Boundary-spanning work demands and their consequences for guilt and psychological distress.
Using data from a national survey of working Americans (Work, Stress, and Health Survey; N = 1,042), the authors examine the associations between boundary-spanning work demands and self-reported feelings of guilt and distress. The authors document gender differences in the emotional and mental health consequences of boundary-spanning work demands, as indexed by the frequency of receiving work-related contact outside of normal work hours. Specifically, the authors observe that frequent work contact is associated with more feelings of guilt and distress among women only. Analyses also demonstrate that guilt accounts for the positive association between the frequency of work contact and distress among women. Statistical adjustments for levels of guilt reduce the positive association between frequent work contact and distress among women to nonsignificance. The findings underscore the importance of focusing on gender and emotions in work-family interface processes, as well as their implications for psychological health.


Workshop on imaging science development for cancer prevention and preemption.
The concept of intraepithelial neoplasm (IEN) as a near-obligate precursor of cancers has generated opportunities to examine drug or device intervention strategies that may reverse or retard the sometimes lengthy process of carcinogenesis. Chemopreventive agents with high therapeutic indices, well-monitored for efficacy and safety, are greatly needed, as is development of less invasive or minimally disruptive visualization and assessment methods to safely screen nominally healthy but at-risk patients, often for extended periods of time and at repeated intervals. Imaging devices, alone or in combination with anticancer drugs, may also provide novel interventions to treat or prevent precancer.


[Interhemispheric EEG coherence in men with abnormal sexual behavior].
Interhemispheric interactions were examined in individuals with abnormal sexual behaviour by means of method of evaluation of coherent EEG properties in symmetric points of two hemispheres in different levels of awakening and in elaboration of cognitive tests related with preferential activation of one of two hemispheres. It was determined that EEG of the individuals who had committed sexual deliquency and had the signs of paraphilia was characterized by diffuse elevation of the degree of interhemispheric coherence in alpha-activity in the state of both passive and active awakening and by its local elevation in theta-activity. The results obtained might testify to the disorders either in cortical-subcortical interactions and in the cortical processes and permitted to suppose the decrease of degree of functional hemispheric lateralization in persons with paraphilias.


Pathogenicity of the pinewood nematode, Bursaphelenchus xylophilus, to Japanese larch, Larix kaempferi, seedlings.
Pathogenicity of the pine wood nematode, Bursaphelenchus xylophilus, to Japanese larch, Larix kaempferi, seedlings was tested with inoculation experiments under nursery conditions. Water suspensions of nematodes (mixed stages cultured on Botrytis cinerea or dispersal fourth-stage juveniles (DJ4) extracted from the adult Japanese pine sawyer, Monochamus alternatus) were injected into the stems of 2- and 3-year-old Japanese larch and Japanese black pine, Pinus thunbergii, seedlings growing in a nursery. In another treatment, Japanese pine sawyer adults holding DJ4 were released under a net that covered the upper half of the seedlings. Regardless of nematode inoculation method, Japanese larch seedlings were as susceptible as Japanese black pine seedlings to B. xylophilus under nursery conditions. The rate of disease development was similar on larch and pine seedlings. Nematode population densities were lower in the stems of dead larch seedlings than in the stems of dead pine seedlings. Histopathological observations revealed that the distribution of nematodes in the stems of dead larch seedlings was mostly limited to the cortex, phloem and cambial zone. Traumatic resin canal formation was one of the most characteristic symptoms in larch seedlings which was dissimilar to that in pine seedlings.


Drug dependence in cancer is exploitable by optimally constructed treatment holidays.
Cancers with acquired resistance to targeted therapy can become simultaneously dependent on the presence of the targeted therapy drug for survival, suggesting that intermittent therapy may slow resistance. However, relatively little is known about which tumours are likely to become dependent and how to schedule intermittent therapy optimally. Here we characterized drug dependence across a panel of over 75 MAPK-inhibitor-resistant BRAFV600E mutant melanoma models at the population and single-clone levels. Melanocytic differentiated models exhibited a much greater tendency to give rise to drug-dependent progeny than their dedifferentiated counterparts. Mechanistically, acquired loss of microphthalmia-associated transcription factor in differentiated melanoma models drives ERK-JunB-p21 signalling to enforce drug dependence. We identified the optimal scheduling of 'drug holidays' using simple mathematical models that we validated across short and long timescales. Without detailed knowledge of tumour characteristics, we found that a simple adaptive therapy protocol can produce near-optimal outcomes using only measurements of total population size. Finally, a spatial agent-based model showed that optimal schedules derived from exponentially growing cells in culture remain nearly optimal in the context of tumour cell turnover and limited environmental carrying capacity. These findings may guide the implementation of improved evolution-inspired treatment strategies for drug-dependent cancers.


Ocular quinine toxicity treated with hyperbaric oxygen.
Ocular quinine toxicity typically involves a partial or total and often permanent loss of vision. Apart from gastric lavage and oral administration of activated charcoal, current treatment modalities are of doubtful efficacy. Two patients with quinine amaurosis were treated with hyperbaric oxygen (HBO2) in an effort to increase oxygen delivery to the retina. Visual outcomes in these patients were evaluated. Two patients had bilateral no light perception vision and dilated, nonreactive pupils within hours of ingesting 13-15 g of quinine in addition to other drugs. Following initial oral charcoal administration, HBO2 therapy was used. Within 17 h after quinine ingestion, both patients underwent HBO2 therapy at 2.4 atm abs with 100% O2 for 90 min. Both patients had return of visual acuity to 20/20 in both eyes less than 24 h after treatment. Follow-up visual fields revealed constriction and paracentral scotomas bilaterally. We conclude that HBO2 may represent an additional or alternative, and perhaps safer, method of treatment for ocular quinine toxicity.


Promising effects of eribulin for cystic lung metastases of epithelioid sarcoma: a case report.
Epithelioid sarcoma (ES) is a rare and aggressive type of soft tissue sarcoma with resistance to systemic chemotherapy. Therefore, new treatment options are required for patients with advanced ES. Eribulin is a novel potential treatment option for patients with inoperative sarcoma. We herein report a case of a 27-year-old Japanese man with cystic lung metastases from an ES in the left forearm, resulting in long-term stable disease. A solid, metastatic sarcomatous nodule appeared in the right lung, as shown by computed tomography, within the first 2 months after surgery. After the lung metastasis was detected, he underwent a total of 37 cycles of chemotherapy in six regimens. However, multiple cystic lung metastases had progressed in segments S6 and S9/10; hence, eribulin treatment was initiated. After two courses of eribulin, the excess fluid density material in the cystic metastases was completely absorbed, and an additional four courses of treatment resulted in shrinkage of the cystic metastases. These effects lasted for 13 months without severe adverse effects. Cystic lung metastases are an extremely rare consequence of soft tissue sarcoma, and eribulin is one of the most promising options for the treatment of advanced ES.


Gender differences in suicide methods.
Gender differences in suicide completion rates have been attributed to the differences in lethality of suicide methods chosen by men and women, but few empirical studies have investigated factors other than demographic characteristics that might explain this differential. Data from the 621 suicides in Summit County, Ohio during 1997-2006 were disaggregated by gender to compare known correlates of suicide risk on three methods of suicide-firearm, hanging and drug poisoning. Compared to women, men who completed suicide with firearms were more likely to be married and committed the act at home. Unmarried men were likelier to hang themselves than married men, but unmarried women were less likely to hang themselves than married women. Men with a history of depression were more likely to suicide by hanging, but women with depression were half as likely to hang themselves compared to the women without a history of depression. Men with a history of substance abuse were more likely to suicide by poisoning than men without such history, but substance abuse history had no influence on women's use of poisoning to suicide. For both sexes, the odds of suicide by poisoning were significantly higher for those on psychiatric medications.


Changes in soil hydraulic conductivity after prescribed fires in Mediterranean pine forests.
Prescribed fire removes or reduces the plant material that is prone to forest fires by creating fuel discontinuity and minimising fire intensity. This forest management tool potentially impacts Mediterranean ecosystems hydrological response by influencing water infiltration into soil. As direct measurements (e.g. by infiltrometers) of unsaturated infiltration in soil subjected to prescribed fires are scarce, this study has evaluated changes in soil hydraulic conductivity (SHC) using Minidisk infiltrometer after prescribed fires in representative plots of forests in the Iberian Peninsula under Mediterranean semi-arid conditions: (i) pure forest of Black pine Arnold ssp salzmannii; (ii) mixed forest of Maritime and Black pine; (iii) mixed forest of Aleppo and Maritime pine. The results have shown that fire reduced the organic layer thickness and its organic matter content. Consequently, after the prescribed fire the water content of burned plots was always lower than in untreated soils; conversely, the reverse soil behaviour was noticed before applying fire. Compared to the untreated soils, and with very few exceptions, prescribed fire did not cause significant changes in SHC. No general patterns in the comparisons between treatments (burned/unburned soils), in time evolution after fires and in the interactions between these effects were detected. This means that the SHC of burned soils followed the temporal variations of untreated soils. The lack of significance of these differences between treatments could be due to the low-fire severity and the limited effect of temperature in the mineral layer on soil hydraulic properties. This effect was expected and agrees with other studies. Overall prescribed fires did not alter SHC in Mediterranean forest ecosystems under unsaturated conditions since fire was of low-severity.


Genome-wide study of an elite rice pedigree reveals a complex history of genetic architecture for breeding improvement.
Improving breeding has been widely utilized in crop breeding and contributed to yield and quality improvement, yet few researches have been done to analyze genetic architecture underlying breeding improvement comprehensively. Here, we collected genotype and phenotype data of 99 cultivars from the complete pedigree including Huanghuazhan, an elite, high-quality, conventional indica rice that has been grown over 4.5 million hectares in southern China and from which more than 20 excellent cultivars have been derived. We identified 1,313 selective sweeps (SSWs) revealing four stage-specific selection patterns corresponding to improvement preference during 65 years, and 1113 conserved Huanghuazhan traceable blocks (cHTBs) introduced from different donors and conserved in >3 breeding generations were the core genomic regions for superior performance of Huanghuazhan. Based on 151 quantitative trait loci (QTLs) identified for 13 improved traits in the pedigree, we reproduced their improvement process in silico, highlighting improving breeding works well for traits controlled by major/major + minor effect QTLs, but was inefficient for traits controlled by QTLs with complex interactions or explaining low levels of phenotypic variation. These results indicate long-term breeding improvement is efficient to construct superior genetic architecture for elite performance, yet molecular breeding with designed genotype of QTLs can facilitate complex traits improvement.


