Liver Imaging Reporting and Data System: Discordance Between Computed Tomography and Gadoxetate-Enhanced Magnetic Resonance Imaging for Detection of Hepatocellular Carcinoma Major Features.
The goal of this study was to compare agreement between computed tomography (CT) and magnetic resonance imaging (MRI) in the evaluation of the major Liver Imaging Reporting and Data System (LI-RADS) features used in assessment of hepatocellular carcinoma: arterial phase hyperenhancement (APHE), portal venous phase washout (WO), capsule appearance (capsule), and largest diameter (diameter). Patients with liver protocol CT and gadoxetate-enhanced MRI within 1 month of each other and at least 1 discrete untreated liver lesion were included. Two readers independently reviewed hepatic arterial phase and portal venous phase of each lesion on both CT and MRI, presented at random. The APHE, WO, capsule, and diameter were assessed for each lesion on CT and MRI. The LI-RADS category was assigned based on the recorded major features. Interobserver agreements between the readers for both imaging modalities and for each of the major features were assessed using κ statistics. Agreement between CT and MRI for each reader and for each feature was assessed using κ statistics. Agreement was interpreted based on κ as follows: 0.20 or less, slight agreement; 0.21 to 0.40, fair agreement; 0.41 to 0.60, moderate agreement; 0.61 to 0.80, substantial agreement; and 0.81 to 1.00, almost perfect agreement. Intraclass correlation coefficient was used to assess concordance of diameter measurements. There were 42 patients (mean age, 62.2 ± 7.0 years; 33 men [78.6%]) with 50 lesions. On CT, the interobserver agreement between the readers was almost perfect for APHE (κ = 0.85), WO (κ = 0.83), and capsule (κ = 0.86). On MRI, the interobserver agreement between the readers was almost perfect for APHE (κ = 0.86) and WO (κ = 0.83) and moderate for capsule (κ = 0.59). Intraclass correlation coefficient for diameter measurement was 0.99 for CT and 0.98 for MRI. For reader 1, the agreement between CT and MRI was fair for APHE (κ = 0.39) and capsule (κ = 0.26) and moderate for WO (κ = 0.49). For reader 2, the agreement between CT and MRI was moderate for APHE (κ = 0.43) and capsule (κ = 0.43) and fair (κ = 0.38) for WO. Agreement between readers for final LI-RADS category was substantial for CT (κ = 0.79) and moderate for MRI (κ = 0.60). Agreement for final LI-RADS categories between MRI and CT was fair for both reader 1 (κ = 0.33) and reader 2 (κ = 0.39). Interobserver agreement for the major LI-RADS features varies from moderate to almost perfect, for both CT and MRI. However, the agreement between CT and MRI for each of the major LI-RADS features is poor, ranging from fair to moderate. This poor agreement contributes to substantial differences between final LI-RADS category assigned on CT versus MRI.


Development of copy number assays for detection and surveillance of piperaquine resistance associated plasmepsin 2/3 copy number variation in Plasmodium falciparum.
Long regarded as an epicenter of drug-resistant malaria, Southeast Asia continues to provide new challenges to the control of Plasmodium falciparum malaria. Recently, resistance to the artemisinin combination therapy partner drug piperaquine has been observed in multiple locations across Southeast Asia. Genetic studies have identified single nucleotide polymorphisms as well as copy number variations in the plasmepsin 2 and plasmepsin 3 genes, which encode haemoglobin-degrading proteases that associate with clinical and in vitro piperaquine resistance. To accurately and quickly determine the presence of copy number variations in the plasmepsin 2/3 genes in field isolates, this study developed a quantitative PCR assay using TaqMan probes. Copy number estimates were validated using a separate SYBR green-based quantitative PCR assay as well as a novel PCR-based breakpoint assay to detect the hybrid gene product. Field samples from 2012 to 2015 across three sites in Cambodia were tested using DNA extracted from dried blood spots and whole blood to monitor the extent of plasmepsin 2/3 gene amplifications, as well as amplifications in the multidrug resistance transporter 1 gene (pfmdr1), a marker of mefloquine resistance. This study found high concordance across all methods of copy number detection. For samples derived from dried blood spots, a success rate greater than 80% was found in each assay, with more recent samples performing better. Evidence of extensive plasmepsin 2/3 copy number amplifications was observed in Pursat (94%, 2015) (Western Cambodia) and Preah Vihear (87%, 2014) (Northern Cambodia), and lower levels in Ratanakiri (16%, 2014) (Eastern Cambodia). A shift was observed from two copies of plasmepsin 2 in Pursat in 2013 to three copies in 2014-2015 (25% to 64%). Pfmdr1 amplifications were absent in all samples from Preah Vihear and Ratanakiri in 2014 and absent in Pursat in 2015. The multiplex TaqMan assay is a robust tool for monitoring both plasmepsin 2/3 and pfmdr1 copy number variations in field isolates, and the SYBR-green and breakpoint assays are useful for monitoring plasmepsin 2/3 amplifications. This study shows increasing levels of plasmepsin 2 copy numbers across Cambodia from 2012 to 2015 and a complete reversion of multicopy pfmdr1 parasites to single copy parasites in all study locations.


Femtosecond Laser-Assisted Lamellar Keratectomy for Corneal Opacities Secondary to Anterior Corneal Dystrophies: An Interventional Case Series.
To report results of femtosecond laser-assisted lamellar keratectomy (FLK) for corneal opacities secondary to anterior corneal dystrophies. Patients with a clinical diagnosis of Reis-Bücklers corneal dystrophy, granular corneal dystrophy, lattice corneal dystrophy, and macular corneal dystrophy were treated. FLK was performed to remove a central corneal free cap of 9.5 mm in diameter at a depth of 110 to 140 μm on which histological analysis was undertaken. Preoperative and postoperative refraction, best spectacle-corrected visual acuity, corneal topography results, and color photographs were recorded. Postoperative in vivo confocal microscopy of the cornea was performed. Changes in uncorrected visual acuity and best spectacle-corrected visual acuity, keratometry, refractive error, corneal irregularity, residual or recurrent central corneal opacities, and corneal haze were used to assess the outcome. Eight eyes of 6 patients were treated. The clinical diagnosis was confirmed histologically in all cases. Visual acuity improved significantly from 0.49 ± 0.2 logMAR to 0.14 ± 0.13 logMAR after a mean follow-up of 29 ± 14 (range 8-54) months. Residual central stromal opacities were noted in 5 of 8 eyes immediately postoperatively. Clinically significant recurrence of disease was noted in 1 eye. Keratometry and refraction remained stable, and no further surgical intervention was needed. Patients with stromal corneal dystrophies had worse outcome than those with Reis-Bücklers corneal dystrophy. In this case series, FLK provided both therapeutic and diagnostic intervention, delaying more invasive surgery. In vivo confocal microscopy showed signs of postoperative corneal stromal neuropathy.


CPS49-induced neurotoxicity does not cause limb patterning anomalies in developing chicken embryos.
Thalidomide notoriously caused severe birth defects, particularly to the limbs, in those exposed in utero following maternal use of the drug to treat morning sickness. How the drug caused these birth defects remains unclear. Many theories have been proposed including actions on the forming blood vessels. However, thalidomide survivors also have altered nerve patterns and the drug is known for its neurotoxic actions in adults following prolonged use. We have previously shown that CPS49, an anti-angiogenic analog of thalidomide, causes a range of limb malformations in a time-sensitive manner in chicken embryos. Here we investigated whether CPS49 also is neurotoxic and whether effects on nerve development impact upon limb development. We found that CPS49 is neurotoxic, just like thalidomide, and can cause some neuronal loss late developing chicken limbs, but only when the limb is already innervated. However, CPS49 exposure does not cause defects in limb size when added to late developing chicken limbs. In contrast, in early limb buds which are not innervated, CPS49 exposure affects limb area significantly. To investigate in more detail the role of neurotoxicity and its impact on chicken limb development we inhibited nerve innervation at a range of developmental timepoints through using β-bungarotoxin. We found that neuronal inhibition or ablation before, during or after limb outgrowth and innervation does not result in obvious limb cartilage patterning or number changes. We conclude that while CPS49 is neurotoxic, given the late innervation of the developing limb, and that neuronal inhibition/ablation throughout limb development does not cause similar limb patterning anomalies to those seen in thalidomide survivors, nerve defects are not the primary underlying cause of the severe limb patterning defects induced by CPS49/thalidomide.


Association between HLA DQB1 * 03 and cervical intra-epithelial neoplasia.
Cervical intraepithelial neoplasia (CIN) and cervical cancer have been shown to be strongly associated with infection by human papillomavirus (HPV). However, other factors may be contributory in the progression from normal epithelium to CIN and cervical cancer, since not all women with HPV infection develop disease. Recently, it was demonstrated that there is a high risk for cervical cancer and CIN in women with HLA DQB1 * 03 (RR = 7.1, p < 0.0009) (1). Subsequent reports have been conflicting, due to sample size, genetic heterogeneity and differences in the techniques employed for the detection of HLA DQB1 * 03. DNA from cervical smears of 178 women with CIN and 420 controls with normal cervical cytology was analyzed by polymerase chain reaction (PCR) with type-specific primers for HPV 16, 18, 31, and 33. The DNA from test and control samples were also analyzed by a novel PCR technique, which mutates the first base of codon 40 (DQ alleles) from T to G to create an artificial restriction site for an enzyme Mlu I that distinguish DQB1 * 03 from other alleles and are confirmed by digestion of amplified DNA with Mlu I. Further analysis of individual DQB1 * 03 alleles was performed using PCR and allele-specific primers. One hundred forty-four (34%) out of 420 controls (all HPV 16, 18, 31, or 33 negative and normal cytology), 37/66 (56%) of CIN I and 72/112 (64%) of CIN III were positive for DQB1 * 03 (trend test, p < 0.001, chi 2 = 37.3). A significant association was observed between DQB1 * 03 and CIN (odds ratio 3.03; 95% CI 2.11-3.45). Of women with CIN, 131/178 (73.5%) had HPV (types 16, 18, 31, or 33) infection. There was a significant association between DQB1 * 03 and presence of HPV (odds ratio 3.43; 95% CI 2.25-5.10). Homozygosity for DQB1 * 03 was more strongly associated with CIN than heterozygosity (odds ratios 4.0 and 2.63, respectively); and for the presence of HPV (odds ratio 4.47; 95% CI 2.58-7.77). HLA DQB1 * 0301 was the most strongly associated allele with CIN and HPV (odds ratios 2.53 and 2.63, respectively). HLA DQB1 * 03 is associated significantly with CIN and may be permissive for HPV infection. Further analysis of class II HLA typing in CIN is necessary to evaluate this association.


