Use of respiratory impedance in prehospital care of hypotensive patients associated with hemorrhage and trauma: a case series.
The respiratory pump can be optimized to enhance circulation in patients with hypotension by having patients spontaneously breathe through a low level of inspiratory resistance. This can be achieved with an impedance threshold device (ITD) designed to provide 7 cm H2O resistance during spontaneous inspiration with minimal resistance during expiration. Little is known about the effects of harnessing this physiological concept to increase blood pressure (BP) in the prehospital setting of care for patients with hypotension caused by blood loss or trauma. In this case series, we report on the feasibility, effectiveness, and safety of rapidly deploying the ITD by first responders to treat hypotension secondary to blood loss and trauma in the urban setting by emergency medical services personnel. Hemodynamic data from hypotensive patients (pretreatment systolic BP [SBP] <100 mm Hg) from 3 U.S. cities where the ITD is deployed were evaluated. The primary end point was maximum change in SBP and diastolic BP (DBP) from before to during ITD use in patients with hypotension secondary to documented blood loss or trauma. Secondary end points were device tolerance, whether the patient felt "better," change in heart rate, O2 saturation, and adverse events. Of the 255 hypotensive patients treated, there were 26 categorized with blood loss and 13 with trauma. In this 39-patient subgroup, the SBP and DBP (mean ± SD) increased from 79 ± 14 mm Hg and 48 ± 12 mm Hg before ITD placement to 110 ± 17 mm Hg and 66 ± 14 mm Hg after ITD placement (p < 0.001). Breathing through the ITD resulted in no reported adverse events, was well tolerated, and resulted in feeling "better" in more than 85% of the patients. Use of an ITD by emergency medical services personnel on hypotensive spontaneously breathing patients secondary to blood loss and trauma increased SBP and DBP and was feasible, well tolerated, and not associated with adverse effects (e.g., increased bleeding).


Increasing cell size remodels the proteome and promotes senescence.
Cell size is tightly controlled in healthy tissues, but it is unclear how deviations in cell size affect cell physiology. To address this, we measured how the cell's proteome changes with increasing cell size. Size-dependent protein concentration changes are widespread and predicted by subcellular localization, size-dependent mRNA concentrations, and protein turnover. As proliferating cells grow larger, concentration changes typically associated with cellular senescence are increasingly pronounced, suggesting that large size may be a cause rather than just a consequence of cell senescence. Consistent with this hypothesis, larger cells are prone to replicative, DNA-damage-induced, and CDK4/6i-induced senescence. Size-dependent changes to the proteome, including those associated with senescence, are not observed when an increase in cell size is accompanied by an increase in ploidy. Together, our findings show how cell size could impact many aspects of cell physiology by remodeling the proteome and provide a rationale for cell size control and polyploidization.


Challenges and prospects about the graphene role in the design of photoelectrodes for sunlight-driven water splitting.
Graphene and its derivatives have emerged as potential materials for several technological applications including sunlight-driven water splitting reactions. This review critically addresses the latest achievements concerning the use of graphene as a player in the design of hybrid-photoelectrodes for photoelectrochemical cells. Insights about the charge carrier dynamics of graphene-based photocatalysts which include metal oxides and non-metal oxide semiconductors are also discussed. The concepts underpinning the continued progress in the field of graphene/photoelectrodes, including different graphene structures, architecture as well as the possible mechanisms for hydrogen and oxygen reactions are also presented. Despite several reports having demonstrated the potential of graphene-based photocatalysts, the achieved performance remains far from the targeted benchmark efficiency for commercial application. This review also highlights the challenges and opportunities related to graphene application in photoelectrochemical cells for future directions in the field.


Who's in, who's out, and who's back: follow-up data on 59 juveniles incarcerated in adult prison for murder or attempted murder in the early 1980s.
Since the mid-1980s, when juvenile arrests for violent crime increased dramatically, interest has focused on juvenile offenders who commit violent acts. Legislatures across the United States have enacted a variety of measures to "get tough" with juveniles in response to escalating crime rates and the perceptions that longer sentences were needed. This manuscript provides follow-up data on 59 juveniles who were committed to the adult Department of Corrections in Florida during the period January 1982 through January 1984 for one or more counts of murder, attempted murder, or, in a few cases, manslaughter. Although many of these adolescents received lengthy prison sentences, more than two-thirds had been released from prison prior to November 1999. This article presents data on amount of time served and recidivism over the 15 to 17 year period. Results indicated that 60 percent of sample subjects released from prison were returned to prison, and most of those who failed did so within the first three years of release. Findings from the present study, when examined in the context of previous comparative follow-up studies of delinquent youths, suggest that the dialogue on how to handle violent youths must be continued if juvenile homicide offenders are going to be released to society at some point in the future.


De novo genome assembly of the tobacco hornworm moth (Manduca sexta).
The tobacco hornworm, Manduca sexta, is a lepidopteran insect that is used extensively as a model system for studying insect biology, development, neuroscience, and immunity. However, current studies rely on the highly fragmented reference genome Msex_1.0, which was created using now-outdated technologies and is hindered by a variety of deficiencies and inaccuracies. We present a new reference genome for M. sexta, JHU_Msex_v1.0, applying a combination of modern technologies in a de novo assembly to increase continuity, accuracy, and completeness. The assembly is 470 Mb and is ∼20× more continuous than the original assembly, with scaffold N50 > 14 Mb. We annotated the assembly by lifting over existing annotations and supplementing with additional supporting RNA-based data for a total of 25,256 genes. The new reference assembly is accessible in annotated form for public use. We demonstrate that improved continuity of the M. sexta genome improves resequencing studies and benefits future research on M. sexta as a model organism.


Femtosecond stimulated Raman spectroscopy using a scanning multichannel technique.
A scanning multichannel technique (SMT) has been implemented in femtosecond stimulated Raman spectroscopy (FSRS). By combining several FSRS spectra detected at slightly different positions of the spectrograph via SMT, we have eliminated the systematic noise patterns ("fixed pattern noise") due to the variation in sensitivity and noise characteristics of the individual charge-coupled device (CCD) pixels. In nonresonant FSRS, solvent subtraction can effectively remove the systematic noise pattern even without SMT. However, in the case of resonant FSRS, we show that a similar solvent subtraction procedure is ineffective at removing the noise patterns without SMT. Application of SMT results in averaged FSRS spectra with improved signal-to-noise ratios that approach the shot-noise limit.


A new gnathosaurine (Pterosauria, Archaeopterodactyloidea) from the Late Jurassic of Portugal.
An incomplete, yet remarkably-sized dentated rostrum and associated partial cervical vertebrae of a pterosaur (ML 2554) were recently discovered from the Late Jurassic (Late Kimmeridgian-Early Tithonian) Lourinhã Formation of Praia do Caniçal, of central west Portugal. This specimen exhibits features such as a spatulated anterior expansion of the rostrum, robust comb-like dentition, and pronounced rims of the tooth alveoli, indicating gnathosaurine affinities. Based on its further unique tooth and dentary morphology, a new genus and species, Lusognathus almadrava gen. et spec. nov., is proposed, making this the first named pterosaur species found within Portugal. The presence of this taxon adds yet another element to the fluvio-deltaic lagoonal environment that has been suggested as representative of the Lourinhã Formation in the Late Jurassic, further contributing to the diversity and distribution of gnathosaurines worldwide.


Cumulative social class and mortality from various causes of adult men.
It is possible that circumstances over the lifecourse contribute to social inequalities in mortality in adulthood. The aim of this study is to assess the cumulative effect of social class at childhood and adulthood on mortality from various causes of death in young adult men. The data consist of census records for all Finnish men born in 1956-60 (112,735 persons and 895,001 person years), and death records (1834 deaths) by cause of death for 1991-98. Mortality from each cause of death increased from the stable non-manual group to mobile groups, and further to the stable manual group. However, mortality in the downwardly mobile group was 150% higher than in the upwardly mobile group. Furthermore, analyses show that mortality was mainly related to current adult social class, though, within each adult social class men with a manual parental background showed slightly increased mortality from cardiovascular disease and from alcohol related causes. In these data the effects of adult social class were stronger than childhood class for all causes of death. It is more useful to differentiate between childhood and adulthood effects than to use a combined measure of social class to assess the contribution of social class at different stages of life on mortality.


Integrated Transcriptome and Pathway Analyses Revealed Multiple Activated Pathways in Breast Cancer.
Breast cancer (BC) is the leading cause of cancer-related death in women. Therefore, a better understanding of BC biology and signaling pathways might lead to the development of novel biomarkers and targeted therapies. Although a number of transcriptomic studies have been performed on breast cancer patients from various geographic regions, there are almost no such comprehensive studies performed on breast cancer from patients in the gulf region. This study aimed to provide a better understanding of the altered molecular networks in BC from the gulf region. Herein, we compared the transcriptome of BC to adjacent normal tissue from six BC patients and identified 1,108 upregulated and 518 downregulated transcripts. A selected number of genes from the RNA-Seq analysis were subsequently validated using qRT-PCR. Differentially expressed (2.0-fold change, adj. p < 0.05) transcripts were subjected to ingenuity pathway analysis, which revealed a myriad of affected signaling pathways and functional categories. Activation of ERBB2, FOXM1, ESR1, and IGFBP2 mechanistic networks was most prominent in BC tissue. Additionally, BC tissue exhibited marked enrichment in genes promoting cellular proliferation, migration, survival, and DNA replication and repair. The presence of genes indicative of immune cell infiltration and activation was also observed in BC tissue. We observed high concordance [43.5% (upregulated) and 62.1% (downregulated)] between differentially expressed genes in our study group and those reported for the TCGA BC cohort. Our data provide novel insight on BC biology and suggest common altered molecular networks in BC in this geographic region. Our data suggest future development of therapeutic interventions targeting those common signaling pathways.


