Protective Effects of Naringenin in a Rat Model of Sepsis-Triggered Acute Kidney Injury via Activation of Antioxidant Enzymes and Reduction in Urinary Angiotensinogen.
BACKGROUND Sepsis is a devastating medical condition. In the USA, about 745 000 people are diagnosed with sepsis annually. Although many anti-inflammatory drugs have been used to manage sepsis, the treatment success rate is very low. This study was undertaken to examine the protective effects of naringenin on sepsis-induced kidney injury in rats. MATERIAL AND METHODS Sepsis was induced in Wistar albino rats by cecal ligation and puncture methods. Histological analysis was performed with hematoxylin and eosin (HE) staining. Reactive oxygen species (ROS) levels were determined by flow cytometery. TUNEL assay was used to demonstrate apoptosis. Sandwich ELISA method was used for the determination of urinary angiotensinogen, and protein expression was determined by Western blot analysis. RESULTS We found that naringenin decreased atrophy in the glomerulus and enabled maintenance of the capsule area and normal tubular cavity of the septic rats. Admistration of naringenin at the dosage of 10 and 20 mg/kg to sepsis rats caused significant reduction in the sepsis-induced apoptosis of kidney cells, accompanied by decrease in Bax and increase in Bcl-2 expression. Moreover, naringenin also decreased the ROS levels in septic rats and downregulated the expression of SOD, CAT, and APX. The effects of naringenin were also examined on the levels of urinary angiotensinogen in sepsis rats. We found that naringenin caused a significant decrease in urinary angiotensinogen levels of septic rats. CONCLUSIONS Naringenin appears to have potential in the treatment of sepsis.


Polymer nanofibers via nozzle-free centrifugal spinning.
We report on the unexpected finding of nanoscale fibers with a diameter down to 25 nm that emerge from a polymer solution during a standard spin-coating process. The fiber formation relies upon the Raleigh-Taylor instability of the spin-coated liquid film that arises due to a competition of the centrifugal force and the Laplace force induced by the surface curvature. This procedure offers an attractive alternative to electrospinning for the efficient, simple, and nozzle-free fabrication of nanoscale fibers from a variety of polymer solutions.


Age Distribution of Epistaxis in Outpatient Pediatric Patients.
Objectives: Little is known about the prevalence of epistaxis in children. Existing reports focus on hospitalized children or those presenting to an emergency department. To better understand pediatric epistaxis in clinical practice, we sought out a searchable, representative outpatient database and examined the incidence of epistaxis in children of different ages. Methods: A cross-sectional analysis of data from the National Hospital Ambulatory Medical Care Survey (NHAMCS) from the years 2007 to 2011 was performed. The NHAMCS is a Centers for Disease Control and Prevention-curated national sample of data from visits to non-federally employed office-based physicians and health centers. We queried the NHAMCS to determine the cumulative incidence of epistaxis in children of different age groups. The International Classification of Diseases Ninth Revision code 784.7 was chosen to identify epistaxis. Comparisons of rates were performed using the chi-squared test. A P-value of <.05 was considered statistically significant. Results: In total, 55,435,691 children [27,816,237 (50.2%) males, 55,435,691 (77.2%) white] were included. The overall cumulative incidence rate of epistaxis was 2.4/1000 children. Children in the 3- to 5-year range had the highest cumulative incidence of epistaxis (5.0/1000), followed by those in the 6 to 8 (3.0/1000), 9 to 11 (2.0/1000), 0 to 2 (1.9/1000), 12 to 14 (1.6/1000), and 15 to 17 (0.5/1000) year ranges (P < .001). Conclusion: Pediatric epistaxis is common in the office setting (2.4 per 1000 children)-and well above emergency department estimates (1.7 per 1000 people). Children between the ages of 3 to 5 years have the highest cumulative incidence. Epistaxis is sufficiently unusual in infants and the late teens that alternative causes for nasal bleeding should be included in the differential diagnosis.


[Stent or not in the case of renal artery stenosis? No clear-cut answer yet].
Recent trials, STAR and ASTRAL, indicate that medicinal treatment alone is as good as stenting plus medicinal treatment in preventing renal events in patients with atherosclerotic renal artery stenosis of more than 50-70%. In this commentary, it is stated that the high number of complications in catheter interventions is related to the generalised atherosclerosis in these patients. In this group of patients intervention is too late to prevent renal events. Stenting in patients with low-grade renal artery stenosis should be considered as an option for treating hypertension and atherosclerotic renal artery stenosis. This should be studied in a prospective randomised controlled trial.


Redox imbalance of red blood cells impacts T lymphocyte homeostasis: implication in carotid atherosclerosis.
Oxidative stress and immune/inflammatory responses are key pathogenetic factors of atherosclerotic disease. In this contest, mechanisms that regulate survival and death of immune cells may be relevant. Previous studies have demonstrated that red blood cells (RBCs) are physiologically able to inhibit apoptosis and to promote proliferation of activated T lymphocytes from healthy subjects. The aim of the present study was to evaluate whether RBCs from patients with carotid atherosclerosis maintain their property to modulate T cell homeostasis. Peripheral blood lymphocytes (PBLs) obtained from healthy subjects were activated in vitro by phytohemagglutinin in the presence/absence of RBCs from patients with carotid atherosclerosis or of in vitro oxidised RBCs from healthy subjects. Levels of reactive oxygen species (ROS) and aging markers of RBCs as well as susceptibility to apoptosis of PBLs were evaluated by flow cytometry. PBL proliferation was evaluated by 3H-methyl-thymidine incorporation assay whereas secretion of cytokines, analysed in view of their key role in T cell function, was assessed by ELISA. Levels of ROS and phosphatidyl-serine externalisation, a sign of RBC aging, resulted significantly higher in RBCs from patients than in those from healthy subjects, whereas surface glycophorin A expression and reduced glutathione content did the opposite. Unlike RBCs obtained from healthy subjects, RBCs from patients and in vitro oxidised RBCs did not protect activated T lymphocytes from apoptosis. Hence, RBCs from patients with carotid atherosclerosis, probably due to their oxidative imbalance, impact T cell integrity and function. Our results suggest a new regulatory role for RBCs in atherosclerosis.


Evaluation of the traditional and revised WHO classifications of Dengue disease severity.
Dengue is a major public health problem worldwide and continues to increase in incidence. Dengue virus (DENV) infection leads to a range of outcomes, including subclinical infection, undifferentiated febrile illness, Dengue Fever (DF), life-threatening syndromes with fluid loss and hypotensive shock, or other severe manifestations such as bleeding and organ failure. The long-standing World Health Organization (WHO) dengue classification and management scheme was recently revised, replacing DF, Dengue Hemorrhagic Fever (DHF), and Dengue Shock Syndrome (DSS) with Dengue without Warning Signs, Dengue with Warning Signs (abdominal pain, persistent vomiting, fluid accumulation, mucosal bleeding, lethargy, liver enlargement, increasing hematocrit with decreasing platelets) and Severe Dengue (SD; dengue with severe plasma leakage, severe bleeding, or organ failure). We evaluated the traditional and revised classification schemes against clinical intervention levels to determine how each captures disease severity using data from five years (2005-2010) of a hospital-based study of pediatric dengue in Managua, Nicaragua. Laboratory-confirmed dengue cases (n = 544) were categorized using both classification schemes and by level of care (I-III). Category I was out-patient care, Category II was in-patient care that did not meet criteria for Category III, which included ICU admission, ventilation, administration of inotropic drugs, or organ failure. Sensitivity and specificity to capture Category III care for DHF/DSS were 39.0% and 75.5%, respectively; sensitivity and specificity for SD were 92.1% and 78.5%, respectively. In this data set, DENV-2 was found to be significantly associated with DHF/DSS; however, this association was not observed with the revised classification. Among dengue-confirmed cases, the revised WHO classification for severe dengue appears to have higher sensitivity and specificity to identify cases in need of heightened care, although it is no longer as specific for a particular pathogenic entity as was the traditional schema.


Public-health impact of outdoor and traffic-related air pollution: a European assessment.
Air pollution contributes to mortality and morbidity. We estimated the impact of outdoor (total) and traffic-related air pollution on public health in Austria, France, and Switzerland. Attributable cases of morbidity and mortality were estimated. Epidemiology-based exposure-response functions for a 10 microg/m3 increase in particulate matter (PM10) were used to quantify the effects of air pollution. Cases attributable to air pollution were estimated for mortality (adults > or = 30 years), respiratory and cardiovascular hospital admissions (all ages), incidence of chronic bronchitis (adults > or = 25 years), bronchitis episodes in children (< 15 years), restricted activity days (adults > or = 20 years), and asthma attacks in adults and children. Population exposure (PM10) was modelled for each km2. The traffic-related fraction was estimated based on PM10 emission inventories. Air pollution caused 6% of total mortality or more than 40,000 attributable cases per year. About half of all mortality caused by air pollution was attributed to motorised traffic, accounting also for: more than 25,000 new cases of chronic bronchitis (adults); more than 290,000 episodes of bronchitis (children); more than 0.5 million asthma attacks; and more than 16 million person-days of restricted activities. This assessment estimates the public-health impacts of current patterns of air pollution. Although individual health risks of air pollution are relatively small, the public-health consequences are considerable. Traffic-related air pollution remains a key target for public-health action in Europe. Our results, which have also been used for economic valuation, should guide decisions on the assessment of environmental health-policy options.


The Root Apex of Arabidopsis thaliana Consists of Four Distinct Zones of Growth Activities: Meristematic Zone, Transition Zone, Fast Elongation Zone and Growth Terminating Zone.
In the growing apex of Arabidopsis thaliana primary roots, cells proceed through four distinct phases of cellular activities. These zones and their boundaries can be well defined based on their characteristic cellular activities. The meristematic zone comprises, and is limited to, all cells that undergo mitotic divisions. Detailed in vivo analysis of transgenic lines reveals that, in the Columbia-0 ecotype, the meristem stretches up to 200 microm away from the junction between root and root cap (RCJ). In the transition zone, 200 to about 520 microm away from the RCJ, cells undergo physiological changes as they prepare for their fast elongation. Upon entering the transition zone, they progressively develop a central vacuole, polarize the cytoskeleton and remodel their cell walls. Cells grow slowly during this transition: it takes ten hours to triplicate cell length from 8.5 to about 35 microm in the trichoblast cell files. In the fast elongation zone, which covers the zone from 520 to about 850 microm from the RCJ, cell length quadruplicates to about 140 microm in only two hours. This is accompanied by drastic and specific cell wall alterations. Finally, root hairs fully develop in the growth terminating zone, where root cells undergo a minor elongation to reach their mature lengths.