Neonatal population pharmacokinetics of phenobarbital derived from routine clinical data.
Routine clinical pharmacokinetic (PK) data collected from 59 preterm infants who received phenobarbital were analyzed to estimate population PK parameters. Birth weights ranged from 600 to 3,620 g (mean 1,520 g), and gestational ages ranged from 24 to 42 weeks (mean 31 weeks). Data were analyzed using NONMEM, a computer program designed for population PK analysis. The mean (+/- CV) clearance of phenobarbital was 0.0047 liter/h/kg (+/- 19%) and the mean (+/- CV) volume of distribution (Vd) was 0.96 liter/kg (+/- 16%). The calculated mean serum half-life was 141 h. Clearance was not affected by gestational age, sex, asphyxia, or duration of therapy. Vd was not affected by gestational age or sex but was increased by 13% in the presence of asphyxia (5-min Apgar score less than 5, p less than 0.05).


Bilateral vessel-outlining carotid artery calcifications in panoramic radiographs: an independent risk marker for vascular events.
In odontology, panoramic radiographs (PRs) are regularly performed. PRs depict the teeth and jaws as well as carotid artery calcifications (CACs). Patients with CACs on PRs have an increased risk of vascular events compared to healthy controls without CACs, but this association is often caused by more vascular events and risk factors at baseline. However, the risk of vascular events has only been analyzed based on the presence of CACs, and not their shape. Thus, this study determined if the shape of CACs in PRs affects the risk of future vascular events. The study cohort included 117 consecutive patients with CACs in PRs and 121 age-matched controls without CACs. CAC shape in PRs was dichotomized into bilateral vessel-outlining CACs and other CAC shapes. Participants were followed prospectively for an endpoint of vascular events including myocardial infarction, stroke, and vascular death. Patients with bilateral vessel-outlining CACs had more previous vascular events than those with other CAC shapes and the healthy controls (p < 0.001, χ2). The mean follow-up duration was 9.5 years. The endpoint was reached in 83 people. Patients with bilateral vessel-outlining CACs had a higher annual risk of vascular events (7.0%) than those with other CAC shapes (4.4%) and the controls (2.6%) (p < 0.001). In multivariate analysis, bilateral vessel-outlining CACs (hazard ratio: 2.2, 95% confidence interval: 1.1-4.5) were independent risk markers for the endpoint. Findings of bilateral vessel-outlining CACs in PRs are independent risk markers for future vascular events.


'PET' vs. 'push-pull' induced ICT: a remarkable coumarinyl-appended pyrimidine based naked eye colorimetric and fluorimetric sensor for the detection of Hg2+ ions in aqueous media with test trips.
A novel colorimetric and fluorescent chemosensor based on 7-(diethylamino)-3-(pyrimidin-4-yl)-2H-chromen-2-one (PYC) has been designed and synthesized for the detection of Hg(2+) in the presence of other competing metals in mixed aqueous media. The PYC exhibits naked eye color change from green to red, and the fluorescence color changes from yellowish green to light orange with Hg(2+). It also shows a red shift in wavelength of about 80 nm in absorption spectra. Test strips based on PYC were fabricated, which could act as convenient and efficient Hg(2+) test kits.


False allegations of satanic abuse: case studies from the witch panic in Rättvik 1670-71.
The creation of false memories, psychiatric symptoms and false allegations of satanic child abuse during an outbreak of witch hysteria in Sweden in the seventeenth century are described and related to contemporary issues in child testimonies. Case studies of 28 children and 14 adults are presented. The mechanisms underlying the spread of these allegations, as well as the reactions and influence of the adult world on the children's testimonies, are discussed.


A novel group 6 LEA protein from diapause embryos of Artemia franciscana is cytoplasmically localized.
The expression of late embryogenesis abundant (LEA) proteins is one mechanism by which anhydrobiotic organisms survive periods of severe water loss. Artemia franciscana is an animal extremophile that uniquely expresses LEA proteins concurrently from Groups 1, 3, and 6. In this study we examine the subcellular localization of AfrLEA6, a Group 6 LEA protein from embryos of A. franciscana. Immunohistochemistry reveals that AfrLEA6 is located in the cytoplasm of diapause embryos and does not co-localize with nuclei or mitochondria. Due to a trace contaminant arising from chitin-based affinity chromatography during AfrLEA6 purification, the primary antiserum displayed affinities for both AfrLEA6 as well as Artemia chitin-binding proteins. This contaminant (fusion protein of intein plus chitin binding domain) co-migrates with AfrLEA6 during SDS-PAGE. Pre-adsorption of the antiserum with dechorionated embryos was required to remove the non-specific fluorescence in the embryonic cuticular membrane. Results of this study are consistent with the apparent importance of distributing multiple types of LEA proteins across many subcellular locations in anhydrobiotic organisms.


Androgens induce prostate cancer cell proliferation through mammalian target of rapamycin activation and post-transcriptional increases in cyclin D proteins.
Androgen receptor (AR) plays a central role in prostate cancer, with most tumors responding to androgen deprivation therapies, but the molecular basis for this androgen dependence has not been determined. Androgen [5alpha-dihydrotestosterone (DHT)] stimulation of LNCaP prostate cancer cells, which have constitutive phosphatidylinositol 3-kinase (PI3K)/Akt pathway activation due to PTEN loss, caused increased expression of cyclin D1, D2, and D3 proteins, retinoblastoma protein hyperphosphorylation, and cell cycle progression. However, cyclin D1 and D2 message levels were unchanged, indicating that the increases in cyclin D proteins were mediated by a post-transcriptional mechanism. This mechanism was identified as mammalian target of rapamycin (mTOR) activation. DHT treatment increased mTOR activity as assessed by phosphorylation of the downstream targets p70 S6 kinase and 4E-BP1, and mTOR inhibition with rapamycin blocked the DHT-stimulated increase in cyclin D proteins. Significantly, DHT stimulation of mTOR was not mediated through activation of the PI3K/Akt or mitogen-activated protein kinase/p90 ribosomal S6 kinase pathways and subsequent tuberous sclerosis complex 2/tuberin inactivation or by suppression of AMP-activated protein kinase. In contrast, mTOR activation by DHT was dependent on AR-stimulated mRNA synthesis. Oligonucleotide microarrays showed that DHT-stimulated rapid increases in multiple genes that regulate nutrient availability, including transporters for amino acids and other organic ions. These results indicate that a critical function of AR in PTEN-deficient prostate cancer cells is to support the pathologic activation of mTOR, possibly by increasing the expression of proteins that enhance nutrient availability and thereby prevent feedback inhibition of mTOR.


Deficient production of cyclic AMP: pharmacologic evidence of an important cause of contractile dysfunction in patients with end-stage heart failure.
We studied the effects of different classes of inotropic drugs on human working myocardium in vitro that was isolated from the hearts of patients with end-stage heart failure, and compared the responses to these drugs with those noted in muscles from nonfailing control hearts. Although peak isometric force generated in response to increased extracellular calcium reached control levels in the muscles from patients with heart failure, the time course of contraction and rate of relaxation were greatly prolonged. The inotropic effectiveness of the beta-adrenergic agonist isoproterenol and the phosphodiesterase inhibitors milrinone, caffeine, and isobutylmethylxanthine was markedly reduced in muscles from the patients with heart failure. In contrast, the effectiveness of inotropic stimulation with acetylstrophanthidin and the adenylate cyclase activator forskolin was preserved. After a minimally effective dose of forskolin was given to elevate intracellular cyclic AMP levels, the inotropic responses of muscles from the failing hearts to phosphodiesterase inhibitors were markedly potentiated. These data indicate that an abnormality in cyclic AMP production may be a fundamental defect present in patients with end-stage heart failure that can markedly diminish the effectiveness of agents that depend on generation of this nucleotide for production of a positive inotropic effect.


Renal effects of angiotensin-converting enzyme inhibition in cardiac failure.
Compensatory mechanisms such as systemic vasoconstriction and sodium retention are activated to various degrees in patients with congestive heart failure (CHF). The renin-angiotensin system (RAS) mediates many of these compensatory responses. Increased angiotensin can result in net vasoconstriction and increase afterload. In addition, it contributes to the avid sodium retention that occurs in patients with CHF. The RAS promotes sodium retention by enhancing proximal tubular reabsorption of sodium and water through changes in intrarenal hemodynamics. In addition, the direct renal tubular influence of angiotensin II and aldosterone contribute to sodium retention. In experimental heart failure in the rat, angiotensin activation leads to reduction in effective plasma flow whereas the glomerular filtration rate was unchanged. In clinical CHF, activation of the RAS may also contribute to azotemia in patients with CHF. Significant correlations between serum creatinine levels and plasma renin activity have been observed. Furthermore, renal plasma flow and the glomerular filtration rate frequently improve after therapy with converting-enzyme inhibitors (CEIs). Occasionally, however, CEIs may produce an excessive fall in systemic BP and, hence, renal perfusion pressure, which may overcome autoregulatory mechanisms and contribute to a deterioration in renal function. CEIs enhance the effectiveness of furosemide in promoting natriuresis and correction of hyponatremia. Diuretic-induced hypokalemia can also be corrected by angiotensin-converting enzyme (ACE) inhibitors due to blockade of aldosterone production. On the basis of these results, we recommend titration of the ACE inhibitors' dose to minimize hypotension and the concurrent use of furosemide in patients with CHF receiving ACE inhibitors, especially for those in whom azotemia, hyponatremia, and hypokalemia complicate the course.