Medication error reporting in a university teaching hospital in Saudi Arabia.
Medication errors are common in the hospital setting. Little is known about these errors in Saudi Arabia. The objective of the current study was to explore the rate of reporting medication errors and factors associated with the root causes of these errors in a large tertiary teaching hospital in Saudi Arabia. This study was conducted at the university teaching hospital in Riyadh, Saudi Arabia. All occurrence/variant reports related to medication errors were documented on a hospital Web-based medication error form that was designed to capture information on all aspects. Medication error reports were reviewed and reported at quarterly intervals over a 1-year period (November 2009 to October 2010). The medication error rate over the 1-year study period was 0.4% (949 medication errors for 240,000 prescriptions). During this period, 14 (1.5%) errors were categorized as resulting in any harm to the patient (all category E). Medication errors were reported predominantly at the prescribing stage of the medication process (89%). The most common types of errors were prescribing (44%) and improper dose/quantity (31%). Antibiotics (12%), antihypertensive agents (10%), and oral hypoglycemic agents (8%) were the pharmacological classes of medication most commonly involved with errors. Nonspecific performance deficit (43%), knowledge deficit (28%), and illegible or unclear handwriting (17%) were the main reported causes of error. Medication errors are underreported in a tertiary teaching hospital in Riyadh, Saudi Arabia. Future studies should evaluate the effectiveness of interventions to stimulate medication errors reporting by health-care providers.


Screening for abdominal aortic aneurysms in Western Australia.
A study of ultrasound screening for abdominal aortic aneurysms (AAA) was performed. During a 6 month period, 1225 men and women aged 60-80 years were screen at a variety of community venues. Screening was well received by the public and logistically simple to perform. Thirty-three AAA were detected with sizes between 30 and 81 mm. In the 60-80 year age group, the prevalence of AAA > 30 mm in diameter was 4.7% in men and 0.35% in women, and the prevalence of AAA > 50 mm was 0.6% in men and 0.17% in women. Cigarette smoking, but not hypertension or diabetes, was found to be a significant risk factor for AAA. This study confirms that screening for AAA is feasible and yields high prevalence rates in major population centres.


A Multidisciplinary Approach to Simultaneously Monitoring Real-Time Neuronal Activity and Pain Behaviors During Optogenetic Stimulation of Brain Neurons in Freely Moving Mice.
Highlighted by the current opioid epidemic, identifying novel therapies to treat chronic trigeminal neuropathic pain is a critical need. To develop these treatments, it is necessary to have viable targets in the brain to act on. Historically, neural tracing studies have been extremely useful in determining connections between brain areas but do not provide information about the functionality of these connections. Combining optogenetics and behavioral observation allows researchers to determine whether a particular brain area is involved in the regulation of such behavior. The addition of multi-channel electrophysiological recording provides information on real-time neuronal activity in the specific neuronal pathway. Male C57/BL/6J mice (8-week-old) underwent either chronic constriction injury of infraorbital nerve (CCI-ION) or a sham surgery and were injected with either channelrhodopsin (ChR2) or a control virus in the hypothalamic A11 nucleus. Two weeks after CCI-ION, they were tested in real-time place preference (RTPP), while neuronal activity in the spinal trigeminal nucleus caudalis (Sp5C) was recorded. Optogenetic excitation of the A11 neurons results in more time spent in the stimulation chamber during RTPP testing. Additionally, stimulation of the A11 results in a greater number of neuronal activity increase in the Sp5C in animals with the injection of AAV carrying ChR2 compared to animals injected with a control virus or that underwent a sham surgery. In vivo multi-channel electrophysiological recording, optogenetic stimulation, and behavioral observation can be combined in a mouse model of chronic trigeminal neuropathic pain to validate brain areas involved in the modulation of such pain.


Insomnia with objective short sleep duration is associated with cognitive impairment: a first look at cardiometabolic contributors to brain health.
Insomnia with objective short sleep duration has been previously associated with adverse cardiometabolic health outcomes as well as poorer cognitive performance in otherwise noncognitively impaired adults. However, studies demonstrating an increased prevalence of cognitive impairment (CI) in this insomnia phenotype are lacking. We analyzed data from Penn State Adult Cohort (N = 1,524; 48.9 ± 13.4 years; 53.4% women). Self-reported sleep difficulty was defined as normal sleep (n = 899), poor sleep (n = 453), and chronic insomnia (n = 172). Objective short sleep duration was defined as less than 6-h of sleep, based on in-lab, 8-h polysomnography. CI (n = 155) and possible vascular cognitive impairment (pVCI, n = 122) were ascertained using a comprehensive neuropsychological battery. Analyses adjusted for age, sex, race, education, body mass index, apnea/hypopnea index, smoking, alcohol, psychoactive medication, and mental and physical health problems. Participants who reported poor sleep or chronic insomnia and slept objectively less than 6 hours were associated with a 2-fold increased odds of CI (OR = 2.06, 95% confidence limits [CL] = 1.15-3.66 and OR = 2.18, 95% CL = 1.07-4.47, respectively) and of pVCI (OR = 1.94, 95% CL = 1.01-3.75 and OR = 2.33, 95% CL = 1.07-5.06, respectively). Participants who reported poor sleep or chronic insomnia and slept objectively more than 6 hours were not associated with increased odds of either CI (OR = 0.72, 95% CL = 0.30-1.76 and OR = 0.75, 95% CL = 0.21-2.71, respectively) or pVCI (OR = 1.08, 95% CL = 0.42-2.74 and OR = 0.76, 95% CL = 0.16-3.57, respectively). Insomnia with objective short sleep duration is associated with an increased prevalence of CI, particularly as it relates to cardiometabolic health (i.e. pVCI). These data further support that this insomnia phenotype may be a more biologically severe form of the disorder associated with cardiovascular, cerebrovascular, and neurocognitive morbidity.


Reactive oxygen species (ROS)-responsive biomaterials mediate tissue microenvironments and tissue regeneration.
Reactive oxygen species (ROS) have been considered the pivotal signaling molecules in many physiological processes, and are usually overproduced in various inflammatory tissues. Overproduction of ROS may disrupt cellular homeostasis, cause non-specific damage to critical components, and lead to a series of diseases. ROS are acknowledged as a type of emerging triggered event similar to acidic pH, overproduced enzymes, temperature and other specific stimuli found in pathological microenvironments. Recently, ROS-responsive biomaterials have been identified as a type of promising therapeutic substance to alleviate oxidative stress in tissue microenvironments, and for use as a vehicle triggered by inflammatory diseases to realize drug release under physiological oxidative microenvironments. In this review, we discuss mainly the mechanisms of ROS-responsive biomaterials with solubility switch and chemical degradation, and those ROS-responsive groups used in ROS-responsive biomaterials. The mechanism of ROS overproduction in pathophysiological conditions is introduced. The various applications of ROS-responsive biomaterials in tissue regeneration and disease therapy, such as cardiovascular diseases, osteoarthritis, chronic diabetic wounds, inflammatory bowel disease and other inflammatory diseases, are summarized.


Fatal Epstein-Barr virus infection in a case of familial hemophagocytic lymphohistiocytosis with syntaxin-11 mutation.
Familial hemophagocytic lymphohistiocytosis (FHL) is a fatal disease of early infancy caused by defective natural killer cell activity and is characterized by fever, organomegaly, pancytopenia, and coagulopathy. Disease-causing mutations have been found in perforin, Munc 13-4 and syntaxin-11 genes. We herein describe a case of late-onset FHL with syntaxin-11 mutation in a six-year-old boy in whom only partial response was obtained by immunochemotherapy (HLH-94 protocol) and who died with persistent Epstein-Barr virus (EBV) infection. The role of EBV infection in the prognosis of FHL is discussed.


Ecological aspects of the heritability of body size in Drosophila buzzatii.
The heritability of thorax length in the cactophilic Drosophila buzzatii was determined for flies from each of 10 rotting cactus cladodes. For each rot, emerging flies were used as parents of progeny reared in the laboratory. The methods used were full sib analysis with the parents mated assortatively and also offspring-parent regression. From this, heritabilities were measured for the laboratory environment and for the natural environment of the rotting cladode. For the laboratory environment, h2 = 0.3770 +/- 0.0203 and for the natural environment h2 = 0.0936 +/- 0.0087 within rots and h2 = 0.0595 +/- 0.0123 for a population drawn randomly from different rots. Because of the possibility of genotype-environment interaction between the laboratory and rot environments, the methods of B. Riska, T. Prout and M. Turelli were used to show it is possible that there is no such interaction, but if there is, the above natural heritabilities are approximate lower bounds. These results are related to the general problem of determining heritabilities in nature where it is impractical to measure both parents and progeny in nature. Determining heritability not only in nature but in relation to subdivision into ephemeral patches (cladodes in this case) has an important bearing on natural selection response and to general theories of stabilizing selection proposed to explain the existence of genetic variation. Attempts were made to detect selection by using the size of emerging adults as an indicator of various levels of larval stress. No selection was detected, but the power to do so was very weak. Differences between progeny means from different rots indicated some genetic differences between rots which can be adequately explained by small numbers of founders. This suggests a random fine scale subdivision amounting to FST = 0.1483 +/- 0.0462.


Epigenetic mechanisms in cardiac development and disease.
During mammalian development, cardiac specification and ultimately lineage commitment to a specific cardiac cell type is accomplished by the action of specific transcription factors (TFs) and their meticulous control on an epigenetic level. In this review, we detail how cardiac-specific TFs function in concert with nucleosome remodeling and histone-modifying enzymes to regulate a diverse network of genes required for processes such as cell growth and proliferation, or epithelial to mesenchymal transition (EMT), for instance. We provide examples of how several cardiac TFs, such as Nkx2.5, WHSC1, Tbx5, and Tbx1, which are associated with developmental and congenital heart defects, are required for the recruitment of histone modifiers, such as Jarid2, p300, and Ash2l, and components of ATP-dependent remodeling enzymes like Brg1, Baf60c, and Baf180. Binding of these TFs to their respective sites at cardiac genes coincides with a distinct pattern of histone marks, indicating that the precise regulation of cardiac gene networks is orchestrated by interactions between TFs and epigenetic modifiers. Furthermore, we speculate that an epigenetic signature, comprised of TF occupancy, histone modifications, and overall chromatin organization, is an underlying mechanism that governs cardiac morphogenesis and disease.


