18.0, 21.2]) of AGYW experienced any IPV ever (physical or sexual) by a partner. The prevalence of any IPV ever among AGYW aged 13 years to 14 years, 15 years to 16 years, and 17 years to 20 years was 10.8%, 17.7%, and 32.1%, respectively. Key variables significantly associated with any IPV ever across all age groups included borrowing money from someone outside the home in the past 12 months, ever having had vaginal sex, ever having had anal sex, and consuming any alcohol. Few statistically significant associations were unique to specific age groups. The history of IPV among the youngest AGYW is a critical finding and should be a focus of prevention efforts.


Magnetic resonance imaging and magnetic resonance spectroscopy of bone tumors and bone marrow disease.
The authors have made use of an integrated magnetic resonance imaging/spectroscopy (MRI/MRS) examination to study seven patients with a variety of bone tumors. The spatial localization method used in the 31P portion of the examination was surface coil localization and a one-dimensional chemical shift imaging method (3 cases). The authors found that the precision of spatial localization was critical in many of these cases, since most of these bone tumors were surrounded by muscle tissue that contained high concentrations of phosphocreatine (PCr). For this reason, they suggest that the metabolite ratios should be referenced to the adenosine triphosphate (beta-NTP) resonance rather than PCr. The phosphate monoester (PME) to beta-NTP ratio was elevated as compared with normal muscle in all of the bone tumors studied. The authors found that all of these tumors exhibited pHs between 7.0 and 7.2, which are similar to the values found for normal muscle. They also show the feasibility of using a line-selective proton chemical shift imaging sequence with high spatial resolution for investigating changes in the fatty composition of bone marrow. This method is illustrated in an example of a patient with advanced avascular necrosis in the femoral heads.


Optimal Regression Method for Near-Infrared Spectroscopic Evaluation of Articular Cartilage.
Near-infrared (NIR) spectroscopy has been successful in nondestructive assessment of biological tissue properties, such as stiffness of articular cartilage, and is proposed to be used in clinical arthroscopies. Near-infrared spectroscopic data include absorbance values from a broad wavelength region resulting in a large number of contributing factors. This broad spectrum includes information from potentially noisy variables, which may contribute to errors during regression analysis. We hypothesized that partial least squares regression (PLSR) is an optimal multivariate regression technique and requires application of variable selection methods to further improve the performance of NIR spectroscopy-based prediction of cartilage tissue properties, including instantaneous, equilibrium, and dynamic moduli and cartilage thickness. To test this hypothesis, we conducted for the first time a comparative analysis of multivariate regression techniques, which included principal component regression (PCR), PLSR, ridge regression, least absolute shrinkage and selection operator (Lasso), and least squares version of support vector machines (LS-SVM) on NIR spectral data of equine articular cartilage. Additionally, we evaluated the effect of variable selection methods, including Monte Carlo uninformative variable elimination (MC-UVE), competitive adaptive reweighted sampling (CARS), variable combination population analysis (VCPA), backward interval PLS (BiPLS), genetic algorithm (GA), and jackknife, on the performance of the optimal regression technique. The PLSR technique was found as an optimal regression tool (R2Tissue thickness = 75.6%, R2Dynamic modulus = 64.9%) for cartilage NIR data; variable selection methods simplified the prediction models enabling the use of lesser number of regression components. However, the improvements in model performance with variable selection methods were found to be statistically insignificant. Thus, the PLSR technique is recommended as the regression tool for multivariate analysis for prediction of articular cartilage properties from its NIR spectra.


[Ophthalmophysiological features of applying soft contact lenses to correct myopia in pilots].
The development of effective methods for preventing and correcting myopia in pilots is one of the most important problems of aeronautic ophthalmology. The majority of Russian and foreign aeronautic ophthalmologists agree that contact correction is the principal method for repair of occupational vision of myopic pilots. Experiments with soft contact lenses with 38 and 70% humor content were carried out. The methodology was based on comprehensive examinations of the vision status and status of the organ of vision under common conditions on the Earth and during simulation of unfavorable flight factors. The results indicate that contact lenses are a safe means for correcting myopia in pilots. This is confirmed by the absence of unfavorable changes in the main visual functions and in the status of the anterior segment of the eye, by a constant level of visual working capacity, and by the absence of negative subjective sensations.