Hospital Readmissions Reduction Program: Safety-Net Hospitals Show Improvement, Modifications To Penalty Formula Still Needed.
Many observers are calling for modification of Medicare's Hospital Readmissions Reduction Program (HRRP) to relieve an unfair burden on safety-net hospitals, which serve low-income populations and consequently have relatively high readmission rates. To broaden the perspective on this issue, we addressed the fundamental question of whether the HRRP has been an effective tool for reducing thirty-day readmissions in safety-net hospitals. In the first three years of the program, these hospitals reduced readmissions for heart attack by 2.86 percentage points, heart failure by 2.78 percentage points, and pneumonia by 1.77 percentage points, and they also reduced the disparity between their readmission rates and those of other hospitals. While the fairness issue remains unresolved, it appears that safety-net hospitals have been able to respond to HRRP incentives.


Effect of oxygen withdrawal on active and passive electrical properties of arterially perfused rabbit ventricular muscle.
Oxygen withdrawal from myocardial cells leads to changes of the transmembrane action potential (mainly action potential shortening), to cellular uncoupling, and to changes of vascular permeability. This study was aimed at the simultaneous measurement of electrical activity and passive electrical properties (extracellular and intracellular longitudinal resistance) in arterially perfused rabbit papillary muscles under different conditions of changed oxygen supply. These included 1) complete anoxia (erythrocyte-free perfusate), 2) hypoxia (PO2 between 23-28 mm Hg, erythrocytes present) in the presence and absence of glucose, and 3) normoxia with erythrocyte-free perfusate. Similarly to myocardial ischemia, rapid cellular uncoupling occurred only after an initial stable period of approximately 17 minutes, and it required complete anoxia. The marked shortening of the action potential developed before cellular uncoupling. In six out of eight experiments, the fibers were inexcitable when uncoupling started. In severe hypoxia, no significant change of internal longitudinal resistance was observed over 35-40 minutes. The time course of the extracellular longitudinal resistance was different from the change in intracellular resistance: A marked decrease occurred almost immediately after the onset of oxygen withdrawal. This decrease was followed by a small increase in conduction velocity, which was most likely due to a change in the interstitial compartment (edema). It was observed during anoxic as well as during hypoxic perfusion. We conclude that 1) cellular uncoupling in arterially perfused tissue requires almost complete oxygen lack and occurs with a delay of more than 10 minutes, 2) marked action potential shortening precedes uncoupling, and therefore can not simply be attributed to an increase in free, intracellular calcium, and 3) vascular endothelial function is more sensitive to oxygen withdrawal than the myocyte.


A method for typing strains of Legionella pneumophila serogroup 1 by analysis of restriction fragment length polymorphisms.
A restriction fragment length polymorphism (RFLP) typing method for Legionella pneumophila serogroup 1 was developed. The method depended upon the use of cloned EcoR1 fragments from L. pneumophila (Knoxville-1) probing Nci1 restriction fragments of chromosomal DNA. Examination of strains of L. pneumophila which were apparently unrelated showed that inter-strain RFLPs were common, and these formed the basis of the typing scheme. The technique was found to be highly reproducible and discriminatory. When the RFLP data were compared to that obtained by monoclonal antibody (MAb) subgrouping both methods of strain differentiation gave consistent results. The isolates examined by either method were also sub-divided by the alternative technique. The analysis of RFLPs by cloned probes should be of considerable epidemiological value.


Telepsychiatry During the COVID-19 Pandemic: Development of a Protocol for Telemental Health Care.
The rapid spread of the Coronavirus disease 2019 (COVID-19) has forced most countries to take drastic public health measures, including the closure of most mental health outpatient services and some inpatient units. This has suddenly created the need to adapt and expand telepsychiatry care across the world. However, not all health care services might be ready to cope with this public health demand. The present study was set to create a practical and clinically useful protocol for telemental health care to be applied in the context of the current COVID-19 pandemic. A panel of psychiatrists from 15 different countries [covering all World Health Organization (WHO) regions] was convened. The panel used a combination of reactive Delphi technique and consensus development conference strategies to develop a protocol for the provision of telemental health care during the COVID-19 pandemic. The proposed protocol describes a semi-structured initial assessment and a series of potential interventions matching mild, moderate, or high-intensity needs of target populations. Telemedicine has become a pivotal tool in the task of ensuring the continuous provision of mental health care for the population, and the outlined protocol can assist with this task. The strength of this protocol lies in its practicality, clinical usefulness, and wide transferability, resulting from the diversity of the consensus group that developed it. Developed by psychiatrists from around the globe, the proposed protocol may prove helpful for many clinical and cultural contexts, assisting mental health care providers worldwide.


Digested Early Preterm Human Milk Suppresses Tumor Necrosis Factor-induced Inflammation and Cytotoxicity in Intestinal Epithelial Cells.
The aim of this study was to determine the effect of digested whole human milk (HM; first sample available after birth from mothers of premature infants) on inflammation, oxidative stress, and cytotoxicity in Caco-2 human intestinal epithelial cells stimulated with lipopolysaccharides or tumor necrosis factor (TNF) to mimic the potential in vivo insults facing the premature infant's gastrointestinal tract. Fully differentiated Caco-2 cells were exposed to digested HM (n = 10; samples from 10 different individuals) before stimulation with lipopolysaccharides, TNF, or no stimulation overnight. Inflammation was determined by production of interleukin-8, oxidative stress by levels of F2-isoprostane, and cytotoxicity by released lactate dehydrogenase. HM significantly suppressed interleukin-8 production and cytotoxicity in TNF-stimulated cells, while also suppressing cell death under baseline conditions. Individual HM samples differed widely in their ability to modulate cellular responses. Results from this study provide evidence that digested HM can reduce both an exaggerated inflammatory response and intestinal damage that contribute to the pathogenesis of necrotizing enterocolitis.


Atlas of Subcellular RNA Localization Revealed by APEX-Seq.
We introduce APEX-seq, a method for RNA sequencing based on direct proximity labeling of RNA using the peroxidase enzyme APEX2. APEX-seq in nine distinct subcellular locales produced a nanometer-resolution spatial map of the human transcriptome as a resource, revealing extensive patterns of localization for diverse RNA classes and transcript isoforms. We uncover a radial organization of the nuclear transcriptome, which is gated at the inner surface of the nuclear pore for cytoplasmic export of processed transcripts. We identify two distinct pathways of messenger RNA localization to mitochondria, each associated with specific sets of transcripts for building complementary macromolecular machines within the organelle. APEX-seq should be widely applicable to many systems, enabling comprehensive investigations of the spatial transcriptome.


Learning from others: shall the last be the first?
The cross-national exchange of ideas and experience in health care reform has, in recent years, reached epidemic proportions. Taking stock of this suggests the need for critical caution. The various participants in the process have different motives; models may be exported before they have been fully tested; information may be sought chiefly as political ammunition; and selective perception often distorts the evidence. A more helpful approach might be to concentrate on evaluating the experience of countries over time, rather than concentrating on the latest fashionable panacea, and to focus more on the process of introducing change.


Radioprotective effect of moderate wine consumption in patients with breast carcinoma.
Given the high cost and side effects of radioprotective agents such as amifostine, attention has been focused on potentially equally effective but less expensive and toxic natural substances. We evaluated the potential radioprotective effects of wine in preventing skin toxicity in patients with breast cancer. Before treatment, the medical history and habits of patients were assessed and the information recorded in their clinical folders. Patients were divided into three groups based on the dose/fractionation scheme used: control group, 60.4 Gy (standard technique); Modulated Accelerated Radiotherapy in Adjuvant treatment of breast cancer (MARA)-1 protocol group, 44 Gy (concomitant boost to tumoral bed); and MARA-2 protocol group, 60 Gy (concomitant boost to tumoral bed). The impact of the following variables on acute skin toxicity was evaluated by chart review: radiotherapy protocol, planning target volume (PTV), comorbidity (e.g., hypertension and diabetes), hemoglobin level before therapy, adjuvant hormone therapy, adjuvant chemotherapy, cigarette smoking, and drinking habits. The study population consisted of 348 patients. More severe skin toxicity was significantly associated with the radiotherapy protocol (p < 0.001) and median PTV (p = 0.005). In addition, the incidence of acute toxicity of Grade 2 or greater was higher in patients without alcohol intake (38.4% vs. 22.3%, p = 0.021). The daily amount of alcohol intake also influenced the incidence of skin toxicity, with an incidence of 38.4% in patients with no wine intake, 31.8% in patients drinking half a glass per day, 13.6% in patients drinking one glass per day, and 35.0% in patients drinking two glasses per day. Multivariate analysis showed that wine intake, PTV, and radiotherapy protocol were all significantly correlated with acute toxicity. Our results indicate that wine may have a radioprotective effect; however, prospective studies are needed to confirm this beneficial effect of wine and its components.


Terrein biosynthesis in Aspergillus terreus and its impact on phytotoxicity.
Terrein is a fungal metabolite with ecological, antimicrobial, antiproliferative, and antioxidative activities. Although it is produced by Aspergillus terreus as one of its major secondary metabolites, not much is known about its biosynthetic pathway. Here, we describe an unexpected discovery of the terrein biosynthesis gene locus made while we were looking for a PKS gene involved in production of conidia coloration pigments common for Aspergilli. The gene, ATEG_00145, here named terA, is essential for terrein biosynthesis and heterologous production of TerA in Aspergillus niger revealed an unusual plasticity in the products formed, yielding a mixture of 4-hydroxy-6-methylpyranone, orsellinic acid, and 6,7-dihydroxymellein. Biochemical and molecular genetic analyses indicate a low extension cycle specificity of TerA. Furthermore, 6-hydroxymellein was identified as a key intermediate in terrein biosynthesis. We find that terrein production is highly induced on plant-derived media, that terrein has phytotoxic activity on plant growth, and induces lesions on fruit surfaces.