Portuguese Validated Versions of the Alcohol Use Disorders Identification Test: A Systematic Review Protocol.
Alcohol consumption ranks among the top ten risk factors contributing to the global disease burden. Several international organizations recommend the use of the Alcohol Use Disorders Identification Test to screen for at-risk drinkers. However, a fully validated Portuguese version of this test is lacking. The aim of this study is to systematically review validated versions of the Alcohol Use Disorders Identification Test in the Portuguese language, the documented problems and solutions in its application and proposed cut-offs to identify at-risk drinkers. A systematic search will be performed in Ovid MEDLINE, CINAHL, PsycINFO, ÍndexRMP, LILACS, African Journals Online and SciELO databases, along with grey literature searches to identify validation studies of the AUDIT in Portuguese. Two authors will independently extract data and assess the studies' methodological quality, using QUADAS-2 and CASP checklists. This review will compare different validation studies of the Alcohol Use Disorders Identification Test in Portuguese-speaking countries, reporting, where possible, the psychometric properties, performance characteristics, suggested cut-offs and any documented limitations and suggestions. The results of this review could be used to propose an update of the alcohol screening and brief intervention guidelines in Portugal. The results could also prove useful to support the implementation of alcohol screening delivery by healthcare providers in Portugal and other official Portuguese-speaking countries. This review will provide important information on the validity of the Alcohol Use Disorders Identification Test as a screening tool for at-risk drinking in Portugal and other official Portuguese speaking countries.


Retinoic acid signaling positively regulates liver specification by inducing wnt2bb gene expression in medaka.
During vertebrate embryogenesis, the liver develops at a precise location along the endodermal primitive gut tube because of signaling delivered by adjacent mesodermal tissues. Although several signaling molecules have been associated with liver formation, the molecular mechanism that regulates liver specification is still unclear. We previously performed a screen in medaka to isolate mutants with impaired liver development. The medaka hio mutants exhibit a profound (but transient) defect in liver specification that resembles the liver formation defect found in zebrafish prometheus (prt) mutants, whose mutation occurs in the wnt2bb gene. In addition to their liver abnormality, hio mutants lack pectoral fins and die after hatching. Positional cloning indicated that the hio mutation affects the raldh2 gene encoding retinaldehyde dehydrogenase type2 (RALDH2), the enzyme principally responsible for retinoic acid (RA) biosynthesis. Mutations of raldh2 in zebrafish preclude the development of pectoral fins. Interestingly, in hio mutants, expression of wnt2bb in the lateral plate mesoderm (LPM) directly adjacent to the liver-forming endoderm was completely lost. Our data reveal the unexpected finding that RA signaling positively regulates the wnt2bb gene expression required for liver specification in medaka. These results suggest that a common molecular mechanism may underlie liver and pectoral fin specification during piscine embryogenesis.


Effects of procaine on the intracellular pH of Chinese hamster ovary cells heated at 42.0 or 45.0 degrees C.
The local anesthetic procaine greatly sensitizes cells to hyperthermia. Though it is generally accepted that procaine is a membrane-active agent that increases membrane fluidity in cells, the mechanism by which it potentiates heat killing is unknown. In this paper we report changes in intracellular pH (pHi) of Chinese hamster ovary (CHO) cells heated at 42.0 or 45.0 degrees C in the presence of procaine. The pHi was measured with flow cytometry using the dye 1,4-diacetoxy-2,3-dicyanobenzene (ADB). Studies were carried out using cells grown at normal pH (7.3) or cells placed in low-pH (6.6) medium 4 h prior to and during heating (acute low-pH treatment). Low-pH-adapted cells (PHV2), which were obtained previously by continuous culture in pH 6.6 medium, were also used. Normal cells heated in the presence of procaine at pH 7.3 underwent a large decrease in pHi compared to cells heated without procaine. Procaine had little additional effect on the intracellular pH of cells in medium with a pH of 6.6 for 4 h before and during 30 min of heating. PHV2 cells exposed to chronic low-pH conditions were resistant to acidification when heated with or without procaine. The surviving fraction of cells heated with procaine was significantly lower under all pH conditions than that of cells heated without procaine. Cells heated at 42.0 degrees C with procaine also became greatly acidified and their survival was reduced. These data suggest that the reduction in pHi caused by procaine may be part of the mechanism of heat sensitization, but cannot account for it entirely. Furthermore, the degree of procaine sensitization and intracellular acidification is dependent on the extracellular pH, with a larger effect occurring at pH 7.3 than at pH 6.6.


Chemotaxis in Bacillus subtilis: how bacteria monitor environmental signals.
Virtually all organisms have means of monitoring their environment and making use of information gained to aid their survival. Many organisms, from bacteria to animals, move from place to place and can alter their movements. Chemotaxis is a signal transduction system found in motile bacteria that allows them to sense changes in the concentrations of various extracellular compounds and change their swimming behavior in a way that moves them toward more favorable environments. Chemotaxis is the most ancient sensory-motor process in nature. For years, studies of enteric bacteria, such as Escherichia coli and Salmonella typhimurium, have served as the paradigm for understanding this process on a molecular level. Recent studies on the gram-positive bacterium, Bacillus subtilis, and other bacteria, suggest that a slightly more complex system may be ancestral to that of the more extensively studied enterics. Aspects of chemotaxis that are unique to B. subtilis include a more complex adaptation system, with protein-protein methyl group transfer, chemotaxis proteins having no counterparts in E. coli, and a very extensive repertoire of repellents that are sensed at very low concentrations by receptors.


Jasmonates are phytohormones with multiple functions, including plant defense and reproduction.
The plant hormones jasmonic acid and methyl jasmonate, along with their intermediate compounds, produced in the octadecanoid pathway, are important signaling molecules that are collectively called jasmonates. These are widespread in the plant kingdom and play crucial roles in biotic/abiotic stress responses, as well as in processes related to plant growth and development. Recently, it has been shown that jasmonates are also involved in reproductive processes. We present the most recent findings related to the biosynthesis, regulation and signaling mechanisms of jasmonates. Additionally, we discuss the identification of [(+)-7-iso-JA-L-Ile] as the active biological hormonal form of jasmonate; this fills the greatest gap in our knowledge about the signaling mechanism that is responsible for the activation of downstream genes in the jasmonate-signaling cascade. The identification of several Arabidopsis thaliana mutants was crucial to the elucidation of the signaling mechanisms involved in jasmonate-mediated responses. Finally, the involvement of jasmonates in the reproductive process of Nicotiana tabacum L. is briefly discussed, since some of the main enzymes of the jasmonic acid biosynthesis pathway were identified in a stigma/style expressed sequence tag database (TOBEST) of this Solanaceae species.


Nanomaterial-based drug delivery systems as promising carriers for patients with COVID-19.
Once the World Health Organization (WHO) declared the COVID-19 outbreak to be pandemic, massive efforts have been launched by researchers around the globe to combat this emerging infectious disease. Here we review the most recent data on the novel SARS-CoV-2 pathogen. We analyzed its etiology, pathogenesis, diagnosis, prevention, and current medications. After that, we summarized the promising drug delivery application of nanomaterial-based systems. Their preparation routes, unique advantages over the traditional drug delivery routes and their toxicity though risk analysis were also covered. We also discussed in detail the mechanism of action for one example of drug-loaded nanomaterial drug delivery systems (Avigan-contained nano-emulsions). This review provides insights about employing nanomaterial-based drug delivery systems for the treatment of COVID-19 to increase the bioavailability of current drugs, reducing their toxicity, and to increase their efficiency.


Nicotine exposure affects mother's and pup's nutritional, biochemical, and hormonal profiles during lactation in rats.
We have shown that maternal nicotine exposure during lactation has long-lasting effects on body adiposity and hormonal status of rat offspring. Here, we studied the nutritional and hormonal profiles in this experimental model. Two days after birth, osmotic minipumps were implanted in lactating rats divided into two groups: NIC - continuous s.c. infusions of nicotine (6 mg/kg per day) for 14 days and C - saline. Dams and pups were killed at 15 and 21 days of lactation. Body weight and food intake were evaluated. Milk, blood, visceral fat, carcass, and adrenal gland were collected. All the significant data were P<0.05. At the end of nicotine exposure (15 days), dams presented higher milk production, hyperprolactinemia, and higher serum high-density lipoprotein cholesterol (HDL-C). Milk from NIC dams had higher lactose concentration and energy content. After nicotine withdrawal (21 days), dams showed lower food intake and hyperleptinemia. The 15-day-old NIC pups presented higher total body fat, higher HDL-C, serum leptin, serum corticosterone, and adrenal catecholamine content, but lower tyrosine hydroxylase protein levels. The 21-day-old NIC pups had higher body protein content and serum globulin. Thus, maternal nicotine exposure during lactation results in important changes in nutritional, biochemical, and hormonal parameters in dams and offspring. The pattern of these effects is clearly distinct when comparing the nicotine-exposed group to the withdrawal group, which could be important for the programming effects observed previously.


Pharmacovigilance in hospice/palliative care: rapid report of net clinical effect of metoclopramide.
Understanding the performance of prescribed medications in day-to-day practice is important to minimize harm, maximize clinical benefits, and, eventually, better target the people who are most likely to benefit, especially in hospice/palliative care where there may be limited time to optimize prescribing. Metoclopramide, a benzamide prokinetic antiemetic, is widely used for a number of indications including nausea, vomiting, hiccups, and reflux. It has recently had a new "black box" warning issued by the Food and Drug Administration in relation to tardive dyskinesia to limit use to 12 weeks. A consecutive cohort of patients from 12 participating centers in two countries who were having metoclopramide initiated had data collected at three time points--baseline, 2 days (clinical benefit), and day 7 (clinical harm). Additionally, harms could be recorded at any time. Of the 53 people included in the cohort, 23 (43%) reported benefit at 48 hours, but only 18 (34%) of these people were still using it one week after commencing it. For the other 5, the medication was ceased due to harms. The most frequent harms were akathisia (n=4), headache (n=4), and abdominal pain (n=4). Nine people (17%) had no clinical benefit and experienced harms. Overall, one in three people gained net clinical benefit at one week. Limiting effects include side-effects that need to be sought actively in clinical care.