Association between a variation in the phosphodiesterase 4D gene and bone mineral density.
Fragility fractures caused by osteoporosis are a major cause of morbidity and mortality in aging populations. Bone mineral density (BMD) is a useful surrogate marker for risk of fracture and is a highly heritable trait. The genetic variants underlying this genetic contribution are largely unknown. We performed a large-scale association study investigating more than 25,000 single nucleotide polymorphisms (SNPs) located within 16,000 genes. Allele frequencies were estimated in contrasting DNA pools from white females selected for low (<0.87 g/cm2, n = 319) and high (> 1.11 g/cm2, n = 321) BMD at the lumbar spine. Significant findings were verified in two additional sample collections. Based on allele frequency differences between DNA pools and subsequent individual genotyping, one of the candidate loci indicated was the phosphodiesterase 4D (PDE4D) gene region on chromosome 5q12. We subsequently tested the marker SNP, rs1498608, in a second sample of 138 white females with low (<0.91 g/cm2) and 138 females with high (>1.04 g/cm2) lumbar spine BMD. Odds ratios were 1.5 (P = 0.035) in the original sample and 2.1 (P = 0.018) in the replication sample. Association fine mapping with 80 SNPs located within 50 kilobases of the marker SNP identified a 20 kilobase region of association containing exon 6 of PDE4D. In a second, family-based replication sample with a preponderance of females with low BMD, rs1498608 showed an opposite relationship with BMD at different sites (p = 0.00044-0.09). We also replicated the previously reported association of the Ser37Ala polymorphism in BMP2, known to interact biologically with PDE4D, with BMD. This study indicates that variants in the gene encoding PDE4D account for some of the genetic contribution to bone mineral density variation in humans. The contrasting results from different samples indicate that the effect may be context-dependent. PDE4 inhibitors have been shown to increase bone mass in normal and osteopenic mice, but up until now there have been no reports implicating any member of the PDE4 gene family in human osteoporosis.


Cell property determination from the acoustic microscope generated voltage versus frequency curves.
Among the methods for the determination of mechanical properties of living cells acoustic microscopy provides some extraordinary advantages. It is relatively fast, of excellent spatial resolution and of minimal invasiveness. Sound velocity is a measure of the stiffness or Young's modulus of the cell. Attenuation of cytoplasm is a measure of supramolecular interactions. These parameters are of crucial interest for studies of cell motility, volume regulations and to establish the functional role of the various elements of the cytoskeleton. Using a phase and amplitude sensitive modulation of a scanning acoustic microscope (Hillman et al., 1994, J. Alloys Compounds. 211/212:625-627) longitudinal wave speed, attenuation and thickness profile of a biological cell are obtained from the voltage versus frequency or V(f) curves. A series of pictures, for instance in the frequency range 980-1100 MHz with an increment of 20 MHz, allows the experimental generation of V(f) curves for each pixel while keeping the lens-specimen distance unchanged. Both amplitude and phase values of the V(f) curves are used for obtaining the cell properties and the cell thickness profile. The theoretical analysis shows that the thin liquid layer, between the cell and the substrate, has a strong influence on the reflection coefficient and should not be ignored during the analysis. Cell properties, cell profile and the thickness of the thin liquid layer are obtained from the V(f) curves by the simplex inversion algorithm. The main advantages of this new method are that imaging can be done near the focal plane, therefore an optimal signal to noise ratio is achieved, no interference with Rayleigh waves occurs, and the method requires only an approximate estimate of the material properties of the solid substratum where the cells are growing on.


Identification of types of wound bed tissue as a percentage and total wound area by planimetry in neuropathic and venous ulcers.
Neuropathic and venous leg ulcers are chronic wounds associated with devitalized tissue and recurrent infection. Management should be guided by accurate tissue assessment, including the use of planimetry, which provides tissue types as a percentage of the total wound bed surface area. This innovative study aimed to assess and identify the wound bed tissues, as a percentage, of neuropathic and venous ulcers using digital planimetry, providing support to nurses optimize the management of necrotic tissues and, consequently, to avoid wound infection. This cross-sectional study enrolled 24 patients with chronic wounds who were assessed from January to March 2021 at the Wound Outpatients Clinic. The wound photographs were analyzed using Image J 1.53e and a smartphone with WoundDoc Plus® 2.8.2 via digital planimetry. Statistical analyses were performed using the binomial test, t-test, and Mann-Whitney. Median wound areas (p=0.3263) did not differ between the group with 2 or 3 risk factors for delayed healing (Md: 31.7) and the group with up to 1 risk factor (Md: 5.3). A low exudate level was associated with the up-to-1-risk-factor-for-delayed-healing group (p=0.0405), while a medium level was associated with the two-or-three-risk-factor group (p=0.0247). A heat map displayed the tissue percentages in the wound bed. In the group with 2 or 3 risk factors for delayed healing, 91.7% (11/12) had less than 70% granulation tissue, which was the primary factor for this group (p<0.0001). Additionally, 66.7% (8/12) of patients with 2 or 3 risk factors for delayed healing exhibited discolored and/or dark red granulation tissue as the primary factor (p=0.0130). This novel identification of wound area and tissue types as a percentage, using digital planimetry, can play a crucial role in assisting nurses in decision-making related to the appropriate management of devitalized tissues. Furthermore, this measurements may facilitate the conducting of virtual wound consultations and offer valuable support in the development of protocols aimed at preventing infection and biofilm formation in the wound bed.


Healthcare professionals' experiences during the initial stage of the COVID-19 pandemic in the intensive care unit: A qualitative study.
The COVID-19 pandemic called for rapidly considerable changes in the healthcare system. Healthcare professionals from different departments within the hospital settings were enrolled in the emergency preparedness. This study, therefore, aimed to explore the healthcare professionals' experiences attending the ICU-preparedness and caring for patients with COVID-19 during the initial stage of the pandemic. A descriptive explorative qualitative study was conducted by interviewing healthcare professionals during spring 2020, exploring their experiences as part of the ICU-preparedness team and caring for patients with COVID-19 in the ICU. Healthcare professionals from different departments were recruited by purposive sampling. The interviews were transcribed verbatim and analysed using content analysis. Sixteen nurses and four physicians from a university hospital in Denmark participated. The analysis revealed three main themes and eight sub-themes. The main themes were (1) Professionalism in work-life (adaption, the patient's welfare, insecurity, and security), (2) Community Spirit (responsibility and contribution), and (3) Institutional organisation (the role of management, loss of freedom, and information). Despite work specialities and professions, the participants reported a uniformity of similar experiences of uncertainties, but also a sense of community arose during the first phase of COVID-19. To ensure resilience and mental health, and well-being for the healthcare professionals, comprehensive support should be provided. Guidelines for interventions and training are necessary to promote preparedness and reduce psychological stress.


[The use of antihomotoxic therapy in dentistry].
Efficacy of traumel C is assessed in patients with various maxillofacial inflammations. Sixty-nine patients with diseases of the buccal mucosa (aphthous stomatitis, herpes labialis, and perlèches), periodontal tissues, and painful syndrome after dental canal filling were treated. Applications of the drug were administered to patients with diseases of the buccal mucosa and slight or medium-severe periodontitis. Patients with grave periodontitis and pain after dental canal filling were injected with traumel solution. Positive shifts were attained in all cases. The duration of treatment depended on the process severity. The results recommend traumel C as an ointment for local applications and an injection solution for combined therapy of diseases of the buccal mucosa and periodontitis. The treatment schemes are convenient for outpatient treatment; there are no contraindications precluding the drug use. No side effects were observed.


Influence of Cognitive Impairment on the Recovery of Subjects with Subacute Stroke Undergoing Upper Limb Robotic Rehabilitation.
Cognitive decline is often present in stroke survivors, with a significant impact on motor recovery. However, how specific cognitive domains could impact motor recovery after robotic rehabilitation in patients with stroke is still not well understood. In this study, we analyzed the relationship between cognitive impairment and the outcome of a robot-mediated upper limb rehabilitation intervention in a sample of 51 subacute stroke patients. Participants were enrolled and treated with a set of robotic and sensor-based devices. Before the intervention, patients underwent a cognitive assessment by means of the Oxford Cognitive Screen. To assess the effect of the 30-session rehabilitation intervention, patients were assessed twice with the following outcome measures: the Fugl-Meyer Assessment for Upper Extremity (FMA-UE), to evaluate motor function; the Upper limb Motricity Index (MI), to evaluate upper limb muscle strength; the Modified Barthel Index (mBI), to evaluate activities of daily living and mobility. We found that deficits in spatial attention and executive functions impacted the mBI improvement, while language, number processing, and spatial attention deficits reduced the gains in the FMA-UE. These results suggest the importance to evaluate the cognitive functions using an adequate tool in patients with stroke undergoing a robotic rehabilitation intervention.


Characterization of an oligopeptide chemoattractant receptor on human blood monocytes using a new radioligand.
The study of chemoattractant receptors on human monocytes had been limited by the lack of a radioligand suitable for use with the small numbers of cells routinely available from human donors. A new synthetic oligopeptide radioligand f[35S]met-leu-phe, with a higher specific radioactivity than was available with the tritiated compound, was used to characterize a chemoattractant receptor on freshly isolated human blood monocytes. These cells bind f[35S]met-leu-phe with a dissociation constant (KD) of 30.2 +/- 5.6 nM and contain 84,000 +/- 11,300 receptors per cell. f[35S]met-leu-phe does not bind specifically to blood lymphocytes. The specificity of the oligopeptide receptor on monocytes is indistinguishable from the oligopeptide chemoattractant receptor on human polymorphonuclear leukocytes. Using f[35S]met-leu-phe, it will now be feasible to study the chemotactic peptide receptor on small numbers of partially purified peripheral blood monocytes from patients with defects of immune function.


Geographic information systems: their use in environmental epidemiologic research.
Advances in geographic information system (GIS) technology, developed by geographers, provide new opportunities for environmental epidemiologists to study associations between environmental exposures and the spatial distribution of disease. A GIS is a powerful computer mapping and analysis technology capable of integrating large quantities of geographic (spatial) data as well as linking geographic with nongeographic data (e.g., demographic information, environmental exposure levels). In this paper we provide an overview of some of the capabilities and limitations of GIS technology; we illustrate, through practical examples, the use of several functions of a GIS including automated address matching, distance functions, buffer analysis, spatial query, and polygon overlay; we discuss methods and limitations of address geocoding, often central to the use of a GIS in environmental epidemiologic research; and we suggest ways to facilitate its use in future studies. Collaborative efforts between epidemiologists, biostatisticians, environmental scientists, GIS specialists, and medical geographers are needed to realize the full potential of GIS technology in environmental health research and may lead to innovative solutions to complex questions.