Lead-free hunting rifle ammunition: product availability, price, effectiveness, and role in global wildlife conservation.
Proposals to end the use of lead hunting ammunition because of the established risks of lead exposure to wildlife and humans are impeded by concerns about the availability, price, and effectiveness of substitutes. The product availability and retail prices of different calibers of lead-free bullets and center-fire rifle ammunition were assessed for ammunition sold in the USA and Europe. Lead-free bullets are made in 35 calibers and 51 rifle cartridge designations. Thirty-seven companies distribute internationally ammunition made with lead-free bullets. There is no major difference in the retail price of equivalent lead-free and lead-core ammunition for most popular calibers. Lead-free ammunition has set bench-mark standards for accuracy, lethality, and safety. Given the demonstrated wide product availability, comparable prices, and the effectiveness of high-quality lead-free ammunition, it is possible to phase out the use of lead hunting ammunition world-wide, based on progressive policy and enforceable legislation.


Risk factors for osteonecrosis of the jaws: a case-control study from the CONDOR dental PBRN.
Case reports and cohort studies have linked bisphosphonate therapy and osteonecrosis of the jaws (ONJ), but neither causality nor specific risks for lesion development have been clearly established. We conducted a 1:3 case-control study with three dental Practice-based Research Networks, using dentist questionnaires and patient interviews for collection of data on bisphosphonate therapy, demographics, co-morbidities, and dental and medical treatments. Multivariable logistic regression analyses tested associations between bisphosphonate use and other risk factors with ONJ. We enrolled 191 ONJ cases and 573 controls in 119 dental practices. Bisphosphonate use was strongly associated with ONJ (odds ratios [OR] 299.5 {95%CI 70.0-1282.7} for intravenous [IV] use and OR = 12.2 {4.3-35.0} for oral use). Risk markers included local suppuration (OR = 7.8 {1.8-34.1}), dental extraction (OR = 7.6 {2.4-24.7}), and radiation therapy (OR = 24.1 {4.9-118.4}). When cancer patients (n = 143) were excluded, bisphosphonate use (OR = 7.2 {2.1-24.7}), suppuration (OR = 11.9 {2.0-69.5}), and extractions (OR = 6.6 {1.6-26.6}) remained associated with ONJ. Higher risk of ONJ began within 2 years of bisphosphonate initiation and increased four-fold after 2 years. Both IV and oral bisphosphonate use were strongly associated with ONJ. Duration of treatment > 2 years; suppuration and dental extractions were independent risk factors for ONJ.


Porokeratotic adnexal ostial naevus: review on the entity and therapeutic approach.
Porokeratotic adnexal ostial nevus (PAON) is a term that encompass porokeratotic eccrine ostial and dermal duct naevus and porokeratotic eccrine and hair follicle naevus. PAON is a rare adnexal hamartoma characterized by the presence of a cornoid lamella exclusively overlying eccrine acrosyringia or over both, eccrine acrosyringia and hair follicles. Most of the articles on PAON report its clinical or histopathological characteristics but literature review offer scarce data on the evolution or treatment. We have reviewed it. We report an 8-year-old boy with presence of bilateral asymtomatic verrucous lesions mostly in left side of the body without other added abnormalities through the twelve-year follow-up. As our case presented cornoid lamella overlying eccrine acrosyringia and hair follicles we diagnosed PEHFN. He received treatment with CO2 ablation in 2004 and in 2012 with marked improvement. Review of the previous reported cases of PAON show that it is mostly unilateral and that some patients have coexisting problems like seizures, left hemiparesis, scoliosis, deafness, developmental delay, palmoplantar keratoderma, psoriasis, hyperthyroidism, polinerupathy, breast hipoplasia and KID. PAON responded with tazaroten gel, dithranol short-contact treatment and topical photodynamic therapy but laser treatments appear to be more efficient in most patients.


The patient-therapist relationship: beliefs that shape care.
The results of a previous inquiry suggest that three images of occupational therapists dominate patients' stories about them: the images of technician, parent, and collaborator or friend. These ways of being in practice can be said to reflect the various understandings that therapists have about how to enact the profession's commitment to both competence and caring. When therapists act as technicians or authoritarian parents, patients register their disappointment over a valuation of competence that excludes caring actions. In a more current inquiry into the climate of caring, patients and caregivers reflect about the current health care system and identify three societal constructs that shape a preference for competence over caring: (a) emphasis on the rational fixing of the health care problem, (b) overreliance on methods and protocols, and (c) a health care system driven by business, efficiency, and profit. Occupational therapists who are concerned about complaints that the health care system is increasingly uncaring might benefit from a consideration of the extent to which societal beliefs shape the manner in which they care.


Prospective study of seaweed consumption and thyroid cancer incidence in women: the Japan collaborative cohort study.
Excess intake of iodine is a suspected risk factor for thyroid cancer. Previous epidemiological research from Japan reported that daily intake of seaweed was associated with a four-fold higher risk in postmenopausal women, whereas others reported a null association. A major source of iodine intake in Japan is from edible seaweeds, and it is reported to be among the highest in the world. We examined the association between seaweed intake frequency and the risk of thyroid cancer in women in the Japan Collaborative Cohort Study followed from 1988 to 2009. Seaweed intake, together with other lifestyle-related information was collected using a self-administered questionnaire at baseline. Seaweed intake frequency was categorized as follows: 1-2 times/week or less, 3-4 times/week, and almost daily. Hazard ratios and the 95% confidence intervals of thyroid cancer incidence according to seaweed intake frequency were estimated using Cox proportional hazards regression. During 447 876 person-years of follow-up (n=35 687), 94 new cases of thyroid cancer were identified. The crude incidence rate was 20.9 per 100 000 person-years. The hazard ratio of thyroid cancer in women who consumed seaweed daily compared with women who ate it 1-2 times/week or less was 1.15 (95% confidence interval: 0.69-1.90, P for trend=0.59). Further analyses did not indicate any association between seaweed intake and the risk of thyroid cancer on statistically adjusting for potential confounding variables as well as on stratification by menopausal status. The present study did not find an association between seaweed intake and thyroid cancer incidence in premenopausal or in postmenopausal women.


Mechanical Stimuli Affect Escherichia coli Heat-Stable Enterotoxin-Cyclic GMP Signaling in a Human Enteroid Intestine-Chip Model.
Modeling host-pathogen interactions with human intestinal epithelia using enteroid monolayers on permeable supports (such as Transwells) represents an alternative to animal studies or use of colon cancer-derived cell lines. However, the static monolayer model does not expose epithelial cells to mechanical forces normally present in the intestine, including luminal flow and serosal blood flow (shear force) or peristaltic forces. To determine the contribution of mechanical forces in the functional response of human small intestine to a virulence factor of a pathogenic intestinal bacterium, human jejunal enteroids were cultured as monolayers in microengineered fluidic-based Organ-Chips (Intestine-Chips) exposed to enterotoxigenic Escherichia coli heat-stable enterotoxin A (ST) and evaluated under conditions of static fluid, apical and basolateral flow, and flow plus repetitive stretch. Application of flow increased epithelial cell height and apical and basolateral secretion of cyclic GMP (cGMP) under baseline, unstimulated conditions. Addition of ST under flow conditions increased apical and basolateral secretion of cGMP relative to the level under static conditions but did not enhance intracellular cGMP accumulation. Cyclic stretch did not have any significant effect beyond that contributed by flow. This study demonstrates that fluid flow application initiates changes in intestinal epithelial cell characteristics relative to those of static culture conditions under both baseline conditions and with exposure to ST enterotoxin and suggests that further investigations of the application of these mechanical forces will provide insights into physiology and pathophysiology that more closely resemble intact intestine than study under static conditions.


Identifying abdominal aortic aneurysm risk factors in postmenopausal women.
Evaluation of: Lederle FA, Larson JC, Margolis KL et al.: Abdominal aortic aneurysm events in the Women's Health Initiative: cohort study. Br. Med. J. 337, A1724 (2008). A linked cohort study of 161,808 postmenopausal women aged 50-79 years enrolled in the Women's Health Initiative was conducted during which participants were followed for the incidence of abdominal aortic aneurysm repair or rupture. This study evaluated the association between potential risk factors and subsequent abdominal aortic aneurysm events in women. A total of 467 women reported a diagnosis of abdominal aortic aneurysm before entering the study or during participation, with 184 aneurysm-related events identified. Abdominal aortic aneurysm events were strongly associated with age and smoking and negatively associated with diabetes and baseline use of postmenopausal hormone supplementation. Previous studies investigating abdominal aortic aneurysm have focused primarily on men, with little reliable information available on women. This study contributes a large female cohort to provide better insight into gender-specific abdominal aortic aneurysm risks and disease associations.


Single-cell cloning of colon cancer stem cells reveals a multi-lineage differentiation capacity.
Colon carcinoma is one of the leading causes of death from cancer and is characterized by a heterogenic pool of cells with distinct differentiation patterns. Recently, it was reported that a population of undifferentiated cells from a primary tumor, so-called cancer stem cells (CSC), can reconstitute the original tumor on xenotransplantation. Here, we show that spheroid cultures of these colon CSCs contain expression of CD133, CD166, CD44, CD29, CD24, Lgr5, and nuclear beta-catenin, which have all been suggested to mark the (cancer) stem cell population. More importantly, by using these spheroid cultures or freshly isolated tumor cells from multiple colon carcinomas, we now provide compelling evidence to indicate that the capacity to propagate a tumor with all differentiated progeny resides in a single CSC. Single-cell-cloned CSCs can form an adenocarcinoma on xenotransplantation but do not generate the stroma within these tumors. Moreover, they can self-renew and are capable of multilineage differentiation. Further analysis indicated that the lineage decision is dictated by phosphoinositide 3-kinase (PI3K) signaling in CSCs. These data support the hypothesis that tumor hierarchy can be traced back to a single CSC that contains multilineage differentiation capacity, and provides clues to the regulation of differentiation in colon cancers in vivo.