STUDY OF THE SUITABILITY OF ISRAELI HOUSEHOLD SALT FOR RETROSPECTIVE DOSIMETRY.
The first results of an in-depth evaluation of the practical potential of common household Israeli salt as a retrospective dosemeter in the event of a nuclear accident or terror attack are presented. Ten brands of salt were investigated with emphasis on four of the bestselling brands that constitute 76 % of the total consumer market. Eight of the ten brands show similar glow curves with two main glow peaks at maximum temperatures of ∼176°C and ∼225°C measured at a heating rate of 1°C s(-1) Chemical analysis of three major brands indicates substantial impurity levels of 200-500 ppm of Ca, K, Mg and S and significant differences of additional ppm trace impurities, which lead to an ∼50 % difference in the TL response of the three major brands. Fading in the dark is in significant but under room light is of the order of 35 % per day. The dose response is linear/supralinear with the threshold of supralinearity at ∼0.01 Gy reaching maximum value of ∼4 at 0.5-1 Gy for two of the major brands. The precision of repeated measurements is ∼10 % (1 SD), but the accuracy of dose assessment under field conditions requires further study.


The spectrum of gastrointestinal functional bowel disorders in joint hypermobility syndrome and in an academic referral center.
Joint hypermobility syndrome (JHS) is a non-inflammatory hereditary disorder of connective tissue with varied clinical presentations, including frequent joint dislocations, hyperextensible skin, easy bruising, and abnormal paper-thin scar formation. Many of these patients have unexplained gastrointestinal (GI) symptoms. Our aim was to evaluate the prevalence of JHS in a tertiary gastroenterology motility clinic and the spectrum of functional bowel disorders in JHS patients. In this retrospective case series, we screened the medical records of 277 patients seen over 4 years at an academic GI Motility Center. The patients who met the criteria for JHS by Beighton hypermobility score were evaluated for the presence of functional GI disorders by Rome IV criteria. They also underwent gastric emptying study and glucose breath testing for small intestinal bacterial overgrowth. The prevalence of JHS in the study population was 9.7%. The mean age was 27 years, and 92.5% were female. The symptoms experienced by these patients include nausea/vomiting (89%), abdominal pain (70%), constipation (48%), and bloating (18.5%). The disorders associated with JHS include gastroparesis (52%), irritable bowel syndrome (55.5%), and gastroesophageal reflux disease (30%). Also, 10 patients (37%) were diagnosed with postural hypotension tachycardia syndrome secondary to autonomic dysfunction. Approximately 10% of patients with suspected functional bowel disorders have hypermobility syndrome. Hence, it is crucial to familiarize gastrointestinal practitioners with the criteria utilized to diagnose JHS and the methods to identify physical examination findings related to this condition.


Review article: the many potential roles of intestinal serotonin (5-hydroxytryptamine, 5-HT) signalling in inflammatory bowel disease.
Serotonin (5-hydroxytryptamine, 5-HT) is an important mediator of every major gut-related function. Recent investigations also suggest that 5-HT can influence the development and severity of inflammation within the gut, particularly in the setting of inflammatory bowel disease (IBD). To review the roles that the intestinal serotonin signalling system plays in gut function, with a specific focus on IBD. We reviewed manuscripts from 1952 to 2017 that investigated and discussed roles for 5-HT signalling in gastrointestinal function and IBD, as well as the influence of inflammation on 5-HT signalling elements within the gut. Inflammation appears to affect every major element of intestinal 5-HT signalling, including 5-HT synthesis, release, receptor expression and reuptake capacity. Importantly, many studies (most utilising animal models) also demonstrate that modulation of selective serotonergic receptors (via agonism of 5-HT4 R and antagonism of 5-HT3 R) or 5-HT signal termination (via serotonin reuptake inhibitors) can alter the likelihood and severity of intestinal inflammation and/or its complicating symptoms. However, there are few human studies that have studied these relationships in a targeted manner. Insights discussed in this review have strong potential to lead to new diagnostic and therapeutic tools to improve the management of IBD and other related disorders. Specifically, strategies that focus on modifying the activity of selective serotonin receptors and reuptake transporters in the gut could be effective for controlling disease activity and/or its associated symptoms. Further studies in humans are required, however, to more completely understand the pathophysiological mechanisms underlying the roles of 5-HT in this setting.


Replication of herpesvirus DNA. V. Maturation of concatemeric DNA of pseudorabies virus to genome length is related to capsid formation.
The maturation of pseudorabies virus DNA from the replicative concatemeric form to molecules of genome length was examined using nine DNA+ temperature-sensitive mutants of pseudorabies virus, each belonging to a different complementation group. At the nonpermissive temperature, cells infected with each of the mutants synthesized concatemeric DNA. Cleavage of the concatemeric DNA to genome-length viral DNA was defective in all the DNA+ ts mutants tested, indicating that several viral gene products are involved in the DNA maturation process. In none of the ts mutant-infected cells were capsids with electron-dense cores (containing DNA) formed. Empty capsids with electron-translucent cores were, however, formed in cells infected with six of the nine temperature-sensitive mutants; in cells infected with three of the mutants, no capsid assembly occurred. Because these three mutants are deficient both in maturation of DNA and in the assembly of viral capsids, we conclude that maturation of viral DNA is dependent upon the assembly of capsids. In cells infected with two of the mutants (tsN and tsIE13), normal maturation of viral DNA occurred after shiftdown of the cells to the permissive temperature in the presence of cycloheximide, indicating that the temperature-sensitive proteins involved in DNA maturation became functional after shiftdown. Furthermore, because cycloheximide reduces maturation of DNA in wild-type-infected cells but not in cells infected with these two mutants, we conclude that a protein(s) necessary for the maturation of concatemeric DNA, which is present in limiting amounts during the normal course of infection, accumulated in the mutant-infected cells at the nonpermissive temperature. Concomitant with cleavage of concatemeric DNA, full capsids with electron-dense cores appeared after shiftdown of tsN-infected cells to the permissive temperature, indicating that there may be a correlation between maturation of DNA and formation of full capsids. The number of empty and full capsids (containing electron-dense cores) present in tsN-infected cells incubated at the nonpermissive temperature, as well as after shiftdown to the permissive temperature in the presence of cycloheximide, was determined by electron microscopy and by sedimentation analysis in sucrose gradients. After shiftdown to the permissive temperature in the presence of cycloheximide, the number of empty capsids present in tsN-infected cells decreased with a concomitant accumulation of full capsids, indicating that empty capsids are precursors to full capsids.


Polychlorinated Biphenyls (PCBs) and Sex Hormone Concentrations in Ringed and Grey Seals: A Possible Link to Endocrine Disruption?
Polychlorinated biphenyls (PCBs) are recognised reproductive and immune system toxicants in marine mammals mediated by endocrine-disrupting mechanisms. As with other predators, seals are exposed to elevated bioaccumulated concentrations of PCBs and other persistent organic pollutants (POPs). Cryopreserved plasma samples from adult ringed (Phoca hispida; n = 39) and grey (Halichoerus grypus; n = 38) seals, sampled between 1998 and 2002 from Baltic Sea, Svalbard, and Sable Island (Canada) were used to investigate relationships between PCB exposure and sex hormone concentrations (progesterone; P4, 17α-hydroxy progesterone; 17α-OH-P4, testosterone; T4, 17β-estradiol; E2, estrone; E3). Immunoassay methods were used for quantification of analytes due to the limited sample volumes available. PCB concentrations were found to be significantly higher in Baltic seals than other sampling locations and were classed as "Exposed" seals while Svalbard and Sable Is seal were classed "Reference" seals (sexes and species separate). Mean hormone concentrations in Exposed seal were lower than Reference seals, and this was statistically significantly for 17α-OH-P4 (both sexes and both species), E2 (ringed and grey seal females), and E3 (grey seal females). Regression analyses (PCB v hormone concentrations) for each sex and species revealed significant correlations for P4 (Sable Is. female grey seals and female ringed seals), 17α-OH-P4 (Sable Is. male grey seals and Svalbard male ringed seals), T4 (Svalbard male ringed seals), E2 (female ringed seals), and E3 (female ringed seals and Baltic female grey seals). Although significant correlations are not evidence of cause and effect, the potential impact of hormone changes on endocrine homeostasis and reproductive health for seal populations warrants further investigation given that PCB concentrations found here are in the same range as those currently reported in seals from these populations.


Skeletal system as a rare metastatic site in patients with ovarian carcinoma.
To evaluate the frequency and predictors of bone metastasis in patients with ovarian cancer and to determine prognostic factors associated with this finding. Patients diagnosed with ovarian cancer between January 2009 and December 2019 were evaluated. Patients with radiologically or pathologically confirmed bone metastasis were included in the study. Survival was analyzed using Kaplan-Meier curves and compared using the log-rank test. Multivariate analysis of prognostic factors related to survival was performed using the Cox proportional hazards model. Nineteen (2.6%) of 736 patients had bone metastases. Patients with clear cell histology had a higher risk of bone metastases than patients with the other epithelial histology groups (12.3% vs 2.1%, p<0.001). Overall survival was significantly lower in patients diagnosed with bone metastasis at the time of cancer diagnosis than in those diagnosed with bone metastasis during the course of the disease (median 63 vs 6.1 months, p<0.001). However, when the survival time after the development of bone metastasis was examined, no difference was found between patients with metastasis at the time of diagnosis and at the time of first or later progression (median 13.6 vs 4 months, p=0.09). In addition, the median survival of patients with clear cell histology after bone metastasis did not differ statistically from that of patients with other epithelial histology (median 22 vs 7.5 months; p=0.13). In the clear cell subgroup, bone metastasis was an independent prognostic factor for survival after multivariate analysis. For all patients, the stage at diagnosis and serum CA125 and alkaline phosphatase levels at the time of bone metastasis were prognostic factors for survival. Bone metastasis is rare in patients with ovarian cancer. However, the risk of bone metastasis is highest in patients with clear cell histology.