[Phytotherapy in combined treatment of chronic infection of the upper urinary tract in patients exposed to extracorporeal shock-wave lithotripsy].
Multicomponent biologically active additive Uroprofit made by Russian company Ecomir is used for phytotherapy of urolithiasis and urogenital infectious and inflammatory processes. Of 60 patients with urolithiasis and chronic pyelonephritis exposed to extracorporeal shock-wave lithotripsy (ECSWL), 30 patients received additional anti-inflammatory therapy with Uroprofit. As shown by reduction of leukocyturia and crystalluria, increased 24-h diuresis in patients with urolithiasis after exposure to ECSWL, a positive effect was achieved on urethral inflammation. Thus, Uroprofit phytotherapy has antiinflammatory, diuretic, lithokinetic effects which makes it appropriate for treatment and prophylaxis of infectious-inflammatory process in the urethra.


[Treatment of psychologically traumatised patients in Norway].
The Norwegian Directorate of Health and Social Affairs is carrying out a project on various aspects of traumatic stress. The present survey was carried out in 2001 in order to gather more knowledge about these patients within the mental health care system. 325 hospitals, outpatient clinics and district psychiatric centres were asked by questionnaire to comment on the types of traumatised patients they treated, their own perceived professional skills, the diagnoses they used and the therapy they offered, and their contact with national and regional specialist units. Refugees and victims of sexual abuse were the groups treated by most units; only a few treated veterans of UN/NATO peacekeeping forces. Half of the units reported lack of professional skills in treating refugees. Only a few units reported changes in diagnosis and treatment as a result of advice from the specialist units. The high frequency of units reporting a need for better skills in treating refugees is a challenge to the mental health care system and the specialist units. The results suggest a need for discussion of the role of the national and regional specialist units as well as a need for closer cooperation between psychiatric and primary health care.


IL-4 enhances keratinocyte expression of CXCR3 agonistic chemokines.
IFN-induced protein of 10 kDa (IP-10), monokine induced by IFN-gamma (Mig), and IFN-inducible T-cell alpha-chemoattractant (I-TAC) belong to the non-glutamate-leucine-arginine motif CXC chemokine family and act solely through the CXCR3 receptor for potent attraction of T lymphocytes. In this study, we evaluated the capacity of the T cell-derived cytokines IL-4, IL-10, and IL-17 to modulate IP-10, Mig, and I-TAC in cultured human keratinocytes and CXCR3 expression in T cells from allergic contact dermatitis (ACD). IL-4, but not IL-10 or IL-17, significantly up-regulated IFN-gamma- or TNF-alpha-induced IP-10, Mig, and I-TAC mRNA accumulation in keratinocytes and increased the levels of IP-10 and Mig in keratinocyte supernatants. Immunohistochemistry of skin affected by ACD revealed that >70% of infiltrating cells were reactive for CXCR3 and that CXCR3 staining colocalized in CD4+ and CD8+ T cells. Nickel-specific CD4+ and CD8+ T cell lines established from ACD skin produced IFN-gamma and IL-4 and expressed moderate to high levels of CXCR3. Finally, CXCR3 agonistic chemokines released by stimulated keratinocytes triggered calcium mobilization in skin-derived nickel-specific CD4+ T cells and promoted their migration, with supernatant from keratinocyte cultures stimulated with IFN-gamma and IL-4 attracting more efficaciously than supernatant from keratinocytes activated with IFN-gamma alone. In conclusion, IL-4 exerts a proinflammatory function on keratinocytes by potentiating IFN-gamma and TNF-alpha induction of IP-10, Mig, and I-TAC, which in turn may determine a prominent recruitment of CXCR3+ T lymphocytes at inflammatory reaction sites.


The impact of stretching on sports injury risk: a systematic review of the literature.
We conducted a systematic review to assess the evidence for the effectiveness of stretching as a tool to prevent injuries in sports and to make recommendations for research and prevention. Without language limitations, we searched electronic data bases, including MEDLINE (1966-2002), Current Contents (1997-2002), Biomedical Collection (1993-1999), the Cochrane Library, and SPORTDiscus, and then identified citations from papers retrieved and contacted experts in the field. Meta-analysis was limited to randomized trials or cohort studies for interventions that included stretching. Studies were excluded that lacked controls, in which stretching could not be assessed independently, or where studies did not include subjects in sporting or fitness activities. All articles were screened initially by one author. Six of 361 identified articles compared stretching with other methods to prevent injury. Data were abstracted by one author and then reviewed independently by three others. Data quality was assessed independently by three authors using a previously standardized instrument, and reviewers met to reconcile substantive differences in interpretation. We calculated weighted pooled odds ratios based on an intention-to-treat analysis as well as subgroup analyses by quality score and study design. Stretching was not significantly associated with a reduction in total injuries (OR = 0.93, CI 0.78-1.11) and similar findings were seen in the subgroup analyses. There is not sufficient evidence to endorse or discontinue routine stretching before or after exercise to prevent injury among competitive or recreational athletes. Further research, especially well-conducted randomized controlled trials, is urgently needed to determine the proper role of stretching in sports.


Guiding tail-anchored membrane proteins to the endoplasmic reticulum in a chaperone cascade.
Newly synthesized integral membrane proteins must traverse the aqueous cytosolic environment before arrival at their membrane destination and are prone to aggregation, misfolding, and mislocalization during this process. The biogenesis of integral membrane proteins therefore poses acute challenges to protein homeostasis within a cell and requires the action of effective molecular chaperones. Chaperones that mediate membrane protein targeting not only need to protect the nascent transmembrane domains from improper exposure in the cytosol, but also need to accurately select client proteins and actively guide their clients to the appropriate target membrane. The mechanisms by which cellular chaperones work together to coordinate this complex process are only beginning to be delineated. Here, we summarize recent advances in studies of the tail-anchored membrane protein targeting pathway, which revealed a network of chaperones, cochaperones, and targeting factors that together drive and regulate this essential process. This pathway is emerging as an excellent model system to decipher the mechanism by which molecular chaperones overcome the multiple challenges during post-translational membrane protein biogenesis and to gain insights into the functional organization of multicomponent chaperone networks.


A meta-analysis of fluctuating asymmetry in relation to heterozygosity.
Fluctuating asymmetry, the random departure from perfect bilateral symmetry, is a common measure of developmental instability that has been hypothesized to be inversely correlated with heterozygosity. Although this claim has been widely repeated, several studies have reported no such association. Therefore, we test the generality of this association, using meta-analysis, by converting test statistics for the relationship between heterozygosity (H) and fluctuating asymmetry (FA) into a common effect size, the Pearson's product-moment correlation coefficient. We have analysed a database containing 41 studies with a total of 118 individual samples. Overall we found an unweighted mean negative effect size; r=-0.09 (i. e. a negative correlation between H and FA). Significant heterogeneity in effect size was mainly caused by a difference between ectothermic and endothermic animals, and to a lesser extent by the use of different study designs (i.e. within-population vs. among-populations). Mean effect size for endothermic animals was positive and significantly different from the mean effect size for ectothermic animals. Only for within-population studies of ectothermic animals did we find a significantly negative effect size (r=-0.23 +/- 0.09). The distribution of effect sizes in relation to sample size provided little evidence for patterns typical of those produced by publication bias. Our analysis suggests, at best, only a weak association between H and FA, and heterozygosity seems to explain only a very small amount of the variation in developmental instability among individuals and populations (r2=0.01 for the total material).


Expected treatment benefits of percutaneous transluminal coronary angioplasty: the patient's perspective.
This study aimed to determine the expected treatment benefits from the patient's perspective before and after percutaneous transluminal coronary angioplasty (PTCA). Descriptive study. The sample consisted of 60 patients that underwent PTCA in the cardiology department of a university hospital in Bolu province in Turkey. The patients were interviewed one hour before the procedure and a week after discharge from hospital. The proportion of patients that perceived their health as good after the PTCA was two-fold in comparison to before (48.3%, 88.3%) (P < 0.05). The rate of patients taking heart disease seriously decreased by half after PTCA compared to before (40.0%, 83.3%) (P < 0.05). Almost all of the patients had the wrong interpretation of complete recovery from the diseases before and after PTCA (100.0%, 96.7%) (P > 0.05). Expectations of not having chest pain (P > 0.05), not having MI (P > 0.05), not having fear of death (P < 0.05), and not having stress (P > 0.05) were lower after PTCA in comparison to before. The expectation of not having chest pain was higher in women (60.9%, 47.8%) (P > 0.05), in those not having MI (56.1%, 48.8%) (P > 0.05), and in those having chronic diseases (59.0%, 43.6%) (P > 0.05) before PTCA compared with after. The majority of the patients had the wrong interpretation regarding their PTCA expectations, which decreased dramatically after the procedure. Continuing individualized health education programs on angioplasty for patients and their families will be an important way of dealing with patients' inaccurate interpretations.


Psychopathology in hospitalized alcoholics.
This study utilized the DSM-III criteria and the National Institute of Mental Health Diagnostic Interview Schedule to assess the prevalence of lifetime psychopathology among hospitalized alcoholics. Antisocial personality (ASP) and substance-use disorder were common psychopathologies among male alcoholics and major depression and phobia were common among female alcoholics. The onset of most psychopathologies preceded the abuse of alcohol among women. In men, however, with the exception of ASP and panic disorder, the onset of psychopathology was subsequent to that of alcohol abuse and/or dependence. Diagnoses of ASP and substance abuse were characterized by early onset of regular intoxication and alcohol abuse. Gender and the presence of specific psychopathology appeared to modify the course and symptom picture of alcoholism. In general, alcoholic women showed a later onset of regular intoxication and a more rapid progression to alcohol abuse and dependence than alcoholic men.