Monitoring epidemic viral respiratory infections using one-step real-time triplex RT-PCR targeting influenza A and B viruses and respiratory syncytial virus.
Rapid and specific diagnosis of influenza A/B and respiratory syncytial virus (RSV) viruses is needed for optimal management of patients with acute respiratory infections. In this study, a one-step triplex real-time RT-PCR assay was developed for rapid diagnosis of influenza A/B and RSV infections to optimize diagnosis efficiency of acute respiratory infections. Cell-culture supernatants and clinical samples were used to evaluate specificity and sensitivity of the assay. The assay was used routinely during two winter epidemics for testing respiratory specimens from 2,417 patients. The limit of detection in cell-culture supernatant was 1-10 plaque forming units/input (influenza A/B) and 2 × 10(-2) 50% tissue culture infectious dose/input (RSV). In clinical samples, the assay was as sensitive as commercial molecular assays for the detection of each influenza A/B and RSV (Flu-A/B and RSV-A/B r-gene™) individually, and far more sensitive than antigen detection. During the winter 2008-2009, the assay identified 145 RSV, 42 influenza A, and one mixed RSV-influenza A infections among 298 patients. The next winter, the assay was used in two independent hospital laboratory settings. 776 patients were tested in one hospital and 1,343 in the other, resulting in 184 and 501 RSV, 133 and 150 influenza A, and 1 and 11 mixed RSV-influenza A infections, respectively, being detected. This new user-friendly assay allows rapid (within hours), effective molecular diagnosis of single or mixed infections involving influenza A (including seasonal A H1N1 and H3N2, and A(H1N1) 2009), influenza B, and RSV(A/B). The assay is very valuable for managing patients during winter epidemics when influenza and respiratory syncytial viruses co-circulate.


Suppression of monocular visual direction under fused binocular stimulation: evoked potential measurements.
Visual evoked potentials (VEPs) were recorded in response to a vernier onset/offset target presented to one eye that was combined with matching static targets in the other eye. The monocular response was dominated by a negative peak at 160 ms that occurred after a set of offsets was introduced into a one-dimensional random bar pattern. The static targets produced no discernible VEP response by themselves, but when fused binocularly with the oscillating vernier target, they produced shifts in perceived visual direction that influenced the VEP response. A vernier target fused with static vertical bars was perceived to alternate in depth between a flat surface and one broken into two interleaved surfaces. The response to this "surface-breaking" was as large or larger than the response to the monocular vernier offset. This response was much reduced when the oscillating vernier was fused with a static offset vernier (5' offset) that produced a percept of segregated regions moving in depth. Apparently, the VEP is strongly driven by shifts in visual direction that alter surface, texture, or contour contiguity.


Metal Concentrations in e-Cigarette Liquid and Aerosol Samples: The Contribution of Metallic Coils.
Electronic cigarettes (e-cigarettes) generate an aerosol by heating a solution (e-liquid) with a metallic coil. Whether metals are transferred from the coil to the aerosol is unknown. Our goal was to investigate the transfer of metals from the heating coil to the e-liquid in the e-cigarette tank and the generated aerosol. We sampled 56 e-cigarette devices from daily e-cigarette users and obtained samples from the refilling dispenser, aerosol, and remaining e-liquid in the tank. Aerosol liquid was collected via deposition of aerosol droplets in a series of conical pipette tips. Metals were reported as mass fractions (μg/kg) in liquids and converted to mass concentrations (mg/m3) for aerosols. Median metal concentrations (μg/kg) were higher in samples from the aerosol and tank vs. the dispenser (all p<0.001): 16.3 and 31.2 vs. 10.9 for Al; 8.38 and 55.4 vs. <0.5 for Cr; 68.4 and 233 vs. 2.03 for Ni; 14.8 and 40.2 vs. 0.476 for Pb; and 515 and 426 vs. 13.1 for Zn. Mn, Fe, Cu, Sb, and Sn were detectable in most samples. Cd was detected in 0.0, 30.4, and 55.1% of the dispenser, aerosol, and tank samples respectively. Arsenic was detected in 10.7% of dispenser samples (median 26.7μg/kg) and these concentrations were similar in aerosol and tank samples. Aerosol mass concentrations (mg/m3) for the detected metals spanned several orders of magnitude and exceeded current health-based limits in close to 50% or more of the samples for Cr, Mn, Ni, and Pb. Our findings indicate that e-cigarettes are a potential source of exposure to toxic metals (Cr, Ni, and Pb), and to metals that are toxic when inhaled (Mn and Zn). Markedly higher concentrations in the aerosol and tank samples versus the dispenser demonstrate that coil contact induced e-liquid contamination. https://doi.org/10.1289/EHP2175.


Delivery and Payment Redesign to Reduce Disparities in High Risk Postpartum Care.
Purpose This paper describes the implementation of an innovative program that aims to improve postpartum care through a set of coordinated delivery and payment system changes designed to use postpartum care as an opportunity to impact the current and future health of vulnerable women and reduce disparities in health outcomes among minority women. Description A large health care system, a Medicaid managed care organization, and a multidisciplinary team of experts in obstetrics, health economics, and health disparities designed an intervention to improve postpartum care for women identified as high-risk. The program includes a social work/care management component and a payment system redesign with a cost-sharing arrangement between the health system and the Medicaid managed care plan to cover the cost of staff, clinician education, performance feedback, and clinic/clinician financial incentives. The goal is to enroll 510 high-risk postpartum mothers. Assessment The primary outcome of interest is a timely postpartum visit in accordance with NCQA healthcare effectiveness data and information set guidelines. Secondary outcomes include care process measures for women with specific high-risk conditions, emergency room visits, postpartum readmissions, depression screens, and health care costs. Conclusion Our evidence-based program focuses on an important area of maternal health, targets racial/ethnic disparities in postpartum care, utilizes an innovative payment reform strategy, and brings together insurers, researchers, clinicians, and policy experts to work together to foster health and wellness for postpartum women and reduce disparities.


Paleogenomics of echinoderms.
Paleogenomics propels the meaning of genomic studies back through hundreds of millions of years of deep time. Now that the genome of the echinoid Strongylocentrotus purpuratus is sequenced, the operation of its genes can be interpreted in light of the well-understood echinoderm fossil record. Characters that first appear in Early Cambrian forms are still characteristic of echinoderms today. Key genes for one of these characters, the biomineralized tissue stereom, can be identified in the S. purpuratus genome and are likely to be the same genes that were involved with stereom formation in the earliest echinoderms some 520 million years ago.


Atomic structure of the DNA repair [4Fe-4S] enzyme endonuclease III.
The crystal structure of the DNA repair enzyme endonuclease III, which recognizes and cleaves DNA at damaged bases, has been solved to 2.0 angstrom resolution with an R factor of 0.185. This iron-sulfur [4Fe-4S] enzyme is elongated and bilobal with a deep cleft separating two similarly sized domains: a novel, sequence-continuous, six-helix domain (residues 22 to 132) and a Greek-key, four-helix domain formed by the amino-terminal and three carboxyl-terminal helices (residues 1 to 21 and 133 to 211) together with the [4Fe-4S] cluster. The cluster is bound entirely within the carboxyl-terminal loop with a ligation pattern (Cys-X6-Cys-X2-Cys-X5-Cys) distinct from all other known [4Fe-4S] proteins. Sequence conservation and the positive electrostatic potential of conserved regions identify a surface suitable for binding duplex B-DNA across the long axis of the enzyme, matching a 46 angstrom length of protected DNA. The primary role of the [4Fe-4S] cluster appears to involve positioning conserved basic residues for interaction with the DNA phosphate backbone. The crystallographically identified inhibitor binding region, which recognizes the damaged base thymine glycol, is a seven-residue beta-hairpin (residues 113 to 119). Location and side chain orientation at the base of the inhibitor binding site implicate Glu112 in the N-glycosylase mechanism and Lys120 in the beta-elimination mechanism. Overall, the structure reveals an unusual fold and a new biological function for [4Fe-4S] clusters and provides a structural basis for studying recognition of damaged DNA and the N-glycosylase and apurinic/apyrimidinic-lyase mechanisms.


Tests of behavioral-economic assessments of relative reinforcer efficacy II: economic complements.
This experiment was conducted to test the predictions of two behavioral-economic approaches to quantifying relative reinforcer efficacy. The normalized demand analysis suggests that characteristics of averaged normalized demand curves may be used to predict progressive-ratio breakpoints and peak responding. By contrast, the demand analysis holds that traditional measures of relative reinforcer efficacy (breakpoint, peak response rate, and choice) correspond to specific characteristics of non-normalized demand curves. The accuracy of these predictions was evaluated in rats' responding for food or water: two reinforcers known to function as complements. Consistent with the first approach, predicted peak normalized response output values obtained under single-schedule conditions ordinally predicted progressive-ratio breakpoints and peak response rates obtained in a separate condition. Combining the minimum-needs hypothesis with the normalized demand analysis helped to interpret prior findings, but was less useful in predicting choice between food and water--two strongly complementary reinforcers. Predictions of the demand analysis had mixed success. Peak response outputs predicted from the non-normalized water demand curves were significantly correlated with obtained peak responding for water in a separate condition, but none of the remaining three predicted correlations was statistically significant. The demand analysis fared better in predicting choice--relative consumption of food and water under single schedules of reinforcement predicted preference under concurrent schedules significantly better than chance.


Identification of microRNAs specifically expressed in hepatitis C virus-associated hepatocellular carcinoma.
Although several studies have investigated the association of miRNAs with hepatocellular carcinoma (HCC), the data published so far are not concordant. A reason for these discrepancies may be the fact that most studies used the nontumorous tissue surrounding the HCC lesion as a control, which is almost invariably affected by cirrhosis or chronic hepatitis, as well as other pathological conditions such as hepatitis B virus (HBV) or hepatitis C virus (HCV) infection. Moreover, HCC is often analyzed as a single group regardless of the different viral etiologies. The miRNAs differentially expressed in HCV-related HCC were investigated by comparing the tumorous tissues to a wide range of liver specimens, including healthy livers obtained from liver donors and patients who underwent liver resection for angioma, in addition to tissues from various acute and chronic liver diseases, including HCV-related cirrhosis not associated with HCC, HCV-related cirrhosis associated with HCC and HBV-associated acute liver failure. The whole set of 2,226 human miRNAs were examined, including 1,121 pre-miRNAs and 1,105 mature miRNAs, available in a microarray platform. Stringent statistical methods were applied to reduce the risk of false discoveries to less than 1%. These data identified 18 miRNAs exclusively expressed in HCV-associated HCC, characterized by high specificity and selectivity versus all other liver diseases and healthy conditions and connected into a regulatory network pivoting on p53, phosphatase and tensin homolog and all-trans retinoic acid signaling.