A functional approach to the body condition assessment of lactating donkeys as a tool for welfare evaluation.
The breeding of lactating donkeys is increasing in Western Europe; with it the evaluation of body condition is growing in importance since it is considered a key principle for their welfare. However, assessment of body condition is a complex task, since several factors are involved. The aim of the present study is to investigate which animal-based indicators are the most reliable to describe the body condition of lactating donkeys. For this purpose, new animal-based indicators, which are easy to measure in field conditions (including body measurements, fatty neck score (FNS), dental score), are recorded and their relationship with BCS (a proxy measure for overall adiposity) was assessed. The ones that reveal an association with the BCS are included in an integrated principal component analysis to understand which are the most related to BCS. Fifty-three healthy lactating donkeys of various breeds, including 7 Martina Franca, 10 Ragusano, 2 Romagnolo and 34 crossbreeds, were evaluated. The animal-based indicators that were recorded were: length (OP, olecranon tuber-pinbone and SH, shoulder-hip), heart girth (HG), abdominal circumference (AC), neck length (NL), neck height (NH) and neck thickness (NT) at 0.50 and neck circumference (NC) at 0.25, 0.50 and 0.75, body condition score (BCS) and fatty neck score (FNS). The owners' evaluation of the BCS was also considered. A dental assessment was performed and the month of lactation and age of each animal was recorded. No correlation was found between BCS and the other morphometric body measurements. On the contrary the FNS was correlated with the morphometric measurements of the neck (positive correlation to 0.50 NH and 0.50 NT, 0.50 NC, 0.75 mean NC, and negative correlation to the mean NC:NH and mean NC:NT, 0.50 NC:NT and 0.50 NC:NH ratios). A significant inverse relationship was identified between BCS and dental score. A Principal Component analysis (PCA) separated the BCS classes on the first principal component (PC1). PC1 revealed a meaningful positive correlation between the BCS and the neck measurements (NT, NH and FNS), with high positive loadings, while a negative correlation was found for dental abnormalities. The owners' evaluation of BCS was different from the expert evaluator' assessment, since they tended to give higher score that was slightly but significantly correlated to AC. A new scoring system, called Fatty Neck Score (FNS), has been proposed for the judgement of the adiposity status of donkey neck. The results suggest that caregivers might use the proposed animal based indicators (BCS, FNS and dental scores) together as a tool for the evaluation of the body condition of lactating donkeys. Our findings highlight that caregivers need to be trained in order to be able to properly record these indicators. Ultimately use of these indicators may help to improve the welfare of lactating donkeys.


Injected with controversy: sales and administration of injectable contraceptives in drug shops in Uganda.
Informal drug shops are the first line of health care in many poor countries. In Uganda, these facilities commonly sell and administer the injectable contraceptive depot medroxyprogesterone acetate (DMPA), even though they are prohibited by law from selling any injectable drugs. It is important to understand drug shop operators' current practices and their potential to provide DMPA to hard-to-reach populations. Between November 2007 and January 2008, 157 drug shops were identified in three rural districts of Uganda, and the operators of the 124 facilities that sold DMPA were surveyed. Data were analyzed with descriptive methods. Only 35% of operators reported that the facility in which they worked was a licensed drug shop and another 9% reported that the facility was a private clinic; all claimed to have some nursing, midwifery, or other health or medical qualification. Ninety-six percent administered DMPA in the shop. Operators gave a mean of 10 injections (including three of DMPA) per week. Forty-three percent of those who administered DMPA reported disposing of used syringes in sharps containers; in the previous 12 months, 24% had had a needle-stick injury and 17% had had a patient with an injection-related abscess. Eleven percent said they had ever reused a disposable syringe. Overall, contraceptive knowledge was low, and attitudes toward family planning reflected common traditional biases. Provision of DMPA is common in rural drug shops, but needs to be made safer. Absent stronger regulation and accreditation, drug shop operators can be trained as community-based providers to help meet the extensive unmet demand for family planning in rural areas.


Iron transport in Escherichia coli: uptake and modification of ferrichrome.
During the transport of iron as ferrichrome complex into cells of Escherichia coli K-12, the ligand was modified and excreted into the medium. The rate of the formation of the modified product corresponded with the rate of iron transport. The modified product showed a decreased affinity for ferric iron and did not serve as an effective iron ionophore. After all of the ferrichrome had been converted, the modified product was taken up into the cell in an iron-free form. The uptake of ferrichrome and of the modified product depended on the transport system specified by the tonA and tonB genes. The modified product could be converted back into ferrichrome by mild acid or alkaline hydrolysis. One mole of acetate was released per mole of ferrichrome. It is proposed that one N-hydroxyl group of ferrichrome is acetylated to explain the low affinity for iron as the N-hydroxyl groups form the ligands for iron (III). A weak ester linkage by which the acetyl group is covalently bonded would account for the easy hydrolysis. The iron-free form of ferrichrome, deferri-ferrichrome, was also rapidly converted when incubated with cells with a functional transport system. It is therefore likely that iron is released from ferrichrome by reduction before modification takes place. The conversion of the ligand could be a mechanism by which cells rid themselves of a potentially deleterious ligand for iron in the cytoplasm. A possible role in ferrichrome transport is discussed.


Effect of pitavastatin on glucose, HbA1c and incident diabetes: A meta-analysis of randomized controlled clinical trials in individuals without diabetes.
Whether adverse effect of statins on glycaemic indices is common to all statins remains controversial and as yet data for pitavastatin are limited. We sought to assess the effects of pitavastatin on glycaemia and new-onset diabetes (NOD) in non-diabetic individuals using data from RCT pooled together by means of a meta-analysis. We searched Medline, Cochrane, Embase and clinical trials registries websites until November-2014 for ≥12-week follow-up placebo or statin-controlled RCT of pitavastatin that included participants without diabetes and reported on fasting blood glucose (FBG), HbA1c or NOD. We additionally sought studies by consulting with Kowa Ph. Ltd. The association of pitavastatin with the outcomes were estimated by random-effects meta-analyses. Heterogeneity was assessed by the I(2) statistic and sensitivity and subgroup analyses, and publication bias with funnel plots and Egger and Harbord Tests. 15 studies (approx. 1600 person-years) were included. No significant differences associated with pitavastatin (vs. control) were observed for FBG (MD -0.01 mg/dL [95%CI -0.77, 0.74], I(2) = 0%), HbA1c (MD -0.03% [95%CI -0.11, 0.05], I(2) = 43%) or NOD (RR 0.70 [95%CI 0.30, 1.61]; RD 0.0 [95%CI -0.004, 0.003]; I(2) = 0%). Sensitivity and subgroup analyses (including type of control [placebo or other statin], pitavastatin dose or follow-up] did not yield significant results. Potential publication bias may occur for NOD. In the present meta-analysis pitavastatin did not adversely affect glucose metabolism or diabetes development compared with placebo or other statins.


Cooperation and conflict in the evolution of individuality. IV. Conflict mediation and evolvability in Volvox carteri.
The continued well being of evolutionary individuals (units of selection and evolution) depends upon their evolvability, that is their capacity to generate and evolve adaptations at their level of organization, as well as their longer term capacity for diversifying into more complex evolutionary forms. During a transition from a lower- to higher-level individual, such as the transition between unicellular and multicellular organisms, the evolvability of the lower-level (cells) must be restricted, while the evolvability of the new higher-level unit (multicellular organism) must be enhanced. For these reasons, understanding the factors leading to an evolutionary transition should help us to understand the factors underlying the emergence of evolvability of a new evolutionary unit. Cooperation among lower-level units is fundamental to the origin of new functions in the higher-level unit. Cooperation can produce a new more complex evolutionary unit, with the requisite properties of heritable fitness variations, because cooperation trades fitness from a lower-level (the costs of cooperation) to the higher-level (the benefits for the group). For this reason, the evolution of cooperative interactions helps us to understand the origin of new and higher-levels of fitness and organization. As cooperation creates a new level of fitness, it also creates the opportunity for conflict between levels of selection, as deleterious mutants with differing effects at the two levels arise and spread. This conflict can interfere with the evolvability of the higher-level unit, since the lower and higher-levels of selection will often "disagree" on what adaptations are most beneficial to their respective interests. Mediation of this conflict is essential to the emergence of the new evolutionary unit and to its continued evolvability. As an example, we consider the transition from unicellular to multicellular organisms and study the evolution of an early-sequestered germ-line in terms of its role in mediating conflict between the two levels of selection, the cell and the cell group. We apply our theoretical framework to the evolution of germ/soma differentiation in the green algal group Volvocales. In the most complex member of the group, Volvox carteri, the potential conflicts among lower-level cells as to the "right" to reproduce the higher-level individual (i.e. the colony) have been mediated by restricting immortality and totipotency to the germ-line. However, this mediation, and the evolution of an early segregated germ-line, was achieved by suppressing mitotic and differentiation capabilities in all post-embryonic cells. By handicapping the soma in this way, individuality is ensured, but the solution has affected the long-term evolvability of this lineage. We think that although conflict mediation is pivotal to the emergence of individuality at the higher-level, the way in which the mediation is achieved can greatly affect the longer-term evolvability of the lineage.


Antigenic and structural properties of the hemagglutinin-neuraminidase glycoprotein of human parainfluenza virus type 3: sequence analysis of variants selected with monoclonal antibodies which inhibit infectivity, hemagglutination, and neuraminidase activities.
The hemagglutinin-neuraminidase (HN) gene sequence was determined for 16 antigenic variants of human parainfluenza virus type 3 (PIV3). The variants were selected by using monoclonal antibodies (MAbs) to the HN protein which inhibit neuraminidase, hemagglutination, or both activities. Each variant had a single-point mutation in the HN gene, coding for a single amino acid substitution in the HN protein. Operational and topographic maps of the HN protein correlated well with the relative positions of the substitutions. There was little correlation between the cross-reactivity of a MAb with the bovine PIV3 HN and the amount of amino acid homology between the human and bovine PIV3 HN proteins in the regions of the epitopes, suggesting that many of the epitopes are conformational in nature. Computer-assisted analysis of the HN protein predicted a secondary structure composed primarily of hydrophobic beta sheets interconnected by random hydrophilic coil structures. The HN epitopes were located in predicted coil regions. Epitopes recognized by MAbs which inhibit neuraminidase activity of the virus were located in a region which appears to be structurally conserved among several paramyxovirus HN proteins and which may represent the sialic cid-binding site of the HN molecule.