Analgesic intolerance with or without bronchial asthma: is there a marker?
The prevalence of analgesic intolerance (AI) is less than 1% in the general population and about 10% in adult asthmatics, in whom the disease tends to be more severe. A possible clinical-laboratory marker was sought that would differentiate patients who have AI with/without asthma, AI with asthma, and AI without asthma from the healthy subjects. In the survey, 66 analgesic-intolerant patients (36 having asthma) were compared with 50 healthy subjects using a nickel patch-test, 65 patients (39 having asthma) with 55 healthy subjects for the presence of a genetic marker (A38G and A444C SNPs in CC16 and LTC4S genes), and 32 patients (14 having asthma) with 118 healthy subjects for presence and frequency of human leukocyte antigens (HLA). The mean age of the patients with AI with/without asthma and the healthy subjects for the nickel patch-test group, genetic marker group, and the HLA group was 39.8 +/- 10.5 and 33.3 +/- 11.1, 41.5 +/- 11.6 and 38.1 +/- 13.4, and 39.4 +/- 12.5 and 41 +/- 2.6, respectively. The frequency of the females in the same groups, in the same order, was 72.7% and 54%, 81.5% and 62%, and 71.9% and 59.3%, respectively. The frequency of positive nickel patch-test results and the A38G and A444C frequency in CC16 and LTC4S genes were not significantly different (p > 0.05). The frequency of HLA antigens HLA-A3, -B52, -DR16, -DQ5, -DQ8 and -DQ9 were significantly higher; and -A24, -B35, -B44, -DQ6 and -DQ7 were significantly lower in the AI group with/without asthma compared to the control group (p < 0.05). As a result, nickel patch-test positivity and the genes which we have studied do not seem to be markers for AI with/without asthma. However, there might be a relation between AI with/without asthma and the types of the HLA system. Further surveys are needed with other genes and possible markers.


Real-time visualization of the dynamic evolution of CS2 4d and 6s Rydberg wave packet components.
The dynamic evolution of CS2 4d and 6s Rydberg wave packet components has been experimentally visualized via femtosecond time-resolved photoelectron imaging coupled with time-resolved mass spectroscopy. The temporal evolution of the four components of the prepared Rydberg wave packet is directly observed as time-dependent changes of the intensities of different parts in the main photoelectron peak. Furthermore, time-resolved photoelectron angular distributions (PADs) clearly reflect the different component characters of 4d and 6s molecular orbitals. The lifetime of Rydberg wave packets is determined to be about 830fs and their decay is attributed to predissociation. Our results suggest the possibility of directly visualizing and determining the amplitudes and relative phases of different electronic and vibrational wave packet components in polyatomic molecules.


Novel strategies for eutherian x marsupial somatic cell hybrids: mapping the genome of Monodelphis domestica.
Two hundred thirty-seven independent somatic cell hybrids have been obtained between opossum (Monodelphis domestica) splenocytes, bone marrow cells, or primary fibroblasts, and HPRT-deficient or TK-deficient Chinese hamster, mouse, American mink, or common vole fibroblast lines. Because extreme segregation and fragmentation of marsupial chromosomes commonly occurs in eutherian x marsupial somatic cells hybrids, we developed a rapid primary screening method that enables the identification of primary clones containing a large amount of opossum DNA 20-25 d after fusion. This method, which depends on in situ hybridization of biotin-labeled total opossum DNA on interphase nuclei of hybrid cells fixed on the bottom of microwell plates, was used to screen the 237 hybrid clones; 52 of them had a substantial amount of opossum DNA. G-banding and in situ hybridization of biotin-labeled total opossum DNA on metaphase spreads of the clones enabled identification of 17 hybrid clones containing from two to seven intact chromosomes of M. domestica on the background of Chinese hamster or vole chromosomes. The hybrid clones with intact opossum chromosomes are used in a panel constructed for mapping the opossum genome. Initial mapping results from these clones have led to the tentative assignment of GPI and GOT1 to chromosome 1; 6PGD to chromosome 4; LDHA to chromosome 5; LDHB to chromosome 8; and PGK and G6PD to the X chromosome. On the basis of indirect evidence we also tentatively assigned HPRT to the X chromosome and TK to chromosome 5 of M. domestica. These are the first tentative chromosomal assignments by any technique for this species.