Penile prosthesis implantation for treatment of postpriapism erectile dysfunction.
Our aim was to evaluate the procedure and outcome of penile prosthesis surgery in the treatment of men with postpriapism erectile dysfunction. During the period between 1997 and 2004, a total of 17 patients with postpriapism erectile dysfunction underwent penile prosthesis implantation at our institution. Prosthesis implantation was done electively 6 to 18 months after priapism, when the patients presented with erectile dysfunction. Of the prosthesis implanted, 11 were malleable, 4 were 2-piece, and 2 were 3-piece prostheses (AMS, Minnetonka, Minnesota, USA). All the 17 patients were successfully implanted with penile prosthesis. Intra-operatively, corporeal dilation was difficult due to extensive corporeal fibrosis, which led to urethral injury in 2 patients. There were no major postoperative complications. The median hospital stay was 5 days. The follow-up period ranged from 2 to 9 years (median, 6 years). All the patients were satisfied with the prosthesis. Penile prosthesis implantation is the modality of treatment for patients with postpriapism erectile dysfunction at our institution. It has a high patient satisfaction rate. Although procedure-related complications are common due to corporeal fibrosis, they were mostly minor ones and did not affect the outcome of the procedure.


Hepatitis C Virus Seroprevalence in Mongolian Women Assessed by a Novel Multiplex Antibody Detection Assay.
Hepatitis C virus (HCV) infection causes hepatocellular carcinoma and is an important cause of mortality in both industrialized and developing countries. We developed a single-step high-throughput multiplex serology assay for HCV antibody detection and determined HCV prevalence in a highly endemic country. Five proteins (Core, NS3, NS4A, NS5A, NS5B) each from the three most common subtypes of HCV (1a, 1b, 2a) were recombinantly expressed and used as antigens in a multiplexed antibody detection assay. Multiplex HCV serology was validated with 432 reference sera whose HCV status was established by commercial ELISA, Western blot, and RNA assays. HCV antibodies were determined in 1,023 sera representative for the adult female population of Mongolia. In reference sera, detection of HCV (mostly Core and NS3) antibodies by multiplex serology showed 100% sensitivity and 99.6% specificity, and was in very good agreement with the commercial diagnostic assays (kappa, 0.96; 95% confidence interval, 0.92-0.99). The role of antibodies to NS4 and NS5 remains to be evaluated. In Mongolia, overall HCV antibody prevalence was 18.9% (17.8% when age-standardized to the world population). HCV seroprevalence increased with age from 10% in women <30 years to 32% in women ≥50 years, but was not related to sexual risk factors. The single-step high-throughput multiplex HCV serology assay performs similarly to conventional HCV antibody screening followed by secondary confirmation assays. A very high HCV seroprevalence was confirmed across all socio-economic groups in the female population of Mongolia. Multiplex HCV serology facilitates large seroepidemiologic studies of HCV infection.


Inhibition by ricin of protein synthesis in vitro. Ribosomes as the target of the toxin.
1. Ricin (a toxic protein from the seeds of Ricinus communis) is a powerful inhibitor of the poly(U)-directed incorporation of phenylalanine into polypeptides catalysed by isolated rat liver ribosomes and elongation factors 1 and 2 (EF 1 and EF 2). The inhibition can be largely overcome by increasing the concentration of ribosomes. 2. The toxin does not affect the binding of phenylalanyl-tRNA to ribosomes catalysed by EF 1, nor does it inhibit the puromycin reaction used as a test for peptide-bond formation catalysed by ribosomes. 3. Ricin inhibits the ribosome-linked GTP hydrolysis catalysed by EF 2. 4. Ribosomes treated with ricin and washed through sucrose gradients containing 0.6m-NH(4)Cl are functionally inactive in those assay systems that are sensitive to the presence of added toxin. 5. It is suggested that ricin brings about an irreversible modification of ribosomes which impairs their ability to interact with EF 2. Since ricin inhibits at a molar concentration much lower than that of ribosomes it probably acts catalytically. No added cofactor is necessary for the inhibitory action of the toxin.


Edema is a sign of early acute myocardial infarction on post-mortem magnetic resonance imaging.
The aim of this study was to investigate if acute myocardial infarction can be detected by post-mortem cardiac magnetic resonance (PMMR) at an earlier stage than by traditional autopsy, i.e., within less than 4 h after onset of ischemia; and if so, to determine the characteristics of PMMR findings in early acute infarcts. Twenty-one ex vivo porcine hearts with acute myocardial infarction underwent T2-weighted cardiac PMMR imaging within 3 h of onset of iatrogenic ischemia. PMMR imaging findings were compared to macroscopic findings. Myocardial edema induced by ischemia and reperfusion was visible on PMMR in all cases. Typical findings of early acute ischemic injury on PMMR consist of a central zone of intermediate signal intensity bordered by a rim of increased signal intensity. Myocardial edema can be detected on cardiac PMMR within the first 3 h after the onset of ischemia in porcine hearts. The size of myocardial edema reflects the area of ischemic injury in early acute (per-acute) myocardial infarction. This study provides evidence that cardiac PMMR is able to detect acute myocardial infarcts at an earlier stage than traditional autopsy and routine histology.


A novel mutation in COL4A1 gene: a possible cause of early postnatal cerebrovascular events.
COL4A1 is located in humans on chromosome13q34 and it encodes the alpha 1 chain of type IV collagen, a component of basal membrane. It is expressed mainly in the brain, muscles, kidneys and eyes. Different COL4A1 mutations have been reported in many patients who present a very wide spectrum of clinical symptoms. They typically show a multisystemic phenotype. Here we report on the case of a patient carrying a novel de novo splicing mutation of COL4A1 associated with a distinctive clinical picture characterized by onset in infancy and an unusual evolution of the neuroradiological features. At three months of age, the child was diagnosed with a congenital cataract, while his brain MRI was normal. Over the following years, the patient developed focal epilepsy, mild diplegia, asymptomatic microhematuria, raised creatine kinase levels, MRI white matter abnormalities and brain calcification on CT. During the neuroradiological follow-up the extension and intensity of the brain lesions progressively decreased. The significance of a second variant in COL4A1 carried by the child and inherited from his father remains to be clarified. In conclusion, our patient shows new aspects of this collagenopathy and possibly a COL4A1 compound heterozygosity.


Specific sensitization of lymphocytes to tumor antigens by co-cultivation with peritoneal cells exposed to such antigens.
Peritoneal cells from BALB/c mice were seeded into 50-mm plastic Petri dishes and exposed either to antigen extracts prepared from individual BALB/c sarcomas having unique tumor-specific antigens, to similar extracts prepared from 14- to 18-day-old mouse embryos, or to culture medium only. Lymphoid cells from BALB/c lymph nodes were than added to the peritoneal cells. Following 4-5 days of co-cultivation, the lymphoid cells were harvested and tested in a 30 h microcytotoxicity assay for reactivity against tumor cells carrying the respective sensitizing antigens as well as against control cells which consisted of either tumor cells lacking the sensitizing antigens or normal skin fibroblasts. By this approach sensitization to individually unique tumor antigens, as well as to antigens shared by mouse embryos and mouse sarcoma cells, was achieved.


Tax treatment of cafeteria plans. Internal Revenue Service (IRS), Treasury. Final regulations.
This document contains final regulations relating to section 125 cafeteria plans. The final regulations clarify the circumstances under which a section 125 cafeteria plan election may be changed. The final regulations permit an employer to allow a section 125 cafeteria plan participant to revoke an existing election and make a new election during a period of coverage for accident or health coverage or group-term life insurance coverage.


Effects of different sources of carbohydrates on intake, digestibility, chewing, and performance of Holstein dairy cows.
To investigate the effects of different sources of carbohydrates on intake, digestibility, chewing, and performance, nine lactating Holstein dairy cows (day in milk= 100±21 d; body weight=645.7 ± 26.5 kg) were allotted to a 3 × 3 Latin square design at three 23-d periods. The three treatments included 34.91% (B), 18.87% (BC), and 18.86% (BB) barley that in treatment B was partially replaced with only corn or corn plus beet pulp in treatments BC and BB, respectively. The concentration of starch and neutral detergent soluble carbohydrate varied (22.2, 20.2, and 14.5; 13.6, 15.9, and 20.1% of DM in treatments B, BC, and BB, respectively). Cows in treatment BB showed a higher DMI and improved digestibility of DM, NDF, and EE compared with treatments B or BC. Ruminal pH was higher in cows fed on BB (6.83) compared with those that received B or BC treatments (6.62 and 6.73, respectively). A lower proportion of propionate accompanied the higher pH in the BB group; however, a greater proportion of acetate and acetate: propionate ratio was observed compared with cows fed either on the B or BC diet. Moreover, cows fed on the BB diet showed the lowest ruminal passage rate and longest ruminal and total retention time. Eating time did not differ among treatments, rumination time was greater among cows fed on the BB diet compared with the others, whereas total chewing activity was greater than those fed on BC, but similar to those fed on B. The treatments showed no effect on milk yield. Partially replacing barley with corn or beet pulp resulted in an increase in milk fat and a lower protein concentration. Changing dietary NFC with that of a different degradability thus altered intake, chewing activity, ruminal environment, retention time or passage rate, and lactation performance. The results of this study showed that beet pulp with a higher NDF and a detergent-soluble carbohydrate or pectin established a more consistent ruminal mat than barley and corn, thus resulting in higher mean retention time and chewing activity, whereas no changes in 3.5% FCM and milk fat were observed.


The challenges of a circumsporozoite protein-based malaria vaccine.
A safe and effective vaccine will likely be necessary for the control or eradication of malaria which kills 400,000 annually. Our most advanced vaccine candidate to date is RTS,S which is based on the Plasmodium falciparum circumsporozoite protein (PfCSP) of the malaria parasite. However, protection by RTS,S is incomplete and short-lived. Here we summarize results from recent clinical trials of RTS,S and critically evaluate recent studies that aim to understand the correlates of protective immunity and why vaccine-induced protection is short-lived. In particular, recent systems serology studies have highlighted a key role for the necessity of inducing functional antibodies. In-depth analyses of immune responses to CSP in both mouse models and vaccinated humans have also highlighted difficulties in generating the maintaining high-quality antibody responses. Finally, in recent years biophysical and structural studies of antibody binding to PfCSP have led to a better understanding of how highly potent antibodies can block infection, which can inform vaccine design. We highlight how both structure-guided vaccine design and a better understanding of the immune response to PfCSP can inform a second generation of PfCSP-based vaccines stimulating a broader range of protective targets within PfCSP.