Dimethyl fumarate increases fetal hemoglobin, provides heme detoxification, and corrects anemia in sickle cell disease.
Sickle cell disease (SCD) results from a point mutation in the β-globin gene forming hemoglobin S (HbS), which polymerizes in deoxygenated erythrocytes, triggering recurrent painful vaso-occlusive crises and chronic hemolytic anemia. Reactivation of fetal Hb (HbF) expression ameliorates these symptoms of SCD. Nuclear factor (erythroid derived-2)-like 2 (Nrf2) is a transcription factor that triggers cytoprotective and antioxidant pathways to limit oxidative damage and inflammation and increases HbF synthesis in CD34+ stem cell-derived erythroid progenitors. We investigated the ability of dimethyl fumarate (DMF), a small-molecule Nrf2 agonist, to activate γ-globin transcription and enhance HbF in tissue culture and in murine and primate models. DMF recruited Nrf2 to the γ-globin promoters and the locus control region of the β-globin locus in erythroleukemia cells, elevated HbF in SCD donor-derived erythroid progenitors, and reduced hypoxia-induced sickling. Chronic DMF administration in SCD mice induced HbF and increased Nrf2-dependent genes to detoxify heme and limit inflammation. This improved hematological parameters, reduced plasma-free Hb, and attenuated inflammatory markers. Chronic DMF administration to nonanemic primates increased γ-globin mRNA in BM and HbF protein in rbc. DMF represents a potential therapy for SCD to induce HbF and augment vasoprotection and heme detoxification.


Is there a common denominator for the antimanic effect of lithium and anticonvulsants?
Starting from the idea that different primary actions of carbamazepine, valproate and lithium might converge into a similar secondary mechanism of action, e.g. a modulation of the functioning of certain transmitter systems, a review of the literature was made on the effects of these and related substances on GABAergic, dopaminergic, cholinergic, noradrenergic and serotonergic transmission, in an attempt to find a common denominator. It was concluded that similarities of the effects of these drugs are most evident with respect to a reduction of dopaminergic transmission. Some similarities also exist with respect to GABAergic and, to a lesser degree, cholinergic transmission. The effects on noradrenergic and serotonergic transmission seem too disparate to be considered as a potential basis for the antimanic effects of these drugs.


Fast shoreline erosion induced by ship wakes in a coastal lagoon: Field evidence and remote sensing analysis.
An investigation based on in-situ surveys combined with remote sensing and GIS analysis revealed fast shoreline retreat on the side of a major waterway, the Malamocco Marghera Channel, in the Lagoon of Venice, Italy. Monthly and long-term regression rates caused by ship wakes in a reclaimed industrial area were considered. The short-term analysis, based on field surveys carried out between April 2014 and January 2015, revealed that the speed of shoreline regression was insignificantly dependent on the distance from the navigation channel, but was not constant through time. Periods of high water levels due to tidal forcing or storm surges, more common in the winter season, are characterized by faster regression rates. The retreat is a discontinuous process in time and space depending on the morpho-stratigraphy and the vegetation cover of the artificial deposits. A GIS analysis performed with the available imagery shows an average retreat of 3-4 m/yr in the period between 1974 and 2015. Digitization of historical maps and bathymetric surveys made in April 2015 enabled the construction of two digital terrain models for both past and present situations. The two models have been used to calculate the total volume of sediment lost during the period 1968-2015 (1.19×106 m3). The results show that in the presence of heavy ship traffic, ship-channel interactions can dominate the morphodynamics of a waterway and its margins. The analysis enables a better understanding of how shallow-water systems react to the human activities in the post-industrial period. An adequate evaluation of the temporal and spatial variation of shoreline position is also crucial for the development of future scenarios and for the sustainable management port traffic worldwide.


Quantitative detection of fecal contamination with domestic poultry feces in environments in China.
Poultry are an important source of fecal contamination in environments. However, tools for detecting and tracking this fecal contamination are in the early stages of development. In practice, we have found that source tracking methods targeting the 16S rRNA genes of poultry-specific microbiota are not sufficiently sensitive. We therefore developed two quantitative PCR assays for detection of poultry fecal contamination, by targeting chicken and duck mitochondrial genes: NADH dehydrogenase subunit 5 (ND5) and cytochrome b (cytb). The sensitivity of both assays was 100% when tested on 50 chicken and duck fecal samples from 10 provinces of China. These assays were also tested in field samples, including soil and water collected adjacent to duck farms, and soils fertilized with chicken manure. Poultry mitochondrial DNA was detected in most of these samples, indicating that the assays are a robust method for monitoring environmental contamination with poultry feces. Complemented with existing indicators of fecal contamination, these markers should improve the efficiency and accuracy of microbial source tracking.


Comparison of BRICS-TM Countries' Biosimilar Regulatory Frameworks With Australia, Canada and Switzerland: Benchmarking Best Practices.
Objectives: The aim of this study was to identify, compare and evaluate regulatory requirements for the biosimilar development and review processes in BRICS-TM (Brazil, Russia, India, China, South Africa, Turkey, Mexico) countries with mature regulatory systems of Australia, Canada, Singapore and Switzerland. It is hoped that this benchmark study provides an opportunity for BRICS-TM agencies to identify the key areas for improvement in their regulatory processes. Materials and Methods: A semi-quantitative questionnaire was developed covering the different criteria used in biosimilar development and registration process. Eleven regulatory agencies from BRICS-TM and ACSS (Australia, Canada, Switzerland and Singapore) countries were invited to take part in this study. Data processing and analysis was carried out using descriptive statistics for quantitative data and content analysis to generate themes for qualitative data. Results and Discussions: Nine of the 11 regulatory agencies recruited for the study completed the questionnaire. China and Singapore did not meet the deadline due to lack of resources. The organisational structure of BRICS-TM agencies revealed support from external assessors by most of these agencies in comparison with ACSS agencies. There was absence of reliance approach and participation in harmonisation activities across most BRICS-TM agencies. Despite alignment over biosimilarity, the mandate for in vivo non-clinical studies and additional local clinical studies in some of the BRICS-TM countries illustrates a lack of effective implementation of a step-wise approach. Adopting flexible regulatory standards in the sourcing of a RBP (Reference Biologic Product) by BRICS-TM similar to ACSS, will facilitate cost-effective development of biosimilar products. Conclusions: Comparative assessment of the biosimilar regulatory framework of BRICS-TM with ACSS agencies reveals the scope for enhancing efficiency of the regulatory approval process. To achieve this, BRICS-TM agencies should consider relying on reference agencies for alternative review mechanisms such as abridged or verification models, streamlined processes for providing scientific advice to developers and for waiving local clinical studies in-lieu of advanced scientific data.


Updated ILAE evidence review of antiepileptic drug efficacy and effectiveness as initial monotherapy for epileptic seizures and syndromes.
The purpose of this report was to update the 2006 International League Against Epilepsy (ILAE) report and identify the level of evidence for long-term efficacy or effectiveness for antiepileptic drugs (AEDs) as initial monotherapy for patients with newly diagnosed or untreated epilepsy. All applicable articles from July 2005 until March 2012 were identified, evaluated, and combined with the previous analysis (Glauser et al., 2006) to provide a comprehensive update. The prior analysis methodology was utilized with three modifications: (1) the detectable noninferiority boundary approach was dropped and both failed superiority studies and prespecified noninferiority studies were analyzed using a noninferiority approach, (2) the definition of an adequate comparator was clarified and now includes an absolute minimum point estimate for efficacy/effectiveness, and (3) the relationship table between clinical trial ratings, level of evidence, and conclusions no longer includes a recommendation column to reinforce that this review of efficacy/evidence for specific seizure types does not imply treatment recommendations. This evidence review contains one clarification: The commission has determined that class I superiority studies can be designed to detect up to a 20% absolute (rather than relative) difference in the point estimate of efficacy/effectiveness between study treatment and comparator using an intent-to-treat analysis. Since July, 2005, three class I randomized controlled trials (RCT) and 11 class III RCTs have been published. The combined analysis (1940-2012) now includes a total of 64 RCTs (7 with class I evidence, 2 with class II evidence) and 11 meta-analyses. New efficacy/effectiveness findings include the following: levetiracetam and zonisamide have level A evidence in adults with partial onset seizures and both ethosuximide and valproic acid have level A evidence in children with childhood absence epilepsy. There are no major changes in the level of evidence for any other subgroup. Levetiracetam and zonisamide join carbamazepine and phenytoin with level A efficacy/effectiveness evidence as initial monotherapy for adults with partial onset seizures. Although ethosuximide and valproic acid now have level A efficacy/effectiveness evidence as initial monotherapy for children with absence seizures, there continues to be an alarming lack of well designed, properly conducted epilepsy RCTs for patients with generalized seizures/epilepsies and in children in general. These findings reinforce the need for multicenter, multinational efforts to design, conduct, and analyze future clinically relevant adequately designed RCTs. When selecting a patient's AED, all relevant variables and not just efficacy and effectiveness should be considered.


Carbonate apatite artificial bone.
Bone apatite is not hydroxyapatite (HAp), it is carbonate apatite (CO3Ap), which contains 6-9 mass% carbonate in an apatitic structure. The CO3Ap block cannot be fabricated by sintering because of its thermal decomposition at the sintering temperature. Chemically pure (100%) CO3Ap artificial bone was recently fabricated through a dissolution-precipitation reaction in an aqueous solution using a precursor, such as a calcium carbonate block. In this paper, methods of fabricating CO3Ap artificial bone are reviewed along with their clinical and animal results. CO3Ap artificial bone is resorbed by osteoclasts and upregulates the differentiation of osteoblasts. As a result, CO3Ap demonstrates much higher osteoconductivity than HAp and is replaced by new bone via bone remodeling. Granular-type CO3Ap artificial bone was approved for clinical use in Japan in 2017. Honeycomb-type CO3Ap artificial bone is fabricated using an extruder and a CaCO3 honeycomb block as a precursor. Honeycomb CO3Ap artificial bone allows vertical bone augmentation. A CO3Ap-coated titanium plate has also been fabricated using a CaCO3-coated titanium plate as a precursor. The adhesive strength is as high as 76.8 MPa, with excellent tissue response and high osteoconductivity.