Structural insights into amino acid binding and gene control by a lysine riboswitch.
In bacteria, the intracellular concentration of several amino acids is controlled by riboswitches. One of the important regulatory circuits involves lysine-specific riboswitches, which direct the biosynthesis and transport of lysine and precursors common for lysine and other amino acids. To understand the molecular basis of amino acid recognition by riboswitches, here we present the crystal structure of the 174-nucleotide sensing domain of the Thermotoga maritima lysine riboswitch in the lysine-bound (1.9 ångström (A)) and free (3.1 A) states. The riboswitch features an unusual and intricate architecture, involving three-helical and two-helical bundles connected by a compact five-helical junction and stabilized by various long-range tertiary interactions. Lysine interacts with the junctional core of the riboswitch and is specifically recognized through shape-complementarity within the elongated binding pocket and through several direct and K(+)-mediated hydrogen bonds to its charged ends. Our structural and biochemical studies indicate preformation of the riboswitch scaffold and identify conformational changes associated with the formation of a stable lysine-bound state, which prevents alternative folding of the riboswitch and facilitates formation of downstream regulatory elements. We have also determined several structures of the riboswitch bound to different lysine analogues, including antibiotics, in an effort to understand the ligand-binding capabilities of the lysine riboswitch and understand the nature of antibiotic resistance. Our results provide insights into a mechanism of lysine-riboswitch-dependent gene control at the molecular level, thereby contributing to continuing efforts at exploration of the pharmaceutical and biotechnological potential of riboswitches.


Ginkgo may prevent genetic-associated ovarian cancer risk: multiple biomarkers and anticancer pathways induced by ginkgolide B in BRCA1-mutant ovarian epithelial cells.
Women carrying BRCA1 mutations have a higher risk of developing ovarian cancers. Options to reduce this risk are largely limited to prophylactic surgery, which leads to a decrease in the quality of life and permanently damages fertility. There is a obvious and an urgent need to identify a noninvasive approach to effectively prevent the ovarian cancer risk, specifically for those women of reproductive age. Our previous studies showed that the use of the herbal remedy Ginkgo biloba may reduce the risk for nonmucinous ovarian cancer. Here, we explored whether Ginkgo biloba might also be an effective agent to reduce BRCA1-associated ovarian cancer (always serous-type) risk. A human ovarian surface epithelial cell line-636 was developed from a BRCA1-mutant carrier. Cells were treated with ginkgolide B (GB) or dimethyl sulfoxide, and protein lysates from the cells were applied to antibody microarrays to determine upregulated or downregulated protein expression patterns. Anticancer activities and the associated networking pathways with the altered proteins were analyzed by using the Pathway Studio software. After GB treatment, 28 proteins were shown to be consistently upregulated (1.5-15.5-fold), and 22 proteins were downregulated (1.5-28.3-fold). Bioinformatics software analysis indicated that multiple mechanisms and signal pathways are involved in anticancer activities in BRCA1-mutant cells induced by GB treatment. These pathways include cell proliferation, tumor suppression, and DNA damage repair. Our study suggested that GB found in the herbal Ginkgo biloba may have cancer-preventive activities in BRCA1-mutant ovarian epithelial cells.


Comparative aerodynamic performance of flapping flight in two bat species using time-resolved wake visualization.
Bats are unique among extant actively flying animals in having very flexible wings, controlled by multi-jointed fingers. This gives the potential for fine-tuned active control to optimize aerodynamic performance throughout the wingbeat and thus a more efficient flight. But how bat wing performance scales with size, morphology and ecology is not yet known. Here, we present time-resolved fluid wake data of two species of bats flying freely across a range of flight speeds using stereoscopic digital particle image velocimetry in a wind tunnel. From these data, we construct an average wake for each bat species and speed combination, which is used to estimate the flight forces throughout the wingbeat and resulting flight performance properties such as lift-to-drag ratio (L/D). The results show that the wake dynamics and flight performance of both bat species are similar, as was expected since both species operate at similar Reynolds numbers (Re) and Strouhal numbers (St). However, maximum L/D is achieved at a significant higher flight speed for the larger, highly mobile and migratory bat species than for the smaller non-migratory species. Although the flight performance of these bats may depend on a range of morphological and ecological factors, the differences in optimal flight speeds between the species could at least partly be explained by differences in their movement ecology.


Long-term outcome after ischaemic stroke/transient ischaemic attack.
During the first 30 days after a stroke, the case fatality is about 25% and the major cause of death is the index stroke and its sequelae. The most consistent predictor of 30-day mortality after stroke is stroke severity. Other predictors include increasing age, a history of previous stroke, cardiac failure, and a high blood glucose concentration and white blood cell count. Other less common, but important, causes of early mortality are recurrent ischaemic stroke and a coronary event. The risk of a recurrent cerebrovascular event is highest in the first month (4%) and year (12%) after a stroke and transient ischaemic attack (TIA), probably reflecting the presence of active, unstable atherosclerotic plaque. Thereafter, the risk of a recurrent cerebrovascular event falls to about 5% per year, similar to the risk of a coronary event. During years 1-5 after a TIA and ischaemic stroke, cardiovascular disease increasingly becomes the major cause of death, reflecting the generalized nature of atherothrombosis, the most common cause of the index stroke. The most robust predictor of death within 1-5 years after stroke is increasing age, closely followed by cardiac failure. Additional baseline predictors of longer-term mortality include a history of previous symptomatic atherothrombosis (TIA, ischaemic stroke, peripheral arterial disease, and early-onset ischaemic heart disease), risk factors for atherothrombosis (smoking), other heart diseases (cardiac failure, atrial fibrillation) and increasing stroke severity. Lacunar syndromes can be predictive of relative longevity. At 5 years after stroke, survival is about 40%, and about half of survivors are disabled and dependent. The most robust predictors of disability at 5 years after stroke are increasing age, stroke severity, and recurrent stroke. The most powerful predictor of early recurrent stroke (within 30 days after stroke) is an atherosclerotic ischaemic stroke caused by large-artery atherosclerosis with >50% stenosis, whereas the strongest predictor of stroke recurrence over 5 years is diabetes. Other predictors of recurrent stroke include increasing age, previous TIA, atrial fibrillation, high alcohol consumption, haemorrhagic index stroke, and hypertension at discharge. The clinical implication of these findings is that strategies for optimizing long-term outcome after TIA and stroke should be directed toward reducing the high risk of recurrent stroke and coronary events by removing/recanalizing the symptomatic atherosclerotic plaque, controlling the underlying causal vascular risk factors, and administering long-term, effective antiplatelet therapy.


Uterine-specific p53 deficiency confers premature uterine senescence and promotes preterm birth in mice.
Many signaling pathways that contribute to tumorigenesis are also functional in pregnancy, although they are dysregulated in the former and tightly regulated in the latter. Transformation-related protein 53 (Trp53), which encodes p53, is a tumor suppressor gene whose mutation is strongly associated with cancer. However, its role in normal physiological processes, including female reproduction, is poorly understood. Mice that have a constitutive deletion of Trp53 exhibit widespread development of carcinogenesis at early reproductive ages, compromised spermatogenesis, and fetal exencephaly, rendering them less amenable to studying the role of p53 in reproduction. To overcome this obstacle, we generated mice that harbor a conditional deletion of uterine Trp53 and examined pregnancy outcome in females with this genotype. These mice had normal ovulation, fertilization, and implantation; however, postimplantation uterine decidual cells showed terminal differentiation and senescence-associated growth restriction with increased levels of phosphorylated Akt and p21, factors that are both known to participate in these processes in other systems. Strikingly, uterine deletion of Trp53 increased the incidence of preterm birth, a condition that was corrected by oral administration of the selective COX2 inhibitor celecoxib. We further generated evidence to suggest that deletion of uterine Trp53 induces preterm birth through a COX2/PGF synthase/PGF(2alpha) pathway. Taken together, our observations underscore what we believe to be a new critical role of uterine p53 in parturition.


Crowdsourcing, the great meteor storm of 1833, and the founding of meteor science.
Yale science professor Denison Olmsted used crowdsourcing to gather observations from across the United States of the unexpected deluge of meteors on 13 November 1833--more than 72,000/h. He used these observations (and newspaper accounts and correspondence from scientists) to make a commendably accurate interpretation of the meteor storm, overturning 2100 years of erroneous teachings about shooting stars and establishing meteor science as a new branch of astronomy. Olmsted's success was substantially based on his use of newspapers and their practice of news pooling to solicit observations from throughout the country by lay and expert observers professionally unaffiliated with Yale College and him. In today's parlance, Olmsted was a remarkably successful early practitioner of scientific crowdsourcing, also known as citizen science. He may have been the first to use mass media for crowdsourcing in science. He pioneered many of the citizen-science crowdsourcing practices that are still in use today: an open call for citizen participation, a clearly defined task, a large geographical distribution for gathering data and a rapid response to opportunistic events. Olmsted's achievement is not just that he used crowdsourcing in 1833 but that crowdsourcing helped him to advance science significantly.


Cancer Gene Discovery by Network Analysis of Somatic Mutations Using the MUFFINN Server.
Identifying genes that are capable of inducing tumorigenesis has been a major challenge in cancer research. In many cases, such genes frequently show somatic mutations in tumor samples; thus various computational methods for predicting cancer genes have been developed based on "significantly mutated genes." However, this approach is intrinsically limited by the fact that there are many cancer genes infrequently mutated in cancer genomes. Therefore, we recently developed MUFFINN (Mutations For Functional Impact on Network Neighbors), a method for cancer gene prediction based not only on mutation occurrences in each gene but also those of neighbors in functional networks. This enables the identification of cancer genes with infrequent mutation occurrence. We demonstrated that MUFFINN could retrieve known cancer genes more efficiently than gene-based methods and predicted cancer genes with low mutation occurrences in tumor samples. Users can freely access a web server ( http://www.inetbio.org/muffinn ) and run predictions with either public or private data of cancer somatic mutations. For given information of mutation occurrence profiles, the MUFFINN server returns lists of candidate cancer genes by four distinct predictions with different combinations between gene networks and scoring algorithms. Stand-alone software is also available, which allows MUFFINN to be run on local machines with a custom gene network. Here, we present an overall guideline for using the MUFFINN web server and stand-alone software for the discovery of novel cancer genes.