The Value of Telehealth and a Team-Based Approach in Improving Developmental and Behavioral Care During the COVID-19 Pandemic.
Billy is a 2.6-year-old boy who presented for evaluation in the developmental-behavioral pediatrics (DBP) clinic 2 weeks before the onset of pandemic-related clinic restrictions. Billy had received early intervention for the past year because of speech and fine motor delays. Billy's parents requested the evaluation in the DBP clinic because his delayed speech and disruptive behaviors had raised concern that he may have autism spectrum disorder. Owing to the onset of the pandemic, subsequent visits were completed through telehealth with a developmental-behavioral pediatrician, psychologist, behavioral clinician, and social workers who developed a collaborative plan of care. Billy was diagnosed with global developmental delay, significant tantrums, and impulsivity but did not meet the criteria for autism spectrum disorder.Billy lives with his parents and 2 sisters in a rural area, 3 hours from the DBP clinic. Both of his parents have been treated for depression in the past and reported that school was difficult for them. His sisters, ages 5 and 6 years, receive speech/language therapy but have not required additional special education services. His family has endured recent stressors including a flooding event that caused significant damage to their home, financial difficulties, and the recent unexpected death of a close family member. Billy's disruptive behaviors have resulted in difficulty finding and maintaining child care, further contributing to parental stress and dysfunction in the home.Despite assistance from the social worker, additional developmental and behavioral support services near the family's home were not identified. Therefore, services were offered to Billy and his parents through telehealth. Billy's parents began behavioral parent training with a clinician embedded within the DBP clinic and, with direct support from his parents, Billy began receiving supplemental speech/language and occupational therapies through telehealth. Through recurrent engagement with Billy's parents and frequent communication among the behavioral clinician, developmental-behavioral pediatrician, psychologist, and social worker, Billy was able to make significant developmental progress, and his parents reported improved ability to manage his difficult behaviors.How can telehealth be used to help families navigate complex systems and obtain optimal care and support?


Dietary Genistein Inhibits Methylglyoxal-Induced Advanced Glycation End Product Formation in Mice Fed a High-Fat Diet.
Methylglyoxal (MGO), an important precursor of advanced glycation end products (AGEs), circulates at high concentrations in diabetic patients' blood and plays an important role in the pathogenesis of diabetes and other chronic diseases. The aim of this study was to determine whether dietary genistein can prevent indicators of metabolic syndrome (MetS) induced by a very-high-fat (VHF) diet or a high-fat (HF) diet plus exogenous MGO, and the accumulation of MGO and AGEs in mice. Male, 6-wk-old C57BL/6J mice (n = 15) were fed a low-fat (LF) diet (10% fat energy) or a VHF diet (60% fat energy) alone or including 0.25% genistein (VHF-G) for 16 wk in study 1. In study 2, 75 similar mice were fed the LF diet (LF) or the HF diet alone (HF) or in combination with up to 0.2% MGO in water (HFM) and 0.067% (HFM-GL) or 0.2% (HFM-GH) dietary genistein for 18 wk. Anthropometric and metabolic data were obtained in both studies to determine the effects of MGO and genistein on variables indicative of MetS. Body weight gain, fat deposits, dyslipidemia, hyperglycemia, and fatty liver were ameliorated by dietary genistein in both studies. The plasma MGO concentration in VHF-G mice was 52% lower than that in VHF mice. Moreover, the AGE concentrations in plasma, liver, and kidney of VHF-G mice were 73%, 52%, and 49%, respectively, lower than in the VHF group (study 1). Similarly, the concentrations of plasma MGO and AGE in plasma, liver, and kidney of HFM-GH mice were 33.5%, 49%, 69%, and 54% lower than in HFM mice (study 2). Genistein inhibited AGE formation by trapping MGO to form adducts and upregulating the expressions of glyoxalase I and II and aldose reductase in liver and kidney to detoxify MGO in both studies. Our data demonstrate for the first time that genistein significantly lowers MGO and AGE concentrations in 2 mouse MetS models via multiple pathways.


Research Progress of Nanocarriers for the Treatment of Alzheimer's Disease.
Currently, many therapeutic drugs are difficult to cross the blood-brain barrier (BBB), making it difficult to reach the site of action and thus fail to achieve the desired efficacy. In recent years, researchers and drug designers have increasingly focused on nanotechnology to break through the difficulty of small molecule inhibitors to cross the blood-brain barrier (BBB) and improve the success rate of drug delivery to the central nervous system. Among the common central neurological diseases, such as encephalitis, Parkinson's, Alzheimer's disease, and epilepsy, Alzheimer's disease has attracted much attention from researchers. Alzheimer's disease is a specific neurodegenerative disease, which causes irreversible degeneration of neurons as well as synapses in the brain, resulting in memory and cognitive dysfunction, along with other psychiatric symptoms and behavioral disorders, which seriously affects people's everyday life. Moreover, nanotechnology has excellent potential for application in AD treatment. Studies have shown that nanocarriers can target the delivery of chemotherapeutic drugs, antioxidants, and other therapeutic substances to brain tissue using existing physiological mechanisms, thus effectively alleviating the disease progression of AD. Therefore, various nanoparticles and nanomedicine have been developed and constructed for diagnosing and treating AD in the past decades, such as nanoparticles, bionanoparticles, liposomes, nano-gel, dendrimers, and self-assembled nanoparticles. This study aims to review the applications and results of nanotechnology in the treatment of Alzheimer's disease in recent years and provide some ideas and clues for future research and development of more effective drug delivery systems.


Second-generation Notch1 activity-trap mouse line (N1IP::CreHI) provides a more comprehensive map of cells experiencing Notch1 activity.
We have previously described the creation and analysis of a Notch1 activity-trap mouse line, Notch1 intramembrane proteolysis-Cre6MT or N1IP::Cre(LO), that marked cells experiencing relatively high levels of Notch1 activation. Here, we report and characterize a second line with improved sensitivity (N1IP::Cre(HI)) to mark cells experiencing lower levels of Notch1 activation. This improvement was achieved by increasing transcript stability and by restoring the native carboxy terminus of Cre, resulting in a five- to tenfold increase in Cre activity. The magnitude of this effect probably impacts Cre activity in strains with carboxy-terminal Ert2 fusion. These two trap lines and the related line N1IP::Cre(ERT2) form a complementary mapping tool kit to identify changes in Notch1 activation patterns in vivo as the consequence of genetic or pharmaceutical intervention, and illustrate the variation in Notch1 signal strength from one tissue to the next and across developmental time.


Bordetella pertussis isolation in general practice: 1977-79 whooping cough epidemic in West Glamorgan.
Some of the factors influencing the isolation rate of Bordetella pertussis during a whooping cough epidemic in West Glamorgan, Wales, are reported. The organism was isolated from 39% of patients with clinical whooping cough, pernasal swabbing being much more successful than cough plates. Isolation rates were increased in the non-immunized, particularly in the first year of life. Erythromycin and co-trimoxazole significantly reduced the isolation rate of B. pertussis but this did not occur with penicillin. In this study 20% of patients were culture positive 6 weeks after the onset of their infection. It is suggested that the Department of Health and Social Security recommendation of a minimum period of three weeks exclusion of children from school is inadequate. During the epidemic, the proportion of strains of B. pertussis containing antigen 2 more than doubled.


Histopathology of salivary and mammary gland tumors in transgenic mice expressing a human Ha-ras oncogene.
Mutated ras genes are powerful transforming agents in vitro and are found in a wide variety of human tumors in vivo. We characterized the histopathology and p21 protein expression associated with tumorigenesis in line 69 transgenic mice carrying an activated, human c-Ha-ras gene on the Y-chromosome (A. C. Andres, C. A. Schonenberger, B. Groner, L. Hennighausen, M. LeMeur, and P. Gerlinger, Proc. Natl. Acad. Sci. USA, 84: 1299-1303, 1987). Male mice developed salivary and/or mammary gland tumors. The salivary tumors were adenosquamous carcinomas arising from serous areas of the submandibular gland. They characteristically exhibited densely packed cords and sheets of moderately anaplastic cells. Tumorigenic tissue had a high mitotic index, and all tumor-bearing animals had an ongoing inflammatory response as evidenced by extensive immune cell infiltration of affected tissue. Half of the mammary gland tumors were adenosquamous carcinomas with multiple foci of squamous metaplasia, while the rest were adenocarcinomas containing glandular tissue. Most tumors had a high mitotic index, and abnormal mitotic figures were common. All tumors produced p21 ras, as confirmed by immunohistochemistry and Western blots. Both tumor types expressed elevated levels of p21 protein. Microscopic lung metastases were present in 5 of 35 animals (14%). Our results suggest that this transgenic mouse will provide a useful model for testing therapies directed against ras-associated tumorigenesis.


Artesunate-amodiaquine and artemether-lumefantrine for the treatment of uncomplicated falciparum malaria in Liberia: in vivo efficacy and frequency of molecular markers.
Artesunate-amodiaquine (ASAQ) and Artemether-lumefantrine (AL) are the recommended treatment for uncomplicated Plasmodium falciparum malaria in Liberia. Intermittent preventive treatment with sulfadoxine/pyrimethamine is also recommended for pregnant women. The therapeutic efficacy of Artesunate-amodiaquine and Artemether-lumefantrine, and the frequency of molecular markers associated with anti-malarial drug resistance were investigated. The therapeutic efficacy of ASAQ and AL was evaluated using the standard World Health Organization protocol (WHO. Methods for Surveillance of Antimalarial Drug Efficacy. Geneva: World Health Organization; 2009. https://www.who.int/malaria/publications/atoz/9789241597531/en/ ). Eligible children were recruited and monitored clinically and parasitologically for 28 days. Polymorphisms in the Pfkelch 13, chloroquine resistance transporter (Pfcrt), multidrug resistance 1 (Pfmdr-1), dihydrofolate reductase (Pfdhfr), and dihydropteroate synthase (Pfdhps) genes and copy number variations in the plasmepsin-2 (Pfpm2) gene were assessed in pretreatment samples. Of the 359 children enrolled, 180 were treated with ASAQ (89 in Saclepea and 91 in Bensonville) and 179 with AL (90 in Sinje and 89 in Kakata). Of the recruited children, 332 (92.5%) reached study endpoints. PCR-corrected per-protocol analysis showed ACPR of 90.2% (95% CI: 78.6-96.7%) in Bensonville and 92.7% (95% CI: 83.4.8-96.5%) in Saclepea for ASAQ, while ACPR of 100% was observed in Kakata and Sinje for AL. In both treatment groups, only two patients had parasites on day 3. No artemisinin resistance associated Pfkelch13 mutations or multiple copies of Pfpm2 were found. Most samples tested had the Pfcrt 76 T mutation (80/91, 87.9%), while the Pfmdr-1 86Y (40/91, 44%) and 184F (47/91, 51.6%) mutations were less frequent. The Pfdhfr triple mutant (51I/59R/108 N) was the predominant allele (49.2%). For the Pfdhps gene, it was the 540E mutant (16.0%), and the 436A mutant (14.3%). The quintuple allele (51I/59R/108 N-437G/540E) was detected in only one isolate (1/357). This study reports a decline in the efficacy of ASAQ treatment, while AL remained highly effective, supporting the recent decision by NMCP to replace ASAQ with AL as first-line treatment for uncomplicated falciparum malaria. No association between the presence of the mutations in Pfcrt and Pfmdr-1 and the risk of parasite recrudescence in patients treated with ASAQ was observed. Parasites with signatures known to be associated with artemisinin and piperaquine resistance were not detected. The very low frequency of the quintuple Pfdhfr/Pfdhps mutant haplotype supports the continued use of SP for IPTp. Monitoring of efficacy and resistance markers of routinely used anti-malarials is necessary to inform malaria treatment policy. Trial registration ACTRN12617001064392.