Combined influences of strategy and selection history on attentional control.
Visual attention is guided by top-down mechanisms and pre-stimulus task preparation, but also by selection history (i.e., the bias to prioritize previously attended items). Here we examine how these influences combine. Two groups of participants completed two intermingled tasks. One task involved categorization of a unique target; one group categorized the target based on color, and the other based on shape. The other task involved searching for a target defined by unique shape while ignoring a distractor defined by unique color. Our expectation was that the search task would be difficult for the color-categorization group because their categorization task required attentional resolution of color, but the search task required that they ignore color. In some experimental blocks, trials from the two tasks appeared predictably, giving the color-categorization group an opportunity to strategically prepare by switching between color-prioritizing and shape-prioritizing attentional templates. We looked to pre-stimulus oscillatory activity as a direct index of this preparation, and to reaction times and post-stimulus ERPs for markers of resultant change in attentional deployment. Results showed that preparation in the color-categorization group optimized attentional templates, such that these participants became less sensitive to the color distractor in the search task. But preparation was not sufficient to entirely negate the influence of selection history, and participants in the color-categorization group continued to show a propensity to attend to the color distractor. These results indicate that preparatory effort can be scaled to the anticipated attentional requirements, but attention is nevertheless considerably biased by selection history.


Early Determination of Prognosis in Neonatal Moderate or Severe Hypoxic-Ischemic Encephalopathy.
Early determination of prognosis is important in neonates with hypoxic-ischemic encephalopathy (HIE). Our objective was to test scoring systems developed earlier (original scoring system) and develop new prognostic models. Secondary analysis of data from the multicenter randomized controlled trial of longer, deeper, or usual care cooling in neonatal HIE (NCT01192776) that enrolled 364 neonates diagnosed with moderate or severe HIE. The primary outcome was death or moderate or severe disability at 18 to 22 months, and secondary outcome was death during initial hospitalization. Testing of early neurologic clinical examination (<6 hours of age) and the original scoring system for prognostic ability was done, followed by development of new scoring systems and classification and regression tree (CART) models by using early clinical variables (<6 hours of age). For death or disability, the original scoring system correctly classified 75% (95% confidence interval: 69%-81%), whereas the new scoring system correctly classified 78% (73%-82%), and the CART model correctly classified 76% (72%-81%). Early neurologic clinical examination also had a correct classification rate of 76% (71%-80%). Depth and duration of cooling did not affect prediction. Only a few components of the early neurologic examination were associated with poor outcome. For death, the original scoring system correctly classified 72% (66%-77%), the new scoring system 68% (63%-72%), the new CART model 87% (83%-90%), and early neurologic evaluation 81% (77%-85%). The 3 models (scoring system, CART, and early neurologic evaluation) were comparable in predicting death or disability. For in-hospital death, CART models were superior to scoring systems and early neurologic examination.


Impact of Induction Therapy with VRD versus VCD on Outcomes in Patients with Multiple Myeloma in Partial Response or Better Undergoing Upfront Autologous Stem Cell Transplantation.
Bortezomib-based triplet regimens-specifically bortezomib, lenalidomide, and dexamethasone (VRD) and bortezomib, cyclophosphamide, and dexamethasone (VCD)-are the 2 most common induction regimens used in transplantation-eligible patients with newly diagnosed multiple myeloma (NDMM), with conflicting data on comparative efficacy and outcomes in this population. We compared long-term outcomes of patients with NDMM receiving VRD induction and those receiving VCD induction prior to autologous stem cell transplantation (ASCT). Patients registered with the Center for International Blood and Marrow Transplant Registry were included if they had undergone ASCT for MM within 6 months of diagnosis between January 2013 and December 2018, received VRD or VCD induction, and achieved a pretransplantation partial or better response. Of 1135 patients, 914 received VRD and 221 received VCD. The patients receiving VCD were more likely to have renal impairment and International Staging System (ISS) stage III disease and less likely to receive full-dose melphalan (200 mg/m2) conditioning (69% versus 80%; P < .001). Very good partial response rates pretransplantation, post-transplantation, and at best response were not significantly different in the 2 groups. Maintenance use was more common after VRD induction (88% versus 76%; P < .001), with lenalidomide the most common agent (80% versus 63%). Patients in the VRD group had a higher rate of renal recovery (74% versus 43%; P < .001), possibly due to a rapid reduction of light chains in the VRD group or improvement in renal function with VCD, which allowed a switch over to VRD, as patients who switched were classified in the VRD group. Patients receiving VRD had better survival on univariate analysis, with a median progression-free survival (PFS) from transplantation of 44.6 months versus 34.1 months (P = .004) and median 5-year overall survival (OS) of 79% versus 60% (P < .001). Multivariate analysis showed no significant survival difference, with a hazard ratio for VCD versus VRD induction of 1.22 (95% CI, 0.96 to 1.55; P = .10) for PFS and 1.33 (95% CI, 0.93 to 1.92, P = .12) for OS. Maintenance use was independently associated with superior PFS and OS, along with ISS stage, cytogenetics, and pretransplantation response (PFS only). In patients with MM undergoing upfront ASCT after VRD or VCD induction, no independent survival difference was seen based on the induction therapy received after adjusting for other prognostic factors. The use of maintenance treatment was uniformly associated with superior outcomes. © 2021 American Society for Transplantation and Cellular Therapy. Published by Elsevier Inc.


Dopamine D4 Receptor Is a Regulator of Morphine-Induced Plasticity in the Rat Dorsal Striatum.
Long-term exposition to morphine elicits structural and synaptic plasticity in reward-related regions of the brain, playing a critical role in addiction. However, morphine-induced neuroadaptations in the dorsal striatum have been poorly studied despite its key function in drug-related habit learning. Here, we show that prolonged treatment with morphine triggered the retraction of the dendritic arbor and the loss of dendritic spines in the dorsal striatal projection neurons (MSNs). In an attempt to extend previous findings, we also explored whether the dopamine D4 receptor (D4R) could modulate striatal morphine-induced plasticity. The combined treatment of morphine with the D4R agonist PD168,077 produced an expansion of the MSNs dendritic arbors and restored dendritic spine density. At the electrophysiological level, PD168,077 in combination with morphine altered the electrical properties of the MSNs and decreased their excitability. Finally, results from the sustantia nigra showed that PD168,077 counteracted morphine-induced upregulation of μ opioid receptors (MOR) in striatonigral projections and downregulation of G protein-gated inward rectifier K+ channels (GIRK1 and GIRK2) in dopaminergic cells. The present results highlight the key function of D4R modulating morphine-induced plasticity in the dorsal striatum. Thus, D4R could represent a valuable pharmacological target for the safety use of morphine in pain management.


Differences in time-dependent mechanical properties between extruded and molded hydrogels.
The mechanical properties of hydrogels used in biomaterials and tissue engineering applications are critical determinants of their functionality. Despite the recent rise of additive manufacturing, and specifically extrusion-based bioprinting, as a prominent biofabrication method, comprehensive studies investigating the mechanical behavior of extruded constructs remain lacking. To address this gap in knowledge, we compared the mechanical properties and swelling properties of crosslinked gelatin-based hydrogels prepared by conventional molding techniques or by 3D bioprinting using a BioBots Beta pneumatic extruder. A preliminary characterization of the impact of bioprinting parameters on construct properties revealed that both Young's modulus and optimal extruding pressure increased with polymer content, and that printing resolution increased with both printing speed and nozzle gauge. High viability (>95%) of encapsulated NIH 3T3 fibroblasts confirmed the cytocompatibility of the construct preparation process. Interestingly, the Young's moduli of extruded and molded constructs were not different, but extruded constructs did show increases in both the rate and extent of time-dependent mechanical behavior observed in creep. Despite similar polymer densities, extruded hydrogels showed greater swelling over time compared to molded hydrogels, suggesting that differences in creep behavior derived from differences in microstructure and fluid flow. Because of the crucial roles of time-dependent mechanical properties, fluid flow, and swelling properties on tissue and cell behavior, these findings highlight the need for greater consideration of the effects of the extrusion process on hydrogel properties.


Relationships Between Body Image, Body Composition, Sexual Functioning, and Sexual Satisfaction Among Heterosexual Young Adults.
This study investigated the association between body image and body-image self-consciousness on sexual satisfaction, accounting for relationships between body fat and body image, and between sexual functioning and sexual satisfaction, while controlling for relationship satisfaction. Participants were 143, 18-25 year-old Caucasian men and women in heterosexual monogamous relationships, recruited from the University of Guelph and surrounding community in Ontario, Canada. Various domains of body image, body-image self-consciousness, sexual satisfaction and functioning, and relationship satisfaction data were collected by questionnaires. Body fat was measured using dual energy X-ray absorptiometry. Among men, body image was positively associated with sexual satisfaction, after controlling for relationship satisfaction. Men with greater body fat were more likely to have poorer behavioral and affective body image. Only body image specific to the sexual encounter influenced sexual functioning. Among women, no domain of body image was associated with sexual satisfaction, after controlling for relationship satisfaction. Women with greater body fat were more likely to have poorer affective and sexual-encounter-specific body image. As percent total fat increased, sexual functioning decreased. Our results suggest a complex pattern of relationships exists among body image and body composition constructs and sexual and relationship variable; and that these relationships are not the same for men and women.