Regulation of the Raf-MEK-ERK pathway by protein phosphatase 5.
The Raf-MEK-ERK pathway couples growth factor, mitogenic and extracellular matrix signals to cell fate decisions such as growth, proliferation, migration, differentiation and survival. Raf-1 is a direct effector of the Ras GTPase and is the initiating kinase in this signalling cascade. Although Raf-1 activation is well studied, little is known about how Raf-1 is inactivated. Here, we used a proteomic approach to identify molecules that may inactivate Raf-1 signalling. Protein phosphatase 5 (PP5) was identified as an inactivator that associates with Raf-1 on growth factor stimulation and selectively dephosphorylates an essential activating site, Ser 338. The PP5-mediated dephosphorylation of Ser 338 inhibited Raf-1 activity and downstream signalling to MEK, an effect that was prevented by phosphomimetic substitution of Ser 338, or by ablation of PP5 catalytic function. Furthermore, depletion of endogenous PP5 increased cellular phospho-Ser 338 levels. Our results suggest that PP5 is a physiological regulator of Raf-1 signalling pathways.


CFTR anion channel modulates expression of human transmembrane mucin MUC3 through the PDZ protein GOPC.
The transmembrane mucins in the enterocyte are type 1 transmembrane proteins with long and rigid mucin domains, rich in proline, threonine and serine residues that carry numerous O-glycans. Three of these mucins, MUC3, MUC12 and MUC17 are unique in harboring C-terminal class I PDZ motifs, making them suitable ligands for PDZ proteins. A screening of 123 different human PDZ domains for binding to MUC3 identified a strong interaction with the PDZ protein GOPC (Golgi-associated PDZ and coiled-coil motif-containing protein). This interaction was mediated by the C-terminal PDZ motif of MUC3, binding to the single GOPC PDZ domain. GOPC is also a binding partner for cystic fibrosis transmembrane conductance regulator (CFTR) that directs CFTR for degradation. Overexpression of GOPC downregulated the total levels of MUC3, an effect that was reversed by introducing CFTR. The results suggest that CFTR and MUC3 compete for binding to GOPC, which in turn can regulate levels of these two proteins. For the first time a direct coupling between mucins and the CFTR channel is demonstrated, a finding that will shed further light on the still poorly understood relationship between cystic fibrosis and the mucus phenotype of this disease.


Differential antagonist activity of alpha-helical corticotropin-releasing factor9-41 in three bioassay systems.
Studies were performed in conscious unrestrained rats to compare the ability of the CRF receptor antagonist, alpha-helical CRF9-41, to inhibit the actions of CRF in three in vivo bioassay systems. When both peptides were administered intracerebroventricularly, an antagonist:agonist ratio between 6:1-12:1 was required to abolish CRF-induced elevations of plasma catecholamine levels. When both peptides were administered iv, CRF-induced hypotension and tachycardia were completely prevented by an antagonist:agonist ratio of 6:1, whereas total blockade of CRF-induced elevations of plasma ACTH and beta-endorphin levels required an antagonist:agonist ratio of 3000:1. These results demonstrate marked differences in the ability of alpha-helical CRF9-41 to antagonize various biological actions of CRF and support the existence of multiple CRF receptor subtypes.


Oesophageal atresia: The growth gap.
Poor growth is an under-recognised yet significant long-term sequelae of oesophageal atresia (OA) repair. Few studies have specifically explored the reasons for growth impairment in this complex cohort. The association between poor growth with younger age and fundoplication appears to have the strongest supportive evidence, highlighting the need for early involvement of a dietitian and speech pathologist, and consideration of optimal medical reflux management prior to referring for anti-reflux surgery. However, it remains difficult to reach conclusions regarding other factors which may negatively influence growth, due to conflicting findings, inconsistent definitions and lack of validated tool utilisation. While swallowing and feeding difficulties are particularly frequent in younger children, their relationship with growth remains unclear. It is possible that these morbidities impact on the diet of children with OA, but detailed analysis of dietary composition and quality, and its relationship with these complications and growth, has not yet been conducted. Another potential area of research in OA is the role of the microbiota in growth and nutrition. While the microbiota has been linked to growth impairment in other paediatric conditions, it is yet to be investigated in OA. Further research is needed to identify the most important contributory factors to poor growth, the role of the intestinal microbiota, and effective interventions to maximise growth and nutritional outcomes in this cohort.


Putamen: activity of single units during slow and rapid arm movements.
The activity of putamen neurons was studied in a monkey during the performance of both slow and rapid arm movements. More than half of all movement-related units discharged preferentially in relation to slow movements and less than 10 percent in relation to rapid movements. These findings indicate that at least a portion of the basal ganglia (the putamen) is primarily involved in the control of slow movements and are consistent with the hypothesis of Kornhuber that the primary motor function of the basal ganglia is to generate slow ("ramp") rather than rapid ("ballistic") movements.