Developmental dyscalculia.
Developmental dyscalculia is a specific learning disability affecting the normal acquisition of arithmetic skills. Genetic, neurobiologic, and epidemiologic evidence indicates that dyscalculia, like other learning disabilities, is a brain-based disorder. However, poor teaching and environmental deprivation have also been implicated in its etiology. Because the neural network of both hemispheres comprises the substrate of normal arithmetic skills, dyscalculia can result from dysfunction of either hemisphere, although the left parietotemporal area is of particular significance. The prevalence of developmental dyscalculia is 5 to 6% in the school-aged population and is as common in girls as in boys. Dyscalculia can occur as a consequence of prematurity and low birthweight and is frequently encountered in a variety of neurologic disorders, such as attention-deficit hyperactivity disorder (ADHD), developmental language disorder, epilepsy, and fragile X syndrome. Developmental dyscalculia has proven to be a persisting learning disability, at least for the short term, in about half of affected preteen pupils. Educational interventions for dyscalculia range from rote learning of arithmetic facts to developing strategies for solving arithmetic exercises. The long-term prognosis of dyscalculia and the role of remediation in its outcome are yet to be determined.


A comparison of visual hallucinations across disorders.
Research into hallucinations typically regards them as single sensory or unimodal experiences leading to a comparative neglect of co-occurring multi-sensory hallucinations (MSH). People with psychosis who have visual hallucinations (VH) report high rates of hallucinations in other senses (auditory, olfactory, tactile). However, it is not known if this is similar to other groups who report VH. Consequently, this study explored MSH in four different patient groups who all had current VH. Archival data from standardised assessments of visual hallucinations in people with psychosis (n = 22), eye disease (ED) (n = 82), Lewy body Dementia (LBD) (n = 41), and Parkinson's disease (PD) (n = 41) determined the presence of MSH. People with psychosis and visual hallucinations reported significantly higher rates of MSH (auditory, 73%; tactile, 82%; olfactory/gustatory hallucinations, 27%) than the LBD group (auditory, 21%; tactile, 28%; olfactory/gustatory, 6%), ED (auditory, 1%; tactile, 11%; olfactory/gustatory, 0%) and PD patients (auditory, 3%; tactile, 8%; olfactory/gustatory, 3%). Regardless of diagnostic grouping, participants with MSH reported greater conviction that the VH were real, and reported greater distress. People with psychosis with VH report high rates of MSH unlike groups of older adults with VH. These between group differences in MSH prevalence have implications for clinical practice and theory.


Mitigating bias from intermittent measurement of time-dependent covariates in failure time analysis.
Cox regression models are routinely fitted to examine the association between time-dependent markers and a failure time when analyzing data from clinical registries. Typically, the marker values are measured periodically at clinic visits with the recorded value carried forward until the next assessment. We examine the asymptotic behavior of estimators from Cox regression models under this observation and data handling scheme when the true relationship is based on a Cox model using the current value of the marker. Specifically, we explore the impact of the marker process dynamics, the clinic visit intensity, and the marginal failure rate on the limiting value of the estimator of the marker effect from the Cox model. We also illustrate how a joint multistate model that accommodates intermittent observation of the time-varying marker can be formulated. Simulation studies demonstrate that the finite sample performance of the naive estimator aligns with the asymptotic results and shows good performance of the estimators from the joint model. We apply both methods to data from a study of bone markers and their effect on the development of skeletal complications in metastatic cancer.


Inactivated polio vaccine: industrial production from micro-carrier Vero cells culture.
In 1980, we presented our preliminary results of large-scale production of Inactivated Polio-Vaccine (IPV). The virus was produced by Vero cells culture grown on micro-carrier in 150 litre tanks; now we produce the poliovirus in 1000 litre tanks. All the tests certifying the 'Cell Bank' have been passed; the quality of purification procedure was demonstrated; additional clinical studies were done; this new reassessed IPV was licensed in France in July 1982.


Constructing covariance functions for random regression models for growth in Gelbvieh beef cattle.
Genetic parameters for a random regression model of growth in Gelbvieh beef cattle were constructed using existing estimates. Information for variances along ages was provided by parameters used for routine Gelbvieh multiple-trait evaluation, and information on correlations among different ages was provided by random regression model estimates from literature studies involving Nellore cattle. Both sources of information were combined into multiple-trait estimates; corrected for continuity, smoothness, and general agreement with literature estimates; and extrapolated to 730 d. Covariance functions using standardized Legendre polynomials were fit for the following effects: additive genetic (direct and maternal), and animal and maternal permanent environment. Residual variances at different ages were fitted using linear splines with three knots. Fit was by least squares. The order of polynomials was varied from third to sixth. Increasing the fit beyond cubic provided small improvements in R2 and increased the number of small eigenvalues of covariance matrices, especially for the additive effect. Parameters for a random regression model in beef cattle can be constructed with negligible artifacts from literature estimates. Formulas can easily be modified for other types of polynomials and splines.


Ocular melanosis in the Cairn Terrier: clinical description and investigation of mode of inheritance.
To describe the clinical features of ocular melanosis in Cairn Terriers. One hundred and fourteen Cairn Terriers diagnosed with ocular melanosis. A complete eye examination was performed on each dog. Four dogs (and two unaffected control dogs) underwent a high frequency ultrasound examination of the anterior segment. The pedigrees of affected dogs were analyzed. Forty-four (38.6%) dogs were male and 67 (58.7%) female; the sex of three dogs (2.6%) was not provided. A four-stage grading system of the ocular changes was developed. There was a variable age of onset, and the earliest change was a dark-colored thickening of the iris root. This was followed by the development of episcleral/scleral pigment plaques, release of pigment into the aqueous and deposition in the drainage apparatus, particularly ventrally. Secondary glaucoma developed in the most severely affected dogs. A slow progression of pigmentation in the tapetal fundus was observed and in some dogs pigment on the surface of the optic nerve head was seen. Three dogs developed uveal melanocytic neoplasms. Pedigree analysis suggested a possible autosomal dominant mode of inheritance. Ocular melanosis is an inherited, probably autosomal-dominant condition with a variable age of onset and rate of progression. It results in a thickening and pigmentation of the iris, release of pigmented material into the aqueous, pigment deposition in the sclera/episclera, and to a lesser extent posterior segment pigment deposition. Following extensive pigment deposition in the aqueous drainage pathways it can result in secondary glaucoma.


CO2 Enrichment Stimulates Dissolved Organic Carbon Release in Coral Reef Macroalgae.
Dissolved organic carbon (DOC) released by macroalgae is important in the context of coral reef degradation as it contributes to coral mortality by promoting bacterial metabolism on the coral surface. Using experimental carbon dioxide (CO2 ) manipulations in outdoor flow-through tanks, we found that seawater CO2 enrichment enhances daily net DOC release in a range of macroalgal species in the Great Barrier Reef (Australia). There was, however, large variability in DOC release among species, light and dark conditions, and CO2 exposure times. Under light conditions, DOC release in the red macroalga Amansia was 15 times higher under high CO2 conditions compared to ambient CO2 , however, CO2 enhancement did not affect DOC production in the other species. Results from the night incubations were more consistent as three of the four species (Amansia, Lobophora, and Sargassum) enhanced DOC release when enriched with CO2 . DOC fluxes shifted from production in the 1-d incubations to consumption in the 19-d experiment under light conditions, suggesting an important role of bacteria in DOC balances. The results suggest that rising CO2 (and ocean acidification) will continue to intensify space competition in favor of the macroalgae, potentially exacerbating reef degradation and ecological phase shifts from coral to macroalgal dominance.


Public expenditure on hospitalizations for COVID-19 treatment in 2020, in Brazil.
Describe the expenditure resulting from hospitalizations for clinical treatment of users diagnosed with COVID-19 in the Unified Health System (SUS) between February and December 2020. This is a descriptive study based on data from the Hospital Information System about government expenditure on hospitalizations for clinical treatment of users diagnosed with COVID-19 and causes included in the ICD-10 chapters. We obtained the number of hospitalizations, average length of stay, lethality rate, and total expenditure considering hospital services, professional services and average expenditure per hospitalization. In the period evaluated, SUS registered 462,149 hospitalizations, 4.9% of them for COVID-19 treatment. Total expenditure exceeded R$ 2.2 billion, with 85% allocated to hospital services and 15% to professional services. Expenditure for treating COVID-19 was distributed differently between the country's regions. The Southeast region had the highest number of hospitalizations, highest total amount spent, highest average length of stay in days, and highest lethality rate; the South region, in turn, recorded the highest percentage of spending on non-profit hospitals (58%) and corporate hospitals (15%). Hospitalizations for clinical treatment of coronavirus infection were more costly compared to those for treatment of acute respiratory failure and pneumonia or influenza. Our results show the disparities in hospitalization expenditure for similar procedures between the regions of Brazil, underlining the vulnerability and the need for strategies to reduce the differences in access, use, and distribution of SUS resources, ensuring equanimity, and considering the unfair inequalities between the country's regions.