Response of the microcirculation in rat cremaster muscle to peripheral and central sympathetic stimulation.
Microvascular diameter and flow responses to peripheral and central sympathetic stimulation were measured in different segments of the arteriolar network in rat cremaster muscle. For peripheral stimulation, a bipolar electrode was placed on the internodal segment of the lumbar sympathetic chain between ganglia L1 and L2. For central stimulation, a bipolar electrode was stereotaxically implanted in the posterior hypothalamus. Inside vessel diameter, red blood cell velocity, and volumetric flow rate were recorded in response to electrical stimulations of varying magnitude in four series-coupled segments of the arteriolar network: 1A, 2A, 3A, and 4A. Systemic arterial pressure was also monitored. The vasoconstriction and flow reduction produced by stimulation of the lumbar chain was graded with the frequency of stimulation over the range of 0.5-16.0 Hz in all arteriolar segments. Examination of the relation between stimulation frequency and vasoconstrictor response measured as percent of control diameter indicated a sequence of responsiveness to peripheral stimulation where 4A = 3A greater than 2A = 1A. No changes in diameter were recorded in the venous microcirculation at any level of stimulation. Stimulation of the posterior hypothalamus with currents of 38-300 microA for 60 seconds produced graded vasoconstriction in only 3A and 4A vessels. Fluorescence histochemistry for biogenic amines was used to examine the distribution of innervation to the microvasculature. All segments of the arteriolar network from 1A to 4A possessed an adrenergic innervation; no vessels of the venous network were found to be innervated. The results indicate that the pattern of response of the arteriolar network in rat cremaster muscle to peripheral and central sympathetic stimulation is segmentally differentiated and consistent with the distribution of the vasomotor innervation.


Occurrence of serious injury in real-world side impacts of vehicles with good side-impact protection ratings.
The Insurance Institute for Highway Safety (IIHS) introduced its side impact consumer information test program in 2003. Since that time, side airbags and structural improvements have been implemented across the fleet and the proportion of good ratings has increased to 93% of 2012-2014 model year vehicles. Research has shown that drivers of good-rated vehicles are 70% less likely to die in a left-side crash than drivers of poor-rated vehicles. Despite these improvements, side impact fatalities accounted for about one quarter of passenger vehicle occupant fatalities in 2012. This study is a detailed analysis of real-world cases with serious injury resulting from side crashes of vehicles with good ratings in the IIHS side impact test. NASS-CDS and Crash Injury Research and Engineering Network (CIREN) were queried for occupants of good-rated vehicles who sustained an Abbreviated Injury Scale (AIS) ≥ 3 injury in a side-impact crash. The resulting 110 cases were categorized by impact configuration and other factors that contributed to injury. Patterns of impact configuration, restraint performance, and occupant injury were identified and discussed in the context of potential upgrades to the current IIHS side impact test. Three quarters of the injured occupants were involved in near-side impacts. For these occupants, the most common factors contributing to injury were crash severities greater than the IIHS test, inadequate side-airbag performance, and lack of side-airbag coverage for the injured body region. In the cases where an airbag was present but did not prevent the injury, occupants were often exposed to loading centered farther forward on the vehicle than in the IIHS test. Around 40% of the far-side occupants were injured from contact with the struck-side interior structure, and almost all of these cases were more severe than the IIHS test. The remaining far-side occupants were mostly elderly and sustained injury from the center console, instrument panel, or seat belt. In addition, many far-side occupants were likely out of position due to events preceding the side impact and/or being unbelted. Individual changes to the IIHS side impact test have the potential to reduce the number of serious injuries in real-world crashes. These include impacting the vehicle farther forward (relevant to 28% of all cases studied), greater test severity (17%), the inclusion of far-side occupants (9%), and more restrictive injury criteria (9%). Combinations of these changes could be more effective.


Impacts of experimental defoliation on native and invasive saplings: are native species more resilient to canopy disturbance?
Many non-native, invasive woody species in mesic forests of North America are both shade tolerant and more productive than their native counterparts, but their ability to tolerate disturbances remains unclear. In particular, complete defoliation associated with herbivory and extreme weather events may have larger impacts on invaders if natives maintain greater resource reserves to support regrowth. On the other hand, invaders may be more resilient to partial defoliation by means of upregulation of photosynthesis or may be better able to take advantage of canopy gaps to support refoliation. Across a light gradient, we measured radial growth, new leaf production, non-structural carbohydrates (NSCs), chlorophyll content and survival in response to varying levels of defoliation in saplings of two native and two invasive species that commonly co-occur in deciduous forests of Eastern North America. Individuals were subjected to one of the four leaf removal treatments: no-defoliation controls, 50% defoliation over three growing seasons, 100% defoliation over one growing season and 100% defoliation over two growing seasons. Contrary to our hypothesis, native and invasive species generally did not differ in defoliation responses, although invasive species experienced more pronounced decreases in leaf chlorophyll following full defoliation and native species' survival was more dependent on light availability. Radial growth progressively decreased with increasing defoliation intensity, and refoliation mass was largely a function of sapling size. Survival rates for half-defoliated saplings did not differ from controls (90% of saplings survived), but survival rates in fully defoliated individuals over one and two growing seasons were reduced to 45 and 15%, respectively. Surviving defoliated saplings generally maintained control NSC concentrations. Under high light, chlorophyll concentrations were higher in half-defoliated saplings compared with controls, which may suggest photosynthetic upregulation. Our results indicate that native and invasive species respond similarly to defoliation, despite the generally faster growth strategy of invaders.


GIS-based estimation of exposure to particulate matter and NO2 in an urban area: stochastic versus dispersion modeling.
Stochastic modeling was used to predict nitrogen dioxide and fine particles [particles collected with an upper 50% cut point of 2.5 microm aerodynamic diameter (PM2.5)] levels at 1,669 addresses of the participants of two ongoing birth cohort studies conducted in Munich, Germany. Alternatively, the Gaussian multisource dispersion model IMMIS(net/em) was used to estimate the annual mean values for NO2 and total suspended particles (TSP) for the 40 measurement sites and for all study subjects. The aim of this study was to compare the measured NO2 and PM2.5 levels with the levels predicted by the two modeling approaches (for the 40 measurement sites) and to compare the results of the stochastic and dispersion modeling for all study infants (1,669 sites). NO2 and PM2.5 concentrations obtained by the stochastic models were in the same range as the measured concentrations, whereas the NO2 and TSP levels estimated by dispersion modeling were higher than the measured values. However, the correlation between stochastic- and dispersion-modeled concentrations was strong for both pollutants: At the 40 measurement sites, for NO2, r = 0.83, and for PM, r = 0.79; at the 1,669 cohort sites, for NO2, r = 0.83 and for PM, r = 0.79. Both models yield similar results regarding exposure estimate of the study cohort to traffic-related air pollution, when classified into tertiles; that is, 70% of the study subjects were classified into the same category. In conclusion, despite different assumptions and procedures used for the stochastic and dispersion modeling, both models yield similar results regarding exposure estimation of the study cohort to traffic-related air pollutants.


Asbestos exposure, pleural mesothelioma, and serum osteopontin levels.
We investigated the presence of osteopontin in pleural mesothelioma and determined serum osteopontin levels in three populations: subjects without cancer who were exposed to asbestos, subjects without cancer who were not exposed to asbestos, and patients with pleural mesothelioma who were exposed to asbestos. A group of 69 subjects with asbestos-related nonmalignant pulmonary disease were compared with 45 subjects without exposure to asbestos and 76 patients with surgically staged pleural mesothelioma. Tumor tissue was examined for osteopontin by immunohistochemical analysis, and serum osteopontin levels were measured by an enzyme-linked immunosorbent assay. There were no significant differences in mean (+/-SE) serum osteopontin levels between age-matched subjects with exposure to asbestos and subjects without exposure to asbestos (30+/-3 ng per milliliter and 20+/-4 ng per milliliter, respectively; P=0.06). In the group with exposure to asbestos, elevated serum osteopontin levels were associated with pulmonary plaques and fibrosis (56+/-13 ng per milliliter) but not with normal radiographic findings (21+/-5 ng per milliliter), plaques alone (23+/-3 ng per milliliter), or fibrosis alone (32+/-7 ng per milliliter) (P=0.004). Serum osteopontin levels were significantly higher in the group with pleural mesothelioma than in the group with exposure to asbestos (133+/-10 ng per milliliter vs. 30+/-3 ng per milliliter, P<0.001). Immunohistochemical analysis revealed osteopontin staining of the tumor cells in 36 of 38 samples of pleural mesothelioma. An analysis of serum osteopontin levels comparing the receiver-operating-characteristic curve in the group exposed to asbestos with that of the group with mesothelioma had a sensitivity of 77.6 percent and a specificity of 85.5 percent at a cutoff value of 48.3 ng of osteopontin per milliliter. Subgroup analysis comparing patients with stage I mesothelioma with subjects with exposure to asbestos revealed a sensitivity of 84.6 percent and a specificity of 88.4 percent at a cutoff value of 62.4 ng of osteopontin per milliliter. Serum osteopontin levels can be used to distinguish persons with exposure to asbestos who do not have cancer from those with exposure to asbestos who have pleural mesothelioma.


A proteomic signature for CNS adaptations to the valence of environmental stimulation.
Environmental Enrichment leads to a significant improvement in long-term performance across a range of cognitive functions in mammals and it has been shown to produce an increased synaptic density and neurogenesis. Nevertheless it is still an open question as to whether some key aspects of spatial learning & memory and procedural learning might be embodied by different molecular pathways to those of social cognition. Associated with synaptic changes and potentially underlying conditions, the Ras-ERK pathway has been proposed to be the primary mediator of in vivo adaptations to environmental enrichment, acting via the downstream Ras-ERK signalling kinase MSK1 and the transcription factor CREB. Herein, we show that valence of environmental stimulation increased social competition and that this is associated with a specific proteomic signature in the frontal lobe but notably not in the hippocampus. Specifically, we show that altering the valence of environmental stimuli affected the level of social competition, with mice from negatively enriched environments winning significantly more encounters-even though mice from positive were bigger and should display dominance. This behavioural phenotype was accompanied by changes in the proteome of the fronto-ventral pole of the brain, with a differential increase in the relative abundance of proteins involved in the mitochondrial metabolic processes of the TCA cycle and respiratory processes. Investigation of this proteomic signature may pave the way for the elucidation of novel pathways underpinning the behavioural changes caused by negative enrichment and further out understanding of conditions whose core feature is increased social competition.