Synergistic action of cavity and catalytic sites in etched Pd-Cu2O octahedra to augment the peroxidase-like activity of Cu2O nanoparticles for the colorimetric detection of isoniazid and ascorbic acid.
Despite the widespread use of nanozyme-based colorimetric assays in biosensing, challenges such as limited catalytic efficiency, inadequate sensitivity to analytes, and insufficient understanding of the structure-activity relationship still persist. Overcoming these hurdles by enhancing the inherent enzyme-like performance of nanozymes using the unique attributes of nanomaterials is still a significant obstacle. Here, we designed and constructed Pd-Cu2O nanocages (Pd-Cu2O NCs) by selectively etching the vertices of the copper octahedra to enhance the peroxidase-like (POD-like) activity of Cu2O nanoparticles. The improved catalytic activity of Pd-Cu2O NCs was attributed to their high specific surface area and abundant catalytic sites. Mechanistic studies revealed that reactive oxygen species (ROS) intermediates (•OH) were generated through the decomposition of H2O2, resulting in POD-like activity of the Pd-Cu2O NCs. The designed Pd-Cu2O NCs can oxidize 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2, producing a blue oxidation product (oxTMB). The oxidation reaction was inhibited and led to a significant bleaching of the blue color in the presence of reducing substances isoniazid (INH) and ascorbic acid (AA). Based on these principles, we developed a colorimetric sensing platform for the detection of INH and AA, exhibiting good sensitivity and stability. This work provided a straightforward approach to the structural engineering of nanomaterials and the enhancement of enzyme-mimicking properties.


Helicobacter pylori FlgR is an enhancer-independent activator of sigma54-RNA polymerase holoenzyme.
Helicobacter pylori FlgR activates transcription with sigma54-RNA polymerase holoenzyme (sigma54-holoenzyme) from at least five flagellar operons. Activators of sigma54-holoenzyme generally bind enhancer sequences located >70 bp upstream of the promoter and contact sigma54-holoenzyme bound at the promoter through DNA looping to activate transcription. H. pylori FlgR lacks the carboxy-terminal DNA-binding domain present in most sigma54-dependent activators. As little as 42 bp of DNA upstream of the flaB promoter and 26 bp of DNA sequence downstream of the transcriptional start site were sufficient for efficient FlgR-mediated expression from a flaB'-'xylE reporter gene in H. pylori, indicating that FlgR does not use an enhancer to activate transcription. Other examples of sigma54-dependent activators that lack a DNA-binding domain include Chlamydia trachomatis CtcC and activators from the other Chlamydia spp. whose genomes have been sequenced. FlgR from Helicobacter hepaticus and Campylobacter jejuni, which are closely related to H. pylori, appear to have carboxy-terminal DNA-binding domains, suggesting that the loss of the DNA-binding domain from H. pylori FlgR occurred after the divergence of these bacterial species. Removal of the amino-terminal regulatory domain of FlgR resulted in a constitutively active form of the protein that activated transcription from sigma54-dependent genes in Escherichia coli. The truncated FlgR protein also activated transcription with E. coli sigma54-holoenzyme in an in vitro transcription assay.


Candida albicans Interaction with Oral Epithelial Cells: Adhesion , Invasion, and Damage Assays.
Microbial interactions with epithelial barriers are important steps preceding disease. Infections with Candida albicans are no exception. This opportunistic fungus, commonly harmlessly residing in close proximity to human epithelia, can shift to a more pathogenic form, can invade tissues, and cause disease. Pathogenesis, in C. albicans as well as in many other microorganisms, is characterized by three important steps: adhesion to-, invasion into-, and damage of host cells. In this book chapter, we describe three well-established protocols that allow us to differentially stain C. albicans cells adhering to and invading into host cells, therefore allowing quantifications of such processes. We also describe a common host cell cytotoxicity assay that employs a commercial kit, adapted to C. albicans.


Recreational Physical Activity Is Associated with Reduced Breast Cancer Risk in Adult Women at High Risk for Breast Cancer: A Cohort Study of Women Selected for Familial and Genetic Risk.
Although physical activity is associated with lower breast cancer risk for average-risk women, it is not known if this association applies to women at high familial/genetic risk. We examined the association of recreational physical activity (self-reported by questionnaire) with breast cancer risk using the Prospective Family Study Cohort, which is enriched with women who have a breast cancer family history (N = 15,550). We examined associations of adult and adolescent recreational physical activity (quintiles of age-adjusted total metabolic equivalents per week) with breast cancer risk using multivariable Cox proportional hazards regression, adjusted for demographics, lifestyle factors, and body mass index. We tested for multiplicative interactions of physical activity with predicted absolute breast cancer familial risk based on pedigree data and with BRCA1 and BRCA2 mutation status. Baseline recreational physical activity level in the highest four quintiles compared with the lowest quintile was associated with a 20% lower breast cancer risk (HR, 0.80; 95% confidence interval, 0.68-0.93). The association was not modified by familial risk or BRCA mutation status (P interactions >0.05). No overall association was found for adolescent recreational physical activity. Recreational physical activity in adulthood may lower breast cancer risk for women across the spectrum of familial risk. SIGNIFICANCE: These findings suggest that physical activity might reduce breast cancer risk by about 20% for women across the risk continuum, including women at higher-than-average risk due to their family history or genetic susceptibility.See related commentary by Niehoff et al., p. 23.


Recent studies on aphrodisiac herbs for the management of male sexual dysfunction--a review.
An aphrodisiac is a type of food or drink that has the effect of making those who eat or drink it more aroused in a sexual way. Aphrodisiacs can be categorized according to their mode of action into three groups: substances that increase libido (i.e., sexual desire, arousal), substances that increase sexual potency (i.e., effectiveness of erection) and substances that increase sexual pleasure. Some well-known aphrodisiacs are Tribulus terrestrins, Withania somnifera, Eurycoma longifolia, Avena sativa, Ginko biloba, and Psoralea coryifolia. Ethnobotanical surveys have indicated a large number of plants as aphrodisiacs. The paper reviews the recent scientific validation on traditionally used herbal plants as aphrodisiac herbs for the management of sexual disorder erectile dysfunction.


Urokinase therapy for Silastic catheter-induced intravascular thrombi in infants and children.
Among the serious complications encountered with long-term, indwelling Silastic central venous catheters are catheter-induced intravascular thrombi. These thrombi are usually treated by removal of the catheter to prevent thrombus propagation, embolization, or infection. We treated ten patients with urokinase infusion who had experienced 12 incidents of induced intravascular thrombi. Catheter phlebography and two-dimensional echocardiography were used for diagnosis and follow-up. Eleven of the 12 episodes were treated successfully, with complete dissolution of the thrombus. One patient with a calcific thrombus had only partial clot lysis and required catheter removal. By utilizing urokinase infusion to treat Silastic catheter-induced intravascular thrombi, nine of ten central venous catheters were preserved and the possible need for thrombectomy was averted. No serious complications were encountered. In our experience, urokinase therapy has been an effective and safe method for treating Silastic catheter-induced intravascular thrombi.


Adolescent vulnerability to posttraumatic stress disorder.
This study found that men who developed posttraumatic stress disorder after combat in Vietnam tended to have been adolescents while in combat. They had formed an intense attachment to other men in their combat unit, which had been disrupted by the death of a buddy. This loss generally was followed by acts of revenge and subsequent feelings of a profound lack of control over their destiny. Adolescents use their peer group as an intermediary stage between dependency on their family and emotional maturity, and the army, particularly under battlefield conditions, maximizes the impact of peer group cohesion. For these younger men, the death of a friend was experienced as the dissolution of the once omnipotent group and as a narcissistic injury. Group psychotherapy for Vietnam veterans allows the partial re-creation of the peer group in the context in which the trauma occurred. The sharing and reliving of common experiences may facilitate entrance into the world of adult relationships, a process that was arrested by the trauma.


Storage temperatures for cold-chain delivery in cell therapy: a study of alginate-encapsulated liver cell spheroids stored at -80°c or -170°c for up to 1 year.
A bioartificial liver comprising alginate-encapsulated liver cell spheroids (ELS) could bridge the gap to transplant or spontaneous recovery in acute liver failure, but will be required for emergency use, necessitating cryopreservation. A cryopreservation protocol has been developed, but beyond this, the feasibility of cold-chain storage is considered here. Cryopreservation will be increasingly required for timely delivery of tissue and bioengineered products, and significant, but often, over-looked factors that impact on cost and ease of clinical application are the storage temperature and useful preservation time. Storage in the vapor phase of liquid nitrogen (∼-170°C) is the gold standard, but for safety and economic purposes, storing ELS in electric freezers at -80°C may be preferable. ELS were cryopreserved using an optimized protocol and stored at either -80°C or at -170°C for up to 1 year. ELS were removed from storage after 1, 2, 3, 6, 9, or 12 months, and recovery was assessed 24 h postwarming. Cell recovery was assessed using viability (fluorescent staining with image analysis), cell number (nuclei count), and functional (hepatospecific protein enzyme-linked immunosorbent assay) assays. Viability, the viable cell number, and function of ELS stored at -170°C were maintained at similar values throughout the year. In contrast, ELS stored at -80°C exhibited decreased viability, viable cell numbers, and function by as early as 1 month. Progressive deterioration was subsequently observed. After 12 months of storage at -80°C, viable cell recovery of ELS was ∼15% that of ELS stored at -170°C. While convenience and cost might support the use of -80°C for storage of multicellular bioengineered products such as ELS, results indicate rapid deterioration in functional recoveries after only a few weeks. This study demonstrates that storage temperature is an important consideration in regenerative medicine and caution should be applied by limiting storage at -80°C to only a few weeks.


New potential eukaryotic substrates of the mycobacterial protein tyrosine phosphatase PtpA: hints of a bacterial modulation of macrophage bioenergetics state.
The bacterial protein tyrosine phosphatase PtpA is a key virulence factor released by Mycobacterium tuberculosis in the cytosol of infected macrophages. So far only two unrelated macrophage components (VPS33B, GSK3α) have been identified as PtpA substrates. As tyrosine phosphatases are capable of using multiple substrates, we developed an improved methodology to pull down novel PtpA substrates from an enriched P-Y macrophage extract using the mutant PtpA D126A. This methodology reduced non-specific protein interactions allowing the identification of four novel putative PtpA substrates by MALDI-TOF-MS and nano LC-MS: three mitochondrial proteins - the trifunctional enzyme (TFP), the ATP synthase, and the sulfide quinone oxidoreductase - and the cytosolic 6-phosphofructokinase. All these proteins play a relevant role in cell energy metabolism. Using surface plasmon resonance, PtpA was found to bind immunopurified human TFP through its catalytic site since TFP-PtpA association was inhibited by a specific phosphatase inhibitor. Moreover, PtpA wt was capable of dephosphorylating immunopurified human TFP in vitro supporting that TFP may be a bona fide PtpA susbtrate. Overall, these results suggest a novel scenario where PtpA-mediated dephosphorylation may affect pathways involved in cell energy metabolism, particularly the beta oxidation of fatty acids through modulation of TFP activity and/or cell distribution.