Financing the Fourth Year: Experiences of Required 4-Year Family Medicine Residency Programs.
The optimal length of family medicine training has been debated since the specialty's inception. Currently there are four residency programs in the United States that require 4 years of training for all residents through participation in the Accreditation Council for Graduate Medical Education Length of Training Pilot. Financing the additional year of training has been perceived as a barrier to broader dissemination of this educational innovation. Utilizing varied approaches, the family medicine residency programs at Middlesex Health, Greater Lawrence Health Center, Oregon Health and Science University, and MidMichigan Medical Center all demonstrated successful implementation of a required 4-year curricular model. Total resident complement increased in all programs, and the number of residents per class increased in half of the programs. All programs maintained or improved their contribution margins to their sponsoring institutions through additional revenue generation from sources including endowment funding, family medicine center professional fees, institutional collaborations, and Health Resources and Services Administration Teaching Health Center funding. Operating expense per resident remained stable or decreased. These findings demonstrate that extension of training in family medicine to 4 years is financially feasible, and can be funded through a variety of models.


Gene-environment interactions in Leber hereditary optic neuropathy.
Leber hereditary optic neuropathy (LHON) is a genetic disorder primarily due to mutations of mitochondrial DNA (mtDNA). Environmental factors are thought to precipitate the visual failure and explain the marked incomplete penetrance of LHON, but previous small studies have failed to confirm this to be the case. LHON has no treatment, so identifying environmental triggers is the key to disease prevention, whilst potentially revealing new mechanisms amenable to therapeutic manipulation. To address this issue, we conducted a large, multicentre epidemiological study of 196 affected and 206 unaffected carriers from 125 LHON pedigrees known to harbour one of the three primary pathogenic mtDNA mutations: m.3460G>A, m.11778G>A and m.14484T>C. A comprehensive history of exposure to smoking, alcohol and other putative environmental insults was collected using a structured questionnaire. We identified a strong and consistent association between visual loss and smoking, independent of gender and alcohol intake, leading to a clinical penetrance of 93% in men who smoked. There was a trend towards increased visual failure with alcohol, but only with a heavy intake. Based on these findings, asymptomatic carriers of a LHON mtDNA mutation should be strongly advised not to smoke and to moderate their alcohol intake.


Atomic Scale Characterization of Fluxional Cation Behavior on Nanoparticle Surfaces: Probing Oxygen Vacancy Creation/Annihilation at Surface Sites.
Oxygen vacancy creation and annihilation are key processes in nonstoichiometric oxides such as CeO2. The oxygen vacancy creation and annihilation rates on an oxide's surface partly govern its ability to exchange oxygen with the ambient environment, which is critical for a number of applications including energy technologies, environmental pollutant remediation, and chemical synthesis. Experimental methods to probe and correlate local oxygen vacancy reaction rates with atomic-level structural heterogeneities would provide significant information for the rational design and control of surface functionality; however, such methods have been unavailable to date. Here, we characterize picoscale fluxional behavior in cations using time-resolved in situ aberration-corrected transmission electron microscopy to locate atomic-level variations in oxygen vacancy creation and annihilation rates on oxide nanoparticle surfaces. Low coordination number sites such as steps and edges, as well as locally strained sites, exhibited the greatest number of cation displacements, implying enhanced surface oxygen vacancy activity at these sites. The approach has potential applications to a much wider class of materials and catalysis problems involving surface and interfacial transport functionalities.


An epitope-based malaria vaccine targeting the junctional region of circumsporozoite protein.
A malaria vaccine that elicits long-lasting protection and is suitable for use in endemic areas remains urgently needed. Here, we assessed the immunogenicity and prophylactic efficacy of a vaccine targeting a recently described epitope on the major surface antigen on Plasmodium falciparum sporozoites, circumsporozoite protein (CSP). Using a virus-like particle (VLP)-based vaccine platform technology, we developed a vaccine that targets the junctional region between the N-terminal and central repeat regions of CSP. This region is recognized by monoclonal antibodies, including mAb CIS43, that have been shown to potently prevent liver invasion in animal models. We show that CIS43 VLPs elicit high-titer and long-lived anti-CSP antibody responses in mice and is immunogenic in non-human primates. In mice, vaccine immunogenicity was enhanced by using mixed adjuvant formulations. Immunization with CIS43 VLPs conferred partial protection from malaria infection in a mouse model, and passive transfer of serum from immunized macaques also inhibited parasite liver invasion in the mouse infection model. Our findings demonstrate that a Qβ VLP-based vaccine targeting the CIS43 epitope combined with various adjuvants is highly immunogenic in mice and macaques, elicits long-lasting anti-CSP antibodies, and inhibits parasite infection in a mouse model. Thus, the CIS43 VLP vaccine is a promising pre-erythrocytic malaria vaccine candidate.


Activation of neurons in the insular cortex and lateral hypothalamus during food anticipatory period caused by food restriction in mice.
Mice fed a single meal daily at a fixed time display food anticipatory activity (FAA). It has been reported that the insular cortex (IC) plays an essential role in food anticipation, and lateral hypothalamus (LH) regulates the expression of FAA. However, how these areas contribute to FAA production is still unclear. Thus, we examined the temporal and spatial activation pattern of neurons in the IC and LH during the food anticipation period to determine their role in FAA establishment. We observed an increase of c-Fos-positive neurons in the IC and LH, including orexin neurons of male adult C57BL/6 mice. These neurons were gradually activated from the 1st day to 15th day of restricted feeding. The activation of these brain regions, however, peaked at a distinct point in the food restriction procedure. These results suggest that the IC and LH are differently involved in the neural network for FAA production.


Lowering the UK domestic radon Action Level to prevent more lung cancers--is it cost-effective?
Case studies have shown that radon gas can accumulate within domestic properties at sufficiently high levels that it can cause lung cancer, and recent studies have suggested that this risk remains significant below the UK domestic Action Level of 200 Bq m(-3). Raised radon levels can be reduced by engineering measures, and it has been shown that domestic radon remediation programmes in UK Affected Areas can result in reduced risks to the population and can be cost-effective. We consider here the benefits and costs of the domestic radon remediation programme in Northamptonshire, UK, and consider the implications for that programme of reducing the UK Action Level below its present value. A radon remediation programme based on an Action Level above 200 Bq m(-3) will cost less and will target those most at risk, but will be less cost-effective and will lead to higher residual dose and greater risk of cancer in the remaining population. Reducing the Action Level below 200 Bq m(-3) will prevent more cancers, but at significantly higher cost. It will also be less cost-effective, because remediation of a significant number of houses with moderate radon levels will provide only a modest health benefit to occupants. Overall, a completed radon remediation programme of the type implemented in Northamptonshire is most cost-effective for an Action Level between 200 and 300 Bq m(-3). The implications for future health policy are discussed.


Electroporation of short hairpin RNAs for rapid and efficient gene knockdown in the starlet sea anemone, Nematostella vectensis.
A mechanistic understanding of evolutionary developmental biology requires the development of novel techniques for the manipulation of gene function in phylogenetically diverse organismal systems. Recently, gene-specific knockdown by microinjection of short hairpin RNA (shRNA) was applied in the sea anemone Nematostella vectensis, demonstrating that the shRNA approach can be used for efficient and robust sequence-specific knockdown of a gene of interest. However, the time- and labor-intensive process of microinjection limits access to this technique and its application in large scale experiments. To address this issue, here we present an electroporation protocol for shRNA delivery into Nematostella eggs. This method leverages the speed and simplicity of electroporation, enabling users to manipulate gene expression in hundreds of eggs or embryos within minutes. We provide a detailed description of the experimental procedure, including reagents, electroporation conditions, preparation of Nematostella eggs, and follow-up care of experimental animals. Finally, we demonstrate the knockdown of several endogenous and exogenous genes with known phenotypes and discuss the potential applications of this method.


Identification of cyclin D1 and other novel targets for the von Hippel-Lindau tumor suppressor gene by expression array analysis and investigation of cyclin D1 genotype as a modifier in von Hippel-Lindau disease.
Germ-line mutations in the von Hippel-Lindau (VHL) tumor suppressor disease are associated with a high risk of retinal and cerebellar hemangioblastomas, renal cell carcinoma (RCC), and, in some cases, pheochromocytoma (PHE). In addition, somatic mutation or epigenetic inactivation of the VHL gene occurs in most clear cell RCCs. VHL protein (pVHL) has a critical role in regulating proteasomal degradation of the HIF transcription factor, and VHL inactivation results in overexpression of many hypoxia-inducible mRNAs including vascular endothelial growth factor (VEGF). To identify novel pVHL target genes we investigated the effect of wild-type (WT) pVHL on the expression of 588 cancer-related genes in two VHL-defective RCC cell lines. Expression array analysis identified nine genes that demonstrated a >2-fold decrease in expression in both RCC cell lines after restoration of WT pVHL. Three of the nine genes (VEGF, PAI-1, and LRP1) had been reported previously as pVHL targets and are known to be hypoxia-inducible. In addition, six novel targets were detected: cyclin D1 (CCND1), cell division protein kinase 6, collagen VIII alpha 1 subunit, CD59 glycoprotein precursor, integrin beta8, and interleukin 6 precursor IFN-beta2. We found no evidence that CCND1, cell division protein kinase 6, CD59, and integrin beta8 expression was influenced by hypoxia suggesting that pVHL down-regulates these targets by a HIF-independent mechanism. A type 2C pVHL mutant (V188L), which is associated with a PHE only phenotype (and had been shown previously to retain the ability to promote HIF ubiquitylation), retained the ability to suppress CCND1expression suggesting that loss of pVHL-mediated suppression of cyclin D1 is not necessary for PHE development in VHL disease. Other studies have suggested that: (a) genetic modifiers influence the phenotypic expression of VHL disease; and (b) polymorphic variation at a CCND1 codon 242 A/G single nucleotide polymorphism (SNP) may influence cancer susceptibility or prognosis in some situations. Therefore, we analyzed the relationship between CCND1 genotype and phenotypic expression of VHL disease. There was an association between the G allele and multiple retinal angiomas (P = 0.04), and risk of central nervous system hemangioblastomas (P = 0.05). These findings suggest that a variety of HIF-independent mechanisms may contribute to pVHL tumor suppressor activity and that polymorphic variation at one pVHL target influences the phenotypic expression of VHL disease.