Development and application of real-time PCR for specific detection of Lepeophtheirus salmonis and Caligus elongatus larvae in Scottish plankton samples.
Lepeophtheirus salmonis and Caligus elongatus are important parasites of wild and cultured salmonids in the Northern Hemisphere. These species, generically referred to as sea lice, are estimated to cost the Scottish aquaculture industry in excess of pound 25 million per annum. There is great interest in countries such as Ireland, Scotland, Norway and Canada to sample sea lice larvae in their natural environment in order to understand lice larvae distribution and improve parasite control. Microscopy is currently relied on for use in the routine identification of sea lice larvae in plankton samples. This method is, however, limited by its time-consuming nature and requirement for highly skilled personnel. The development of alternative methods for the detection of sea lice larvae which might be used to complement and support microscopic examinations of environmental samples is thus desirable. In this study, a genetic method utilising a real-time PCR Taqman-MGB probe-based assay targeting the mitochondrial cytochrome oxidase I (mtCOI) gene was developed, which allowed species-specific detection of L. salmonis and C. elongatus larvae from unsorted natural and spiked plankton samples. Real-time PCR is a rapid, sensitive, highly specific and potentially quantitative technique. This study demonstrated its suitability for the routine identification of L. salmonis and C. elongatus in mixed plankton samples. The real-time PCR assay developed has considerable potential for use in complementing, supporting and reducing reliance on time-consuming conventional microscopic examination for the specific identification of sea lice larvae in plankton samples.


Participation of bacteriorhodopsin active-site lysine backbone in vibrations associated with retinal photochemistry.
Bacteriorhodopsin (bR) has been biosynthetically prepared with lysine deuterated at its alpha carbon (C alpha--H). The labeled membranes containing bR were investigated by difference Fourier transform infrared (FTIR) spectroscopy. It has been derived from K/bR and M/bR difference spectra (K and M are photocycle intermediates) that several bands previously assigned to the retinal chromophore are coupled to the C alpha--H. The vibrational modes that exhibit this coupling are principally associated with C15--H and N--H vibrations. [C alpha--2H]Lysine-labeled bR was fragmented enzymatically, and bR structures were regenerated with the C alpha--2H label either on lysine-216 and -172 or on the remaining five lysine residues of the protein. FTIR studies of the regenerated bR system, together with methylation of all lysines except the active-site lysine, reveal that the changes observed due to backbone labeling arise from the active-site lysine. The intensity of the C15--H out-of-plane wag is interpreted as a possible indication of a twist around the C15 = N bond.


Who's minding the shop? The role of Canadian research ethics boards in the creation and uses of registries and biobanks.
The amount of research utilizing health information has increased dramatically over the last ten years. Many institutions have extensive biobank holdings collected over a number of years for clinical and teaching purposes, but are uncertain as to the proper circumstances in which to permit research uses of these samples. Research Ethics Boards (REBs) in Canada and elsewhere in the world are grappling with these issues, but lack clear guidance regarding their role in the creation of and access to registries and biobanks. Chairs of 34 REBS and/or REB Administrators affiliated with Faculties of Medicine in Canadian universities were interviewed. Interviews consisted of structured questions dealing with diabetes-related scenarios, with open-ended responses and probing for rationales. The two scenarios involved the development of a diabetes registry using clinical encounter data across several physicians' practices, and the addition of biological samples to the registry to create a biobank. There was a wide range of responses given for the questions raised in the scenarios, indicating a lack of clarity about the role of REBs in registries and biobanks. With respect to the creation of a registry, a minority of sites felt that consent was not required for the information to be entered into the registry. Whether patient consent was required for information to be entered into the registry and the duration for which the consent would be operative differed across sites. With respect to the creation of a biobank linked to the registry, a majority of sites viewed biobank information as qualitatively different from other types of personal health information. All respondents agreed that patient consent was needed for blood samples to be placed in the biobank but the duration of consent again varied. Participants were more attuned to issues surrounding biobanks as compared to registries and demonstrated a higher level of concern regarding biobanks. As registries and biobanks expand, there is a need for critical analysis of suitable roles for REBs and subsequent guidance on these topics. The authors conclude by recommending REB participation in the creation of registries and biobanks and the eventual drafting of comprehensive legislation.


Evaluation of nine popular de novo assemblers in microbial genome assembly.
Next generation sequencing (NGS) technologies are revolutionizing biology, with Illumina being the most popular NGS platform. Short read assembly is a critical part of most genome studies using NGS. Hence, in this study, the performance of nine well-known assemblers was evaluated in the assembly of seven different microbial genomes. Effect of different read coverage and k-mer parameters on the quality of the assembly were also evaluated on both simulated and actual read datasets. Our results show that the performance of assemblers on real and simulated datasets could be significantly different, mainly because of coverage bias. According to outputs on actual read datasets, for all studied read coverages (of 7×, 25× and 100×), SPAdes and IDBA-UD clearly outperformed other assemblers based on NGA50 and accuracy metrics. Velvet is the most conservative assembler with the lowest NGA50 and error rate.


Lives saved by helicopter emergency medical services: an overview of literature.
The objective of this review is to give an overview of literature on the survival benefits of Helicopter Emergency Medical Services (HEMS). The included studies were assessed by study design and statistical methodology. A literature search was performed in the National Library of Medicine's Medline database, extending from 1985 until April 2007. Manuscripts had to be written in English and describe effects of HEMS on survival expressed in number of lives saved. Moreover, analysis had to be performed using adequate adjustment for differences in case-mix. Sixteen publications met the inclusion criteria. All indicated that HEMS assistance contributed to increased survival: Between 1.1 and 12.1 additional survivors were recorded for every 100 HEMS uses. A combination of four reliable studies shows overall mortality reduction of 2.7 additional lives saved per 100 HEMS deployments. Literature shows a clear positive effect on survival associated with HEMS assistance. Efforts should be made to promote consistent methodology, including uniform outcome parameters, in order to provide sufficient scientific evidence to conclude the ongoing debate about the beneficial effects of HEMS.


Comparison of Offender and Non-offender Young Men to Setting Goals for Life and Attributing Meaning to Life.
The main goal of the present study is to compare male offenders and non-offenders in terms of how they attribute meaning to life and set life goals. The samples were chosen from among offenders (n = 50) and non-offenders (n = 50) who were between ages 19-26. Mixed method was used in this study. The scale of setting life goals and interview form were used to collect data. To analysis of the data, the content analysis technique and t test and were used. Findings of the present study show that offenders tend to have troubles setting life goals when compared to non-offenders. Furthermore, the results of this study could be used in rehabilitating offenders and preventing those at risk of committing criminal acts from doing so.


Pharmacokinetic modulation of irinotecan and metabolites by cyclosporin A.
The focus of this investigation was to modulate the pharmacokinetics of irinotecan and its metabolites, SN-38 and SN-38G, by possibly reducing biliary excretion, which in turn could lower irinotecan toxicity. We determined the effect of a known cholestatic agent, cyclosporin A (CsA), which is transported across the bile canalicular membrane by P-glycoprotein, on the biliary excretion of irinotecan and its metabolites. Wistar rats were pretreated with 60 mg/kg CsA 5 min before an i.v. dose of irinotecan at dose levels of 6, 10, and 20 mg/kg. The control groups received irinotecan only. CsA pretreatment resulted in an average increase of 339, 361, and 192% in the area under the plasma concentration-time curve of irinotecan, SN-38, and SN-38G, respectively. Analysis of clearance (CL) of irinotecan indicated a 55 and 81% reduction in the average renal and nonrenal CLs, respectively, in the pretreated groups. The nonrenal CL, which is the primary component of irinotecan CL, includes protein and tissue binding as well as the metabolic and biliary CL of irinotecan. There was no change in the volume of distribution at steady state (indicative of unchanged binding) and in the metabolic conversion of irinotecan to SN-38 due to pretreatment. Therefore, the significant reduction in the systemic CL of irinotecan due to CsA pretreatment was primarily due to lowered biliary excretion. Studies using a photoaffinity analogue of verapamil, [125I]NAS-VP, and membrane vesicles from the multidrug-resistant cell line, MCF-7/Adr, revealed that irinotecan and metabolites had moderate interaction with P-glycoprotein. Further studies are required to determine the mechanism of inhibitory effect of CsA on the biliary excretion of irinotecan and its metabolites.


Non-contact vital-sign monitoring of patients undergoing haemodialysis treatment.
A clinical study was designed to record a wide range of physiological values from patients undergoing haemodialysis treatment in the Renal Unit of the Churchill Hospital in Oxford. Video was recorded for a total of 84 dialysis sessions from 40 patients during the course of 1 year, comprising an overall video recording time of approximately 304.1 h. Reference values were provided by two devices in regular clinical use. The mean absolute error between the heart rate estimates from the camera and the average from two reference pulse oximeters (positioned at the finger and earlobe) was 2.8 beats/min for over 65% of the time the patient was stable. The mean absolute error between the respiratory rate estimates from the camera and the reference values (computed from the Electrocardiogram and a thoracic expansion sensor-chest belt) was 2.1 breaths/min for over 69% of the time for which the reference signals were valid. To increase the robustness of the algorithms, novel methods were devised for cancelling out aliased frequency components caused by the artificial light sources in the hospital, using auto-regressive modelling and pole cancellation. Maps of the spatial distribution of heart rate and respiratory rate information were developed from the coefficients of the auto-regressive models. Most of the periods for which the camera could not produce a reliable heart rate estimate lasted under 3 min, thus opening the possibility to monitor heart rate continuously in a clinical environment.


Treatment of tuberculosis in patients with advanced human immunodeficiency virus infection.
Infection with the human immunodeficiency virus (HIV) increases the risk of tuberculosis and may interfere with the effectiveness of antituberculosis chemotherapy. To examine the outcomes in patients with both diagnoses, we conducted a retrospective study of all 132 patients listed in both the acquired immunodeficiency syndrome (AIDS) and tuberculosis case registries in San Francisco from 1981 through 1988. At the time of the diagnosis of tuberculosis, 78 patients (59 percent) did not yet have a diagnosis of AIDS, 18 patients (14 percent) were given a concomitant diagnosis of AIDS (as determined by the presence of an AIDS-defining disease other than tuberculosis), and the remaining 36 patients (27 percent) already had AIDS. The manifestations of tuberculosis were entirely pulmonary in 50 patients (38 percent), entirely extrapulmonary in 40 patients (30 percent), and both pulmonary and extrapulmonary in 42 patients (32 percent). The treatment regimens were as follows: isoniazid and rifampin supplemented by ethambutol for the first two months, 52 patients; isoniazid and rifampin supplemented by pyrazinamide and ethambutol for the first two months, 39 patients; isoniazid and rifampin, 13 patients; isoniazid and rifampin supplemented by pyrazinamide for the first two months, 4 patients; and other drug regimens, 17 patients. The intended duration of treatment for patients whose regimen included pyrazinamide was six months, and for patients who did not receive pyrazinamide, nine months. Seven patients received no treatment because tuberculosis was first diagnosed after death. Sputum samples became clear of acid-fast organisms after a median of 10 weeks of therapy. Abnormalities on all chest radiographs taken after three months of treatment were stable or improved except for those of patients who had new nontuberculous infections. The only treatment failure occurred in a man infected with multiple drug-resistant organisms who did not comply with therapy. Adverse drug reactions occurred in 23 patients (18 percent). For all 125 treated patients, median survival was 16 months from the diagnosis of tuberculosis. Tuberculosis was a major contributor to death in 5 of the 7 untreated patients and 8 of the 125 treated patients. Three of 58 patients who completed therapy had a relapse (5 percent); compliance was poor in all 3. Tuberculosis causes substantial mortality in patients with advanced HIV infection. In patients who comply with the regimen, conventional therapy results in rapid sterilization of sputum, radiographic improvement, and low rates of relapse.