Monoclonal antibody treatment exposes three mechanisms underlying the clinical course of multiple sclerosis.
The elective treatment of patients with multiple sclerosis, using a humanized anti-leukocyte (CD52) monoclonal antibody (Campath-1H), has illuminated mechanisms that underlie the clinical course of the disease. Twenty-seven patients were studied clinically and by magnetic resonance imaging (MRI) before and for 18 months after a single pulse of Campath-1H. The first dose of monoclonal antibody was associated with a transient rehearsal of previous symptoms caused by the release of mediators that impede conduction at previously demyelinated sites; this effect remained despite selective blockade of tumor necrosis factor-alpha. Disease activity persisted for several weeks after treatment but thereafter radiological markers of cerebral inflammation were suppressed for at least 18 months during which there were no new symptoms or signs. However, about half the patients experienced progressive disability and increasing brain atrophy, attributable on the basis of MRI spectroscopy to axonal degeneration, which correlated with the extent of cerebral inflammation in the pretreatment phase. These data support the formulation that inflammation and demyelination are responsible for relapses of multiple sclerosis; that inflammatory mediators, but not tumor necrosis factor-alpha, cause symptomatic reactivation of previously demyelinated lesions; and that axonal degeneration, conditioned by prior inflammation but proceeding despite its suppression, contributes to the progressive phase of disability. These results provide evidence supporting the emerging view that treatment in multiple sclerosis must be given early in the course, before the consequences of inflammation are irretrievably established.


Identification of novel risk loci for restless legs syndrome in genome-wide association studies in individuals of European ancestry: a meta-analysis.
Restless legs syndrome is a prevalent chronic neurological disorder with potentially severe mental and physical health consequences. Clearer understanding of the underlying pathophysiology is needed to improve treatment options. We did a meta-analysis of genome-wide association studies (GWASs) to identify potential molecular targets. In the discovery stage, we combined three GWAS datasets (EU-RLS GENE, INTERVAL, and 23andMe) with diagnosis data collected from 2003 to 2017, in face-to-face interviews or via questionnaires, and involving 15 126 cases and 95 725 controls of European ancestry. We identified common variants by fixed-effect inverse-variance meta-analysis. Significant genome-wide signals (p≤5 × 10-8) were tested for replication in an independent GWAS of 30 770 cases and 286 913 controls, followed by a joint analysis of the discovery and replication stages. We did gene annotation, pathway, and gene-set-enrichment analyses and studied the genetic correlations between restless legs syndrome and traits of interest. We identified and replicated 13 new risk loci for restless legs syndrome and confirmed the previously identified six risk loci. MEIS1 was confirmed as the strongest genetic risk factor for restless legs syndrome (odds ratio 1·92, 95% CI 1·85-1·99). Gene prioritisation, enrichment, and genetic correlation analyses showed that identified pathways were related to neurodevelopment and highlighted genes linked to axon guidance (associated with SEMA6D), synapse formation (NTNG1), and neuronal specification (HOXB cluster family and MYT1). Identification of new candidate genes and associated pathways will inform future functional research. Advances in understanding of the molecular mechanisms that underlie restless legs syndrome could lead to new treatment options. We focused on common variants; thus, additional studies are needed to dissect the roles of rare and structural variations. Deutsche Forschungsgemeinschaft, Helmholtz Zentrum München-Deutsches Forschungszentrum für Gesundheit und Umwelt, National Research Institutions, NHS Blood and Transplant, National Institute for Health Research, British Heart Foundation, European Commission, European Research Council, National Institutes of Health, National Institute of Neurological Disorders and Stroke, NIH Research Cambridge Biomedical Research Centre, and UK Medical Research Council.


Intraoral Excision of a Huge Cheek Lipoma.
Lipomas are benign tumors of mature adipocytes unusual in the oral and maxillofacial region. The average size of cheek lipomas in the literature ranges from 1.5 to 2.5 cm, with the maximum size of 5 cm. Their etiology remains unclear. Lipomas present, clinically, as well circumscribed, slow growing, painless masses, usually treated by complete excision. The aim of this paper is to present a 78-year-old Caucasian male patient with a huge cheek lipoma compromising facial esthetics and treated through an intraoral excision. Postoperative period was uneventful with no signs of recurrence. Concluding, the intraoral approach is a relatively simple technique that should be taken into account when considering the surgical removal of cheek lipomas.


Design scaling rules for 2R-optical self-phase modulation-based regenerators.
We present simple scaling rules to optimize the design of 2R optical regenerators relying on Self-Phase Modulation in the normal dispersion regime and associated offset spectral filtering. A global design map is derived which relates both the physical parameters of the regenerator and the properties of the incoming signal to the regeneration performance. The operational conditions for optimum noise rejection are identified using this map and a detailed analysis of the system behavior under these conditions presented. Finally, we demonstrate application of the general design map to the design of a regenerator for a specific 160 Gb/s system.


Reorganized stores and impaired calcium handling in skeletal muscle of mice lacking calsequestrin-1.
Calsequestrin (CS), the major Ca(2+)-binding protein in the sarcoplasmic reticulum (SR), is thought to play a dual role in excitation-contraction coupling: buffering free Ca(2+) increasing SR capacity, and modulating the activity of the Ca(2+) release channels (RyRs). In this study, we generated and characterized the first murine model lacking the skeletal CS isoform (CS1). CS1-null mice are viable and fertile, even though skeletal muscles appear slightly atrophic compared to the control mice. No compensatory increase of the cardiac isoform CS2 is detectable in any type of skeletal muscle. CS1-null muscle fibres are characterized by structural and functional changes, which are much more evident in fast-twitch muscles (EDL) in which most fibres express only CS1, than in slow-twitch muscles (soleus), where CS2 is expressed in about 50% of the fibres. In isolated EDL muscle, force development is preserved, but characterized by prolonged time-to-peak and half-relaxation time, probably related to impaired calcium release from and re-uptake by the SR. Ca(2+)-imaging studies show that the amount of Ca(2+) released from the SR and the amplitude of the Ca(2+) transient are significantly reduced. The lack of CS1 also causes significant ultrastructural changes, which include: (i) striking proliferation of SR junctional domains; (ii) increased density of Ca(2+)-release channels (confirmed also by (3)H-ryanodine binding); (iii) decreased SR terminal cisternae volume; (iv) higher density of mitochondria. Taken together these results demonstrate that CS1 is essential for the normal development of the SR and its calcium release units and for the storage and release of appropriate amounts of SR Ca(2+).


An adaptable neural-network model for recursive nonlinear traffic prediction and modeling of MPEG video sources.
Multimedia services and especially digital video is expected to be the major traffic component transmitted over communication networks [such as internet protocol (IP)-based networks]. For this reason, traffic characterization and modeling of such services are required for an efficient network operation. The generated models can be used as traffic rate predictors, during the network operation phase (online traffic modeling), or as video generators for estimating the network resources, during the network design phase (offline traffic modeling). In this paper, an adaptable neural-network architecture is proposed covering both cases. The scheme is based on an efficient recursive weight estimation algorithm, which adapts the network response to current conditions. In particular, the algorithm updates the network weights so that 1) the network output, after the adaptation, is approximately equal to current bit rates (current traffic statistics) and 2) a minimal degradation over the obtained network knowledge is provided. It can be shown that the proposed adaptable neural-network architecture simulates a recursive nonlinear autoregressive model (RNAR) similar to the notation used in the linear case. The algorithm presents low computational complexity and high efficiency in tracking traffic rates in contrast to conventional retraining schemes. Furthermore, for the problem of offline traffic modeling, a novel correlation mechanism is proposed for capturing the burstness of the actual MPEG video traffic. The performance of the model is evaluated using several real-life MPEG coded video sources of long duration and compared with other linear/nonlinear techniques used for both cases. The results indicate that the proposed adaptable neural-network architecture presents better performance than other examined techniques.


Transcriptional regulation and organization of the dcuA and dcuB genes, encoding homologous anaerobic C4-dicarboxylate transporters in Escherichia coli.
The dcuA and dcuB genes of Escherichia coli encode homologous proteins that appear to function as independent and mutually redundant C4-dicarboxylate transporters during anaerobiosis. The dcuA gene is 117 bp downstream of, and has the same polarity as, the aspartase gene (aspA), while dcuB is 77 bp upstream of, and has the same polarity as, the anaerobic fumarase gene (fumB). To learn more about the respective roles of the dcu genes, the environmental and regulatory factors influencing their expression were investigated by generating and analyzing single-copy dcuA- and dcuB-lacZ transcriptional fusions. The results show that dcuA is constitutively expressed whereas dcuB expression is highly regulated. The dcuB gene is strongly activated anaerobically by FNR, repressed in the presence of nitrate by NarL, and subject to cyclic AMP receptor protein (CRP)-mediated catabolite repression. In addition, dcuB is strongly induced by C4-dicarboxylates, suggesting that dcuB is under the control of an uncharacterized C4-dicarboxylate-responsive gene regulator. Northern blotting confirmed that dcuA (and aspA) is expressed under both aerobic and anaerobic conditions and that dcuB (and fumB) is induced anaerobically. Major monocistronic transcripts were identified for aspA and dcuA, as well as a minor species possibly corresponding to an aspA-dcuA cotranscript. Five major transcripts were observed for dcuB and fumB: monocistronic transcripts for both fumB and dcuB; a dcuB-fumB cotranscript; and two transcripts, possibly corresponding to dcuB-fumB and fumB mRNA degradation products. Primer extension analysis revealed independent promoters for aspA, dcuA, and dcuB, but surprisingly no primer extension product could be detected for fumB. The expression of dcuB is entirely consistent with a primary role for DcuB in mediating C4-dicarboxylate transport during anaerobic fumarate respiration. The precise physiological purpose of DcuA remains unclear.