A Pilot Study of Neonatologists' Decision-Making Roles in Delivery Room Resuscitation Counseling for Periviable Births.
Relatively little is known about neonatologists' roles in helping families navigate the difficult decision to attempt or withhold resuscitation for a neonate delivering at the threshold of viability. Therefore, we aimed to describe the "decision-making role" of neonatologists in simulated periviable counseling sessions. We conducted a qualitative content analysis of audio-recorded simulation encounters and post-encounter debriefing interviews collected as part of a single-center simulation study of neonatologists' resuscitation counseling practices in the face of ruptured membranes at 23 weeks gestation. We trained standardized patients to request a recommendation if the physician presented multiple treatment options. We coded each encounter for communication behaviors, applying an adapted, previously developed coding scheme to classify physicians into four decision-making roles (informative, facilitative, collaborative, or directive). We also coded post-simulation debriefing interviews for responses to the open-ended prompt: "During this encounter, what did you feel was your role in the management decision-making process?" Fifteen neonatologists (33% of the division) participated in the study; audio-recorded debriefing interviews were available for 13. We observed 9 (60%) take an informative role, providing medical information only; 2 (13%) take a facilitative role, additionally eliciting the patient's values; 3 (20%) take a collaborative role, additionally engaging the patient in deliberation and providing a recommendation; and 1 (7%) take a directive role, making a treatment decision independent of the patient. Almost all (10/13, 77%) of the neonatologists described their intended role as informative. Neonatologists did not routinely elicit preferences, engage in deliberation, or provide treatment recommendations-even in response to requests for recommendations. These findings suggest there may be a gap between policy recommendations calling for shared decision making and actual clinical practice.


Elastic fibres in health and disease.
Elastic fibres are a major class of extracellular matrix fibres that are abundant in dynamic connective tissues such as arteries, lungs, skin and ligaments. Their structural role is to endow tissues with elastic recoil and resilience. They also act as an important adhesion template for cells, and they regulate growth factor availability. Mutations in major structural components of elastic fibres, especially elastin, fibrillins and fibulin-5, cause severe, often life-threatening, heritable connective tissue diseases such as Marfan syndrome, supravalvular aortic stenosis and cutis laxa. Elastic-fibre function is also frequently compromised in damaged or aged elastic tissues. The ability to regenerate or engineer elastic fibres and tissues remains a significant challenge, requiring improved understanding of the molecular and cellular basis of elastic-fibre biology and pathology, and ability to regulate the spatiotemporal expression and assembly of its molecular components.


Does emotional reasoning change during cognitive behavioural therapy for anxiety?
Emotional reasoning refers to the use of subjective emotions, rather than objective evidence, to form conclusions about oneself and the world. It is a key interpretative bias in cognitive models of anxiety disorders and appears to be especially evident in individuals with anxiety disorders. However, the amenability of emotional reasoning to change during treatment has not yet been investigated. We sought to determine whether emotional reasoning tendencies change during a course of routine cognitive-behavioural therapy (CBT). Emotional reasoning tendencies were assessed in 36 individuals with a primary anxiety disorder who were seeking treatment at an outpatient clinic. Changes in anxiety and depressive symptoms as well as emotional reasoning tendencies after 12 sessions of CBT were examined in 25 individuals for whom there was complete data. Emotional reasoning tendencies were evident at pretreatment assessment. Although anxiety and depressive symptoms decreased during CBT, only one of six emotional reasoning interpretative styles (pertaining to conclusions that one is incompetent) changed significantly during the course of therapy. Attrition rates were high and there was not enough information regarding the extent to which therapy specifically focused on addressing emotional reasoning tendencies. Individuals seeking treatment for anxiety disorders appear to engage in emotional reasoning, however routine individual CBT does not appear to result in changes in emotional reasoning tendencies.


Measuring management practices in India's district public health bureaucracy.
Weak management is widely recognised as a key impediment to scaling-up coverage of health interventions and ensuring health systems are responsive to population needs. Yet there is scant evidence linking management practices in the public administration to effective health service delivery. We report on the development of a tool to measure management practices in India's district health bureaucracy. We first developed a conceptual framework based on a review of the literature and qualitative interviews with district public health managers. Across 16 management practices, we then drafted and piloted questions to be used with a scoring grid to evaluate process-orientated management practices. We implemented the tool in 34 districts of Maharashtra between April and July 2016, interviewing up to three district public health managers per district (n = 99). Using rigorous psychometric methods, we assessed the acceptability, reliability and validity of the tool. We present three key findings. First, the tool was feasible to implement, response rates were high, and there were no missing data. Second, internal consistency of the tool was high and test-retest reliability was comparable with other management tools used in the literature. Third, there was evidence of validity. The number of staff with a management qualification was positively associated with better management practices. Factor analysis showed that one principal component loaded positively on all the management practices although there was little support for management sub-scales. These findings provide novel evidence on the psychometric properties of a tool designed to measure management practices in the public administration of a developing country. Our framework and tool provide the basis to examine associations between district health management practices and health service delivery, and test the effectiveness of management strengthening interventions in India's public health sector.


Identification of reaction products and intermediates of aromatic-amine dehydrogenase by 15N and 13C NMR.
13C- and 15N-NMR studies of the reaction of aromatic amine dehydrogenase (AADH) with methylamine demonstrated that the products of the reductive half-reaction are an equivalent of formaldehyde hydrate and a reduced aminoquinol form of the tryptophan tryptophylquinone (TTQ) cofactor which contains covalently bound substrate-derived N. These data are consistent with the Ping Pong kinetic mechanism and aminotransferase-type chemical reaction mechanism which have been previously proposed for AADH. Comparison of the 15N-NMR spectra of the aminoquinol TTQ intermediates of AADH and methylamine dehydrogenase (MADH) revealed that the substrate-derived aminoquinol N of AADH and MADH exhibited distinct 15N chemical shifts which are separated by approx. 7 p.p.m. In each case, the signal for the substrate-derived aminoquinol N appears optimally with short pulse delay and exhibits a relaxation time and chemical shift which are consistent with 15N covalently bound to an aromatic ring (i.e. aminoquinol) which is attached to a rigid protein matrix. The aminoquinol of AADH is less stable against reoxidation than that of MADH. These data suggest that differences in the active-site mediated electrostatic environments of the aminoquinol N in the respective enzymes may influence both the observed 15N chemical shift and the relative reactivities of the TTQ aminoquinols towards oxygen. These data also demonstrate the utility of 13C- and 15N-NMR spectroscopy as a tool for monitoring the intermediates and products of enzyme-catalysed transformations.


Differential susceptibilities of murine hepatoma 1c1c7 and Tao cells to the lysosomal photosensitizer NPe6: influence of aryl hydrocarbon receptor on lysosomal fragility and protease contents.
Irradiation of murine hepatoma 1c1c7 cultures presensitized with N-aspartyl chlorin e6 (NPe6) caused lysosomal disruption and apoptosis. Tao cells, a variant of the 1c1c7 line having lower aryl hydrocarbon receptor (AhR) contents, were resistant to the pro-apoptotic effects of NPe6 in the same photodynamic therapy protocol. Colony-forming assays were used to establish light dose-dependent and NPe6 concentration-dependent cytotoxicity curves. Lysosomal breakage and cell survival paralleled one another in both cell types. When analyzed at comparable lethal dose conditions, the onset of apoptosis was delayed, and the magnitude of the apoptotic response was muted in Tao cells, as assessed by morphology, annexin V binding, caspase-3 activities, and analyses of Bid, procaspase-9, and pro-caspase-3 cleavage. In contrast, the kinetics/magnitude of pro-caspase-3 activation in the two cell lines were identical after exposure to HA14 -1 or Jo2 antibody, inducers of the intrinsic and extrinsic apoptotic pathways, respectively. Tao endosomal/lysosomal extracts contained approximately 50%, 35%, and 55% of the Bid cleavage and cathepsin B and D activities of 1c1c7 endosomes/lysosomes, respectively. Western blot analyses confirmed reduced cathepsin B/D contents in Tao cells. Analyses of 1c1c7/Tao variants engineered to express antisense/sense AhR constructs suggested that endosomal/lysosomal cathepsin B and D content, but not whole cell content, correlated with AhR expression. These studies provide a mechanism for the resistance of Tao cultures to the proapoptotic effects of a protocol causing targeted disruption of lysosomes. They also suggest that the AhR, in the absence of exogenous ligand, may affect the trafficking/processing of proteases normally found in endosomes/lysosomes.


Accounting for multimorbidity can affect the estimation of the Burden of Disease: a comparison of approaches.
Various Burden of Disease (BoD) studies do not account for multimorbidity in their BoD estimates. Ignoring multimorbidity can lead to inaccuracies in BoD estimations, particularly in ageing populations that include large proportions of persons with two or more health conditions. The objective of this study is to improve BoD estimates for the Netherlands by accounting for multimorbidity. For this purpose, we analyzed different methods for 1) estimating the prevalence of multimorbidity and 2) deriving Disability Weights (DWs) for multimorbidity by using existing data on single health conditions. We included 25 health conditions from the Dutch Burden of Disease study that have a high rate of prevalence and that make a large contribution to the total number of Years Lived with a Disability (YLD). First, we analyzed four methods for estimating the prevalence of multimorbid conditions (i.e. independent, independent age- and sex-specific, dependent, and dependent sex- and age-specific). Secondly, we analyzed three methods for calculating the Combined Disability Weights (CDWs) associated with multimorbid conditions (i.e. additive, multiplicative and maximum limit). A combination of these two approaches was used to recalculate the number of YLDs, which is a component of the Disability-Adjusted Life Years (DALY). This study shows that the YLD estimates for 25 health conditions calculated using the multiplicative method for Combined Disability Weights are 5 % lower, and 14 % lower when using the maximum limit method, than when calculated using the additive method. Adjusting for sex- and age-specific dependent co-occurrence of health conditions reduces the number of YLDs by 10 % for the multiplicative method and by 26 % for the maximum limit method. The adjustment is higher for health conditions with a higher prevalence in old age, like heart failure (up to 43 %) and coronary heart diseases (up to 33 %). Health conditions with a high prevalence in middle age, such as anxiety disorders, have a moderate adjustment (up to 13 %). We conclude that BoD calculations that do not account for multimorbidity can result in an overestimation of the actual BoD. This may affect public health policy strategies that focus on single health conditions if the underlying cost-effectiveness analysis overestimates the intended effects. The methodology used in this study could be further refined to provide greater insight into co-occurrence and the possible consequences of multimorbid conditions in terms of disability for particular combinations of health conditions.