PTK787/ZK 222584, a novel and potent inhibitor of vascular endothelial growth factor receptor tyrosine kinases, impairs vascular endothelial growth factor-induced responses and tumor growth after oral administration.
PTK787/ZK 222584 (1-[4-chloroanilino]-4-[4-pyridylmethyl] phthalazine succinate) is a potent inhibitor of vascular endothelial growth factor (VEGF) receptor tyrosine kinases, active in the submicromolar range. It also inhibits other class III kinases, such as the platelet-derived growth factor (PDGF) receptor beta tyrosine kinase, c-Kit, and c-Fms, but at higher concentrations. It is not active against kinases from other receptor families, such as epidermal growth factor receptor, fibroblast growth factor receptor-1, c-Met, and Tie-2, or intracellular kinases such as c-Src, c-Abl, and protein kinase C-alpha. PTK787/ZK 222584 inhibits VEGF-induced autophosphorylation of kinase insert domain-containing receptor (KDR), endothelial cell proliferation, migration, and survival in the nanomolar range in cell-based assays. In concentrations up to 1 microM, PTK787/ZK 222584 does not have any cytotoxic or antiproliferative effect on cells that do not express VEGF receptors. After oral dosing (50 mg/kg) to mice, plasma concentrations of PTK787/ZK 222584 remain above 1 microM for more than 8 h. PTK787/ZK 222584 induces dose-dependent inhibition of VEGF and PDGF-induced angiogenesis in a growth factor implant model, as well as a tumor cell-driven angiogenesis model after once-daily oral dosing (25-100 mg/kg). In the same dose range, it also inhibits the growth of several human carcinomas, grown s.c. in nude mice, as well as a murine renal carcinoma and its metastases in a syngeneic, orthotopic model. Histological examination of tumors revealed inhibition of microvessel formation in the interior of the tumor. PTK787/ZK 222584 is very well tolerated and does not impair wound healing. It also does not have any significant effects on circulating blood cells or bone marrow leukocytes as a single agent or impair hematopoetic recovery after concomitant cytotoxic anti-cancer agent challenge. This novel compound has therapeutic potential for the treatment of solid tumors and other diseases where angiogenesis plays an important role.


Mandatory retirement and labor-force participation of respondents in the Retirement History Study.
The Age Discrimination in Employment Act (ADEA) was amended in 1978 to prohibit mandatory retirement before age 70 in most occupations. The impact of this legislation on the probability of older persons remaining in the labor force is the primary concern of this article. Specifically, questions concerning which older workers are affected by mandatory-retirement provisions and the extent to which they are forced to retire and leave the labor force are examined. Tabular analysis of data from the Retirement History Study on persons aged 62-63 in 1969 shows significant variation in mandatory-retirement coverage between the public and private sectors and across industries, occupations, and demographic groups. Until age 65, the labor-force participation rate of those facing compulsory retirement is higher than or equal to that of those not covered but it drops significantly below the noncovered rate after 65. Logit analysis of the labor-force participation of persons before and after age 65 indicates that mandatory retirement at that age reduces the probability of retirement by approximately 16.7 percentage points for white men wage earners. This results in a decline in the labor-force participation rate of all men aged 66-67 of approximately 4 percentage points.


Matrix Mechanotransduction via Yes-Associated Protein in Human Lamina Cribrosa Cells in Glaucoma.
Extracellular matrix stiffening is characteristic of both aging and glaucoma, and acts as a promoter and perpetuator of pathological fibrotic remodeling. Here, we investigate the role of a mechanosensitive transcriptional coactivator, Yes-associated protein (YAP), a downstream effector of multiple signaling pathways, in lamina cribrosa (LC) cell activation to a profibrotic, glaucomatous state. LC cells isolated from glaucomatous human donor eyes (GLC; n = 3) were compared to LC cells from age-matched nonglaucomatous controls (NLC; n = 3) to determine differential YAP expression, protein levels, and proliferation rates. NLC cells were then cultured on soft (4 kPa), and stiff (100 kPa), collagen-1 coated polyacrylamide hydrogel substrates. Quantitative real-time RT-PCR, immunoblotting, and immunofluorescence microscopy were used to measure the expression, activity, and subcellular location of YAP and its downstream targets, respectively. Proliferation rates were examined in NLC and GLC cells by methyl thiazolyl tetrazolium salt assays, across a range of incrementally increased substrate stiffness. Endpoints were examined in the presence or absence of a YAP inhibitor, verteporfin (2 µM). GLC cells show significantly (P < 0.05) increased YAP gene expression and total-YAP protein compared to NLC cells, with significantly increased proliferation. YAP regulation is mechanosensitive, because NLC cells cultured on pathomimetic, stiff substrates (100 kPa) show significantly upregulated YAP gene and protein expression, increased YAP phosphorylation at tyrosine 357, reduced YAP phosphorylation at serine 127, increased nuclear pooling, and increased transcriptional target, connective tissue growth factor. Accordingly, myofibroblastic markers, α-smooth muscle actin (α-SMA) and collagen type I, alpha 1 (Col1A1) are increased. Proliferation rates are elevated on 50 kPa substrates and tissue culture plastic. Verteporfin treatment significantly inhibits YAP-mediated cellular activation and proliferation despite a stiffened microenvironment. These data demonstrate how YAP plays a pivotal role in LC cells adopting a profibrotic and proliferative phenotype in response to the stiffened LC present in aging and glaucoma. YAP provides an attractive and novel therapeutic target, and its inhibition via verteporfin warrants further clinical investigation.


Optimality principles explaining divergent responses of alpine vegetation to environmental change.
Recent increases in vegetation greenness over much of the world reflect increasing CO2 globally and warming in cold areas. However, the strength of the response to both CO2 and warming in those areas appears to be declining for unclear reasons, contributing to large uncertainties in predicting how vegetation will respond to future global changes. Here, we investigated the changes of satellite-observed peak season absorbed photosynthetically active radiation (Fmax ) on the Tibetan Plateau between 1982 and 2016. Although climate trends are similar across the Plateau, we identified robust divergent responses (a greening of 0.31 ± 0.14% year-1 in drier regions and a browning of 0.12 ± 0.08% year-1 in wetter regions). Using an eco-evolutionary optimality (EEO) concept of plant acclimation/adaptation, we propose a parsimonious modelling framework that quantitatively explains these changes in terms of water and energy limitations. Our model captured the variations in Fmax with a correlation coefficient (r) of .76 and a root mean squared error of .12 and predicted the divergent trends of greening (0.32 ± 0.19% year-1 ) and browning (0.07 ± 0.06% year-1 ). We also predicted the observed reduced sensitivities of Fmax to precipitation and temperature. The model allows us to explain these changes: Enhanced growing season cumulative radiation has opposite effects on water use and energy uptake. Increased precipitation has an overwhelmingly positive effect in drier regions, whereas warming reduces Fmax in wetter regions by increasing the cost of building and maintaining leaf area. Rising CO2 stimulates vegetation growth by enhancing water-use efficiency, but its effect on photosynthesis saturates. The large decrease in the sensitivity of vegetation to climate reflects a shift from water to energy limitation. Our study demonstrates the potential of EEO approaches to reveal the mechanisms underlying recent trends in vegetation greenness and provides further insight into the response of alpine ecosystems to ongoing climate change.


Adaptively Evolving Bacterial Communities for Complete and Selective Reduction of Cr(VI), Cu(II), and Cd(II) in Biocathode Bioelectrochemical Systems.
Bioelectrochemical systems (BESs) have been shown to be useful in removing individual metals from solutions, but effective treatment of electroplating and mining wastewaters requires simultaneous removal of several metals in a single system. To develop multiple-reactor BESs for metals removal, biocathodes were first individually acclimated to three different metals using microbial fuel cells with Cr(VI) or Cu(II) as these metals have relatively high redox potentials, and microbial electrolysis cells for reducing Cd(II) as this metal has a more negative redox potential. The BESs were then acclimated to low concentrations of a mixture of metals, followed by more elevated concentrations. This procedure resulted in complete and selective metal reduction at rates of 1.24 ± 0.01 mg/L-h for Cr(VI), 1.07 ± 0.01 mg/L-h for Cu(II), and 0.98 ± 0.01 mg/L-h for Cd(II). These reduction rates were larger than the no adaptive controls by factors of 2.5 for Cr(VI), 2.9 for Cu(II), and 3.6 for Cd(II). This adaptive procedure produced less diverse microbial communities and changes in the microbial communities at the phylum and genus levels. These results demonstrated that bacterial communities can adaptively evolve to utilize solutions containing mixtures of metals, providing a strategy for remediating wastewaters containing Cr(VI), Cu(II), and Cd(II).


Shut up and pet me! Domestic dogs (Canis lupus familiaris) prefer petting to vocal praise in concurrent and single-alternative choice procedures.
The nature of the interactions that maintain the social behavior of dogs toward humans and which interactions dogs prefer have not been thoroughly investigated. We focused here on dogs' preference for petting and vocal praise, and the influence that familiarity (owner vs. stranger) has on that preference. We first used concurrent choice to evaluate dogs' preference for petting or vocal praise and measured the initial choice, the time spent with each alternative, and the number of within-session alternations. We assessed dogs' preference for petting or vocal praise in (1) shelter dogs, (2) owned dogs with strangers providing both interactions, and (3) owned dogs with the dog's owner providing the interactions. Across all experimental groups, dogs preferred petting to vocal praise. We next assessed time spent with each alternative when only one alternative was available at a time in shelter dogs and owned dogs (Experiment 2). Shelter dogs were tested with a stranger and owned dogs were tested with their owners providing the interaction. Dogs alternated between petting and vocal praise, vocal praise and no interaction, or received only petting for eight 3-min sessions of each comparison. Both shelter and owned dogs spent significantly longer in proximity to the experimenter when the interaction was petting compared to vocal praise. Vocal praise produced as little proximity-seeking behavior as did no interaction. Additionally, dogs did not show any sign of satiation with petting across all eight sessions. Overall, petting seems to be an important interaction between dogs and humans that might maintain inter-specific social behavior but vocal praise likely has to be specifically conditioned. This article is part of a Special Issue entitled: Canine Behavior.


Microchip-based cellular biochemical systems for practical applications and fundamental research: from microfluidics to nanofluidics.
By combining cell technology and microchip technology, innovative cellular biochemical tools can be created from the microscale to the nanoscale for both practical applications and fundamental research. On the microscale level, novel practical applications taking advantage of the unique capabilities of microfluidics have been accelerated in clinical diagnosis, food safety, environmental monitoring, and drug discovery. On the other hand, one important trend of this field is further downscaling of feature size to the 10(1)-10(3) nm scale, which we call extended-nano space. Extended-nano space technology is leading to the creation of innovative nanofluidic cellular and biochemical tools for analysis of single cells at the single-molecule level. As a pioneering group in this field, we focus not only on the development of practical applications of cellular microchip devices but also on fundamental research to initiate new possibilities in the field. In this paper, we review our recent progress on tissue reconstruction, routine cell-based assays on microchip systems, and preliminary fundamental method for single-cell analysis at the single-molecule level with integration of the burgeoning technologies of extended-nano space.