Magneto-optic spatial light modulator with submicron-size magnetic pixels for wide-viewing-angle holographic displays.
Spatial light modulators (SLMs) with submicron-size pixels are promising devices for use in wide-viewing-angle glasses-free holographic 3D displays. In accordance with estimations, an SLM with 1 μm pitch pixels can display holographic images over viewing angles of 30 deg. In the present work, we have studied magneto-optic SLMs (MOSLMs) with submicron-size pixels for wide-viewing-angle holographic displays that are driven by thermomagnetic recording. Amorphous TbFe films were used as a magneto-optic material. The MOSLM with an 800 nm diameter pixels array displayed wide-viewing-angle holographic images. These results demonstrated that MOSLMs with submicron-size pixels have promise for glasses-free holographic 3D displays.


Annotation of genes involved in glycerolipid biosynthesis in Chlamydomonas reinhardtii: discovery of the betaine lipid synthase BTA1Cr.
Lipid metabolism in flowering plants has been intensely studied, and knowledge regarding the identities of genes encoding components of the major fatty acid and membrane lipid biosynthetic pathways is very extensive. We now present an in silico analysis of fatty acid and glycerolipid metabolism in an algal model, enabled by the recent availability of expressed sequence tag and genomic sequences of Chlamydomonas reinhardtii. Genes encoding proteins involved in membrane biogenesis were predicted on the basis of similarity to proteins with confirmed functions and were organized so as to reconstruct the major pathways of glycerolipid synthesis in Chlamydomonas. This analysis accounts for the majority of genes predicted to encode enzymes involved in anabolic reactions of membrane lipid biosynthesis and compares and contrasts these pathways in Chlamydomonas and flowering plants. As an important result of the bioinformatics analysis, we identified and isolated the C. reinhardtii BTA1 (BTA1Cr) gene and analyzed the bifunctional protein that it encodes; we predicted this protein to be sufficient for the synthesis of the betaine lipid diacylglyceryl-N,N,N-trimethylhomoserine (DGTS), a major membrane component in Chlamydomonas. Heterologous expression of BTA1Cr led to DGTS accumulation in Escherichia coli, which normally lacks this lipid, and allowed in vitro analysis of the enzymatic properties of BTA1Cr. In contrast, in the bacterium Rhodobacter sphaeroides, two separate proteins, BtaARs and BtaBRs, are required for the biosynthesis of DGTS. Site-directed mutagenesis of the active sites of the two domains of BTA1Cr allowed us to study their activities separately, demonstrating directly their functional homology to the bacterial orthologs BtaARs and BtaBRs.


Student perception of group dynamics predicts individual performance: Comfort and equity matter.
Active learning in college classes and participation in the workforce frequently hinge on small group work. However, group dynamics vary, ranging from equitable collaboration to dysfunctional groups dominated by one individual. To explore how group dynamics impact student learning, we asked students in a large-enrollment university biology class to self-report their experience during in-class group work. Specifically, we asked students whether there was a friend in their group, whether they were comfortable in their group, and whether someone dominated their group. Surveys were administered after students participated in two different types of intentionally constructed group activities: 1) a loosely-structured activity wherein students worked together for an entire class period (termed the 'single-group' activity), or 2) a highly-structured 'jigsaw' activity wherein students first independently mastered different subtopics, then formed new groups to peer-teach their respective subtopics. We measured content mastery by the change in score on identical pre-/post-tests. We then investigated whether activity type or student demographics predicted the likelihood of reporting working with a dominator, being comfortable in their group, or working with a friend. We found that students who more strongly agreed that they worked with a dominator were 17.8% less likely to answer an additional question correct on the 8-question post-test. Similarly, when students were comfortable in their group, content mastery increased by 27.5%. Working with a friend was the single biggest predictor of student comfort, although working with a friend did not impact performance. Finally, we found that students were 67% less likely to agree that someone dominated their group during the jigsaw activities than during the single group activities. We conclude that group activities that rely on positive interdependence, and include turn-taking and have explicit prompts for students to explain their reasoning, such as our jigsaw, can help reduce the negative impact of inequitable groups.


Humoral immune responses in volunteers immunized with irradiated Plasmodium falciparum sporozoites.
Volunteers immunized with gamma-irradiated Plasmodium falciparum sporozoites serve as the gold standard for protective immunity against mosquito-borne malaria transmission and provide a relevant model for studying protective immune effector mechanisms. During a 7-12 month period, we immunized four volunteers via the bites of irradiated, infected mosquitoes. Following these exposures to attenuated sporozoites, all four volunteers developed antibodies to sporozoites as measured by an immunofluorescence assay and by an enzyme-linked immunosorbent assay using the circumsporozoite (CS) protein repeat-based molecule R32LR as capture antigen. Three volunteers also developed antibodies against the nonrepeating (flanking) regions of the CS protein; the level of these antibodies paralleled the serum activity to inhibit sporozoite invasion of hepatoma cells in vitro. These three volunteers were protected against malaria transmitted by the bites of five infected mosquitoes. Two of these protected volunteers received additional immunizing doses of irradiated sporozoites and were subsequently protected against challenge with a heterologous P. falciparum clone. No detectable fluctuations were observed in circulating levels of tumor necrosis factor, interferon-gamma, or interleukin-6 during the course of this study. Analysis of the humoral and cellular immune responses of these protected volunteers is expected to yield important clues to additional targets of immunity against the pre-erythrocytic stages of malaria parasites.


Improving the experience of obtaining repeat complex paediatric prescriptions in the UK.
In the UK, medicines for chronic conditions in children and young people (CYP) are typically initiated within secondary or tertiary care, with responsibility for ongoing supply often then passed to the child's general practitioner (GP) and community pharmacist. The patient should then be reviewed in regular specialist clinics, with two-way communication for any changes in medications or clinical status undertaken between primary and secondary/tertiary care. This arrangement allows long-term medications to be obtained close to home.Although this is what parents expect, the reality is often messy, with families regularly needing to source some medicines from the GPs and others via hospitals or homecare services. In addition, these arrangements are not uniform, they vary across different areas of the UK and depend on individual GP or hospital prescriber acceptance. When neither primary, secondary or tertiary care accepts it is their responsibility to prescribe, or patients are under multiple specialists, families often feel left to navigate this complex and variable supply system themselves. Obtaining a prescription is only the start of the process for families as dispensing from a community pharmacy can also be challenging.In this article, we set out the barriers and potential solutions to this complex issue. We use the term specialist prescribers to include not only paediatricians but all other specialists looking after CYP including child and adolescent psychiatrists, ophthalmologists, dermatologists, surgeons, etc, as well as non-medical prescribers.


Herkogamy and mate diversity in the wild daffodil Narcissus longispathus: beyond the selfing-outcrossing paradigm in the evolution of mixed mating.
Spatial separation of male and female reproductive structures (herkogamy) is a widespread floral trait that has traditionally been viewed as an adaptation that reduces the likelihood of self-pollination. Here we propose that increased herkogamy may also influence another important aspect of plant mating: the diversity of pollen donors siring seeds within fruits. We test this hypothesis in Narcissus longispathus, a wild daffodil species with extensive variation in anther-stigma separation. To study the morphological basis of variation in herkogamy, floral measurements were undertaken in 16 populations of N. longispathus. We then quantified multilocus outcrossing rates and the correlation of outcrossed paternity in three of these populations sampled over several years. Mating system estimates were calculated for each population and year, and also separately for groups of plants that differed markedly in herkogamy within each population and year. In N. longispathus herkogamy was much more variable than other floral traits, and was more closely related to style length than to anther position. Averaged across populations and years, plants with high herkogamy had similar outcrossing rates (0.683) to plants with intermediate (0.648) or low herkogamy (0.590). However, a significant linear trend was found for correlation of outcrossed paternity, which increased monotonically from high herkogamy (0.221), through intermediate herkogamy (0.303) to low herkogamy (0.463) plants. The diversity of pollen donors siring seeds of high herkogamy Narcissus flowers was thus consistently greater than the diversity of pollen donors siring seeds of low herkogamy flowers. Results of this study contribute to the emerging consensus that floral traits can simultaneously influence several aspects of plant mating system in complex ways, thus extending the traditional focus centred exclusively on patterns and relative importance of self- and cross-fertilisation.


Prognostic factors related to add-on dendritic cell vaccines on patients with inoperable pancreatic cancer receiving chemotherapy: a multicenter analysis.
Dendritic cell (DC)-based cancer vaccines may have a significant benefit to patients with advanced pancreatic cancer. However, variations among clinical studies make it difficult to compare clinical outcomes. Here, we identified factors that determined the clinical benefits by analyzing data obtained at seven Japanese institutions that employed the same DC preparation and treatment regimens. Of 354 patients who met the inclusion criteria, 255 patients who received standard chemotherapy combined with peptide-pulsed DC vaccines were analyzed. The mean survival time from diagnosis was 16.5 months (95 % CI 14.4-18.5) and that from the first vaccination was 9.9 months (95 % CI 8.0-12.9). Known prognostic baseline factors related to advanced pancreatic cancer, namely ECOG-PS, peritoneal metastasis, liver metastasis, and the prognostic nutrition index, were also representative. Importantly, we found that erythema reaction after vaccination was an independent and treatment-related prognostic factor for better survival and that OK-432 might be a good adjuvant enhancing the antitumor immunity during DC vaccination. This is the first report of a multicenter clinical study suggesting the feasibility and possible clinical benefit of an add-on DC vaccine in patients with advanced pancreatic cancer who are undergoing chemotherapy. These findings need to be addressed in well-controlled prospective randomized trials.


[Etoposide and cisplatin for non-small cell lung cancer: trial at the Soroka Medical Center 1986-1994].
Etoposide and cisplatin (EP) has been the standard therapy for non-small cell lung cancer (NSCLC) at many cancer centers for over a decade. We analyzed our experience with EP in NSCLC to provide a baseline for comparison with new drugs. From 1986 through 1994, 46 of our patients with NSCLC received EP as first-line chemotherapy. Radiation therapy was administered to 25 of them, including 20 who received it immediately before or concomitantly with chemotherapy. Toxicity was mild and included only 1 episode of neutropenic fever and 1 case of reversible renal failure. Overall response was 22%. In 3, response was complete (pathologically documented in 2 of them) and in 7 partial. Median survival in locally advanced (stage III) and metastatic disease (stage IV and recurrent) were 12 months and 7 months, respectively. 2 patients are alive and free of disease more than 6 years after diagnosis. Our results are consistent with other published studies of EP in NSCLC. EP provides modest benefit in locally advanced NSCLC, with minimal toxicity.