Alternative splicing of dystrobrevin regulates the stoichiometry of syntrophin binding to the dystrophin protein complex.
Dystrophin coordinates the assembly of a complex of structural and signalling proteins that is required for normal muscle function. A key component of the dystrophin-associated protein complex (DPC) is alpha-dystrobrevin, a dystrophin-related and -associated protein whose absence results in muscular dystrophy and neuromuscular junction defects [1,2]. The current model of the DPC predicts that dystrophin and dystrobrevin each bind a single syntrophin molecule [3]. The syntrophins are PDZ-domain-containing proteins that facilitate the recruitment of signalling proteins such as nNOS (neuronal nitric oxide synthase) to the DPC [4]. Here we show, using yeast two-hybrid analysis and biochemical binding studies, that alpha-dystrobrevin in fact contains two independent syntrophin-binding sites in tandem. The previously undescribed binding site is situated within an alternatively spliced exon of alpha-dystrobrevin, termed the variable region-3 (vr3) sequence, which is specifically expressed in skeletal and cardiac muscle [5,6]. Analysis of the syntrophin-binding region of dystrobrevin reveals a tandem pair of predicted alpha helices with significant sequence similarity. These alpha helices, each termed a syntrophin-binding motif, are also highly conserved in dystrophin and utrophin. Together these data show that there are four potential syntrophin-binding sites per dystrophin complex in skeletal muscle: two on dystrobrevin and two on dystrophin or utrophin. Furthermore, alternative splicing of dystrobrevin provides a mechanism for regulating the stoichiometry of syntrophin association with the DPC. This is likely to have important consequences for the recruitment of specific signalling molecules to the DPC and ultimately for its function.


Biological and Genomic Sequence Characterization of Maize streak virus Isolates from Wheat.
ABSTRACT Maize streak virus (MSV) is best known as the causal agent of maize streak disease. However, only a genetically uniform subset of the viruses within this diverse species is actually capable of producing severe symptoms in maize. Whereas these "maize-type" viruses all share greater than 95% sequence identity, MSV strains isolated from grasses may share as little as 79% sequence identity with the maize-type viruses. Here, we present the complete genome sequences and biological characterization of two MSV isolates from wheat that share approximately 89% sequence identity with the maize-type viruses. Clonal populations of these two isolates, named MSV-Tas and MSV-VW, were leafhopper-transmitted to Digitaria sanguinalis and a range of maize, wheat, and barley genotypes. Whereas the two viruses showed some differences in their pathogenicity in maize, they were both equally pathogenic in D. sanguinalis and the various wheat and barley genotypes tested. Phylogenetic analyses involving the genome sequences of MSV-Tas and MSV-VW, a new maize-type virus also fully sequenced in this study (MSV-VM), and all other available African streak virus sequences, indicated that MSV-Tas and MSV-VW are close relatives that together represent a distinct MSV strain. Sequence analyses revealed that MSV-VM has a recombinant genome containing MSV-Tas/VW-like sequences within its movement protein gene.


The cloned neurotensin receptor mediates cyclic GMP formation when coexpressed with nitric oxide synthase cDNA.
Rat neurotensin (NT) receptor (NTR) cDNA was subcloned into the pRC-CMV expression vector and transfected into 293 cells, and cellular clones that stably expressed the NTR were isolated and characterized. [3H]NT binding to membranes prepared from the NTR cDNA-transfected cells displayed specificity and saturability, with an apparent Kd of 1.25 nM and a Bmax of 43.4 pmol/mg of protein (approximately 3.5 x 10(6) binding sites/cell). NT stimulated an increase in [3H]inositol phosphate levels in the NTR-expressing cells up to 2500% of basal levels. The response was time and dose dependent, with an EC50 of 10.4 nM. NT also stimulated cAMP formation in these cells, with an EC50 of 27.0 nM. In addition, NT evoked an increase in the level of intracellular calcium. Approximately 60% of the calcium rise was attributable to the release of intracellular stores and 40% was attributable to calcium influx. Although NTR occupancy has been shown to stimulate cGMP formation in several brain preparations and cell lines, NT was unable to mediate cGMP synthesis in the NTR-expressing 293 cells. We found that 293 cells have guanylate cyclase activity but have undetectable levels of nitric oxide synthase (NOS) activity. Because it was possible that the production of nitric oxide is required as the mediator of NT-induced cGMP synthesis, we subcloned NOS cDNA into the pCEP4 expression vector and transiently expressed it in the NTR cells. We report that NT increased cGMP levels up to 375% of basal levels when NOS cDNA was coexpressed and that the increase was completely inhibited by the NOS inhibitor N omega-nitro-L-arginine. NT-induced cGMP accumulation was time and dose dependent, with an EC50 of 1.7 nM. To our knowledge, this is the first report of NT mediating cGMP formation with a cloned receptor and the first evidence that NT-induced cGMP accumulation requires the production of nitric oxide.


'I have to pretend that I don't care': Stigma management among unmarried young mothers in South-Western Nigeria.
Young mothers often encounter stigma and discrimination, affecting their lives and that of their children. This paper explores stigma management strategies and their effectiveness for young mothers in rural Nigeria. Ten key informants and 24 young mothers were recruited from Ife-East in South-Western Nigeria. Data from semi-structured interviews showed that societal disapproval of pregnant teenagers and young mothers were common experiences. Women used a range of strategies to actively cope with stigma including: belief in predestination, avoidance, concealment, and cohabitation. These strategies could be seen as tools to mitigate negative stereotypes and discrimination. However, they also had the unintended consequences of compounding many young women's difficult circumstances and exposing them to adverse outcomes, including gender-based violence, repeat pregnancies, poor mental health, and low uptake of services. The results show the need for policy frameworks to actively combat stigma by addressing the negative framing of early pregnancy and motherhood and promoting supportive environments for young mothers. Health professionals need to be trained to offer de-stigmatising services to encourage young mothers to seek help and reduce pre-existing inequities in access to services, and policies need to include measures that address the rights of young mothers and protect them from violence and abuse.


Self-assembly and binding of a sorting nexin to sorting endosomes.
The fate of endocytosed membrane proteins and luminal contents is determined by a materials processing system in sorting endosomes. Endosomal retention is a mechanism that traps specific proteins within this compartment, and thereby prevents their recycling. We report that a sorting nexin SNX1, a candidate endosomal retention protein, self-assembles in vitro and in vivo, and has this property in common with its yeast homologue Vps5p. A comparison of SNX1 expressed in bacterial and in mammalian systems and analyzed by size-exclusion chromatography indicates that in cytosol SNX1 tetramers are part of a larger complex with additional proteins. An endosomal retention function would require that SNX1 bind to endosomal membranes, yet the complexes that we analyzed were largely soluble and little SNX1 was found in pellet fractions. Using green fluorescent protein fusions, endocytic compartment markers and fluorescence recovery after photobleaching, we found that there is an equilibrium between free cytoplasmic and early/sorting endosome-bound pools of green fluorescent protein-SNX1. Fluorescence resonance energy transfer indicated that spectral variants of green fluorescent protein-SNX1 were oligomeric in vivo. In cell extracts, these green fluorescent protein-SNX1 oligomers corresponded to tetrameric and larger complexes of green fluorescent protein-SNX1. Using video microscopy, we observed small vesicle docking and tubule budding from large green fluorescent protein-SNX1 coated endosomes, which are features consistent with their role as sorting endosomes. http://www.biologists.com/JCS/movies/jcs2058.html


Subjective Cognitive Complaints Given in Questionnaire: Relationship With Brain Structure, Cognitive Performance and Self-Reported Depressive Symptoms in a 25-Year Retrospective Cohort Study.
Subjective cognitive complaints are common but it is unclear whether they indicate an underlying pathological process or reflect affective symptoms. 800 community-dwelling older adults were drawn from the Whitehall II cohort. Subjective cognitive complaint inquiry for memory and concentration, a range of neuropsychological tests and multimodal MRI were performed in 2012-2016. Subjective complaints were again elicited after 1 year. Group differences in grey and white matter, between those with and without subjective complaints, were assessed using voxel-based morphometry and tract-based spatial statistics, respectively. Mixed effects models assessed whether cognitive decline or depressive symptoms (over a 25-year period) were associated with later subjective complaints. Analyses were controlled for potential confounders and multiple comparisons. Mean age of the sample at scanning was 69.8 years (±5.1, range: 60.3-84.6). Subjective memory complaints were common (41%) and predicted further similar complaints later (mean 1.4 ± 1.4 years). There were no group differences in grey matter density or white matter integrity. Subjective complaints were not cross-sectionally or longitudinally associated with objectively assessed cognition. However, those with subjective complaints reported higher depressive symptoms ("poor concentration": odds ratio = 1.12, 95% CI 1.07-1.18; "poor memory": odds ratio = 1.18, 1.12-1.24). In our sample subjective complaints were consistent over time and reflected depressive symptoms but not markers of neurodegenerative brain damage or concurrent or future objective cognitive impairment. Clinicians assessing patients presenting with memory complaints should be vigilant for affective disorders. These results question the rationale for including subjective complaints in a spectrum with Mild Cognitive Impairment diagnostic criteria.