Modeling the Cellular Uptake of Functionalized Carbon Nanohorns Loaded with Cisplatin through a Breast Cancer Cell Membrane.
The cisplatin encapsulation into carbon nanohorns (CNH) is a promising nanoformulation to circumvent the drug dissipation and to specifically accumulate it in tumor sites. Herein, biased molecular dynamics simulations were used to analyze the transmembrane transport of the CNH loaded with cisplatin through a breast cancer cell membrane prototype. The simulations revealed a four-stage mechanism: approach, insertion, permeation, and internalization. Despite the lowest structural disturbance of the membrane provided by the nanocarrier, the average free energy barrier for the translocation was 55.2 kcal mol-1, suggesting that the passive process is kinetically unfavorable. In contrast, the free energy profiles revealed potential wells of -6.8 kcal mol-1 along the insertion stage in the polar heads region of the membrane, which might enhance the retention of the drug in tumor sites; therefore, the most likely cisplatin delivery mechanism should involve the adsorption and retention of CNH on the surface of cancer cells, allowing the loaded cisplatin be slowly released and passively transported through the cell membrane.


preCICE v2: A sustainable and user-friendly coupling library.
preCICE is a free/open-source coupling library. It enables creating partitioned multi-physics simulations by gluing together separate software packages. This paper summarizes the development efforts in preCICE of the past five years. During this time span, we have turned the software from a working prototype -- sophisticated numerical coupling methods and scalability on ten thousands of compute cores -- to a sustainable and user-friendly software project with a steadily-growing community. Today, we know through forum discussions, conferences, workshops, and publications of more than 100 research groups using preCICE. We cover the fundamentals of the software alongside a performance and accuracy analysis of different data mapping methods. Afterwards, we describe ready-to-use integration with widely-used external simulation software packages, tests, and continuous integration from unit to system level, and community building measures, drawing an overview of the current preCICE ecosystem.


Effects of repeated administration of N-acetyl-L-cysteine on sulfhydryl levels of different tissues and bleomycin-induced lung fibrosis in hamsters.
N-Acetyl-L-cysteine (NAC), 50, 100, 200, or 400 mg/kg, was injected intraperitoneally once a day for 13 days. No change was seen in the total sulfhydryl (TSH) and nonprotein sulfhydryl (NPSH) contents of the liver, kidney, and plasma at any dose. The heart TSH level remained unchanged, but the NPSH level was increased from the control value of 16 nmol/mg to 18, 19, and 18 nmol/mg protein at 50, 100, and 200 mg/kg, respectively. The lung TSH and NPSH levels both were increased from the control values of 65 and 8 nmol/mg to 80 and 16 nmol/mg protein, respectively, at 200 mg/kg. The lung TSH level at 400 mg/kg NAC was not changed, but the NPSH level increased to 13.5 nmol/mg protein. The ratio of TSH to NPSH levels in the liver and kidney was 4:1, whereas in the lung and heart it was 7:1 and 8:1, respectively. Based on amount per milligram of protein, TSH and NPSH levels were highest in the liver, followed by the amounts in the kidney, heart, and lung. The lung had the lowest level of TSH and NPSH. The daily treatment with NAC (200 mg/kg) for 13 days after and 2 days before intratracheal injection of bleomycin (7.5 U/kg) had little effect on lung collagen accumulation. The lung collagen level measured as hydroxyproline in bleomycin and in NAC plus bleomycin was significantly increased to 175% and 183% of the control levels, respectively. There was no difference in the lung hydroxyproline content between the control and NAC groups. The histopathology study also revealed no marked difference between the bleomycin and bleomycin plus NAC groups. Alternatively, treatment with NAC (200 mg/kg) for 13 days before bleomycin made the animals more susceptible to bleomycin toxicity and tended to add to the bleomycin-induced accumulation of collagen in the lung. NAC per se caused no mortality at any dose. The lung TSH and NPSH levels in bleomycin-treated (7.5 U/kg) hamsters were increased to 136% and 111% of control, respectively, whereas the TSH and NPSH levels both were increased to 155% of the levels of their respective controls in hamsters in the NAC plus bleomycin group. The differential effects of NAC treatment on the sulfhydryl content of tissues, the treatment's inability to alter the course of bleomycin-induced lung inflammation and collagen accumulation, and the potential for exacerbation of lung toxicity in response to repeated administration of NAC before exposure to fibrogenic agents are discussed.


Non-Classical Self-Assembly of Anisotropic Magneto-Organosilica Janus Particles Possessing Surfactant Properties and the Field-Triggered Breakdown of Surface Activity and Amphiphilic Properties.
Using colloidal particles as models to understand processes on a smaller scale is a precious approach. Compared to molecules, particles are less defined, but their architecture can be more complex and so is their long-range interaction. One can observe phenomena that are unknown or much more difficult to realize on the molecular level. The current paper focuses on particle-based surfactants and reports on numerous unexpected properties. The main goal is creating an amphiphilic system with responsiveness in surface activity and associated self-organization phenomena depending on applying an external trigger, preferably a physical field. A key step is the creation of a Janus-type particle characterized by two types of dipoles (electric and magnetic) which geometrically stand orthogonal to each other. In a field, one can control which contribution and direction dominate the interparticle interactions. As a result, one can drastically change the system's properties. The features of ferrite-core organosilica-shell particles with grain-like morphology modified by click chemistry are studied in response to spatially isotropic and anisotropic triggers. A highly unusual aggregation-dissolution-reaggregation sequence w as discovered. Using a magnetic field, one can even switch off the amphiphilic properties and use this for the field-triggered breaking of multiphase systems such as emulsions.


VEGF regulates TRPC6 channels in podocytes.
Both, increased plasma concentrations of vascular endothelial growth factor (VEGF) and increased expression of transient receptor potential canonical type 6 (TRPC6) channels in podocytes have been associated with proteinuric kidney diseases. Now, we investigated the hypothesis that VEGF regulates TRPC6 in podocytes. TRPC6 messenger RNA (mRNA) and TRPC6 protein expression were analyzed in cultured podocytes after administration of VEGF165 using quantitative real-time reverse transcription-polymerase chain reaction and immunoblotting, respectively. YFP-tagged TRPC6 in podocytes was analyzed using confocal laser scanning microscopy. TRPC6-associated calcium influx was measured fluorometrically. Both, immunofluorescence and immunohistochemistry were performed in renal tissue from patients with diabetes mellitus and controls. Administration of VEGF165 to podocytes significantly increased TRPC6 mRNA expression and TRPC6 protein levels. The effects of VEGF165 were dose dependent and could be blocked by phosphoinositide-3-kinase inhibitors. In the presence of cycloheximide, an inhibitor of protein biosynthesis, we did not observe an effect of VEGF on TRPC6 protein levels, indicating the requirement of de novo protein synthesis. VEGF165 significantly increased TRPC6-mediated calcium influx in podocytes. Calcium influx was significantly lower in podocytes after gene knockdown using siRNA against TRPC6. Immunohistochemistry showed both increased TRPC6 channel protein and VEGF receptor type 2 (VEGFR-2) protein in podocytes from patients with diabetic nephropathy compared to control subjects. There was a significant association between VEGFR-2 mRNA and TRPC6 mRNA (n = 48; r(2) = 0.585; P < 0.0001) in human renal cortex. VEGF regulates TRPC6 in podocytes.


Association Between Palliative Care and Death at Home in Adults With Heart Failure.
Background Palliative care is associated with improved symptom control and quality of life in people with heart failure. There is conflicting evidence as to whether it is associated with a greater likelihood of death at home in this population. The objective of this study was to describe the delivery of newly initiated palliative care services in adults who die with heart failure and measure the association between receipt of palliative care and death at home compared with those who did not receive palliative care. Methods and Results We performed a population-based cohort study using linked health administrative data in Ontario, Canada of 74 986 community-dwelling adults with heart failure who died between 2010 and 2015. Seventy-five percent of community-dwelling adults with heart failure died in a hospital. Patients who received any palliative care were twice as likely to die at home compared with those who did not receive it (adjusted odds ratio 2.12 [95% CI, 2.03-2.20]; P<0.01). Delivery of home-based palliative care had a higher association with death at home (adjusted odds ratio 11.88 [95% CI, 9.34-15.11]; P<0.01), as did delivery during transitions of care between inpatient and outpatient care settings (adjusted odds ratio 8.12 [95% CI, 6.41-10.27]; P<0.01). Palliative care was most commonly initiated late in the course of a person's disease (≤30 days before death, 45.2% of subjects) and led by nonspecialist palliative care physicians 61% of the time. Conclusions Most adults with heart failure die in a hospital. Providing palliative care near the end-of-life was associated with an increased likelihood of dying at home. These findings suggest that scaling existing palliative care programs to increase access may improve end-of-life care in people dying with chronic noncancer illness.


Adenine Enrichment at the Fourth CDS Residue in Bacterial Genes Is Consistent with Error Proofing for +1 Frameshifts.
Beyond selection for optimal protein functioning, coding sequences (CDSs) are under selection at the RNA and DNA levels. Here, we identify a possible signature of "dual-coding," namely extensive adenine (A) enrichment at bacterial CDS fourth sites. In 99.07% of studied bacterial genomes, fourth site A use is greater than expected given genomic A-starting codon use. Arguing for nucleotide level selection, A-starting serine and arginine second codons are heavily utilized when compared with their non-A starting synonyms. Several models have the ability to explain some of this trend. In part, A-enrichment likely reduces 5' mRNA stability, promoting translation initiation. However T/U, which may also reduce stability, is avoided. Further, +1 frameshifts on the initiating ATG encode a stop codon (TGA) provided A is the fourth residue, acting either as a frameshift "catch and destroy" or a frameshift stop and adjust mechanism and hence implicated in translation initiation. Consistent with both, genomes lacking TGA stop codons exhibit weaker fourth site A-enrichment. Sequences lacking a Shine-Dalgarno sequence and those without upstream leader genes, that may be more error prone during initiation, have greater utilization of A, again suggesting a role in initiation. The frameshift correction model is consistent with the notion that many genomic features are error-mitigation factors and provides the first evidence for site-specific out of frame stop codon selection. We conjecture that the NTG universal start codon may have evolved as a consequence of TGA being a stop codon and the ability of NTGA to rapidly terminate or adjust a ribosome.