Successful xenotransplantation of microencapsulated newborn pig parathyroid cells in the treatment of hypoparathyroidism in rats.
To study the effect of xenotransplantation with pig parathyroid cells, which was prepared using cell microencapsulation technique, on the treatment of hypoparathyroidism in rats without immunosuppressor. Parathyroid cells were isolated from 10 healthy newborn pigs and encapsulated in alginate-polylysine-alginate (APA) membranes. Thirty-two aparathyroid Wistar rats were randomly allocated to microcapsule, non-microcapsule, empty microcapsule, and control groups. Each rat was injected intraperitoneally with encapsulated porcine parathyroid cells, free porcine parathyroid cells, empty capsules or 0.9% NaCl, respectively. Total serum calcium and parathyroid hormone levels were monitored continuously for 40 weeks. And then, the transplant beds were retrieved and subjected to morphologic and electron microscopic examination. In those animals xenotransplanted with microencapsulated porcine parathyroid cells, the calcium and PTH levels were consistently within the normal range during the 40 weeks. In contrast, no therapeutic effects were observed in rats in the non-microcapsule group. Furthermore, neither empty capsules nor 0.9% NaCl were shown to have any effect on the recipient's serum calcium or PTH levels. After 40 weeks, electron microscopic examination demonstrated that the parathyroid cells within the microcapsules had survived well in vivo. Xenotransplantation of microencapsulated newborn pig parathyroid cells can successfully treat hypoparathyroidism in rats without using immunosuppressive drugs. The results of this study show the possible clinical use of microencapsulated porcine parathyroid cells.


The clinical impact of (18)F-FDG PET in patients with suspected or confirmed recurrence of colorectal cancer: a prospective study.
This prospective study aimed to confirm, in a clinical setting, the benefits suggested by earlier retrospective studies of (18)F-FDG PET scanning for the evaluation of patients with suspected recurrence of colorectal cancer. The referring oncologist was asked to prospectively assign a treatment plan for 102 consecutive patients being evaluated by (18)F-FDG PET for suspected or confirmed recurrence of colorectal cancer and without evidence of unresectable disease on conventional staging investigations, including CT. This treatment plan was then compared with that based on incremental information supplied by PET. Management changes were validated by follow-up. For 6 patients, the oncologist would not commit to a management plan without access to PET information, and for all these patients, PET correctly guided management. Of the remaining 96 patients, the management plan for 54 (56%) was altered as a direct result of unexpected PET findings. Thus, PET directly influenced management in 60 (59%) of 102 patients. The discrepant PET results could be validated in 57 patients and were correct for both the presence and the extent of malignant disease in 52 (91%) of these patients but were false-positive in 1 patient because of a pelvic abscess and underestimated the extent of metastatic disease in 4 (7%). Relapse was confirmed in 49 (98%) of 50 evaluable patients with positive PET findings. Significantly, planned surgery was abandoned in 26 (60%) of 43 patients because of incremental PET findings. Of the 42 patients for whom management was not changed by PET findings, false-negative PET findings were documented for 5 (4 with metastases < 1 cm), and the PET findings for 1 were presumed to be false-positive because of sarcoidosis. This prospective study confirms the high impact, suggested by previous retrospective analyses, of (18)F-FDG PET on management of patients with suspected recurrent colorectal cancer. The major benefit of PET is avoidance of inappropriate local therapies by documentation of widespread disease.


14-3-3ε Is required for germ cell migration in Drosophila.
Although 14-3-3 proteins participate in multiple biological processes, isoform-specific specialized functions, as well as functional redundancy are emerging with tissue and developmental stage-specificity. Accordingly, the two 14-3-3ε proteins in Drosophila exhibit functional specificity and redundancy. Homozygotes for loss of function alleles of D14-3-3ε contain significantly fewer germ line cells (pole cells) in their gonads, a phenotype not shared by mutants in the other 14-3-3 gene leo. We show that although D14-3-3ε is enriched within pole cells it is required in mesodermal somatic gonad precursor cells which guide pole cells in their migration through the mesoderm and coalesce with them to form the embryonic gonad. Loss of D14-3-3ε results in defective pole cell migration, reduced pole cell number. We present evidence that D14-3-3ε loss results in reduction or loss of the transcription factor Zfh-1, one of the main regulatory molecules of the pole cell migration, from the somatic gonad precursor cells.