Applying Antibodies Inside Cells: Principles and Recent Advances in Neurobiology, Virology and Oncology.
To interfere with cell function, many scientists rely on methods that target DNA or RNA due to the ease with which they can be applied. Proteins are usually the final executors of function but are targeted only indirectly by these methods. Recent advances in targeted degradation of proteins based on proteolysis-targeting chimaeras (PROTACs), ubiquibodies, deGradFP (degrade Green Fluorescent Protein) and other approaches have demonstrated the potential of interfering directly at the protein level for research and therapy. Proteins can be targeted directly and very specifically by antibodies, but using antibodies inside cells has so far been considered to be challenging. However, it is possible to deliver antibodies or other proteins into the cytosol using standard laboratory equipment. Physical methods such as electroporation have been demonstrated to be efficient and validated thoroughly over time. The expression of intracellular antibodies (intrabodies) inside cells is another way to interfere with intracellular targets at the protein level. Methodological strategies to target the inside of cells with antibodies, including delivered antibodies and expressed antibodies, as well as applications in the research areas of neurobiology, viral infections and oncology, are reviewed here. Antibodies have already been used to interfere with a wide range of intracellular targets. Disease-related targets included proteins associated with neurodegenerative diseases such as Parkinson's disease (α-synuclein), Alzheimer's disease (amyloid-β) or Huntington's disease (mutant huntingtin [mHtt]). The applications of intrabodies in the context of viral infections include targeting proteins associated with HIV (e.g. HIV1-TAT, Rev, Vif, gp41, gp120, gp160) and different oncoviruses such as human papillomavirus (HPV), hepatitis B virus (HBV), hepatitis C virus (HCV) and Epstein-Barr virus, and they have been used to interfere with various targets related to different processes in cancer, including oncogenic pathways, proliferation, cell cycle, apoptosis, metastasis, angiogenesis or neo-antigens (e.g. p53, human epidermal growth factor receptor-2 [HER2], signal transducer and activator of transcription 3 [STAT3], RAS-related RHO-GTPase B (RHOB), cortactin, vascular endothelial growth factor receptor 2 [VEGFR2], Ras, Bcr-Abl). Interfering at the protein level allows questions to be addressed that may remain unanswered using alternative methods. This review addresses why direct targeting of proteins allows unique insights, what is currently feasible in vitro, and how this relates to potential therapeutic applications.


Elongation index of erythrocytes, study of activity of chosen erythrocyte enzymes, and the levels of glutathione, malonyldialdehyde in polycythemia vera (PV).
The principal aim of the study was to investigate rheological properties of erythrocytes obtained from patients admitted to the clinic, and diagnosed with polycythemia vera. The polycythemia vera diagnosis was based on the WHO criteria for polycythemia vera. Using a laser rheometer SSD Rheometer-Rheodyn, the elongation index of erythrocytes was determined, indicating an increased rigidity of the erythrocytes in this disease compared with the erythrocytes in healthy people. In order to explain (albeit partially) the reason for reduced elasticity, the erythrocytes of patients with polycythemia were studied for the activity of enzymes - glucose-6-phosphate dehydrogenase and acetylcholinesterase membrane enzyme, as well as the levels of glutathione and malonyldialdehyde. The elevated activities of these enzymes, the glutathione level, and elevated ‰ of reticulocytes, indicated an increased pool of juvenile erythrocyte forms; furthermore, the elevated value of malonyldialdehyde may suggest a lipid peroxidative damage in certain pool of the erythrocyte membrane in blood circulation.


Mutation Rate Evolution in Partially Selfing and Partially Asexual Organisms.
Different factors can influence the evolution of the mutation rate of a species: costs associated with DNA replication fidelity, indirect selection caused by the mutations produced (that should generally favor lower mutation rates, given that most mutations affecting fitness are deleterious), and genetic drift, which may render selection acting on weak mutators inefficient. In this paper, we use a two-locus model to compute the strength of indirect selection acting on a modifier locus that affects the mutation rate toward a deleterious allele at a second, linked, locus, in a population undergoing partial selfing or partial clonality. The results show that uniparental reproduction increases the effect of indirect selection for lower mutation rates. Extrapolating to the case of a whole genome with many deleterious alleles, and introducing a direct cost to DNA replication fidelity, the results can be used to compute the evolutionarily stable mutation rate, U In the absence of mutational bias toward higher U, the analytical prediction fits well with individual-based, multilocus simulation results. When such a bias is added into the simulations, however, genetic drift may lead to the maintenance of higher mutation rates, and this effect may be amplified in highly selfing or highly clonal populations due to their reduced effective population size.


Isometric and isovelocity contractile performance of red muscle fibres from the dogfish Scyliorhinus canicula.
Maximum isometric tetanic force produced by bundles of red muscle fibres from dogfish, Scyliorhinus canicula (L.), was 142.4+/-10.3 kN m(-2) (N=35 fibre bundles); this was significantly less than that produced by white fibres 289.2+/-8.4 kN m(-2) (N=25 fibre bundles) (means +/- S.E.M.). Part, but not all, of the difference is due to mitochondrial content. The maximum unloaded shortening velocity, 1.693+/-0.108 L(0) s(-1) (N=6 fibre bundles), was measured by the slack-test method. L(0) is the length giving maximum isometric force. The force/velocity relationship was investigated using a step-and-ramp protocol in seven red fibre bundles. The following equation was fitted to the data: [(P/P(0))+(a/P(0))](V+b)=[(P(0)(*)/P(0))+(a/P(0))]b, where P is force during shortening at velocity V, P(0) is the isometric force before shortening, and a, b and P(0)(*) are fitted constants. The fitted values were P(0)(*)/P(0)=1.228+/-0.053, V(max)=1.814+/-0.071 L(0) s(-1), a/P(0)=0.269+/-0.024 and b=0.404+/-0.041 L(0) s(-1) (N=7 for all values). The maximum power was 0.107+/-0.005P(0)V(max) and was produced during shortening at 0.297+/-0.012V(max). Compared with white fibres from dogfish, the red fibres have a lower P(0) (49%) and V(max) (48%), but the shapes of the force/velocity curves are similar. Thus, the white and red fibres have equal capacities to produce power within the limits set by the isometric force and maximum velocity of shortening of each fibre type. A step shortening of 0.050+/-0.003L(0) (N=7) reduced the maximum isometric force in the red fibres' series elasticity to zero. The series elasticity includes all elastic structures acting in series with the attached cross-bridges. Three red fibre bundles were stretched at a constant velocity, and force (measured when length reached L(0)) was 1.519+/-0.032P(0). In the range of velocities used here, -0.28 to -0.63V(max), force varied little with the velocity.


CB2 Cannabinoid Receptor Knockout in Mice Impairs Contextual Long-Term Memory and Enhances Spatial Working Memory.
Neurocognitive effects of cannabinoids have been extensively studied with a focus on CB1 cannabinoid receptors because CB1 receptors have been considered the major cannabinoid receptor in the nervous system. However, recent discoveries of CB2 cannabinoid receptors in the brain demand accurate determination of whether and how CB2 receptors are involved in the cognitive effects of cannabinoids. CB2 cannabinoid receptors are primarily involved in immune functions, but also implicated in psychiatric disorders such as schizophrenia and depression. Here, we examined the effects of CB2 receptor knockout in mice on memory to determine the roles of CB2 receptors in modulating cognitive function. Behavioral assays revealed that hippocampus-dependent, long-term contextual fear memory was impaired whereas hippocampus-independent, cued fear memory was normal in CB2 receptor knockout mice. These mice also displayed enhanced spatial working memory when tested in a Y-maze. Motor activity and anxiety of CB2 receptor knockout mice were intact when assessed in an open field arena and an elevated zero maze. In contrast to the knockout of CB2 receptors, acute blockade of CB2 receptors by AM603 in C57BL/6J mice had no effect on memory, motor activity, or anxiety. Our results suggest that CB2 cannabinoid receptors play diverse roles in regulating memory depending on memory types and/or brain areas.


Keeping the secrecy aspect in mass e-voting.
Two years of the COVID-19 pandemic have pushed society's digitalization forward like nothing ever before. Activities that weren't achievable without personal contact, became present online. Currently, when the COVID-19 restrictions are being lifted and personal meetings are becoming possible, in some cases, it still appeared to be more convenient to "meet" virtually than physically. Some actions proved to be more effective when performed online. That is why some people didn't want to come back to the previous form of communication anymore. Many decision-makers who see all the advantages of online communication have turned to the new possibilities that are served by Internet systems. Hence, they intend to keep it virtual even now, when pandemic restrictions have been lifted in most cases. This situation implies lots of great ideas for virtualization. One of those is online voting and polling conducted on many voters - groups larger than just board members or delegates. Such balloting could be performed using a voting system. Unfortunately, in the case of the decision-making process, which should be treated as a one-time event, using typical solutions is mostly very ineffective. Observations revealed that e-voting procedures conducted on a large number of electors may be challenging for many, not computer-experienced users. The procedure of authentication has to stay present in order to protect the votings against ballot stuffing. Particularly in secret voting, the problem remains the same: how to verify the user for a secret ballot without revealing his data. In mass voting, additional issues appear such as how to complete the procedure on multiple (often not ICT-experienced) voters and how to make the system accessible and credible at the same time.


The Soybean Rhg1 locus for resistance to the soybean cyst nematode Heterodera glycines regulates the expression of a large number of stress- and defense-related genes in degenerating feeding cells.
To gain new insights into the mechanism of soybean (Glycine max) resistance to the soybean cyst nematode (Heterodera glycines), we compared gene expression profiles of developing syncytia in soybean near-isogenic lines differing at Rhg1 (for resistance to Heterodera glycines), a major quantitative trait locus for resistance, by coupling laser capture microdissection with microarray analysis. Gene expression profiling revealed that 1,447 genes were differentially expressed between the two lines. Of these, 241 (16.8%) were stress- and defense-related genes. Several stress-related genes were up-regulated in the resistant line, including those encoding homologs of enzymes that lead to increased levels of reactive oxygen species and proteins associated with the unfolded protein response. These results indicate that syncytia induced in the resistant line are undergoing severe oxidative stress and imbalanced endoplasmic reticulum homeostasis, both of which likely contribute to the resistance reaction. Defense-related genes up-regulated within syncytia of the resistant line included those predominantly involved in apoptotic cell death, the plant hypersensitive response, and salicylic acid-mediated defense signaling; many of these genes were either partially suppressed or not induced to the same level by a virulent soybean cyst nematode population for successful nematode reproduction and development on the resistant line. Our study demonstrates that a network of molecular events take place during Rhg1-mediated resistance, leading to a highly complex defense response against a root pathogen.