Charge transfer and recombination at the metal oxide/CH3NH3PbClI2/spiro-OMeTAD interfaces: uncovering the detailed mechanism behind high efficiency solar cells.
In recent years, organometal halide perovskite-based solid-state hybrid solar cells have attracted unexpected increasing interest because of their high efficiency (the record power conversion efficiency has been reported to be over 15%) and low fabrication cost. It has been accepted that the high efficiency was mainly attributed to the strong optical absorption (absorption coefficient: 15,000 cm(-1) at 550 nm) over a broader range (up to 800 nm) and the long lifetimes of photoexcited charge carriers (in the order of 10 ns - a few 100 ns) of the perovskite absorbers. However, much of the fundamental photophysical properties of perovskite relating to the high photovoltaic performance are remained to be investigated. The charge separation and recombination processes at the material interfaces are particularly important for solar cell performances. To better understand the high efficiency of perovskite solar cells, we systematically investigated the charge separation (electron and hole injection) and charge recombination dynamics of CH3NH3PbClI2 hybrid solar cells employing TiO2 nanostructures as the electron transfer material (ETM) and spiro-OMeTAD as the hole transfer material (HTM). The measurements were carried out using transient absorption (TA) techniques on a time scale from sub-picoseconds to milliseconds. We clarified the timescales of electron injection, hole injection, and recombination processes in TiO2/CH3NH3PbClI2/spiro-OMeTAD solar cells. Charge separation and collection efficiency of the perovskite-based solar cells were discussed. In addition, the effect of TiO2 size on the charge separation and recombination dynamics was also investigated. It was found that all TiO2-based perovskite solar cells possessed similar charge separation processes, but quite different recombination dynamics. Our results indicate that charge recombination was crucial to the performance of the perovskite solar cells, which could be effectively suppressed through optimising nanostructured TiO2 films and surface passivation, thus pushing these cells to even higher efficiency.


Preparation and characterization of plasma membrane-enriched fractions from rat pancreatic islets.
Methods have been developed for the isolation on a semi-micro scale of a plasma membrane-enriched fraction from rat islets of Langerhans. An important feature of these experiments is the use of 125I-labeled wheat germ agglutinin as a specific probe for plasma membrane-containing fractions. The partly purified plasma membrane fraction had a density in sucrose of about 1.10 and was enriched in the activities of 5'-nucleotidase, alkaline phosphatase, sodium-potassium, and magnesium-dependent ATPase and adenylate cyclase. It contained only very low levels of acid phosphatase, cytochrome c oxidase, insulin, and RNA. Further purification was hampered by the relatively small amounts of fresh plasma membrane material that could be obtained from 16-24 rats in each experiment. When islets were prelabeled with radioactive fucose, the plasma membrane-enriched fraction contained radioactivity at a four- to fivefold higher specific acivity than the whole islet homogenate. Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis of plasma membrane-enriched fractions pooled from several experiments revealed a distinctive pattern of protein bands as compared with other less pure fractions. With respect to rapidity, apparent specificity, and easy reversibility of the labeling of the plasma membrane fraction, 125I-wheat germ agglutinin provides a highly useful tool for the detection of microgram quantities of plasma membrane components which should be applicable to many other systems as well.


Effects of illusory kinesthesia by tendon vibratory stimulation on the postoperative neural activities of distal radius fracture patients.
Activation of motor-related regions with illusory kinesthesia had been suspected to contribute toward pain alleviation. We used electroencephalography (EEG) to assess the sensorimotor cortex activation during illusory kinesthesia by vibratory tendon stimulation in patients who were in pain. We investigated the relationship between the sensorimotor cortex and postoperative pain to clarify the analgesia mechanism. Patients who had undergone distal radius fracture surgery were studied. A hand massager provided the vibration stimulus for the illusory kinesthesia on the extensor digitorum muscle at the nonaffected hand's wrist joint on 7 consecutive days starting 1 day after surgery. The main outcomes were pain intensity and EEG activity. EEG activity was recorded during illusory kinesthesia. We also analyzed the high α rhythm (10-12 Hz) event-related desynchronization/event-related synchronization. The pain intensity was reduced. Attenuation of the high α wave was observed in the bilateral sensorimotor cortex (C3ch, C4ch) areas during illusory kinesthesia. A significant negative correlation was also observed in the C3 and the C4 of event-related desynchronization/event-related synchronization% values and the variation of pain amelioration at rest. Activation of the sensorimotor cortex by illusory kinesthesia might have influenced parts of the descending pain-inhibitory system. The postoperative pain in the patients might have been alleviated by the descending pain-inhibitory system.


A high-power and fast charging Li-ion battery with outstanding cycle-life.
Electrochemical energy storage devices based on Li-ion cells currently power almost all electronic devices and power tools. The development of new Li-ion cell configurations by incorporating innovative functional components (electrode materials and electrolyte formulations) will allow to bring this technology beyond mobile electronics and to boost performance largely beyond the state-of-the-art. Here we demonstrate a new full Li-ion cell constituted by a high-potential cathode material, i.e. LiNi0.5Mn1.5O4, a safe nanostructured anode material, i.e. TiO2, and a composite electrolyte made by a mixture of an ionic liquid suitable for high potential applications, i.e. Pyr1,4PF6, a lithium salt, i.e. LiPF6, and standard organic carbonates. The final cell configuration is able to reversibly cycle lithium for thousands of cycles at 1000 mAg-1 and a capacity retention of 65% at cycle 2000.


DJ-1: A promising therapeutic candidate for ischemia-reperfusion injury.
DJ-1 is a multifaceted protein with pleiotropic functions that has been implicated in multiple diseases, ranging from neurodegeneration to cancer and ischemia-reperfusion injury. Ischemia is a complex pathological state arising when tissues and organs do not receive adequate levels of oxygen and nutrients. When the blood flow is restored, significant damage occurs over and above that of ischemia alone and is termed ischemia-reperfusion injury. Despite great efforts in the scientific community to ameliorate this pathology, its complex nature has rendered it challenging to obtain satisfactory treatments that translate to the clinic. In this review, we will describe the recent findings on the participation of the protein DJ-1 in the pathophysiology of ischemia-reperfusion injury, firstly introducing the features and functions of DJ-1 and, successively highlighting the therapeutic potential of the protein.


Morphological Phylogenetics Evaluated Using Novel Evolutionary Simulations.
Evolutionary inferences require reliable phylogenies. Morphological data have traditionally been analyzed using maximum parsimony, but recent simulation studies have suggested that Bayesian analyses yield more accurate trees. This debate is ongoing, in part, because of ambiguity over modes of morphological evolution and a lack of appropriate models. Here, we investigate phylogenetic methods using two novel simulation models-one in which morphological characters evolve stochastically along lineages and another in which individuals undergo selection. Both models generate character data and lineage splitting simultaneously: the resulting trees are an emergent property, rather than a fixed parameter. Standard consensus methods for Bayesian searches (Mki) yield fewer incorrect nodes and quartets than the standard consensus trees recovered using equal weighting and implied weighting parsimony searches. Distances between the pool of derived trees (most parsimonious or posterior distribution) and the true trees-measured using Robinson-Foulds (RF), subtree prune and regraft (SPR), and tree bisection reconnection (TBR) metrics-demonstrate that this is related to the search strategy and consensus method of each technique. The amount and structure of homoplasy in character data differ between models. Morphological coherence, which has previously not been considered in this context, proves to be a more important factor for phylogenetic accuracy than homoplasy. Selection-based models exhibit relatively lower homoplasy, lower morphological coherence, and higher inaccuracy in inferred trees. Selection is a dominant driver of morphological evolution, but we demonstrate that it has a confounding effect on numerous character properties which are fundamental to phylogenetic inference. We suggest that the current debate should move beyond considerations of parsimony versus Bayesian, toward identifying modes of morphological evolution and using these to build models for probabilistic search methods. [Bayesian; evolution; morphology; parsimony; phylogenetics; selection; simulation.].


Resistin levels in gingival crevicular fluid of patients with chronic periodontitis and type 2 diabetes mellitus.
Resistin is associated with local and systemic inflammatory conditions with a direct correlation with type 2 diabetes mellitus (T2DM). The aim of this clinico-biochemical study is to estimate and compare the levels of resistin in the gingival crevicular fluid (GCF) in health, chronic periodontitis (CP), and T2DM. Sixty patients (aged >35 years) who participated in this study were divided into four groups of 15 patients each: healthy individuals (group 1), patients with CP (group 2), patients with T2DM (group 3), and patients with T2DM and CP (group 4). The parameters assessed included plaque index (PI), gingival index (GI), probing depth (PD), periodontal index, body mass index, random blood sugar (RBS), and glycated hemoglobin (HbA1c). GCF (4 μL) was collected and analyzed for resistin levels using an enzyme-linked immunosorbent assay. Resistin was detected in the GCF of all patients. A significant difference was observed in GCF resistin concentrations from group 1 versus group 2 (P = 0.0093), group 3 (P = 0.0341), and group 4 (P = 0.0002); in group 2 versus group 4 (P = 0.0032); and in group 3 versus group 4 (P = 0.0008). When all the samples were analyzed together, GCF resistin levels positively correlated with GI, PD, PI, RBS, and HbA1c and were predictable with PD and HbA1c. Resistin levels are increased in CP and T2DM. Hence, GCF resistin levels may be considered as a potential inflammatory marker for periodontitis with T2DM.


Fluid transport by gallbladder epithelium.
The absorption of fluid by epithelial tissues is thought to be due to the existence of hypertonic regions within the epithelium. The magnitude of the required hypertonicity as well as its localization have been the subject of considerable experimental and theoretical effort. Model calculations demonstrated the need for knowledge of the water permeability of the membranes of epithelial cells for the purpose of estimation of the osmotic gradients required for fluid absorption. We measured the hydraulic water permeability of the individual cell membranes of Necturus gallbladder by quantitative light microscopy. The water permeabilities were sufficiently high so that small osmotic gradients were required to achieve normal rates of fluid transport. The cell osmolality was calculated to exceed that of the mucosal bathing solution by about 2 mosmol kg-1, and the basolateral interstitial osmolality was calculated to be about 1 mosmol kg-1 greater than that of the cell. The fluid absorbed by the epithelium must be slightly hypertonic to the bathing solutions. Knowledge of the apical cell membrane water permeability and the relative area of the cell and tight junction allow a calculation of the relative flow of fluid across both pathways. It can be readily shown that osmotically induced flow across the epithelium occurs predominantly transcellularly because of the small area of the junctional pathway and the high water permeability of the cell membranes.